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ncl-talk 2013 archive: Re: reading raw calipso data in NCL Re: reading raw calipso data in NCL From : Mary Haley < haley_at_nyahnyahspammersnyahnyah > Date : Tue Jul 23 2013 - 21:45:36 MDT Hi Matt (and Joe), I didn't see this message before I sent you the other response. I think you want to check out our "dim_gbits" function, which allows you to extract bits from an NCL data variable. After reading the description below, maybe Joe will have a better idea of how to extract the bits that you need. We might be able to add some examples of this on our applications page. Please look at example "hdf4eos_5.ncl" at: http://www.ncl.ucar.edu/Applications/HDF.shtml This example shows how to extra the bits you want and generate a contour plot. Here's the "dim_gbits" function documentation: http://www.ncl.ucar.edu/Document/Functions/Built-in/dim_gbits.shtml The dim_gbits function requires that you know how the bits are written. If you do an ncl_filedump on the HDF file, you should see a description of howthe bits are laid out. The description looks something like this: % ncl_filedump MOD05_L2.A2002161.1830.004.2003221153410.hdf | less . . . byte Cloud_Mask_QA ( Cell_Along_Swath_1km_mod05, Cell_Across_Swath_1km_mod05 )long_name : MODIS Cloud Mask, First Byteunit : nonescale_factor : 1add_offset : 0Parameter_Type : MODIS InputCell_Along_Swath_Sampling : ( 1, 2030, 1 )Cell_Across_Swath_Sampling : ( 1, 1354, 1 )Geolocation_Pointer : External MODIS geolocation productdescription :bit field Description Key--------- ----------- ---0 Cloud Mask Flag 0 = not determined1 = determined1-2 Unobstructed FOV Quality Flag 00 = cloud01 = 66% prob.clear10 = 95% prob.clear11 = 99% prob.clearPROCESSING PATH FLAGS---------------------3 Day / Night Flag 0 = Night / 1 = Day4 Sunglint Flag 0 = Yes / 1 = No5 Snow / Ice Background Flag 0 = Yes / 1 = No6-7 Land / Water Flag 00=Water/01=Coastal10=Desert/11=Landvalid_range : ( 0, -1 )_FillValue : 0hdf_name : Cloud_Mask_QA. . . --Mary On Jul 23, 2013, at 9:10 PM, Matthew Fearon <[email protected]> wrote: > Hi, Joe > Thanks for your reply. At this time, I'm most interested in Calipso Vertical Feature Mask (VFM) and reading the associated variable 'Feature_Classification_Flags' from the hdf file, eg., CAL_LID_L2_VFM-ValStage1-V3-02.2011-12-31T23-18-11ZD.hdf . NCL reads this variable as an unsigned short. However, in order to extract the correct flag, which should be a single value, it appears the data need to be bit interpreted. I'm not sure how to do this correctly in NCL. If you have any luck unpacking and or plotting up the data in NCL, please let me know. Right now, I'm trying to setup a temporary workaround with Matlab, via the code released by NASA, to extract the data correctly -- not an ideal solution. > > thanks again, > Matt > > > ________________________________________ > From: H. Joe Lee [hyoklee@hdfgroup. org] > Sent: Tuesday, July 23, 2013 7:13 PM > To: Matthew Fearon > Subject: Re: reading raw calipso data in NCL > > Hi, Matt! > > I'm also interested in plotting HDF CALIPSO data and post it under > our website [1] but I'm wondering if you could provide the specific > NASA HDF product name (e.g., > CAL_IIR_L1-Prov-V1-11.2008-08-20T01-40-12ZD.hdf) that you're mostly > interested in. > > Thanks, > > > [1] http://hdfeos.org/zoo > -- > HDF: Software that Powers Science > > > On Mon, Jul 22, 2013 at 2:56 PM, Matthew Fearon <Matthew.Fearon@dri. edu> wrote: >> Dear NCL users: >> >> Just curious if anyone has developed an NCL script to read raw Calipso data, >> particularly vertical feature mask, in hdf format and make a plot? I see >> there are a couple examples here: >> http://www.ncl.ucar.edu/Applications/calipso.shtml , but they are using a >> processed form of the data. Any assistance would be much appreciated. >> >> thanks, >> Matt >> >> >> _______________________________________________ >> ncl-talk mailing list >> List instructions, subscriber options, unsubscribe: >> http://mailman.ucar.edu/mailman/listinfo/ncl-talk >> > _______________________________________________ > ncl-talk mailing list > List instructions, subscriber options, unsubscribe: > http://mailman.ucar.edu/mailman/listinfo/ncl-talk _______________________________________________ncl-talk mailing listList instructions, subscriber options, unsubscribe:http://mailman.ucar.edu/mailman/listinfo/ncl-talk Received on Tue Jul 23 21:45:45 2013 This message : [ Message body ] Next message : Jeff: "Does NCL v 5.2.1 support vis5d" Previous message : Mary Haley: "Re: Questions about draworder" In reply to : Matthew Fearon: "Re: reading raw calipso data in NCL" Maybe reply : Matthew Fearon: "Re: reading raw calipso data in NCL" Reply : Matthew Fearon: "Re: reading raw calipso data in NCL" Contemporary messages sorted : [ By Date ] [ By Thread ] [ By Subject ] [ By Author ] [ By messages with attachments ] This archive was generated by hypermail 2.1.8 : Thu Jul 25 2013 - 21:02:42 MDT

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Constructing the image of the teacher on Reddit: Best and worst teachers | Request PDF Request PDF | Constructing the image of the teacher on Reddit: Best and worst teachers | In addressing the public image of the teacher, this study reports on the analysis of 600 discussion entries on Reddit, a social media platform, in... | Find, read and cite all the research you need on ResearchGate Constructing the image of the teacher on Reddit: Best and worst teachers May 2017 Teaching and Teacher Education 64:43-51 DOI: 10.1016/j.tate.2017.01.019 Abstract In addressing the public image of the teacher, this study reports on the analysis of 600 discussion entries on Reddit, a social media platform, in which respondents described memories of their best and worst teachers. Results of inductive analyses suggest: (1) a model of teacher, student, and teacher-student relationship for characterizing and reflecting on teachers in teacher education programs; (2) the relevance of social media in contributing to the public image of the teacher in an information technology age, as well as its anonymous and, arguably, more authentic nature; and (3) the need to address gender in teacher memory studies. ... Among these, we identified r/Adderall, part of the website Reddit. Reddit can be defined as a massive online sharing and discussion platform (Chang-Kredl and Colannino, 2017) . It houses numerous discussion fora known as subreddits, which vary in membership size and area of interest, known as subreddits. ... ... It houses numerous discussion fora known as subreddits, which vary in membership size and area of interest, known as subreddits. Members can share text, images, links and videos (Chang-Kredl and Colannino, 2017) . Subreddits are created by Reddit users themselves and to register to the website all that is required is an e-mail address and a chosen pseudonym. ... ... However, we also consider that certain members would prefer not to be identified. Reddit fosters the anonymity of its members through the use of pseudonyms, posing a challenge to the identification of users (Chang-Kredl and Colannino, 2017) . Still, we must also be mindful of the stigma that may be associated with psychostimulant use, potentially placing members at risk if the posts were to circulate to a larger audience (Kozinets, 2015). ... Networked psychostimulants: a web-based ethnographic study Article Full-text available Purpose The purpose of this paper is to understand contemporary psychostimulant use among members of online discussion fora. Two objectives are addressed: to describe accounts of practices related to psychostimulant use, and to examine how these pharmaceuticals may shape contemporary subjectivities. Design/methodology/approach This paper presents a qualitative analysis of three online discussion fora belonging to Reddit. Drawing on actor-network theory, psychostimulants are envisaged as networked actants to understand the underlying logics related to their use. Non-participant observation of r/Adderall was carried out over an 18-month period. A qualitative analysis of postings on the three selected fora was also performed. Findings For each discussion forum, a network comprised of human and non-human actors was observed: members of the forum, psychostimulants as objects and subreddits as agentic spaces. This study reveals the emergence of multiple subjectivities associated with psychostimulant use: productivity, wellness and enhancement‐related. Practical implications Findings open to a wider debate regarding public health’s and healthcare professionals’ understanding of psychostimulant use outside of the clinical setting and how this may contrast with how psychostimulant use is practiced in context. Originality/value This research shows new online socio-cultural spaces formed around psychostimulant use. Calling upon a web-based ethnographic approach, this research presents a new perspective on the contemporary use of psychostimulants. ... Among these, we identified r/Adderall, part of the website Reddit. Reddit can be defined as a massive online sharing and discussion platform (Chang-Kredl and Colannino, 2017) . It houses numerous discussion fora known as subreddits, which vary in membership size and area of interest, known as subreddits. ... ... It houses numerous discussion fora known as subreddits, which vary in membership size and area of interest, known as subreddits. Members can share text, images, links and videos (Chang-Kredl and Colannino, 2017) . Subreddits are created by Reddit users themselves and to register to the website all that is required is an e-mail address and a chosen pseudonym. ... ... However, we also consider that certain members would prefer not to be identified. Reddit fosters the anonymity of its members through the use of pseudonyms, posing a challenge to the identification of users (Chang-Kredl and Colannino, 2017) . Still, we must also be mindful of the stigma that may be associated with psychostimulant use, potentially placing members at risk if the posts were to circulate to a larger audience (Kozinets, 2015). ... Networked psychostimulants: a web-based ethnography of online discussion fora Presentation Psychostimulant use is on the rise, notably in North America, referring to pharmaceuticals such as Adderall, Ritalin and Vyvanse. Current understandings of this phenomenon focus on epidemiological or bioethical perspectives that largely put aside socio-cultural dimensions. Drawing on Actor-Network Theory (ANT), we envisage psychostimulants as networked actants to understand the underlying logics related to their use. ANT allows thinking about psychostimulants as agentive entities contributing to socio-cultural transformations. Through this lens, we can shed light on how these pharmaceuticals may shape contemporary subjectivities. Our paper presents a qualitative analysis of one online discussion forum, r/Adderall, belonging to the Reddit community. We utilize a web-based ethnographic approach to access accounts of psychostimulant use as expressed by members of the forum. Exploring online fora affords new opportunities to understand practices such as drug use that lie below-the-radar and are associated with stigma. We performed non-participant observation of r/Adderall over an 18-month period, observing exchanges between members, interactions with moderators, and collected 149 key posts for qualitative analysis (514 pages, double spaced). We observe a network comprised of human and non-human actants: members of r/Adderall, psychostimulants as objects, and r/Adderall as an agentic space, which fosters the emergence of psychostimulant use. Sharing common experiences on the online forum nurtures a sense of belonging among members and leads some to offer guidance and promote what is considered healthy psychostimulant use, even if sometimes contrary to prescribers’ directives. Accounts of drug effects show that members’ subjectivities lie on a continuum between positive, often described as life-changing, and negative, often described as becoming parallel to oneself. Sustained use seems related to the intensity of the sense of attachment one has for these pharmaceuticals, in line with what aspirations can be reached through their use, for example, obtaining a diploma. Our findings open to a wider debate regarding public health’s understanding of psychostimulant use outside of the clinical setting and how this may contrast with how it is being put into practice. View Show abstract ... We contend that these two key translations can shed light on the emergence of contemporary psychostimulant use. (Robards 2018; Chang-Kredl and Colannino 2017) . At the time of writing, there are over 1.2 million, encompassing a wide variety of topics, for example, r/Olympics, r/atheism, or r/fountainpens (Reddit Metrics 2018). ... ... Redditors must also follow general rules of conduct, or "reddiquette" (reddiquette 2018). Each subreddit also sets specific rules, commonly concerning structure and content of posts, the application of which is monitored by moderators (Robards 2018; Chang-Kredl and Colannino 2017) . ... ... We acknowledge that Reddit users are not representative of Internet users or the general population, and that the grassroots and anonymous nature of this website may attract a specific subset of individuals wishing to participate in the discussions taking place there (Robards 2018; Chang-Kredl and Colannino 2017) . Demographic data on Reddit users are scarce. ... “This drug turned me into a robot”: an actor–network analysis of a web-based ethnographic study of psychostimulant use Objectives This study aims to understand contemporary psychostimulant use within the socio-cultural context of Western societies. Two objectives are addressed: to describe accounts of practices related to psychostimulant use among members of selected online fora and to examine how these are related to representations of the self. Methods This research is a qualitative study of psychostimulant use among members of selected online fora. Drawing on actor–network theory (ANT), this study focuses on three publicly accessible online discussion fora belonging to the Reddit website. Non-participant observation was performed over a period of 18 months to observe exchanges between members, interactions with moderators, and esthetic elements. In total, 331 postings were collected from the selected fora for qualitative analysis. We present our analysis of one discussion forum, corresponding to 149 posts (515 pages, double spaced). Results Our study reveals the emergence of an online socio-cultural space formed around psychostimulant use. Members share their experiences openly and some offer guidance, sometimes contrary to prescribers’ directives. An ANT analysis points to translations fostering positive or negative subjectivities, related to psychostimulant effects, and to translations fostering healthy or harmful practices, related to r/Adderall as an agentic space. Conclusion These findings may contribute to shaping public health policies and interventions that acknowledge the values of the individuals they seek to help, and that attempt to reduce the potential harms associated with these pharmaceuticals as an alternative to more prohibitive approaches. ... Indeed, there has been little prior work from any discipline exploring teaching and learning topics on Reddit. We found only four noteworthy prior studies: work by Haythornthwaite et al. (2018), Chang-Kredl and Colannino (2017) , Tannebaum (2018), and Carpenter, McDade, et al. (2018). Haythornthwaite et al. (2018) investigated the informal learning that occurred specifically within "Ask" subreddits-that is, discussions that "tap into crowd knowledge, covering topics from science to professional practice" (Haythornthwaite et al., 2018, p. 221). ... ... Following the precedent of earlier Reddit research (Carpenter, McDade, et al., 2018; Chang-Kredl & Colannino, 2017; Haythornthwaite et al., 2018), we examined educators' uses of Reddit through digital traces-the "machine-readable traces that our increasingly self-documenting and selfarchiving world leaves behind" (Lee, Fielding, & Blank, 2017, p. 5). In this study, we focused on digital traces associated with contributions to subreddits-that is, evidence of subreddit participation including the content of posts and responses, as well as upvotes and downvotes those contributions received. ... Teachers on Reddit? Exploring contributions and interactions in four teaching-related subreddits The website Reddit has received scant attention from educational researchers despite the presence of numerous teaching-related subreddits that host thousands of threaded conversations. The purpose of this study was to provide an initial comparison of discussions occurring in several different affinity spaces on Reddit. Because of the lack of relevant prior research, there was no baseline for what to expect in Reddit discussions, so all posts and responses in four different teaching-related subreddits over the course of a full year were analyzed to answer three research questions: How much did individuals contribute to different subreddits? What content interactions occurred in different subreddits? What social interactions occurred in different subreddits? Comparative quantitative analysis revealed substantial differences among the subreddits in terms of levels of voting and network characteristics. Results suggest that Reddit hosts diverse spaces that could present benefits and challenges for educators. These findings are addressed in relation to the extant literature and implications for educational practice and future research. ... Moreover, these good teachers usually do not agree with the policy of the school administration and adapt the curriculum to the needs of their students. Good students in a similar way demonstrate creative inspiration, responsibility and creative attitude to learning, competitive spirit and friendly support [Chang-Kredl & Colannino, 2017; Dalton, 1995;Gregory, 2007;Marcus & Stoddard, 2007]. A devoted teacher can even sometimes sacrifice one's family interests or health for the sake of his/her students, and often uses non-traditional means to achieve his/her teaching goals. ... ... Bad teachers in audiovisual media texts are portrayed as unpleasant, authoritarian, heartless, strange, boring, unfriendly, unfair, incompetent, corrupt, evil, manipulative. The media images of bad students are built on the deviant behavior, propensity to lie, sexual challenges, and often in the total absence of any cognitive interests [Chang-Kredl & Colannino, 2017; Delamarter, 2015;Dalton, 1995;2013;Gregory, 2007;Marcus & Stoddard, 2007]. Of course, in the Soviet media texts, the images of good and bad students had their own peculiarities associated with accentuation of their commitment to communist and socialist values, atheism (for positive characters) and, on the contrary, ignoring them (for negative characters, for example, school bullies). ... The Structural Model of the Contents of Audiovisual Media Texts on School and University Topic Based on the results of a comparative hermeneutic, anthropological and gender analysis of audiovisual media texts on the theme of the school and university, their classification, content models, modifications of the genre, ideology and stereotypes, the authors synthesized a structural model for the content of audiovisual media texts on the theme of the school and university: The historical period of events in a media text: can be any time period but mostly is contemporary to film production. Location, environment, everyday items: as a rule, the action takes place in the country where the media text is created; furnishings, household items (of varying degrees of quality) are school, vocabulary in this case can be any, although for students, the rough vocabulary and evocative appearance predominate. A crucial change in the characters' life: a) a new teacher / student comes to school / university and faces opposition (including physical) from class / group of students, parents of students, other teachers; with lies, blackmail, aggression, authoritarian domination; b) in an ordinary learning environment, an unexpected event occurs (authoritarian domination, non-ordinary behavior, an act of violence, aggression, deception, suicide, blackmail, including sexual, etc.); c) a teacher has an affair with a student; d) students begin love relationships. The problem that has arisen: reputation, health (and sometimes life) of a teacher / a student is under threat; a pupil/ a student becomes an outcast in a class / group, is alienated; characters feel uncomfortable because of certain characteristics of love relationships. The search for a solution to the problem: the teacher / student struggle (often with colleagues and friends) with non-ordinary behavior, opposition (including physical) from the student, class / group of students, parents of students, other teachers; with lies, blackmail, aggression, authoritarian domination; an attempt of the teacher / student to establish good relations with other participants in the educational process and parents; an attempt by a teacher / student to hide his/her love feelings from outsiders (if it is a school context) or an attempt to adapt to each other (if it is a university). Problem solutions: teachers / students (often with the help of colleagues and friends) win in the struggle against antagonists, class / group of students, parents of students, other teachers (relatively rare option: positive teachers / pupils lose in this fight); educators / students (often with the help of colleagues and friends) expose lies, blackmail, on the part of the student, class / group of students, parents of students, other teachers; teachers / students manage to establish good relations with other participants in the educational process and the parents of students, as a result of which they improve their academic performance; the relationship between a teacher and a high school student is usually broken up; in a teacher / university student's love relationship, harmony is possible, as well as, in the relationships between two students. View ... Более того, эти хорошие учителя обычно не согласны с политикой школьной администрации и адаптируют учебную программу к потребностям учащихся. Хорошие учащиеся аналогичным образом демонстрируют творческое вдохновение, ответственное и творческое отношение к учебе, соревновательный дух и дружескую поддержку [Chang-Kredl & Colannino, 2017; Dalton, 1995;Gregory, 2007;Marcus & Stoddard, 2007]. Хороший педагог иногда может принести в жертву свой брак или здоровье ради своих учеников, и часто использует нетрадиционные средства для достижения своих целей. ... ... Плохие учителя / преподаватели в аудиовизуальных медиатекстах изображаются как неприятные, авторитарные, бессердечные, странные, скучные, недружелюбные, неквалифицированные, несправедливые, некомпетентные, коррумпированные, злые манипуляторы. Медийные образы плохих учащихся построены на девиантности поведения, склонности ко лжи, сексуальным вызовам и часто на полном отсутствии какого-либо интереса к учебе [Chang-Kredl & Colannino, 2017; Delamarter, 2015;Dalton, 1995;2013;Gregory, 2007;Marcus & Stoddard, 2007]. Конечно, в советских медиатекстах образы хороших и плохих учащихся имели свои особенности, связанные с акцентированием их приверженности коммунистическим и социалистическим ценностям, атеизму (для положительных персонажей) и, напротив, игнорирования оных (для персонажей отрицательных, например, школьников-хулиганов). ... Структурная модель содержания аудиовизуальных медиатекстов на тему школы и вуза Федоров А.В., Левицкая А.А., Горбаткова О.И. Структурная модель содержания аудиовизуальных медиатекстов на тему школы и вуза // Медиаобразование. 2018. № 1. Abstract. Исходя из результатов сравнительного герменевтического, антропологического и гендерного анализа аудиовизуальных медиатекстов на тему школы и вуза, их классификации, структурирования моделей содержания, модификаций жанра, идеологии и стереотипов, авторами статьи синтезирована структурная модель содержания аудиовизуальных медиатекстов на тему школы и вуза: Исторический период событий в медиатексте: любой, но в основном – период, современный времени создания медиатекста. Страна действия, обстановка, предметы быта: как правило, действие происходит в стране, где создан медиатекст; обстановка, предметы быта (разной степени качества): школьные классы, аудитории вуза, коридоры, жилища учащихся и педагогов. Приемы изображения действительности: как правило, реалистичные, без гротеска (в телепередачах, в драме, мелодраме, детективе, триллере); гротескные, шаржированные (в комедии, в жанрах фантастики / ужасов). Положительные персонажи, их ценности, идеи, одежда, лексика, мимика, жесты: педагоги и учащиеся с гуманистическими (социалистическими – в советских медиатекстах) ценностями, как правило, аккуратно одетые, обладающие приятной внешностью, артистизмом мимики и жестов, богатой лексикой. Отрицательные персонажи, их ценности, идеи, одежда, телосложение, лексика, мимика, жесты: педагоги и учащиеся с антигуманными наклонностями; одежда, внешность, лексика в данном случае могут быть любыми, хотя преобладает (у учащихся) грубая лексика и вызывающий внешний вид. Существенное изменение в жизни персонажей: новый педагог / учащийся приходит в школу/вуз и сталкивается с неординарностью, противостоянием (в том числе и физическим) со стороны класса / группы учащихся, родителей учащихся, иных педагогов; с ложью, шантажом, агрессией, авторитарным доминированием; в привычной учебной среде возникает неожиданное событие (авторитарное доминирование, неординарность поведения, акт насилия, агрессия, обман, суицид, шантаж, в том числе и сексуальный и т.д.); у педагога завязывается роман со школьником(-цей) / студентом(-кой); любовные отношения завязываются у учащихся. Возникшая проблема: репутация, здоровье (а иногда и жизнь) педагога/учащегося оказывается под угрозой; школьник(-ца) / студент(-ка) становится изгоем в классе/группе, попадает в ситуацию отчуждения; персонажи испытывают дискомфорт из-за тех или иных особенностей любовных связей. Поиски решения проблемы: борьба педагога / учащегося (часто им помогают коллеги и друзья) с неординарностью поведения, противостоянием (в том числе и физическим) со стороны учащегося, класса / группы учащихся, родителей учащихся, иных педагогов; с ложью, шантажом, агрессией, авторитарным доминированием; попытка педагога / учащегося наладить хорошие отношения с иными участниками учебного процесса и родителями; попытка педагога/учащегося скрыть свои любовные чувства от посторонних (если речь идет о школе), или попытка приспособиться друг к другу (если речь идет о вузе). Решения проблемы: педагоги / учащиеся (часто с помощью коллег и друзей) выигрывают в борьбе (в том числе и физической) с отрицательными учащимися, классом / группой учащихся, родителями учащихся, иными педагогами (относительно редкий вариант: положительные педагоги/учащиеся проигрывают в этой борьбе); педагоги / учащиеся (часто с помощью коллег и друзей) разоблачают ложь, шантаж, со стороны учащегося, класса / группы учащихся, родителей учащихся, иных педагогов; педагогам / учащимся удается наладить хорошие отношения с иными участниками учебного процесса и родителями учащихся, в результате чего улучшается успеваемость; в любовных отношениях педагог/школьник, как правило, наступает разрыв; в любовных отношениях педагог/студент возможна гармония, как, впрочем, и в романах учащихся. Keywords: media text, school, university, teacher, educator, school, university, student, education. ... In fact, many students like funny atmosphere in their classes. When Reddit, a social media on internet, presented topic about 'best teacher' to its users, the users agreed that humorous teachers could also be categorized as best teachers [8] . However, joyful elements implementation in learning is not identical with jokes. ... Joyful elements implementation on online learning: How it should be? Conference Paper Jan 2023 Online learning has been applied all over the world during COVID-19 outbreak. Until today, that kind of learning has been about one and a half year conducted in schools. This paper wanted to reveal how joyful elements had been implemented on online learning during that time. Hence, we can learn some results which would show us how joyful elements should be implemented on online learning. Qualitative-descriptive method was used in this study. A questionnaire, which related to joyful elements on online learning, had been shared to 2nd semester college students of 2 departments (Natural Science and Physics Education) and 3 classes (A, B and C), but only 39 students sent back their answers. The students’ answers based on their experience during online learning in this outbreak time. The data was processed and the result consisted of: 1) Jokes and appealing stories were the most effective online learning tactics (as a note, images and videos presentations had almost the same number of voters), 2) Learning in the form of assignments from teachers or lecturers were the most discouraged thing for them, 3) Oral lecturing was the most proper learning method for them, 4) Relaxed feeling because no need to attend formally in real classes was the most thing they liked. Based on the result, it was concluded that on online learning students need teachers’ or lecturers’ roles greater than when face-to-face learning in order to gain joyful elements during learning. Besides, students actually enjoy their houses as learning classes but hope that independent learning methods implementations (such as assignments) are reduced during learning. ... Social networks have been used to create influence networks and find support and professional mentoring (Minor et al, 2019). In this way, social networking sites form real learning communities and meaningful education networks, whether between peers (teachers or students) or between teachers and students, as pointed out by several authors Chang-Kredl & Colannino, 2017; Dennen et al., 2020;Greenhalgh et al., 2020;Hasiloglu et al., 2020;Wickramanayake & Jika, 2018). This, in turn, has a beneficial impact on the teaching-learning process (Torphy et al., 2020a, b, c). ... Pre-service Teachers' Use of General Social Networking Sites Linked to Current Scenarios: Nature and Characteristics Social networking sites form part of everyday life in classrooms at all educational levels. Within these, general social networking sites (GSNSs) offer pre-service teachers flexibility, versatility and the possibility of forming educational communities by connecting formal, non-formal and informal settings. This research analyses the nature, intensity, and type of pre-service teachers’ use of such for educational purposes in their initial training in order to detect the most important aspects for improvement. Possible factors shaping behaviour were gender, whether individuals belonged to universities operating online or in person, differences in the types of studies they were undertaking, and the time at which the questionnaire was administered, before or after the COVID-19 health crisis. To this end, we studied how much and with what aims these students use the most widely used GSNSs for educational purposes. To do so, we administered a questionnaire to a total of 812 students from 6 Spanish universities. The results show a preference for WhatsApp, YouTube, and Instagram. In addition, it was found that undergraduate students used them more intensively than postgraduate students. In the case of online universities, there was a greater need to cover affective and emotional aspects than in in-person universities. As in almost all areas, the situation caused by COVID-19 changed the way social networks were used. The findings also show that pre-service teachers consumed more information on social media than what they produced, which leads to a failure to fully exploit social capital and potential job or academic opportunities that could be generated through their own creations. ... Demostró que la relación a través de esta red aumentó las percepciones favorables de los alumnos. También realizaron un ejercicio de reflexión Chang & Colannino (2017) , que abordaron el tema de la imagen proyectada por los profesores ante sus alumnos en Reddit. Establecieron diversos modelos a través de los recuerdos percibidos por los alumnos y concluyeron que las redes tenían una importancia vital a la hora de contribuir a la imagen pública de los docentes. ... The use of social media in education: a systematic review of the scientific literature Teaching-learning strategies have undergone changes in recent years due to the emergence of digital technology and the emergence of social media as mediators and facilitators of new contexts. A review of the scientific literature that has dealt with the use of generic social media in different educational settings during the last ten years is carried out in this work. For this, a documentary analysis of the articles indexed in the Journal Citation Reports (JCR) of the Web of Science has been carried out. For this purpose, a documentary analysis of the articles indexed in the Journal Citation Reports of the Web of Science was carried out, which, after selection, resulted in 160 papers. To do their analysis they have been divided into five areas: digital competence and teaching digital competence; social media as a vehicle for the creation of virtual environments and educational communities; the teaching-learning process mediated by social media as a didactic resource at various educational levels; problems and addictions derived from the use of social media and social movements and demands in the field of education through social media. The results reveal that, although new environments and contexts are being established in this sense and that research on them is intensified, in all categories of analysis there is a wide margin for implementation. © 2021 Research Group Education and Virtual Learning (GREAV). All rights reserved. ... In educational research, investigations of what people discuss on social media platforms, and how they discuss it, have lent critical insight into the methods by which individuals solicit social support, seek information, and develop opinions on educational systems (Relucio & Palaoag, 2018), teacher quality (Chang-Kredl & Colannino, 2017) , and the implementation of curriculum policies (Y. Wang & Fikis, 2019). ... Gifted Education on Reddit: A Social Media Sentiment Analysis Social media continues to be an ever-present part of people’s lives. One of the largest social media websites, Reddit has more than 430 million unique visitors monthly. What is unknown to scholars is how gifted education fits into this modern form of communication. In our research, we examined how gifted education is discussed over Reddit using text mining in combination with sentiment analysis. In addition, our research conditioned sentiment on variables such as self-disclosure, interest, and controversy. We found that, overall, discussion of giftedness was neutral in terms of sentiment and used common language across subreddits. We also found that the language used and distribution of sentiment were similar between self-disclosures and non-self-disclosures. ... Demostró que la relación a través de esta red aumentó las percepciones favorables de los alumnos. También realizaron un ejercicio de reflexión Chang & Colannino (2017) , que abordaron el tema de la imagen proyectada por los profesores ante sus alumnos en Reddit. Establecieron diversos modelos a través de los recuerdos percibidos por los alumnos y concluyeron que las redes tenían una importancia vital a la hora de contribuir a la imagen pública de los docentes. ... El uso de las redes sociales en educación: una revisión sistemática de la literatura científica Las estrategias de enseñanza-aprendizaje han experimentado cambios en los últimos años debido a la irrupción de la tecnología digital y a la aparición de las redes sociales (RRSS) como mediadoras y facilitadoras de nuevos contextos. En este trabajo, se realiza una revisión de la literatura científica que ha tratado el uso de las RRSS genéricas en diferentes ámbitos educativos durante los últimos diez años. Para ello se ha realizado un análisis documental de los artículos indexados en la Journal Citation Reports (JCR) de la Web of Science que, tras proceder a su selección, han dado como resultado 160 trabajos. Para analizarlos se han divido en cinco ámbitos: competencia digital y competencia digital docente; RRSS como elemento vehicular para la creación de entornos virtuales y comunidades educativas; el proceso de enseñanza-aprendizaje mediado por RRSS como recurso didáctico; dilemas éticos, problemas y adicciones derivados del uso de las RRSS y movimientos y reivindicaciones sociales en el ámbito de la educación a través de RRSS. Los resultados revelan que, si bien se están estableciendo nuevos entornos y contextos en este sentido y que se intensifica la investigación sobre los mismos, en todas las categorías de análisis existe un amplio margen de implementación. ... Отже, студіювання українських і зарубіжних психолого-педагогічних розвідок (Ашиток, 2018;Будник, Васянович, Мікулець, 2021;Дудчак, 2020;Куземко, 2020;Савченко, 2018;Fung, Lina, 2020;O'Brien, 2014; Chang-Kredl, Colannino, 2017; Lewis, Holloway, 2019;Palmér, 2016;Arnon, Reichel, 2007 ... PROFESSIONAL IMAGE OF A FUTURE TEACHER OF FOREIGN LITERATURE: FROM LEARNING TO PRACTICE Full-text available Наталія Грицак The article deals with the formation of the professional image of the future foreign literature teacher. The semantic content of the concept of professional image of a teacher is analyzed and supplemented. The structural components influencing the effective formation of the professional image of the future teacher are highlited and described. It is shown that pedagogical skills, visual image, internal appearance are important components in the structure of the professional image of the future teacher. With the peculiarities of the school subject «Foreign Literature», the specifics of forming the professional image of the future foreign literature teacher are justified. The essence of the concept of professional image of the future foreign literature teacher is revealed. The structure of professional image of future foreign literature teachers, which consists of personal, professional, socio-cultural, communicative, motivational-target, result-analytical components is developed. The basic and specific abilities that contribute to the productive formation of the professional image of the future foreign literature teacher are considered. A survey of fourth-year students of the Faculty of Foreign Languages of Volodymyr Hnatyuk Ternopil National Pedagogical University was conducted. The aim of the poll was to find out the attitude of students to future pedagogical activities, to identify their vision of the concepts of «professional image of a teacher», «professional image of a foreign literature teacher». The results showed that the most students consider the concept of professional image of a foreign literature teacher as a set of external, personal and professional qualities that permit the most effectively realise the educational process. The obtained results confirmed that the productive formation of the professional image of the future foreign literature teacher is based on interrelated abilities, particulary basic, linguistic, specific. ... Результаты показали, что на создание положительного имиджа педагога оказывает влияние не только модель отношений между педагогом и воспитанником, но и активность и информационность педагогического работника в социальных сетях [5] . ... POSITIVE IMAGE OF A FUTURE TEACHER PRESCHOOL EDUCATIONAL ORGANIZATION ... Parents are influenced by what he paints and students as well. ' Chang-Kredl and Colannino (2017) affirmed that adults in general tended to have uninformed preconceptions of teaching and teachers. Figure 2 depicts a male teacher clinging for safety to a logo for 'World Teachers' Day' while a male student waits below wielding a bat to attack the teacher. ... Pre-service teachers' reflections on teaching images in Saudi media This inaugural Saudi Arabian-based (SA) study explored how social media images and cartoons can influence the professional identity of pre-service teachers (PSTs) measured by their reflections on self-selected images of teachers and teaching in Saudi media. PSTs (n = 30) were enrolled in a teacher education program in a faculty of education in a public university in the Eastern province of SA (convenience sampling). Findings from thematically analyzing 30 reflective assignments, nine semi-structured interviews, and a focus group (n = 9) revealed four themes: (a) a pervasive negative stereotype; (b) violence associated with male teachers and students; (c) criticism of the education and administrative system; and (d) suggestions of eroding teacher authority. Findings affirm the imperative that teacher education programs intentionally sensitize PSTs to the benefits of critically deconstructing media images. This will help stave off negative connotations of teachers and make teaching become part of future teachers’ professional identities and the SA collective memory. ... Therefore, the internet is the optimal context in which to study this group. Past research has used data obtained from Reddit, the social media platform used in this study, to provide insight into a range of topics including forums dedicated to teachers, gout, feminism, sexuality, and eating disorders (Chang-Kredl & Colannino, 2017; Derksen et al., 2017;Massanari, 2019;Robards, 2018;Sowles et al., 2018). Reddit is a social media and content sharing website in which individual users can join subsections of the website (termed "subreddits") that include forums relevant to their interests. ... Taking the black pill: An empirical analysis of the “Incel”. Article Jan 2021 Tessa L Dover Men who perform hybrid masculinities claim distance from masculine hegemony while actually reproducing patriarchal power. In this article, we analyze the Incel community through a lens of hybrid masculinities, as originally proposed by Ging. The Incels are a self-proclaimed group of beta males who claim a position at the bottom of the social hierarchy due to their continued romantic and sexual rejection by women. We performed a content analysis of a sample of 400 top-rated posts from the Incel forum on Reddit called r/Braincels. Incels performed a specific version of hybrid masculinity in which they claim lower status than women while also calling other men weak and powerless using comparisons to women. Incels also showed negative outcomes associated with these masculinities, as evidenced by posts reporting violent hatred of both themselves and women. This research provides insight into the ways that masculinities develop in online contexts. (PsycInfo Database Record (c) 2021 APA, all rights reserved) Public Significance Statement: Posters on a Reddit forum dedicated to the involuntarily celibate (“Incel”) community used the language of social justice to claim that they do not have access to traditional masculine status. However, they also looked down on other men who do not conform to these same masculine standards. Individuals in this community also discussed feelings of violence and hatred directed at women and themselves. (PsycInfo Database Record (c) 2021 APA, all rights reserved) ... The findings have several implications for teachers' education programs and point to the potential utility of applying psychological theory when examining the processes by which student teachers develop a professional teaching identity. Chang-Kredl, S. and Colannino, D. (2017) highlight the role of social media in constructing the public image of the teacher. Navrotska, M. (2019) studies the problem of developing the professional image of the teacher in the system of post-graduate pedagogical education. ... Future English Teachers’ Professional Image Forming Full-text available The aim of the research is to substantiate, design pedagogical system of future English teachers’ professional image forming at pedagogical institutions of higher education and verify experimentally its effectiveness. The research was conducted within the mixed methodology in two stages: theoretical and empirical. In the context of theoretical stage the essence of the professional image of English teacher was defined, based on personality’s role theory, its structure was clarified and pedagogical system of its targeted forming at pedagogical institutions of higher education was developed. Research findings made it possible to identify pedagogical factors influencing the future English teachers’ professional image forming as well as pedagogical conditions, which allow achieving positive results in the process of its formation at pedagogical higher educational institutions. Obtained theoretical results have been introduced into the educational process of pedagogical higher educational institutions of Ukraine within the pedagogical experiment (empirical stage) in order to be experimentally testified. Statistical processing of obtained experimental data ensured the validity and objectivity of the research results and verified the effectiveness of the proposed pedagogical system. The research results point to the fact that the proposed pedagogical system provides the high quality of image training of future English teachers’ at higher pedagogical educational institutions. Keywords: teacher; image; professional image; professional training; pedagogical system. ... Existing research on Reddit has explored a variety of topics, including Redditors' self-described best and worst memories of their teachers (Chang-Kredl and Colannino 2017) , analyses of the vernacular criticism of memes on r/MemeEconomy (Literat and van den Berg 2019), and studies that showed that participation on Reddit tends to be motivated by social and entertainment factors associated with the site (Bogers and Wernersen 2014). ... Continuing a community of practice beyond the death of its domain: examining the Tales of Link subreddit Article Jeyoung Oh Communities of practice (COPs) have become increasingly important across a wide range of casual and business contexts, including innovative technical product development, refining skill sets for competitions, or establishing corporate strategies. Members of such communities forge interpersonal relationships and develop sophisticated practices around a shared domain, either interdependently contributing to a joint product or advancing as individuals. However, prior research has not examined attempts to sustain a COP whose domain has been terminated. Such cases have become increasingly common, especially as COPs emerge around ephemeral media products such as mobile games. A recent example, Tales of Link, based on the popular ‘Tales of’ Japanese role-playing game series, shut down on March 2018, but members of the subreddit devoted to it have attempted to keep the community active. As such, a computational text analysis of 33,476 comments and 1,000 posts on the r/talesoflink subreddit was conducted to assess how community members attempt to sustain COPs beyond the lifespan of their original domains. The findings offer insight into the discourse within COPs, the strategies and success rates of survival that may vary by such organisations, conversations surrounding the ever-changing gacha game model, and the potential changes in members’ messages and behaviours. ... An educator is who will do anything to help their students whether it is a positive or negative feedback one who will transform the students learning. In terms of qualities, a teacher's description reflects as admirable in terms of intelligence ("knowing about the teaching field", engaging "able to capture the attention of the learner", "funny, made-up story problems", "storytelling",) dedicated, easy-going, strict but fair (Chang-Kredl, Colannino, 2017) . ... Philosophy of Learning Full-text available Education today is challenging. The classroom is no longer the primary learning place, and knowledge is obtained now from anywhere and everywhere. Technology has changed how people learn, the way the learner wants to learn, and the environment they prefer to learn. The educator’s role has also shifted from who provides resources, monitors progress, and encourages students to problem solve; hence education environment needs to be creative, inventive and room for discoverers. ... The image of a teacher is a concept, influenced by many factors: social processes, economic and political events, and cultural development of a certain state. Image forming in society is the subject of a number of researches, in which the authors have proved the influence of social processes and cultural development on the requirements for a teacher (the works by U. Ansari 11 , S. Arnon [1], D.P. Britzman [2], S. Chang-Kredl [3] , P.A. Ertmer [4], N.L. Louie [11] -personal experience, human biography; in culture this aspect is shown in created art works (in sculpture, painting, literature); ... Chang-Kredl & Colannino, 2017)

https://www.researchgate.net/publication/316606354_Constructing_the_image_of_the_teacher_on_Reddit_Best_and_worst_teachers

(PDF) Special Section: The Bioarchaeology of Postmortem Agency The Bioarchaeology of Postmortem Agency: Integrating Archaeological Theory with Human Skeletal Remains PDF | Taking the social agency of dead bodies as its main theme, this introduction discusses the articles in this special section on the bioarchaeology... | Find, read and cite all the research you need on ResearchGate Special Section: The Bioarchaeology of Postmortem Agency The Bioarchaeology of Postmortem Agency: Integrating Archaeological Theory with Human Skeletal Remains January 2014 Cambridge Archaeological Journal 24(3):429-435 DOI: 10.1017/S0959774314000584 Authors: Taking the social agency of dead bodies as its main theme, this introduction discusses the articles in this special section on the bioarchaeology of post-mortem agency and discusses theoretical concerns relevant to the (bio)archaeological analysis of agency. In particular, the argument that only biological living persons have social agency or impact the direction of social processes, or the decisions of others is challenged. The challenges of defining and archaeologically detecting such agency of past humans as well as socially alive entities such as ghosts, corpses, relics and totem heads are discussed. A review of the ways investigations of the agency of the dead might unify bioarchaeologists and further integrate social archaeology into future research is also presented. It is argued that a holistic anthropological approach to humans and their ability to impact their surroundings is not complete if the symbolic and material effects of the dead are not considered. Special Section: The Bioarchaeology of Postmortem Agency (Guest Editors: John J. Crandall and Debra L. Martin) The Bioarchaeology of Postmortem Agency: Integrating Archaeological Theory with Human Skeletal Remains John J. Crandall & Debra L. Martin T aking the social agency of dead bodies as its main theme, this introduction discusses the articles in this special section on the bioarchaeology of post-mortem agency and discusses theoretical concerns relevant to the (bio)archaeological analysis of agency . In particular , the argument that only biological living persons have social agency or impact the direction of social processes, or the decisions of others is challenged. The challenges of defining and archaeologically detecting such agency of past humans as well as socially alive entities such as ghosts, corpses, relics and totem heads are discussed. A review of the ways investiga- tions of the agency of the dead might unify bioarchaeologists and further integrate social archaeology into future resear ch is also presented. It is argued that a holistic anthropological symbolic and material effects of the dead are not considered. Bridging social theory and bioarchaeology Bioarchaeological methodologies, conceptual frame- works and approaches to ancient human remains have become increasingly sophisticated in terms of adding breadth and scope to the kinds of questions that can be answered with skeletal data. Combining overlap- ping fields such as forensic sciences, clinical radiogra- phy , skeletal biology , biochemistry , archaeology and history , bioarchaeology has emerged as a robust spe- cialty that integrates methods, data and theory drawn from across the social and natural sciences (Buikstra &B e c k 2009 ;M a r t i n et al. 2013 ). Squarely anthropological in its aims and ap- proaches, bioarchaeology seeks to understand the hu- man condition in all its diversity and use knowledge of such variation to explain past human behaviour. The use of contextualized data derived from human remains has focused on two major types of questions. First, osteological data has been marshalled to un- derstand the causes and consequences of major so- cial and/or technological shifts. The often cited ex- opathology at the Origins of Agriculture , edited by Cohen and Armelagos ( 2013 [1984]). In this foundational vol- ume, nineteen studies systematically investigated the consequences of transitioning from foraging to farm- ing on human health and nutrition. This approach, focusing on comparative population-level data sets with temporal variability , has also generated a large amount of data regarding the impacts of other adap- tive transitions in human history . Scholars have inves- tigated, for example, the consequences of colonialism and globalism (e.g. Baker & Kealhofer 1996 ; Hutchin- son 2007 ; Larsen & Milner 1993 ) and the impact of imperialism and state development (T ung 2012 ). These studies demonstrate how powerfully bioarchaeology can synthesize a vast array of data sets in order to examine the consequences of large- scale social and technological shifts in human his- tory . They provide temporal depth to contemporary ethnographic data. These studies can be integrated alongside other kinds of data derived from fields such as public health, environmental science, political Cambridge Archaeological Journal 24:3, 429–435 C doi: 10.1017/S0959774314000584 Received 5 Jul 2013; Accepted 5 Oct 2013; Revised 26 Jan 2014 John J. Crandall and Debra L. Martin science and other issues facing us today , and used to model how human communities can best address large-scale changes facing our species. New studies of this kind continue to emerge, addressing issues like climate change (Robbins Schug 2011 ) and the evo- lution of communicable disease (e.g. Powell & Cook 2005 ; Roberts & Buikstra 2008 ), which are only becom- ing more pressing as globalization and industrialism intensify . Scholarship in bioarchaeology has also begun to draw more explicitly on social theory . Rather than gen- erate large, comparative data sets of population-level studies, many scholars are integrating contextual in- formation drawn from history and sociology as well as other fields within anthropology , such as material cul- ture studies and ethnography , to produce rich contex- tual analyses of individuals or subgroups within re- gional and local contexts. These studies, which are of- ten termed ‘social bioarchaeology’ (Agarwal & Glen- ncross 2011 ), highlight social actors more explicitly than the population-level studies mentioned above (Stodder & Palkovich 2012 ). Population-level health data, mortuary data and information regarding pat- terns of injury , social norms and other data garnered from larger-scale analysis are woven into a contextual mosaic against which particular subgroups or indi- viduals can be examined. These studies often address issues of identity (Knudson & Stojanowski 2009 ), per- sonhood and embodiment (Sofaer 2006 ). In explicitly ‘peopling the past’, they provide insight into how ancient communities operated on a more day-to-day level. Examples of such work include studies focused on gender roles (Grauer & Stuart-Macadam 1998 ), children’s social roles (Thompson et al. 2014 ), inequal- ity (Goodman 1998 ), and how occupation or social identity impacts one’s risk of death (Crandall et al. 2013 ; DeWitte 2010 ; Mathena et al. 2013 ). These two approaches, however , are by no means opposed and much research seeks to use data obtained from hu- man skeletal remains to address many of these issues in combination. At the heart of these two major research foci in bioarchaeology is the issue of agency . For schol- ars investigating large-scale population-level health, it is difficult to consider individual actors or the mi- crocosms of a particular community . As Hodder has noted, the processes being investigated in such stud- ies, ‘are the processes of the long term, the rise and fall of complex political systems, the slow transfor- mation of subsistence technologies, the longue dur ´ ee of mentalit ´ es , the battle-ship curve of styles, and so on’ (Hodder 2000 , 21). For social bioarchaeologists, how- ever, the inter est is in people, their experiences, and how they cope and behave in response to such pro- cesses. Both sorts of research, as well as projects meld- ing the two scalar approaches are invested in humans, how they take action, what constrains how they en- gage social change or cultural transformation. As we pondered the nature of human action, we realized that bioarchaeologists were ideally suited to investi- gate agency owing to the nature of our data sets and our ability to highlight the experiences of individuals in the context of social change. This desire to bring bioarchaeology to the table in discussing agency and its utility inspired these papers. In thinking about agency , the group of authors in this special section shared an interest in the potential agency of the dead – whether as bony objects manipu- lated by the living or spirits seen as having their own agendas. Thinking of the active social lives of memento mori for example, saints’ relics, or the active roles that ancient Chinchorro people invested in the mummies they fed, cared for and interacted with reminded us that W estern dualities of life and death are blurred in other cultures and times (e.g. Rakita et al. 2008 ). W e were moved, at this same time, also to question whether corpses were necessarily even seen as indi- vidual people in other cultures and became interested in the materiality of dead bodies and the political af- terlives of human bone and other tissues as they be- come entangled in larger political struggles. In think- ing of these issues, we borrowed the term postmortem agency from cultural anthropologist W ilreker ( 2007 ), and invited other bioarchaeologists to join us in think- ing through how we might bioarchaeologically iden- tify and theorize how dead bodies often shape social relations, political and social behaviour and influence agency of the dead, we hoped to enrich studies of ancient politics, inequality , warfare and social change by considering ideology , living and non-living actors and the ways bodies become social tools. In short, we sought to infuse new social theory and models of hu- man behavior into bioarchaeology through focusing on the paradox of life after death. The studies in this special section deal specif- ically with post-mortem agency and they demon- strate that the living are not the only social agents influencing the direction of social processes, the fate of social systems and the evolution of communities. Cross-cultural and temporal mortuary studies in re- cent years have revealed the variety of ways that the border between the living and dead, corpse and per- son, social agent and biological entity are fluid, some- times liminal and often subject to constant cultural negotiation. Recent scholarship across bioarchaeol- ogy , medical anthropology and ethnohistory high- lights the cross-cultural flexibility between life and 430 The Bioarchaeology of Postmortem Agency quant 2002 ). Further, such studies highlight the r eality that the agency of the dead is as present as the self- determination and social change created by the living. Such post-mortem agency is seen in ancestor venera- tion, the power of saints’ relics, and the ways in which dead bodies and their parts fuel black markets, alter- native economies, political change, religious systems and living communities (e.g. V erdery 1999 ). The case studies presented here seek to under- stand the consequences of beliefs in and sustained interaction with the supernatural, the afterlife and the undead. These studies interrogate how humans make choices and take action. These studies most im- portantly provide a diverse set of frameworks for how bioarchaeology can integrate data and theories generated from both large- and small-scale projects to advance explanations for human behaviour. Ad- ditionally , papers in this special section seek to en- gage bioarchaeology with ongoing theorizing of the social lives of non-human actors such as spirits and objects. Increasingly , anthropology has seen spirits, objects, ghosts and ancestors not only as symbols but ırito Santo 2014 ; Hodder 2012 ; papers in Krmpotich et al. 2010 ). By asking bioarchaeology to engage theories of social agency , embodiment, and materiality , we hoped to help bring bioarchaeology into further conversation with disciplinary advancements in archaeology and elsewhere. While each paper in this section may differ in how it defines and operationalizes the concept of agency , each is careful to define agency by both draw- ing on and advancing models proposed in archaeology and elsewhere (e.g. Crossland 2009 ; Dobres & Robb 2000 ; Hallam 2010 ; Krmpotich et al. 2010 ). Our intent is to further the development of bioarchaeology in- formed by social theory as well as to challenge our readers to consider the agency of the dead in their own reconstructions of the past. Through peopling the past, and challenging the very definition of per- son by blurring the lines between living and ‘dead’ social agents, we hope to show the potential of theory- infused bioarchaeology and to enliven a diverse array of anthropological research. Additionally , we see these nature and utility of the agency concept in anthropol- ogy more broadly (e.g. Ortner 2006 ). In light of this, we welcomed an array of approaches and case studies that utilized data seen as often tangential to bioarchae- ology , such as architectural data (V alasco, this issue), historical records (Crandall & Harrod, this issue), oral history (Novak, this issue) alongside data from prior work and new analyses. Conclusion: post-mortem agency In archaeology , as in the social sciences broadly , agency is at once ‘everything and nothing’ (Dobres & Robb 2000 , 3). As a theory , a buzzword and the foundation of an approach, it remains one of the most commonly used constructs in recent social archaeolog- ical and post-processual projects (see Dobres & Robb 2000 for a review). In seeking to engage bioarchaeolo- gists around agency , we encouraged each contribution in this section to define and detect the agency of the dead differently in each context. This enabled a con- versation to emerge between the articles in this special section that show various models of how consider- Bioarchaeology has been increasingly influenced by social theory , research on identity and political economy , and post-processual theory in archaeology (see Agarwal & Glenncross 2011 ). This engagement has been accompanied by a re-evaluation of what ex- actly archaeologists, and by extension osteologists, study . Hodder ( 2012 ), Olivier ( 2011 ), Olsen and col- leagues ( 2012 ), and Sofaer ( 2006 ) have provided sum- maries of this shift in which archaeologists have in- creasingly seen archaeology as the study of things, or objects engaged in webs of power. This shift, as Sofaer summarizes, includes a reorientation towards ancient remains as not only persons but objects . The move towards seeing past peoples as objects that might im- pact the action of others through their ‘thingness’ is nicely summarized in this issue by T ung as one reason why considering the agency of dead bodies is timely in terms of both increasingly theorizing bioarchaeol- ogy but also in advancing archaeological theory more broadly . Y et, this move towards the study of the materi- ality of bodies, or bodies as artefacts, is not an aban- human culture, behaviour and experience (see also Geller & Suri, this issue). Indeed, social bioarchae- ology would not be social unless it emphasized the study of bodies as objects who continued to engage, influence, confine or structure other social agents. This assertion is at the heart of the articles in this spe- cial section. Thus, at the most basic level, postmortem agency is defined as the ability of dead bodies (in their new guises as objects, spirits, relics or other symbols, forms or identities) to engage, influence, confine or structure the behaviour of the living whether directly or indirectly . Some scholars contributing to this spe- cial section closely follow Bourdieu ( 1977 ) in seeing agency as the ability of subjects to resist existing social structures and act outside of them. Others, following 431 John J. Crandall and Debra L. Martin Gell ( 1998 ) and Robb ( 2004 ), see agency as forming two major categories: primary (or conscious )a n ds e c - ondary (or effective ) agency . Here, a distinction is made between actors who can act on their own versus ob- jects or actors who only serve to extend the agency theorized through the lens of this second model of agency (Harper 2010 ). T ung’s article, influenced by this work, provides an example of the application of this model in a bioarchaeological context. She re- engages debates about the importance of intention- ality among actors and draws distinctions between Bourdieu’s practice and the kinds of agency defined in archaeological models. Arguments about whether bodies, as objects, have their own inherent primary agency remain a theoretical exercise that is dependent on one’s per- spective. As T ung notes in her piece on dismembered Andean bodies and their social lives, from an etic perspective, dead bodies only have the agency that the living afford them. Y et, from an emic perspective, and from the standpoint of materiality theory , bodies, their presence, absence and spectre may exhibit pri- mary agency . For Crandall and Harrod, and others in this special section, bodies have an inherent ability to shift the emotions of the living in their presence. Their very presence transforms landscapes, requires that the living approach their resting places differently , ies can alter human behaviour. These scholars draw implicitly on Foucault ( 1995 [1977]) and Bourdieu ( 1977 ) in their arguments by implying that it is impos- sible to fully distinguish whether human behaviour is what produces the social agency of dead bodies or whether dead bodies are the anchors of fields of power/social influence that shape human action. Largely , it seems that this relationship is recursive and even Gell noted that objects can indeed be primary agents (1998, 17–19; see also Novak, this issue). Another challenge for contributors in this sec- tion was that not only is agency hard to define, it is hard to identify archaeologically (Barrett 2000 ). Each contribution in this piece identifies the material con- sequences of post-mortem agency differently . Each scholar, however , draws greatly on contextual infor - mation, temporal perspectives and models from past archaeologies of agency to track the impacts of dead bodies in each case study . In most cases, the bod- ies that ‘speak’ to the living, act through them and around them, and are important figures in any be- havioural model, we would argue, are often the ex- hated or unique in life or comprise a class of un- happy spirits, these dead are given the room to per- form largely because of the way they are seen by the living. Whether any corpse has the agential potential seen among the characters in these articles remains a question for bioarchaeology as theories of agency become a part of the toolkit used in our research (see Arnold, this issue). What emerges from the articles presented here is the realization that dead bodies can have mul- tiple, often unexpected effects, on the behaviour of the living. Insights from V erdery’s work on the post- mortem lives of eastern European corpses (1999) are useful here. Bodies, as objects, are silent. Bodies ‘don’t talk much on their own’ (V erdery 1999 , 29) and thus can become easy tools through which the living can achieve their ends. Corpses do, however, “speak” in predictable ways (whether they are the tools of others or not) once an emic perspective is taken. Through integrating ethnographic, historical and other contex- tual data, the articles in this special section highlight these messages and the ways in which the living re- The articles in this special section The eight articles in this special section each uniquely illustrate the utility of the post-mortem agency con- cept in bioarchaeology . T iffiny T ung, in opening the special section, explores the agency of beaten and dis- membered bodies in the ancient Andes. Her analysis of Andean mummies, mutilated bodies and trophies illustrates the many ways that body parts, and whole bodies, both had rich social and political afterlives in among the W ari and their successors. Her careful ar- ticulation of agency orients us towards the ways that bodies serve as tools in political conflict. Y et, the ways in which bodies may have re-oriented communities and subjugated communities in the past are difficult to predict and demonstrate that even as tools the bod- ies of the dead act outside the will of those who seek to use them. Matthew V elasco, also working in an- cient Peru, focuses on the ways that bodies and their resting places serve to unify communities and signal, if not produce, community membership. Emphasized in this piece is the need to situate agency in temporal, social and spatial contexts. This work thus extends 2000 ) into mortuary contexts. Both articles highlight the need to consider spatial context in bioarchaeolog- ical reconstructions. This is a trend carried through- out the special section and certainly reflects the in- creasing incorporation of contextual information into bioarchaeology more broadly . Betsinger and Scott investigate the multifaceted agency of vampires in post-medieval Poland. Care- fully examining how the spectre of the vampire was 432 The Bioarchaeology of Postmortem Agency used by the church, by villagers and also how vam- pire corpses demanded post-mortem ritual engage- ment, they illustrate how dead bodies can be the simultaneous tools of many competing parties. No- vak approaches the bodies and personal effects of the dead as objects that co-construct personhood, action and social structures alongside human actors. Seeing the dead as only one type of artefact which circulate between and around people, as well as anchor the movements of humans, here brings bioarchaeology into conversation with anthropology’s latest engage- analyses of the 1857 Mountain Meadows Massacre (Novak 2008 ) as well as new ethnographic and life history data, Novak also provides a model of how ethnography and bioarchaeology can be paired to ex- amine the long-term effects of massacres and violence. Her attention to the absence as well as presence of hu- man bodies in social contexts provides great insight into how bioarchaeologists can only benefit from con- sidering objects and bodies together rather than in isolation. Crandall and Harrod also investigate the social afterlives of individuals in the historic USA. T ak- ing as their focus the death and social afterlives of two of V egas’ earliest settlers, the notoriously mur- dered Kiel brothers, they investigate the ways that the various spectres of the body , the brother’s status as ghosts and the absence of their bodies at the remain- ing ranch property have impacted city planning in contemporary north Las V egas. Their analysis focuses on how dead bodies haunt places as well as political debates. This haunting they argue produces death- scapes (Maddrell & Sidaway 2010 ; Y oung & Light shape the outcomes of political struggles amongst the living. Their attempt to focus attention towards how the dead are thus mobilized in place-making seeks to demonstrate how the dead can live quite lengthy social afterlives in which they may be refash- ioned as the living contest the ‘truth’ of their former lives. The final case study by Pamela Geller and Mi- randa Suri focuses on the ethical questions raised by considering the obligations that bioarchaeologists do, or perhaps should, have towards the dead. Geller and Suri examine the messy temporalities of the bones that bioarchaeological research examines. Consider- ing both life and ‘death histories’ (Geller 2012 )t h a t bodies leave open to research, the authors present a reflexive critique of bioarchaeological practice. Build- ing on their own work in Mesoamerica and contem- porary politics in the region, they emphasize the ways bodies are sites of memory-making, identity work and a space for descendant communities to engage their past and present social and political positions through the dead. Geller and Suri remind us that bioarchaeo- bodies of the dead but actively constructs, returns and re-shapes the meaning, and subjectivity , of the formerly living. They close their meditations on bioar- chaeological practice, by asking us to consider the eth- ical imperatives that this raises. Asking bioarchaeol- ogists to consider the political effects of their work, the nature of the data we generate and trans-temporal ways the dead emotionally move the living, this piece provides scholars with a provocative call towards a more ethical, positioned bioarchaeology . This special section comes to a close with com- mentaries by Molly Zuckerman, Sarah Marthena and Kelly Kamnikar, and Bettina Arnold. In ‘Recovering the Body Politic’, Zuckerman and colleagues follow Geller and Suri in considering the ethical and profes- sional implications that working with the dead have for bioarchaeologists. They argue that bodies are a unique source of data regarding political processes and power relations that are often obscured in other sources. Thus, bodies, as social subjects embedded in historically unique political economies, demand a relational ethics that other data sources in archaeol- bodies might help us bridge the present and past in ways that realistically impact people, particularly oppressed communities, today . Drawing on embod- iment theory , and advancing discussions of ethics across anthropology , they present a model for un- derstanding how bioarchaeology can speak truth to power and recover the agency and subjectivity of the dead through an examination of a sample of remains from a historical insane hospital in Mississippi. Their example provides a model for future bioarchaeolog- ical research that can help uncover , and redress, his- torical inequalities. T ogether, the articles in this special section raise the question as to whether the dead ever die (socially- speaking) or whether most dead are, as Bettina Arnold notes in her closing remarks, simply only ‘mostly dead’. Indeed, as objects, actors, ghosts, vampires, data, objects, or symbols, the articles in this special section show that the dead have a very active hand in the ways we live, think and even conduct archaeologi- cal research. Arnold reminds us that investigating the moves of the dead requires contextual, theoretically- ing the cases in this special section with other exam- ples of the dead in global politics and recent archae- ological work, Arnold reminds us of the complexities of our collective work. 433 John J. Crandall and Debra L. Martin In sum, this special section emphasizes that bioarchaeology has an active, unique role to play in theorizing human behaviour. Thr ough examinations of the social histories of the dead, these articles seek to inspire a more anthropological bioarchaeology and il- lustrate the growing diversity of methodological and theoretical approaches increasingly being applied in bioarchaeology and its sister disciplines, mortuary ar- chaeology and forensic anthropology . Through case studies derived from across space and time, these ar- ticles reveal that a truly holistic anthropological ap- proach to humans and our ability to transform our worlds is not complete if we do not consider the very real and symbolic ways that the dead influence, and sometimes control, the living. Acknowledgements W e would like to thank all of the participants of the 2012 Department of Anthropology University of Nevada, Las V egas Las Vegas, NV 89154-5003 USA 4505 S. Maryland Pkwy Mailstop 455003 Department of Anthropology University of Nevada, Las V egas Las Vegas, NV 89154-5003 USA Email: [email protected] References Agarwal, S.C. & B.A. Glenncross, 2011. Social Bioarchaeology . New Y ork (NY): Wiley-Blackwell. Baker, B.J. & L.Kealhofer, 1996. Bioarchaeology of Native Amer- ican Adaptation in the Spanish Borderlands . Gainesville (FL): University Press of Florida. Barrett, J.C., 2000. A thesis on agency, in Agency in Archae- ology , eds. M.A. Dobres & J. Robb. New Y ork (NY): Routledge, 61–8. B l a n e s ,R .&D .E s p ´ ırito Santo, 2014. The Social Life of Spirits . Chicago (IL): The University of Chicago Press. Bourdieu, P ., 1977. Outline of a Theory of Practice , trans. R. Nice. Cambridge: Cambridge University Press. Buikstra, J.E. & L.A. Beck, 2009. Bioarchaeology: the Contextual Analysis of Human Remains . New Y ork (NY): Academic Press. Cohen, M.N. & G.J. Armelagos, 2013 [1984]. Paleopathology of the Origins of Agriculture . Reprint edition. Gainesville (FL): University Press of Florida. Crandall, J.J., D.L. Martin & J.L. Thompson, 2013. Illness, identity and the Mesoamerican infant: a regional perspective. American Journal of Physical Anthropology 150(S56), 105 [abstract]. Crossland, Z., 2009. Of clues and signs: the dead body and its evidentiary traces. American Anthropologist 111(1), 69–80. DeWitte, S.N., 2010. Sex differ entials and frailty in me- dieval England. American Journal of Physical Anthro- pology 143(2), 285–97. Dobres, M.A. & J. Robb, 2000. Agency in Archaeology .N e w Y ork (NY): Routledge. Foucault, M., 1995 [1977]. Discipline and Punish: the Birth of the Prison , trans. A. Sheridan. New Y ork (NY): Vintage. Gell, A., 1998. Art and Agency: an Anthropological Theory .O x - ford: Oxford University Press. Geller, P .L., 2012. Parting (with) the Dead: body partibility as evidence of ancestor veneration. Ancient Mesoamerica 23(1), 115–30. Goodman, A.H., 1998. The biological consequences of inequality in antiquity , in Building a New Biocul- tural Synthesis: Political-economic Perspectives on Hu- man Biology , eds. A.H. Goodman & T.L. Leather - man. Ann Arbor (MI): University of Michigan Press, 147–69. Grauer, A.L. & P . Stuart-Macadam, 1998. Sex and Gender in Paleopathological Perspective . Cambridge: Cambridge University Press. Material Culture 15(4), 465–92. Harper, S.A., 2010. The social agency of dead bodies. Mor- tality 15(4), 308–22. Hodder, I., 2000. Agency and individuals in long-term pro- cesses, in Agency in Archaeology , eds. M.A. Dobres & J. Robb. New Y ork (NY): Routledge, 21–33. Hodder, I., 2012. Entangled: an Ar chaeology of the Relationships between Humans and Things . New Y ork (NY): Wiley and Sons. Hutchinson, D.L., 2007. T atham Mound and the Bioarchaeology of European Contact: Disease and Depopulation in Central Gulf Coast Florida . Gainesville (FL): University Press of Florida. Knudson, K.J. & C.M. Stojanowski, 2009. Bioarchaeology and Identity in the Americas . Gainesville (FL): University Press of Florida. Krmpotich, C., J. Fontein & J. Harries, 2010. The substance of bones: the emotive materiality and affective presence of human remains. Journal of Material Culture 15(4), 371–84. 434 The Bioarchaeology of Postmortem Agency ological Responses to Conquest . New Y ork (NY): Wiley- Liss. Maddrell, A. & D. Sidaway, 2010. Deathscapes: Spaces for Death, Dying, Mourning and Remembrance . Burlington: Ashgate Publishing. Martin, D.L., R.P . Harrod & V . Per ´ ez, 2013. Bioarchaeology: an Integrated Approach to Working with Human Remains . New Y ork (NY): Springer. Mathena, S.A., M.K. Zuckerman & N.P . Hermann, 2013. Frailty , social identity and treponemal disease in the southeastern US. American Journal of Physical Anthro- pology 150(S56), 192 [abstract]. Novak, S.A., 2008. House of Mourning: a Biocultural History of the Mountain Meadows Massacre . Salt Lake City (UT): Utah University Press. Olivier,L., 2011. The Dark Abyss of Time: Archaeology and Mem- ory , trans. A Greenspan. Plymouth: AltaMira Press. Olsen, B., M. Shanks, T. W ebmoor & C. W itmore, 2012. Ar- chaeology: the Discipline of Things . Berkeley (CA): Uni- versity of California Press. Ortner, S.B., 2006. Anthr opology and Social Theory: Culture, versity Press. Powell, M.L. & D.C. Cook, 2005. The Myth of Syphilis: the Natural History of T reponematosis in North America . Gainesville (FL): University Press of Florida. Rakita, G.F .M., J.E. Buikstra, L. Beck & S.R. Williams, 2008. Interacting with the Dead: Perspectives on Mortuary Ar- chaeology for the New Millennium . Gainesville (FL): Uni- versity of Florida Press. Robb, J., 2004. The extended artefact and the monumental economy: a methodology for material agency, in Re- thinking Materiality: the Engagement of Mind with the Material World ,e d s .C .G o s d e n ,C .R e n f r e w&E .D e - Marrais. (McDonald Institute Monographs.) Cam- bridge: McDonald Institute for Archaeological Re- search, 131–9. Robbins Schug, G., 2011. The Bioarchaeology of Climate Change: a View fr om South Asian Prehistory . Gainesville (FL): University Press of Florida. Roberts, C.A. & J.E. Buikstra, 2008. The Bioarchaeology of T uberculosis: a Global V iew on a Reemerging Disease . Gainesville (FL): University Press of Florida. Scheper-Hughes, N. & L. W acquant, 2002. Commodifying Sofaer, J.R., 2006. The Body as Material Cultur e: a Theoretical Osteoarchaeology . Cambridge: Cambridge University Press. Stodder, A.L.W . & A. Palkovich, 2012. The Bioarchaeology of Individuals . Gainesville (FL): University Press of Florida. Thompson, J.L., M.P . Alfonso-Durruty & J.J. Crandall, 2014. T racing Childhood: Bioarchaeological Investigations of Early Lives in Antiquity . Gainesville (FL): University Press of Florida. T ung, T.A., 2012. V iolence, Ritual and the Wari Empire: a Social Bioarchaeology of Imperialism in the An- cient Andes . Gainesville (FL): University Press of Florida. V erdery , K., 1999. The Political Lives of Dead Bodies: Reburial and Postsocialist Change . New Y ork (NY): Columbia University Press. Wilreker , B.C., 2007. Living and Dying in an American Neo- pagan Community . Unpublished MA dissertation, Department of Anthropology , University of Nevada, Las V egas. Y oung, C. & D. Light, 2013. Corpses, dead body politics and Dr Petru Groza. T ransactions of the Institute of British Geographers 38(1), 135–48. Author biography John J. Crandall is a PhD Student in the Department of An- thropology , University of Nevada, Las V egas. His research interests in historical inequality , identity and violence have resulted in the publication of T racing Childhood: Bioarchae- ological Investigations of Early Lives in Antiquity (University Press of Florida) which he co-edited. Debra L. Martin is the Lincy Foundation Professor of An- thropology at the University of Nevada Las V egas. Her in- terests in violence and social control have resulted in the recent publication of Bioarchaeology of V iolence (University Press of Florida), and Bioarchaeological and Forensic Perspec- tives on Violence (Cambridge University Pr ess), both of which she co-edited. 435 ... Interaction with the dead can take place 'face to face' or over long distances, mediated by tangible or intangible agents. As a consequence, the study of postmortem interaction is related to a vast field of interconnected research strands: dying, care, and death (Das & Han, 2016;Farman, 2020;Green, 2008;Laqueur, 2015;Robben, 2018;Troyer, 2020); memory and commemoration (Hallam & Hockey, 2001;Jones, 2007;Koselleck, 2002;Williams, 2006); postmortem agency (Crandall & Martin, 2014; Crossland, 2017;Harper, 2010); bodies and embodiment (Hamilakis et al., 2002;Houston et al., 2006;Scheper-Hughes & Lock, 1987;Sofaer, 2006); personhood, ontologies, and assemblages (Alberti, 2016;Carr, 2021;Fahlander, 2020;Fowler, 2004Fowler, , 2013Harrison-Buck & Hendon, 2018); emotion and politics (Ahmed, 2004;Mark, 2010;Stavrakopoulou, 2010;Tarlow, 2012;Verdery, 1999); formation processes, object biographies, and object networks (Appadurai, 1986;Knappett, 2011;Meskell, 2004;Schiffer, 2010;Walker, 1995); landscape and monuments (Bradley, 1998;Díaz-Guardamino et al., 2015;Holtorf, 1998;Tilley, 1994); the agency of objects and materials (DeMarrais et al., 2004;Dobres & Robb, 2000a;Fowler & Harris, 2015;Gell, 1998;Hodder, 2012;Jones & Boivin, 2010;Joyce & Gillespie, 2015;Knappett & Malafouris, 2008;Latour, 2005); digital postmortem identity and legacy on social media (Brubaker, 2015;Maciel & Pereira, 2013;Walter, 2019); and inquiries into multitemporality and non-linear chronology (Dinshaw, 2012;Harris, 2009;Koselleck, 2004;Nagel & Wood, 2010). Since many of the aforementioned themes and research strands are beyond the scope of this book, while others are mentioned only occasionally, in this section we would like to single out the most salient ones. ... ... The agency of the dead is a relatively young topic of study (Arnold, 2014;Crossland, 2017;Harper, 2010) that has been closely positioned to approaches to the agency and properties of things, objects, and materials. The term 'postmortem agency' was coined by sociocultural anthropologist Benjamin Wilreker (2007) and introduced into archaeology in a special section on "The bioarchaeology of postmortem agency" in the Cambridge Archaeological Journal (Crandall & Martin, 2014) . As observed by numerous scholars, 'agency' per se is an ambiguous concept (Emirbayer & Mische, 1998, p. 962;Robb, 2013, p. 448). ... Interdisciplinary Explorations of Postmortem Interaction: An Introduction Chapter Full-text available Jun 2022 Estella Weiss-Krejci Sebastian Niklas Becker Philip Schwyzer In the present as in the past, the dead have been deployed to promote visions of identity, as well as ostensibly wider human values. Through a series of case studies from ancient Egypt through prehistoric, historic, and present-day Europe, this book discusses what is constant and what is locally and historically specific in our ways of interacting with the remains of the dead, their objects, and monuments. Postmortem interaction encompasses not only funerary rituals and intergenerational engagement with forebears, but also concerns encounters with the dead who died centuries and millennia ago. Drawing from a variety of disciplines such as archaeology, bioarchaeology, literary studies, ancient Egyptian philology and literature, and sociocultural anthropology, this volume provides an interdisciplinary account of the ways in which the dead are able to transcend temporal distances and engender social relationships. In this introduction, we highlight some unifying aspects of this book. ... In these cases, the suspicion of unnatural death would have been an indirect motive for burying the individuals in less respected places. This treatment highlights the postmortem agency (Crandall and Martin 2014; Tarlow and Battell Lowman 2018) of autopsied individuals. The individuals with disturbing deaths, the ones who were not even supposed to be buried in the consecrated ground, were incorporated in the folklore of the powerful dead. ... Unusual Death and Memorialization - Burial, Space, and Memory in the Post-Medieval North Book Jan 2022 Titta Kallio-Seppä Sanna Lipkin Tiina Väre Annemari Tranberg Kallio-Seppä, T., Lipkin, S., Väre, T., Moilanen, U. & Tranberg, A. 2022. Unusual Death and Memorialization - Burial, Space, and Memory in the Post-Medieval North. Berghahn Books, New York & Oxford. ... Identifying hormonal fluctuations from bones and estimating the timing of menopause would also assist in studies of fertility. Moreover, the life course does not always end with the physical death of an individual; agency may extend beyond death (Buikstra, 2019b; Crandall & Martin, 2014) . ... Twenty‐first century bioarchaeology: Taking stock and moving forward Article Full-text available Mar 2022 Jane E. Buikstra Sharon N. DeWitte Sabrina C. Agarwal Sonia R Zakrzewski This article presents outcomes from a Workshop entitled “Bioarchaeology: Taking Stock and Moving Forward,” which was held at Arizona State University (ASU) on March 6–8, 2020. Funded by the National Science Foundation (NSF), the School of Human Evolution and Social Change (ASU), and the Center for Bioarchaeological Research (CBR, ASU), the Workshop's overall goal was to explore reasons why research proposals submitted by bioarchaeologists, both graduate students and established scholars, fared disproportionately poorly within recent NSF Anthropology Program competitions and to offer advice for increasing success. Therefore, this Workshop comprised 43 international scholars and four advanced graduate students with a history of successful grant acquisition, primarily from the United States. Ultimately, we focused on two related aims: (1) best practices for improving research designs and training and (2) evaluating topics of contemporary significance that reverberate through history and beyond as promising trajectories for bioarchaeological research. Among the former were contextual grounding, research question/hypothesis generation, statistical procedures appropriate for small samples and mixed qualitative/quantitative data, the salience of Bayesian methods, and training program content. Topical foci included ethics, social inequality, identity (including intersectionality), climate change, migration, violence, epidemic disease, adaptability/plasticity, the osteological paradox, and the developmental origins of health and disease. Given the profound changes required globally to address decolonization in the 21st century, this concern also entered many formal and informal discussions. ... However, within these assemblages the dead body is not simply a passive subject to which meaning is attributed. Though this point is debated (see Crandall and Martin 2014) , many disciplines have argued that human remains exhibit agency, in the sense that their presence and materiality can be "the anchors of fields of power/social influence that shape human action" (Crandall and Martin 2014: 432). As Young and Light (2016: 68) suggest, "The corpse can be seen as playing an active role in a range of performances, practices and rituals incorporating and informed by material culture". ... The corpse, heritage, and tourism Chapter Nov 2017 Craig Young Duncan Light Cooperation and Hierarchy in Ancient Bolivia: Building Community with the Body Book Feb 2023 Sara L. Juengst Historical Bioarchaeology Chapter Jan 2023 Kathryn Marklein Sketchbook archaeology: Bodies multiple and the archives they create Article May 2022 J Soc Archaeol Shannon A. Novak Archaeological bodies and their afflictions have multiplied in recent years, along with the specialists who study them. The result is a cascade of data, much of it difficult to reconcile. I argue that variable enactments of disease, rather than reflecting an epistemological disconnect or difference in scale, engender ontological gaps. To pursue these malleable matters, I trace the proliferation of “cancer” from the Spring Street Presbyterian Church burial vaults (1820–1850) in Manhattan. To explore the struggles involved in making many things one, I consider emergent multiplicities of this “disease” within specialists’ laboratories, archival records, and the writing process. Rather than force these different cancers to cohere, or make one “win” based on disciplinary domain (science/humanities) or hierarchy of substance (bone/paper), I rely on Stengers’s (2018) ecology of partial connects. The outcome is not a rubric of knowledge gained, but a sketchbook of lessons learned with bodies multiple along the way. Catholicism and Taki Onqoy in the Early Colonial Period: Colonial Entanglements of Church Interments at Iglesiachayoq (Chicha‐Soras Valley, Ayacucho, Peru) Article Full-text available May 2021 Scotti Norman Archaeologists of colonial encounters utilize the concept of entanglement to explore shifting cultural relationships between European and Native populations during colonial encounters. In the chaotic decades after Spanish conquest of Peru (1532), Andeans of the south‐central highlands practiced a revitalization cult known as Taki Onqoy (Quechua: “dancing/singing sickness”). Taki Onqoy preachers advocated for a rejection of Spanish food, culture, and religion in favor of a return to veneration of the local deities practiced prior to Spanish invasion. Based on results from excavation of the Catholic church at the site of Iglesiachayoq (Chicha‐Soras Valley, Ayacucho, Peru), this article explores the burial practices of individuals interred beneath the church floor. Contrary to primary documents, which portray Andeans as idolatrous rebels or pious converts, interments displayed an array of entangled practices. Within the milieu of contrasting religious mandates (Catholic and Taki Onqoy), individuals interred their deceased kin in ways that intertwined both traditions. Arqueólogos de encuentros coloniales utilizan el concepto de entrelazamiento para explorar relaciones culturales cambiantes entre poblaciones europeas y nativas durante encuentros coloniales. En las décadas caóticas después de la conquista española del Perú (1532), los andinos de las tierras altas sudcentrales practicaron un culto de revitalización conocido como Taki Onqoy (quechua: “enfermedad del baile/canto”). Los predicadores de Taki Onqoy propugnaron por un rechazo a la comida, la cultura y la religión españolas en favor de un retorno a la veneración de deidades locales practicado con anterioridad a la invasión española. Basado en los resultados de la excavación de la iglesia católica en el sitio de Iglesiachayoq (Valle de Chicha‐Soras, Ayacucho, Perú), este artículo explora las prácticas de sepultura de individuos enterrados bajo el piso de la iglesia. Contrario a documentos primarios, los cuales retratan los andinos como rebeldes idólatras o conversos piadosos, las inhumaciones exhiben una variedad de prácticas entrelazadas. Dentro del ámbito de mandatos religiosos contrastantes (católico y Taki Onqoy), individuos sepultaron a sus parientes fallecidos en formas que entrelazan las dos tradiciones. [revitalización, sepultura, Taki Onqoy, colonialismo español, Andes] Power, Mediation, and Transformation: Dismembered Heads from Uraca (Majes Valley, Peru) and the Andean Feline-Hunter Myth Chapter Nov 2020 Beth Scaffidi Unidentified deceased persons Chapter Feb 2020 Roberto C. Parra Élisabeth Anstett Pierre Perich Jane Buikstra This chapter considers a humanitarian approach to mitigate the disruption in continuity of identity that may lead to social death of the unidentified deceased persons. It argues that an adjustment in the forensic approach is required, and that this adjustment implies prioritizing the rescue of bodies and their belongings from those places where they remain buried and in anonymity. The dead permanently influence the thoughts and behavior of the living, therefore maintaining an active social life. Claire Moon has pointed out that a dead body has a social life, playing several special roles as elements of social contact, confluence and conflict. The bodies of unidentified deceased persons may be deposited in various places such as morgues, cemeteries, or clandestine deposit sites. In cases of massive disaster, corpses and body parts are recovered, transferred to specialized forensic centers where their identification is prioritized, while unidentified corpses and body parts are sent to specific cemeteries. Jan 1998 Alan Goodman In the context of current debates about global warming, archaeology contributes important insights for understanding environmental changes in prehistory, and the consequences and responses of past populations to them. In Indian archaeology, climate change and monsoon variability are often invoked to explain major demographic transitions, cultural changes, and migrations of prehistoric populations. During the late Holocene (1400-700 B.C.), agricultural communities flourished in a semiarid region of the Indian subcontinent, until they precipitously collapsed. Gwen Robbins Schug integrates the most recent paleoclimate reconstructions with an innovative analysis of skeletal remains from one of the last abandoned villages to provide a new interpretation of the archaeological record of this period. Robbins Schug's biocultural synthesis provides us with a new way of looking at the adaptive, social, and cultural transformations that took place in this region during the first and second millennia B.C. Her work clearly and compellingly usurps the climate change paradigm, demonstrating the complexity of human-environmental transformations. This original and significant contribution to bioarchaeological research and methodology enriches our understanding of both global climate change and South Asian prehistory. Anthropology and Social Theory: Culture, Power, and the Acting Subject Nov 2006 Sherry B. Ortner Social Bioarchaeology Sabrina C. Agarwal Bioarchaeology and Identity in the Americas Book Jan 2009 Kelly Knudson Christopher M Stojanowski Bioarchaeology and Identity in the Americas represents an important shift in the interpretation of skeletal remains in the Americas. Until recently, bioarchaeology has focused on interpreting and analyzing populations. The contributors here look to examine how individuals fit into those larger populations. The overall aim is to demonstrate how bioarchaeologists can uniquely contribute to our understanding of the formation, representation, and repercussions of identity. The contributors combine historical and archaeological data with population genetic analyses, biogeochemical analyses of human tooth enamel and bones, mortuary patterns, and body modifications. With case studies drawn from North, Central, and South American mortuary remains from AD 500 to the Colonial period, they examine a wide range of factors that make up identity, including ethnicity, age, gender, and social, political, and religious constructions. By adding a valuable biological element to the study of culture--a topic traditionally associated with social theorists, ethnographers, and historical archaeologies--this volume highlights the importance of skeletal evidence in helping us better understand our past. © 2009 by Kelly J. Knudson and Christopher M. Stojanowski. All rights reserved. Deathscapes: Spaces for Death, Dying, Mourning and Remembrance Book Jan 2012 Avril Maddrell James D. Sidaway Death is at once a universal and everyday, but also an extraordinary experience in the lives of those affected. Death and bereavement are thereby intensified at (and frequently contained within) certain sites and regulated spaces, such as the hospital, the cemetery and the mortuary. However, death also affects and unfolds in many other spaces: the home, public spaces and places of worship, sites of accident, tragedy and violence. Such spaces, or Deathscapes, are intensely private and personal places, while often simultaneously being shared, collective, sites of experience and remembrance; each place mediated through the intersections of emotion, body, belief, culture, society and the state. Bringing together geographers, sociologists, anthropologists, cultural studies academics and historians among others, this book focuses on the relationships between space/place and death/bereavement in 'western' societies. Addressing three broad themes: the place of death; the place of final disposition; and spaces of remembrance and representation, the chapters reflect a variety of scales ranging from the mapping of bereavement on the individual or in private domestic space, through to sites of accident, battle, burial, cremation and remembrance in public space. The book also examines social and cultural changes in death and bereavement practices, including personalisation and secularisation. Other social trends are addressed by chapters on green and garden burial, negotiating emotion in public/private space, remembrance of violence and disaster, and virtual space. A meshing of material and 'more-than-representational' approaches consider the nature, culture, economy and politics of Deathscapes - what are in effect some of the most significant places in human society. The Body as Material Culture: A Theoretical Osteoarchaeology Joanna R. Sofaer Violence, ritual, and the Wari Empire. A social bioarchaeology of imperialism in the ancient Andes Jan 2012 The Wari Empire thrived in the Peruvian Andes between AD 600 and 1000. This study of human skeletons reveals the biological and social impact of Wari imperialism on people's lives, particularly its effects on community organization and frequency of violence of both ruling elites and subjects. The Wari state was one of the first politically centralized civilizations in the New World that expanded dramatically as a product of its economic and military might. Tiffiny Tung reveals that Wari political and military elites promoted and valorized aggressive actions, such as the abduction of men, women, and children from foreign settlements. Captive men and children were sacrificed, dismembered, and transformed into trophy heads, while non-local women received different treatment relative to the men and children. By inspecting bioarchaeological data from skeletons and ancient DNA, as well as archaeological data, Tung provides a better understanding of how the empire's practices affected human communities, particularly in terms of age/sex structure, mortuary treatment, use of violence, and ritual processes associated with power and bodies. Agency and individuals in long-term process The substance of bones: The emotive materiality and affective presence of human remains Dec 2010

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(PDF) U-Net convolutional networks for mining land cover classification based on high-resolution UAV imagery PDF | Mining activities are the leading cause of deforestation, land-use changes, and pollution. Land use/cover mapping in Vietnam every five years is... | Find, read and cite all the research you need on ResearchGate U-Net convolutional networks for mining land cover classification based on high-resolution UAV imagery Abstract and Figures Mining activities are the leading cause of deforestation, land-use changes, and pollution. Land use/cover mapping in Vietnam every five years is not useful to monitor land covers in mining areas, especially in the Central Highland region. It is necessary to equip managers with a better tool to monitor and map land cover using high-resolution images. Therefore, the authors proposed using the U-Net convolutional network for land-cover classification based on multispectral Unmanned aerial vehicle (UAV) image in a mining area of Daknong province, Vietnam. An area of 0.5kmx0.8km was used for training and testing seven U-Net models using seven optimizer function types. The final U-Net model can interpret six land cover types: (1) open-case mining lands, (2) old permanent croplands, (3) young permanent croplands, (4) grasslands, (5) bare soils, (6) water bodies. As a result, two models using Nadam and Adadelta optimizer function can be used to classify six land cover types with accuracy higher than 83%, especially in open-case mining lands and polluted streams flowed out from the mining areas. The trained U-Net models can potentially update new land cover types in other mining areas towards monitoring land cover changes in real-time in the future. The architecture of a U-Net network for mining land cover classification. … Figures - uploaded by Van-Manh Pham This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License. For more information, see https://creativecommons.org/licenses/by-nc-nd/4.0/ This article has been accepted for publication in a future issue of this journal, but has not been fully edited. Content may change prior to final publication. Citation information: DOI 10.1109/ACCESS.2020.3030112, IEEE Access VOLUME XX, 20 20 A. Date of publication xxxx 00, 0000, date of current version xxxx 00, 0000. Digital Object Identifier 10.1109/ACCESS.20 20 .Doi Number U-Net convolutional networks for mining land cover classification based on high-resolution UAV imagery Tuan Linh Giang 1 , K inh Bac Dang 2* , Qua ng Toan Le 3 , Vu Giang N guyen 3 , Si Son Tong 4 and Van-Manh Pham 2 1 SKYMAP High Technology Co., Ltd., No.6, 40/2/1, Ta Quan g Buu, Hai Ba Trung, Hanoi, Vietnam 2 Faculty of Geography, VNU University of Science, 334 N guyen Trai, Thanh Xuan, Hanoi, Vietnam 3 Space Technology Institute, Vietnam Academy of Science and Tech nology, 18 Hoang Quoc Viet, Cau Giay, Ha Noi, Vietn am 4 University of Science and Technology of Hanoi, Vietnam Acad emy of Science and Technology , 18 Hoang Quoc Viet, Cau Giay, Ha Noi, Vietnam Corresponding author: Kinh Bac Dang (e -mail: da [email protected] ). This work wa s supported in part by the National Program on Space Scien ce and Technology, Ministry of Scienc e and Te chnology of Vietnam under Grant No. VT-UD.06/18- 20 . ABSTRACT Mining activities are the leading cause of deforestation, land-use chang es, and pollution. Land use/cover mapping in Vietn am every five years is not useful to monitor land covers in mining are as, espec ially in the Central Highland region. It is neces sary to equip managers with a better tool to monitor and map land cover using high-resolution i mages. Therefore, the au thors proposed using the U-Net convolu tional network for land-cover classification base d on multispectral Unmann ed aerial vehicle (UAV) image in a mining area of Daknong province, Vietnam. An area of 0.5 km x0.8km was used for training and testing seven U -Net models using seven optimizer function types. The final U -Net model can interpret six l and cover types: (1) open-case mining lands , (2) old per manent croplands, (3) young permanent croplands, (4) grassla nds, (5) bare soils, (6) water bod ies. As a result , two models using Nadam and Adade lta optimizer funct ion can be used to classify six land cover types with accura cy higher than 8 3%, especially in open-case mining lands and polluted streams flowed out from the mining areas. The trained U-Net models can potentially update new land cover types in other mining areas towards monitoring land cover changes in real-time in the future. INDEX TERMS U-Net conv olutional network, Unmanned aerial vehicle, Deep learning, Daknong, Segmentation, Permanent cropland, Open-cast mining, Loss function, Optimiza tion. I. INTRODU CTION The Ce ntral H ighland s of V ietnam has m ore th an 450, 000 hectar es (ha) of p erenni al crops , acc ountin g for nearly 90% o f the coffee area and 2 6% of the rubber area [1]. More than 15,792 ha of natural forests in 1990s were destroyed for the constru ction an d mining pr ojects , of which 5,75 5 ha of fores t land has been conver ted into h ydroe lectrici ty t ill 2 015 [2 ] – [4] . G. K issinge r et al. [5] and P . T. Thuy et al. [6] emph asized that mining and land c over conve rsion are two of the six leading causes of deforesta tion and forest degradati on. Although enviro nmenta l impact as sessm ents hav e become a mandator y requir ement sinc e 19 93, illeg al mining a ctivitie s h ave not been though tfully manage d [2]. A real-time land use/co ver monitor ing mach ine is neede d, replaci ng the nation al manual monitor ing activi ties every five years, to quickly assess the enviro nmenta l quality [7] – [10 ]. Accordi ngly, it ensures legal minera l activities in mineral exploi tation areas of the Central Highla nds. Deep learn ing is a branc h of art ificial intel ligenc e in whi ch compu ters generate rules based on the raw input data [11], [12]. Base d on deep learning , models can improv e their perfor mance based on past results or new data sources [13] . Deep lear ning pro vides man y advan tages fo r humans for predict ing natural hazar ds [ 14], [15], and helpi ng to make intell igent d ecisions in real- ti me with out hum ans inte rventio n [16]. In additi on to the ir application s in soc ial technology , deep learning has been used in various remote sensin g analys es, such as det ecting s hips, t urtles, o r houses [17] – [19 ] . Primar ily, m ost o f the stud ies co mmonly used satel lite i mages with low to medium reso lution im ages such as MODIS, Author Name : Preparation of Papers for IEEE Ac cess (February 2017) VOLUME XX, 20 20 Landsa t, and Sentinel - 2 [20] – [22 ] . However , t he applicat ion of deep learning to d etect land c over from high-resolut ion UAV i mages is rare, especi ally in mining areas . Recent ly, the artifi cial neu ral ne twork (A NN) has becom e a useful tool to deal with comp lex non-li near regr ession betwe en inpu t dat a that h as no t yet been optimi zed fo r anc ient remote sensi ng processi ng techniques such as unsupe rvised learnin g, Ran dom For est, pix el-base d and Su pport Vector Machin e [23] – [25]. For examp le, F. Pu et al. [2 6] and K. B. Dang et al. [2 7] used Convolutio nal Neural Networ k (CNN) to class ify (1 ) water quality of inl and lak es based on Land sat- 8 imag es; and (2) co astal types i n Vietn am bas ed on Sentin el- 2 imag es. W. G e et al . [28] us ed ANN models for li thologi cal classif ication based on v arious moder ate-reso lution satelli te images. Instead of analyz ing satellit e images based on pixel- based approa ches, the AN N mo dels c onsider land uses/ covers as objects t o d isting uish the ir pixels , sc ale, spat ial relat ions and texture inform ation for gen erating homog eneous objects [29] . Therefo re, develop ers can opti mize the perf ormanc e and accura cy of land cover class ificat ion models in real-time and space from big datase t, reducing the computa tional costs of traditi onal p hysica l-based mode ls. In additi on, t he mod els can learn or upgrad e from pr evious comput ation if the deve lopers provid e new i nput da ta fro m the r eal tim e [30]. The U-Net concept s ugges ted by O. Ronneberg er et al. [31] i s an u pdated Fully Convol utional Netw ork mod el with a symme trical U-shaped design includi ng a symmetrica l contrac ting and expanding paths. W hile the contracti ng path acquire s the con text, the expanding path pred ict accur ate locali zation of segm ented obje cts. The U-Net int egrates low- level feat ures with comp rehens ive spatial informatio n (in the contrac ting path) with high- level features with sema tic inform ation (in th e expand ing path) to b oost seg mentatio n precis ion [ 32] . A ccordi ngly, succ essive convo lution laye rs a re trained to create a more re liable output based on this knowled ge. Base d on semanti c segmentat ion appro aches, the U-Net models can learn effect ively spatia l relat ions between land-co ver classes [33] . Ther efore, th ey can bette r classif y objects in wh ole im ages, es pecial ly in t he cas e of l abeled data scarcit y, compar ed to the pixel-b ased models . Addition ally, once the predi cted latter are dis continu ities due to nois es in th e mark data and these models implic itly run a regularizat ion process as a post-pr ocessin g smoo thing s tep [34] . Althoug h the first U- Net mo dels w ere in troduced in 20 15, the first models for land-cover classificat ion have been publish ed sin ce 2019, s uch as l and covers in urb an [35] – [37 ] , forest [38] , coas tal ar eas [3 9] a nd sea surf ace. Most of s tudies develop ed the origina l U-Net with the use of secondary satelli te images [40]. For example, A. Stoian et al. [41] have started to use 10m-reso lution Sentinel-2 satellite images for land use/cover mapping in many polygons in France. Their input image data is commonly dependent o n the satellites , leading to difficult ies to monitor land covers in a particula r mining region. An active i mage-rec order machine such as Unmann ed aeri al vehic le (UAV) will be an altern ative opt ion to classif y mining land c overs in the re al t ime, espe cially if th is machin e can integrate with a d eep learning model. In this resear ch, th e autho rs us e the U -Net framew ork to train a mining lan d cover predi ction mod el based on the mul ti- spectr al UAV im age i n a parti cular area o f Da knong prov ince, Vietnam . This study will p rove the p otent ial of the U-N et model and UAV data i n mon itoring mining land c overs. The follow ing resear ch question s - releva nt to land cover classif ication – will make this st udy cl earer: • What are the advantages o f i ntegr ation between deep learnin g and high-res oluti on images for monitorin g mining areas? • How are U-Net models for land cover classif ication on UAV i mages mo re eff ective than b enchma rk mode ls? • How do land cover types distrib ute in a mining area of Daknong provi nce, V ietnam? In this study, 4-band UA V i mages wer e used as inpu t variabl es for land-cov er classifi cation. An area of (0.5x0.8 ) km 2 was used to dev elop a U-N et mode l for land cover classif ication . Aft erward , the trained model was used to interpr et another area of (2x2 ) km 2 in a mining are a of Daknong provin ce, Vietn am. Befo re expla ining da ta collec tion and mode l develo pment ( from s ection II .D to II .E) , the a uthors pres ented a des cripti on of land cove r cl assific ation systems in the research area (secti on II .A). Notably, all process es to pr epare U AV i mage, prepro cess and trainin g models will be exp lained in detai l in sect ion II.B . The t rained U-Net models were compar ed with two benchmar k models using Rand om Forest (RF ) and Support Vector Machin e (SVM) – pres ented in sectio n II. G - to c heck the imp rovemen t. Results con cerning the class ificat ion of min ing land cover and model develop ment will be compared with recent studies in section III . II. MATER IAL AND METH ODS A. RESEARCH A REA To eval uate the ability to monitor minerals explo itation wh en using the remote sensi ng technol ogy UAV, the authors select ed Tan An quarry in Da k R'moa n commune , Gia Ngh ia City, Dak Lak province as the ma in researc h field. The experim ental mine is an ope n-pit mine in operati on, and witness ed significa nt changes in t opogr aphy due to mi ning activi ties. It has an area of 50 ha adja cent to the banks of Dak Tí Hydropower lake and Dak R’Moan inter -commu ne road. The size of the m in ing area meets the requi rements for close - freque ncy flight by UAV technology with ideal cond itions such as lying on a mounta in peak with wide viewing angles and cov ering the ent ire fie ld. Additi onally, the hard groun d of the mine can ensure tha t ground co ntrol poi nts are immo bile dur ing the pro cess of fligh t contro l and measur ement. Ther e was little dus t in the mining object , which provid ed clear images for build ing graph and digital model of surfa ce with high resoluti on. The flight was conduc ted on June 28, 2019. On that day, dry weathe r, This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License. For more information, see https://creativecommons.org/licenses/by-nc-nd/4.0/ VOLUME XX, 20 20 moder ate wind and little dust mining subjects all contribute d to the develo pment of clear aerial photos with optimal imag e quality . T hese photos w ere used to bui ld a d igital surf ace model of high quality lat er on . To asses s the ove rall p rocess of chang es in the st atus of soil cover in the quarry and surrou nding areas , the scop e of experime ntal stud y expanded to an area of 400 ha, tak ing th e quarr y as t he cen ter. Th e min e is an area with the co ordina tes of 11°5 9'55" - 12°00' 14" N orth latitud e, 104 °06'11" - 104 °06'41" East l ongitud e. Tan An quarry area is covered with basalt soil with the thickne ss of 2-5m. Before the ex ploita tion, mo st of the qua rry area was u nused hilly land and peren nial indust rial c rops land. During the explo itation process, the q uarry area has been gradua lly expanded and some areas of perennial crop land have also been exploited. Moreov er, activities such a s dumpin g and unlo ading of surface soil, mak ing sp ace fo r process ing and gatherin g of finis hed ston e produ cts are respons ible f or changes in other soil types . In 2006 , the qua rry was ju st an open- pit exp loitati on are a with a sm all ar ea of less than 1 ha. The stone is mined int o the cliff ar ea. At tha t time, there w as no s ign of expans ion min ing. By 2012, a part of the rock was exploite d in the dir ection of mine exp ansion, a road leading to th e quarr y was a lso fo rmed. Some of the mine's wester n perennial industr ial land has been conv erted an d r emoved s tones. Ho wever, the min ing area was not yet put int o ind ustr ial e xploita tion. By 2014 , after only two years, Tan An q uarry has been expanded , with all the corresp onding pr ocessi ng and sto rage areas. From 20 16 to 2018, a large area of perenni al industria l land was converted to serve th e pu rpose of min e expans ion. The extr action pr ocess becomes stabl e. The six typ es of so il in Tan An quarry can be visual ly obser ved thro ugh UA V imag es shown in Figu re 1. A part o f th e quar ry are a has also been accret ed by new layers of soil. FIGURE 1. The research area with six samples for six land -cover types in a mining area of Daknong p rovince, Vietnam. PRE-PROCES SING OF UAV IMAGE AND SAMPLE COLLECTION UAV technological advancement is m ost b eneficial when applied to monitoring mining are a. Due to the fact that mines are densely located over small areas, it i s very efficient to collect data for UAV processing. For the purpose of monitoring the mining progress, the experi mental mine must be an open-pit one which is still under operation and experiences topography changes ca used by exploitation. In this st udy, image data is taken from UA V d atabase from senseFly of Tan An quarry in Dak Nong province, Vi etnam. The data is collected by a Phantom 4 RTK and underwent a 6-step process o f OrthoEngine Tool in PCI Geomatica Banff Service Pack 1 (Fully Fun ctional Trial). There are 768 JPEG images used f or inp ut da ta. T he crit ical st eps in the pre- processing of UAV data will be introduced briefly as follows: • Step 1 (Set camera calibr ation): This ste p helps to identif y and correct the distort ions of the image which occurr ed due to curvatur e of the lens, the focal length, and the perspe ctive eff ects. In this step , parameters (Focal len gth; Principal-po int offset; Chip Width a nd Height ; Y-scale factor; Radi al-Lens coefficie nts; Decen tering c oeffici ents) a re ent ered ma nually. • Step 2 (Collect tie points ): Tie point (TPs), a sp ecific locati on, can be recognized visua lly in the overlap area among two or more ima ges. By applying image- correla tion t echniqu e a tot al of 2 3,040 op timal tie poi nts were found in 768 images. This resul t has an importan t role to play in comb ining all im ages tog ether. • Step 3 (Poin t refineme nt): in this step, 23,040 tie points found in step 2 were refine d to elimin ate the ones with high values of Ro ot mean square (RMS: the r esidua l values of t he x and y per point). Tie points with RMS higher than 0.5 wer e removed from the calcul ation model. • Step 4 (GCPs corre ction): 12 G CP (G round Contro l Point) from field with an accuracy of ±0.5 pixels were collec ted to impro ve geo referen cing o f the datase t. The y are clear ly recogn ized in the raw images wi th identif ied ground coor dinates . A ll d ata w ere o rthor ectified wit h the WGS84 /UTM ZO NE-48N . • Step 5 (G enerate ortho images base d on DEM) : This step generat ed 768 ortho-image s by usin g raw images, epipola r pair ima ges and a Digital Elev ation Model (DEM) of the research area scann ed during flying process . This helps to enhance the performan ce of the correla tion proce ss and to reduce th e probabil ity of incorr ect matches of the raw images . The spat ial resolu tion of the ou tcome DEM is 0.35 m . • Step 6 (Mosaic king): In this step , 768 ort ho-image s w ere manual ly edit ed to f orm a s ingle an d unifo rm co lor balanc ing image. Because 80% of images overlap , all errors re lated to cam era and coor dinates w ere elim inated . The mosai cking process was conducted precisely with the use of Mos aic Tool of th e P CI Geomatica Banff softwar e. After the merged image had been generate d, the Tan An m ining area was presented for the U-Net develop ment t hat wi ll be e xplaine d in se ction II .D. C. PROCESSIN G DATA Sampl es of six LULC types were taken in the area of (2×2) km 2 around the main mining area. The authors measured in detail the area and the bo undary of e ach LULC in November 2019. Firstly, althou gh the coffee trees in the resear ch area covered a larg e are a, some coffe e areas have been m ixed w ith some pepper trees. The density of coff ee trees is denser than two other agricul tural trees. Therefore, i t is c hallengi ng to separa te coffee and pepper trees in the UAV image. Authors combine d coffee a nd mixed croplands into one l and-cover type, so-called permanen t croplands . However , they will be separa ted into old and young per manent cropla nds depen ding on how large t he canopy of the trees coverin g the land. The canopy of the old coffee trees covers a larger area t han the young one. Therefore, the second land cover type that was separa ted is th e “young perman ent cro plands ”. Grassl and and bare soil respective ly, are third and fourth land cover types were s eparat ed (Figu re 1 ). The grasslan ds in the resear ch area have been left fallow during five year s prepar ing for mining activiti es in the future, whereas t he bare soil are as have bee n l eft fallow af ter finish ing mining activiti es [42] . The whole area used for natura l resourc e min ing w ill be sepa rated i nto th e “op en - cast minin g” land cover. It includes all mining areas and houses used for explor ation activities . Like the last land c over, water bodies were separated and repres ented small lakes an d t wo small streams t hat flow ou t f rom the mi ning area to t he outsid e region . The boundary of all agri cultur al cropl ands will be formatt ed in a polygon file and converted to an integer- 8bit raster file (s o- call ed as “mask”) . The s ize of the m ask is equal to the size of t he in put i mage. D ue t o the large imag e siz e, it is not f easibl e to read the entire image into memo ry and th en us e it f or U-Net dev elopme nt. I n order to incre ase the perfo rmanc e of the training process , the original UAV image was spitted into 12,00 0 sub-ima ges and resiz ed to sub-imag es of 128×128 -pixe l size. Each sub-i mage w as in putted as a samp le to train the U-Net mod els. Afterw ard, they were sep arated int o two tr aining a nd tes ting gro ups of 80% (or 9,600 s ub-imag es) and 2 0% (or 2.400 sub-i mages) for us ing f or the U-Ne t model develop ment. The r atio of land c over ty pes in the tr aining and testing data is equa l. D. BACKGROUND OF MODEL VA LIDATI ON Once a predictio n model is trained, it always needs to be assess ed its pe rforman ce/qua lity, es pecially with th e deep learnin g models , avoiding overfitting and underfitt ing cases [13] . This sectio n will explain all model validat ion methods used i n this stu dy. Th e valid ation process was perfor med through out the U-Net, Random Fores t, and S uppor t Vector Machin e mod el deve lopment (see from s ectio n II-E to se ction study, includin g (1) intersectio n over union value (IoU), (2) class user’s acc uracy (UA) , (3) o veral l accu racy (OA), and (4) Kappa Co effic ient . The IoU valu e was us ed duri ng the U-N et develop ment (see section 2-E), wh ereas three evaluation values , includ ing UA, OA, and K appa, wer e used to c ompare the accur acy between all trained U-Net models and two bench mark models (see section 2-G). The IoU value repres ents the proportion of the combined area of referen ce and predicte d bounding images (so-called as the intersect ion region ) per the union region [43] . The IoU v alue is des cribed in the followi ng for mula:          (1) The IoU value has been commonly used in compu ter vision and deep- learnin g studies [4 3] . D uring th e U-Net dev elopmen t process , the IoU value, th erefor e was calcu lated c ontinuou sly for both training and testing image data in each epoch. Howev er, in compari ng the p redic tion-pow er bet ween th e tr ain ed U-Net models with two benchmar k models, the UA, OA, and Kappa are an alternati ve option to check all trained models ’ ac curacy at p ixel scal e. A ccording ly, these valu es ca n land- co ver class “i” compare d with ground truth/inp ut class “j”, “ N ” is the to tal numbe r of samp les, “m” i s the nu mber of classes , M j is the total n umber of sample in th e c lass “ j ” record ed in m ask d ata, and R i is the tot al num ber of samp le in The UA, OA, and Kapp a va lues were onl y use d on ce all models tha t w ere tr ained . The UA value was use d to ass ess the accura cy of each land cov er type pred icted fro m trained models . It expla ins the p ositive proport ion of the correct classif ied s amples to th e pred icted s amples [43] in a partic ular land-co ver type in this stud y . In additi on to the overa ll classif ication accu racy ( OA), the Ka ppa v alue accounts for all true and false da ta of the model validat ion proc ess based on a confusi on matr ix [44]. Th erefore , t he use of bo th OA and Kappa values can make the selecti on of the best model for mining l and-c over cl assifica tion mo re accu rate. Th e inpu t samples will be expl ained in det ail in s ection 2- G. E. U-NET ARCHI TECTURE FOR LAND-COVER DETECTION The U -Net m odel w as fi rst dev eloped for segmen tation on medica l and geologic al imag es [45], [46] . It includes a contrac ting path (in the le ft s ide) and an ex pansiv e path (in the right side) . The cont ractin g pat h can be consi dered as a typ ical extrac tor like conv ention al convol ution neu ral netwo rk (CNN) models . The right- hand side perform s up-samp ling steps, or conver t predi ction va lues ba ck to th e orig inal im age size. The U-Net archit ecture is s hown in Fi gure 2 . Like a Convolu tional unnec essary neurons can be deact ivated to reduce overfi tting and g eneraliz ation err ors. A drop out valu e is common ly us ed as a probab ility of 0.5 t o k eep th e ou tput of ev ery n ode in a hidd en la yer. In co ntrast , a value clos e to 1.0 is co mmonly us ed to keep inputs fro m the visib le layers [48]. The blocks in blue color shown in Figure 2 are the input and output of each cal culation la yer, w hereas th e pro cessing lay ers are prese nted in arr ows. Exce pting th e l ast convol ution al layer, the other 18 CONV layers are always processed before the batch normali zation layers (combi ned and shown in 18 blue arrows ). In the contr acting path, the PO OL layers run befor e the Dropout layers to downscale data. Meanwhi le, the transpos ed convolu tion matrix es integrat ed with the Dropout layers ar e used for th e u pscalin g p rocess in the expansive layers . They are repres ented in four green arrows . Lastly , the concat enate l ayers a re repr esented in bro wn arr ows. TABLE I M ATHEMATICAL STRUCTURE OF THE DEVELOPED U-N ET FOR LAND COVER CLASSIFICATION . No. Parameter InputLayer BatchNormalization_1 128, 128, 16 Activation_1 128, 128, 16 MaxPooling2D_1 Conv2D_4 64, 64, 32 BatchNormalization_4 64, 64, 32 18 Conv2D_5 32, 32, 64 18 , 496 19 BatchNormalization_5 32, 32, 64 256 10.1109/ACCESS.2020.3030112, IEEE Access Author Name : Preparation of Papers for IEEE Ac cess (February 2017) VOLUME XX, 20 20 No. Layer Output shape Parameter 20 Activation_5 32, 32, 64 0 21 32, 32, 64 36 , 928 22 BatchNormalization_6 32, 32, 64 256 23 Activation_6 32, 32, 64 0 24 MaxPooling2D_3 16, 16, 64 0 25 Dropout_3 16, 16, 64 0 26 Conv2D_7 16, 16, 128 BatchNormalization_7 16, 16, 128 512 28 Activation_7 16, 16, 128 0 29 Conv2D_8 16, 16, 128 147 , 584 30 BatchNormalization_8 16, 16, 128 512 31 Activation_8 16, 16, 128 0 32 MaxPooling2D_4 33 Dropout_4 8, 8, 128 0 34 Conv2D_9 8, 8, 256 295 , 168 35 BatchNormalization_9 8, 8, 256 1, 024 36 Activation_9 8, 8, 256 0 37 Conv2D_10 8, 8, 256 590 , 080 38 18 , 464 62 BatchNormalization_15 64, 64, 32 128 63 Activation_15 64, 64, 32 0 64 Conv2D_16 64, 64, 32 9, 248 65 BatchNormalization_16 64, 64, 32 128 66 Activation_16 64, 64, 32 0 67 128, 128, 16 4, 624 68 Concatenate_4 128, 128, 32 0 69 Dropout_8 128, 128, 32 0 70 Conv2D_17 128, 128, 16 4, 624 71 BatchNormalization_17 128, 128, 16 64 72 Activation_17 128, 128, 16 Conv2D_18 128, 128, 16 2, 320 74 BatchNormalization_18 128, 128, 16 64 75 Activation_18 128, 128, 16 0 76 Conv2D_19 128, 128, 6 102 Total parameters 2,164,822 Table I shows the U-Net architectur e image processing proced ure, with 76 layer s. The ou tput of t he other l ayer is th e input dat a of the following layer . The first 32 layers perfor m the contracting path, where as the last 36 layers perfor m the expans ive pa th . The numb er of filt ers in the follow ing l ayer i n contrac ting path and one half of the precedin g layer in t he expans ive path (Figur e 2), co mpared t o CNN m odels . In contras t, the width and he ight of the prec eding layer is on e h alf of the p recedi ng layer in the contr acting pat h and do uble of the precedi ng lay er in the expa nsive one. The number of parameters of t he first Conv2D layer is calcul ated as follow ing:                 (9) w here ‘H’ is height, ‘W’ is width, ‘D’ is depth and ‘N Filter’ is number of fi lters. Ea ch f ilter has on e e xtra param eter to stor e the bi as valu e. For exampl e, the first Conv2D-1 layer has (3 x 3 x 4 + 1) * 1 6 = 592 para meters . Due to the batch norm alization generat e four paramet ers for each CONV layer, the numbe r of paramet ers in the batch normali zation layer is cal culated as fol lowing :         ( 10 ) where N i is the depth in the input CON V layer. For exampl e, the BatchNo rmaliza tion_1 8 has 4 *16 = 6 4 parameter . The final Conv 2D layer ’s output is a vector wit h six values , corresp onding to 6 l and cov er typ es. Bas ed on 76 lay ers 18xBat ch-Nor malizat ion, 4xConca tenate , 18x Acti vation , and 8x Dropout layers), the trained U-Net transfo rmed the initia l pixel values in 12 ,000 UAV sub-i mages to t he land co ver classes . 23 CONV and 18 Batch Norm alizatio n lay ers con tain parame ters that can be optimiz ed to improve U- Net model’s perfor mance and accur acy. The paramet ers in th e CONV and Batch Normali zation layers will be chang ed with alternative choices of activation and optimi zer functions . It wil l be explain ed in detail in sec tion II. E. The accur acy of both trainin g and testin g data was checked during U-Net deve lopment to avoid overfitting and underf itting is sues. The best U-Net wi ll be chos en if it is land cover pre diction is compati ble with th e labels assi gned in the input im age from th e trainin g and testi ng data. Th erefore , the inters ection o ver union value (IoU ) was used t o assess the accura cy o f all traine d models . During t he trainin g p rocess , th e min and max Io U values of trainin g and testing data w ere record ed t o obs erve the flu ctuation of t his v alue in 100 epoc hs. The IoU value using the trai ning data will be definitely optimi zed in the train ed mod els, therefo re th e IoU value using the testing data w ill be observ ed to check the a ccuracy o f outco me models. In combination with the IoU, to choose the best model for class ifying min ing e cosyste m ty pes, the tr ained model need to have the low est v alues of all loss functio ns. T he modific ation of the lo ss fun ctions will be explai ned in s ection 2- F. The U-Net model is develop ed based on Keras in Python languag e, as an API design ed for easy man ipulat ion w ith Tensorf low develop ed by Google [38], [49]. The parameters observe d d uring the train ing proces s i nclude acc uracy and loss This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License. For more information, see https://creativecommons.org/licenses/by-nc-nd/4.0/ This article has been accepted for publication in a future issue of this journal, but has not been fully edited. Content may change prior to final publication. Citation information: DOI 10.1109/ACCESS.2020.3030112, IEEE Access Author Name : Preparation of Papers for IEEE Ac cess (February 2017) VOLUME XX, 20 20 of testing and validation data. The U-Net training process is limited to 1 00 lo ops ( epoch) , but the pr ocess can b e stop ped if the coefficien t on the set of testing data converges . In other words, all val ues do not i mprove after t en epoc hs. F. ALTERNATIVE OPTIONS T O DEVELOP TH E U-NET Accord ing to the arch itectur e of the U-Net for the land-c over classif ication , to develop the U-Net, three types of functions , namely activatio n functio n, loss function, and optimize r method can be sele cted. These fun ctions ser ve as opt imal parame ters for filters contained in hidden layers . Which functio n and method to be used depends on the kind of input data and output lab els, and th e accurac y/ loss of tr ained models . Thes e will be dis cussed i n the follow ing sec tion. 1) ACTIVATION FUNCTIONS To optimi ze th e con vergen ce sp eed of the U -Net in the CONV layer, an act ivation function shou ld be ch osen. B inary S tep or Linear A ctivatio n Functio ns were no t selected b ecause of the differ ences in the appearance of six land-cover types in four UAV spectral ba nds. Therefo re, one in fiv e non-linear activa tions, which are TanH/Hyperb olic Tangent, ReLU (or Rectif ied Linear Unit), leaky ReLU, Parametric ReLU, and Swish types will be chosen as the m ost s uitabl e for optim izing U-Net mod els [50], [51 ]. The ReLU fun ction poss esses cer tain advanta ges ov er the others . Using max (0 , x) - t hresho lding a t zero, the ReLU function – can keep the considera ble size of the images (128x12 8x4) and speed-up the convergen c e process of U-Net models. The othe r functions fail to achieve such optim al advantages . Furth ermor e, this function also enables b ackpropa gation in the tra ining process to happen [52]. Bes ides, th e re liabili ty o f results f rom the U-Net a pplying ReLU funct ion is al so b etter than whi ch of th e oth er fu nctions . For these above reasons , the ReLU functi on was select ed for four U-N et la yers. Two a ctivat ion functio ns n amely Sigmo id/Log istic and Softma x can be applied for the POOL layer [53]. However, the la tter fun ction was c hosen rather than th e form er. Be cause Softma x function is common ly used to nor malize the outputs to cl asses betwe en 0 and 1 and provi des predict ion pr obabi lity in a specific type [54]; while the Sigmoid /Logist ic function generat es un certaint ies r elated to va nishing gradi ent pro blems if the maximu m and minimum values of input data are too high. In this study, all POOL laye rs, there fore, used Soft max functio n as th e bes t optio n. 2) LOSS FUNCT IONS To narrow the gap between the predicted and actual outputs for th e trained U-Net, a minimi zed Cost functi on (C), or Loss functio n i s needed as convex funct ions based on choosing optimi zed values for weights [55]. Due to the depen dence of the loss function on weights, input imag es, and out put labels , the weight s of the trained netw orks can help to minimize the loss fun ction , which then c ontribut es to a better pred iction fo r new input data. This optimizatio n process was explaine d in detail by J. Redmon et al. [56 ]. The aver age loss val ues were compu ted using en tire trai ning sub-i mage data an d repres ented by th e follow ing fun ction:           17 RMSprop               where  is parame ter value; 󰀪 is the learning rate s; t is time step ;  = 10 - 8 ;   is the gradient; E[g] — mov ing average of squa red gradients; m, v are estimates of first and seco nd moments;   - the ma x operation;  - moving average p arameter (good default value — 0.9);  – step size. G . EXPERIMENTAL SETUP In th is sec tion, the autho rs compar ed th e perf orman ce of other traditi onal classif iers with the train ed U-N et m odels . Two classif iers, incl uding random forests (RF) and suppo rt vector machin es (SVM), w ere chose n. A fter com pleting mi ning land- cover maps in the research area based on the RF, SVM and with the mask. Due to the models using the Random Forest and SVM methods wer e not trained in epochs, the trainin g process of these two models provid e one accura cy value of each model. The cross-v alidati on process used 1000 random points in the test ing group (in 20% of the test ing data) in the U-Net developm ent to compare with the mask. Three evalua tion metrics that were chosen for this step include the UA, OA , and K appa c oeffici ent. T he opt imal model wi ll achiev e higher OA and Kappa values. T he resul ts of this ste p will be presented in section III.C. The developmen t of these classif iers c an be ex plaine d as fo llows: 1) RANDOM F OREST (RF) RF was develop ed in 2001 by L. Breiman [66] and beca me more and more popu lar used deep learnin g ensembles , especia lly r emote sensing . A s et of decisi on tre es as a fore st is trained randomly from training data . Then, the forest, which contain s many d ecision trees, is d evelop ed a s a compos ite classif ier mod el [23] . Once the for est is created , users can use the RF mo del to pre dict acr oss each tree with new inpu t data . Three parameters can be modifi ed to opt imize a RF model includi ng the number of trees in the forests , the number of feature s (cal culated as the squ are ro ot of the r eflecta nce b ands this st udy) and the numb er of s amples tak en at ea ch leaf ( was set at “1”) [67]. In this study, the numb er of trees was tested with 10, 100, 500, and 1000, a nd finally with 100 trees the results ob tained a rela tively high er acc uracy com pared to thre e other n umbers . 2) SUPPORT VECTOR MACHINE (SVM) SVM has known as a supe rvised learning model to analyze and classify ima ge da ta, es pecial ly w ith sm all d atasets [29] . In two-di mension al data, a hyperpl ane was identified based on the S VM m odel to s eparate effe ct th e data in t o tw o cate gories . In m ulti-d imens ional data, as i n this study, the data is conver ted to correspo nding dimen sional spa ce and a plane is id ent ified to se parate data to categ ories [68 ] . Two para meters can be adjus ted to opti mize the SVM m odels , inc luding K ernel coeffic ient “ gamma ” an d error term “C” values. The higher gamma v alue c an make the SVM model s fitted with th e trainin g dataset . Although the error is reduced, it can make over- f itting iss ues. M eanwhil e, the high er “C” v alue can help the SVM models to use more trainin g datas ets for the suppor ting process but it can be less accurate if the selected trainin g datas et is t oo larg e [69] . Therefo re, the “gamm a” and 10.1109/ACCESS.2020.3030112, IEEE Access Author Name : Preparation of Papers for IEEE Ac cess (February 2017) VOLUME XX, 20 20 “C” val ues were selec ted base d on th e OA and Ka ppa valu es. The op timal values were found for t he “ga mma” v alue at 0.25 and “C ” value at 100 . H. APPLICATI ON OF TRAINED U-NET MODELS F OR NEW LAND COVE R CLASSIFICA TION Once the best U-Net for the land cover classific ation from high-r esolut ion image s was d evelop ed, its most import ant functio n is to interpr et new UAV images. In this study, the authors focus ed on land cover types in rock mining in Daknong prov ince, Vietn am. Th erefor e, a new U AV im age i n a min ing a rea of (2 ×2) km 2 was taken in 2019. The image data collec tion and pre-proces sing were also do ne, as exp lained in section II.B and II .C. Once the n ew imag e was inputt ed to th e trained U-Net, the model accesses the trained paramete rs to transfo rm the new images into specific spatial matrices, generat e intermed iate matrix es in 76 layers, and inter pret the final l and cover classes for ev ery pixe l within t he new im age. All th ese pr ediction pro cesses do no t requ ire an y new trainin g samples . III. RESULTS A. U-NET MODEL PER FORMANCE Figure 3 describ es t he loss functio n values of seven models using seven optimizat ion methods. The i ndicat ors used fo r compari son are the "loss valu e" of "traini ng and validation data" over epo chs. In general, validation and train ing loss values were opt imized after abou t 10 to 15 epochs in all methods . The U- Net model’s training process using the Adamax was stop ped a fter ten epochs, and the best model was found in the first epoch. It yields IoU result s with a mi n of 81.5, a m ea n of 82 .3, and a max of 82.5 . Its stand ard d eviatio n of 0.3 i s better than other ones (Tabl e III). After about 3 0 epochs , the models using Adam and RMSprop only have an accura cy of ab out 81% , where as the models us ing th e Nadam and Adadelta achieved the higher accuracy values of nearly 84%. FIGURE 3. The loss function values of seven mo dels using seven optimization m ethods. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License. For more information, see https://creativecommons.org/licenses/by-nc-nd/4.0/ This article has been accepted for publication in a future issue of this journal, but has not been fully edited. Content may change prior to final publication. Citation information: DOI 10.1109/ACCESS.2020.3030112, IEEE Access Author Name : Preparation of Papers for IEEE Ac cess (February 2017) VOLUME XX, 20 20 The m odel using the SG D o ptimize r has the accur acy v alue of 68%, the tr aining an d validat ion loss values are too diffe rent. The predicti on works in detail at the local scale – looks like the outco me using the pixel-base d methods . It can be easy to observe in the location of young permanent cropl ands. The model using the SGD optimizer did not identify spatia l distrib ution of y oung cro p lands a nd bare s oils among the croplan ds. It is dif ferent with t he larg e regio n of th e bare soils in brow n color (Figur e 4 and 5). Th e distri bution of each land cover will be expla ined as followi ng se ctions. TABLE III T HE I O U VALUES OF SEVEN MODELS USING SEVEN OPTIMIZER ALGORITHMS . No. Model using optimizer algorithm IoU (%) Min Mean Max Standard deviation 1 Adam 18.3 74.6 81.2 14.3 2 Adamax 81.5 82.3 82.5 0.3 3 Nadam 18.4 74.2 83.2 16.9 4 Adagrad 24.8 72.3 79.0 15.5 5 Adadelta 31.7 73.5 83.6 16.7 6 SGD 27.9 63.4 67.5 10.0 7 RMSprop 26.2 70.6 81.1 16.4 B. LAND-COVER INTERPRETATI ON IN THE M INING AREA OF DAKN ONG PROVINCE Figure 4 shows the land-cover map based on manual image interpr etation and segmen tation me thods. Th is map was use d as a mask to train all U-Net mod els. Accord ing to this result, the a rea of o ld p ermanen t cro plands (16 hec tares) is four times higher than t hose of the young perman ent cr oplands . The area of gr assland s is abo ut 1 4 he ctares, whe reas the open-c ase mining areas cov ered more than 9 hectar es. As show n in Figure 4, th e open mine i s surrou nded b y the p erman ent croplan ds with v arious species of co mposing trees such as cashew , coffee and durian. These specie s are not separately cultiv ated in di fferent lan d pa rcels but they are mixe d bet ween cashew , durian and c offee tree. The interposing cultivation makes the im age class ificat ion con fused . The a rea o f the open rock mine is about 17.7 ha counted in June 2020, characteri zed by two discrete areas including a exploit ing core and a taking-off soil region . The exploiting area has a roc ky su rface in a gra y cov er, a nd the t aking off so il layer is in yellow-red due to the concentra tion of v olcan ic materi al. In addition, service areas taking parts of the mine, occupy 4.8 ha for the storage, product processing , and transpo rt. The servic e areas are clearly observ ed on sate llite image with br ight gr ay of concret e surf ace. The s outh and sout hwest side of the min e is the ban k of the Dak Tik lake which i s submerg e in a short time during the rainy season . Thus, the bank is covered by grass most of the rest of year. Grass cover presents as a smooth, bright green region , and c losed w ater s urface i n sat ellite i mages . FIGURE 4. A land cover classification in the mining area of Daknong province, Vietnam based on manual image int erpretation and segmentat ion met hod. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License. For more information, see https://creativecommons.org/licenses/by-nc-nd/4.0/ This article has been accepted for publication in a future issue of this journal, but has not been fully edited. Content may change prior to final publication. Citation information: DOI 10.1109/ACCESS.2020.3030112, IEEE Access Author Name : Preparation of Papers for IEEE Ac cess (February 2017) VOLUME XX, 20 20 TABLE IV T HE C ROSS - VALIDATION OF NINE MODELS FOR MINING LAND COVER CLASSIFICATION . Sample Distribution Class accuracy (% ) Class Samples Adam Adamax Nadam Adagrad Adadelta SGD RMSprop RF SVM Open-case mining lan d 141 80.6 80.9 85.1 79.5 82.3 76.7 74.5 81.8 80.5 Old permanent cro pland 252 89.7 83.3 88.9 92.5 90.1 89.3 93.6 90.9 82.9 Young permanent crop land 164 69.5 82.6 85.8 72.7 89.4 20.5 80.3 76.9 79.5 Grassland 208 70.7 72.1 75.5 73.6 75.9 71.8 74.6 71.2 72.7 Bare soils 123 83.4 85.4 85.4 81.3 82.9 73.1 78.9 77.2 80.5 Water bodies 112 87.5 89.3 83.1 84.8 88.4 81.8 92.8 82.1 89.3 TOTAL 1000 Overall accuracy (%) 80.2 82.3 84.0 80.7 84.8 68.7 82.4 80.0 81.0 Kappa Coefficient 0.78 0.80 0.82 0.79 0.83 0.66 0.82 0.78 0.79 FIGURE 5. Land cover interpretation based on Random Forest, Support Vector Machine and the seven U-Net mo dels using seven optimizer functions in the mining area of Daknong province. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License. For more information, see https://creativecommons.org/licenses/by-nc-nd/4.0/ This article has been accepted for publication in a future issue of this journal, but has not been fully edited. Content may change prior to final publication. Citation information: DOI 10.1109/ACCESS.2020.3030112, IEEE Access Author Name : Preparation of Papers for IEEE Ac cess (February 2017) VOLUME XX, 20 20 The d iagrams in Figu re 5 des cribe the image interp retati on results base d on seven U-Net models using seven different optimi zer o ptions . In gener al, all models can effec tively detec t the old perm anent croplands, open-cast mining areas, water bodies , bar e soi ls, an d g rasslan ds. H owever , thre e mo dels using the Adam, Adagr ad, and RMSprop opt imizers coul d no t detect the vast a rea of the young permane nt croplands in the eastern side of the resear ch area. The predict ion process was done after 12 0 s econds . In ter ms o f accuracy , th e m odels usin g the Nadam and Adadelta optim izers are still the best ones. Using the Adam optim izer, the U-Net model can detect the differ ences betwe en the young perm anent crop lands wit h the bare soils that distribu te among crops. Whereas , the model using the Adadel ta learn thorough ly from the mask and then combine the narr ow ar ea of bar e soils a mong the young perman ent cro plands with t he crop s into one ob ject. C. ACCURACY COMPARISION AMONG THE TRAINED MODELS The ac curacy diffe rences betw een ni ne tra ined models (includ ing seven U-Net models a nd two benchmark models) are presen ted in Table IV. The predict ion maps based on the RF and S VM models can be s een in Figure 5. Due to the testing sampl es are a part o f valid ation d ataset, the OA values of sev en models are n early similar to the results show n in Table III. A s sh own in Figur e 5, the O A and Kappa valu es the model using the SGD optimi zer have the lowes t values , compar ed to other models . The results predict ed from the RF and S VM m odels have the OA and K appa values sign ifican tly lower than most of U-Net mode ls. Compa re with the manua l interpr etation mask, the model using the Adadelt a optim izer provid es the best pr edictio n. IV. DISCUSSI ON A. COMPARIS ON WITH FORMAL LAND COVER CLASSIFICAT ION SYSTEMS Classif icatio n of land cover is closely linked to ecologi cal conditi ons on the Earth's surfa ce and partly r epresen ts the global ecosys tem health, quality of the water, and useful for sustain able land manag ement . Most land-cover class ificatio n studies typically use low/med ium-spa tial resolu tion satellit e Mansara y et al. [ 71 ] and A. S. Had eel et al. [72]. Comm only, they are l ack of spatial information ; making di fficul ties in identif ying land covers in high heterogen eous regions accura tely. This study took full advantage of feature s of the high-r esolut ion images take n from UAV images such as rich texture , shap e, and spatia l distr ibution infor mation to the train land-us e pre diction mode ls. T he im age s egmenta tion s tep w as done to generat e a sampl e map or a mask to training mode ls [73] – [7 5]. To make a mask for traini ng the U-Net mod els, in this stu dy, the authors also used image segmen tation . Howev er, the results fro m the image segmenta tion step were ineffec tive to use dir ectly as a mask. An ob ject gen eraliza tion was made as a mask to train the model (shown in Figure 4). Once the U-Net models were trained , this step can be elimin ated. Ac cording to our resu lts, it can be argued that the modifie d U-N et arch itecture is capable of pro ducing land cover maps that impro ve the bench mark class ificatio n produc ed by former Random Forest and SVM models . The land-co ver class ificati on models c an b e attache d d irectly to th e UAV vehicle to detect the land covers quickly without post data a nalysis . Import antly, the time to predic t lan d-use types from dig ital signifi cantly , co mpared to the tra ditiona l me thods. Common ly, s cient ists h ave to take v arious gr ound contro l points as a mask to interpret land-use types from UAV and other s atelli te imag es. Who le inte rpretat ion pro cess req uires a hard work both indoor and outdoor. Sometimes , it can take a month to h ave a land-us e map for a par ticular t ime. The gro und contro l points cannot be used for the new data or new areas . With the us e of deep learn ing metho ds appl ied for train ed U- Net models , n ow users can ge nerate la nd-use map , p articula rly in the mini ng areas in a shor t time (abou t 120s in this study). Additi onally, th e t rained models are no t only use d for the resear ch area , but it can als o be us ed for differ ent are as wher e are mo nitored by the UAV t echnolo gy. Compar ed to th e land-co ver classifica tion in ur ban and agricul tural low- lands usi ng d eep lear ning from for mer studi es [35], [37], [76] – [78], this s tudy chos e a typical minin g ar ea on basalt highland area of Vietn am. Except for the agricultur al lands, two land cover types wer e not train ed in forme r stu dies, includi ng the open-cast mini ng areas and polluted water bodies . Du ring the ro ck minin g a ctiviti es, basalt s oils was extrac ted and left on the top soil surfaces. It leads to the specif ic texture and homoge neous spatial distribut ion of the in the research area were pollu ted by the mining activiti es, leading to the high re flection of sunlight; whereas the freshw ater stream will absorb energy from sunligh t. Therefo re, the po llution le vel of strea m flowing f rom the op en- case minin g areas also can monit or if the UAV images are taken f or a lon g ti me. B. IMPROVEME NT OF LAND C OVER CLASS IFICATI ON Four tr ained U-N et mod els usin g Adam, Adagrad , RMSpro p, and SGD optimi zer functions (Group 1) cannot separ ate old and youn g permanen t cropla nds due to the sp ectral similarit y these two land covers . Whereas , the three models using Adamax , Nada m and Adadelt a (Group 2) can separate these two a gricult ural la nds bas ed on the d ifferen ces in their textur e and sh ape. T he di fferen ces be tween the t wo typ es of crops are their canopy . In the s ame area of cro plands , the old perm anent crops have a m ore mass ive ca nopy than the young on es. Sm all bare-so il regions that are distributed among the youn g perman ent croplan ds c an be elim inated by using the mode ls in Group 1. Meanwh ile, the models in Group 2 can detect both the small bare-s oil regions and young perman ent crops as an object that is diff erent from an ob ject covered by ful l old This article has been accepted for publication in a future issue of this journal, but has not been fully edited. Content may change prior to final publication. Citation information: DOI 10.1109/ACCESS.2020.3030112, IEEE Access Author Name : Preparation of Papers for IEEE Ac cess (February 2017) VOLUME XX, 20 20 perman ent crop s. It e xplains why th e models in Gr oup 2 ha ve higher accuracy values than those in Group 1. It is crucial if develop ers in the futur e want to detect agricultur al lands in differ ent growing periods. New sampl es of agricultur al land cover types can be automa tically updat ed in next UA V flig hts to increas e t he accurac y of t he trained U-N et model . It als o c an update the sample from high-re solutio n image from Google earth d ata sou rces. It shoul d be d one in furth er studi es. The U-Net developmen t for land cover classificat ion requir es a cost and time-con suming dedica tion of dev elopers , especia lly of s cientis ts. T o m onitor mini ng la nd covers in real- time, the managers have t o be equipped with a high- perfor mance machine to (1) transmit data direct ly from the UAV machin e to se rver with a high-s peed conn ection , (2) pre- process UAV images , and (3) interp ret the land cov ers based on the train ed deep-le arning mod el. In this stud y, the authors u sed a CPU Intel (R) Core ™ i7 -97 50H CPU @ 2.60GH z C PU with 16GB RAM and GPU NVI DIA GeFor ce RTX 2 060. The improv ed with a superco mputer , especiall y in the case of the rapid land-cover classific ation base d on the UAV data. Additi onally, the trained model has to b e updated from the new d ata. In stead of us ing s even o ptimiz ers to impro ve t he U- Net models, various optim ization techniques such as

https://www.researchgate.net/publication/344662205_U-Net_convolutional_networks_for_mining_land_cover_classification_based_on_high-resolution_UAV_imagery

EFFECT OF CONSUMING TEMULAWAK (CURCUMA XANTHORRHIZA ROXB.) EXTRACT ON BREAST MILK PRODUCTION IN POSTPARTUM MOTHERS | ScienceGate Background: The dominant factor inhibiting breastfeeding is the lack of milk production. The extract of temulawak (curcuma xanthorrhiza Roxb) ... EFFECT OF CONSUMING TEMULAWAK (CURCUMA XANTHORRHIZA ROXB.) EXTRACT ON BREAST MILK PRODUCTION IN POSTPARTUM MOTHERS Belitung Nursing Journal ◽ 10.33546/bnj.188 ◽ 2017 ◽ Vol 3 (5) ◽ pp. 603-611 Author(s): Chyntia Desbriyani ◽ Soeharyo Hadisaputro ◽ Ari Suwondo ◽ Supriyana Supriyana Keyword(s): Breast Milk ◽ Milk Production ◽ Dominant Factor ◽ Control Group ◽ Control Group Design ◽ Breast Milk Production ◽ Equivalent Control ◽ Increase Milk Production ◽ Curcuma Xanthorrhiza ◽ Postpartum Mothers Background: The dominant factor inhibiting breastfeeding is the lack of milk production. The extract of temulawak (curcuma xanthorrhiza Roxb) is considered having an effect to increase breast milk production.Objective: To examine the effect of temulawak (curcuma xanthorrhiza Roxb) extract to increase milk production in postpartum mothers in the working area of Ambarawa Community Health Center.Methods: This was a quasy-experimental study with non-equivalent control group design conducted on October-December 2016. There were 38 respondents included using consecutive sampling, with 19 assigned in the intervention and control group. Paired t-test and independent test were used for data analysis.Results: Findings showed that there were significant increases of prolactin hormone (p = 0.000), breast milk volume (p = 0.001), baby’s urinary frequency (p = 0.001), baby’s defecation frequency (p = 0.000), and baby’s sleep duration (p = 0.000) after given temulawak (curcuma xanthorrhiza Roxb) extract.Conclusion: Temulawak (curcuma xanthorrhiza Roxb) extract has a significant effect in increasing breast milk production and prolactin levels in postpartum mothers. Thus, it is recommended that temulawak (curcuma xanthorrhiza Roxb) extract can be an option for postpartum mother to increase milk production. Jurnal Keperawatan Silampari ◽ 10.31539/jks.v2i2.529 ◽ 2019 ◽ Vol 2 (2) ◽ pp. 66-86 Author(s): Tabita Mariana Doko ◽ Kun Aristiati ◽ Suhoryo Hadisaputro Keyword(s): Breast Milk ◽ Experimental Method ◽ Milk Production ◽ Design Approach ◽ Control Group ◽ Control Group Design ◽ Group Design ◽ Breast Milk Production ◽ Equivalent Control ◽ Postpartum Mothers The purpose of this study was to prove and verify the effect of oxytocin massage by the husband on increasing milk production in postpartum mothers. The method in this study uses a quasy experimental method with a non equivalent control group design approach. The results of the study, the administration of oxytocin massage by the husband affected the increased production of breast milk (ASI) with indicators of infant weight (p <0.05), frequency of breastfeeding (p <0.05), length of sleep of infants (p <0.05), frequency of discharges baby defecate (BAB) (p <0.05), frequency of urination of infants (BAK) (p <0.05), and resting mother's sleep (p <0.05). Conclusion, giving oxytocin massage by the husband can increase the production of Breast Milk (ASI) in postpartum mothers seen by the weight of the baby today, frequency of breastfeeding, length of sleep of infants, frequency of defecation of infants (BAB), frequency of defecation of infants (BAK) ), and resting mother's sleep.   Keywords: Postpartum Mother, Oxytocin Massage by Husband, Breast Milk Production (ASI) Coping: Community of Publishing in Nursing ◽ 10.24843/coping.2020.v08.i04.p10 ◽ 2020 ◽ Vol 8 (4) ◽ pp. 416 Author(s): Anita Liliana ◽ Melania Wahyuningsih Keyword(s): Breast Milk ◽ Milk Production ◽ Post Partum ◽ Control Group ◽ P Value ◽ Breast Milk Production ◽ Significant Difference ◽ Before And After ◽ Increase Milk Production ◽ Abstract Adequacy of milk production in the newborns will affect the success of exclusive breastfeeding. Not all postpartum mothers are able to increase milk production in the postpartum period. Acupressure at the meridian points is one way to increase milk production in postpartum mothers. This study aims to determine the effect of acupressure therapy on increasing breast milk production in postpartum mothers at PKU Muhamadiyah Bantul. This study was a quantitative study  a quasi-experimental research design with pre-test and post-test nonequivalent control group. The number of respondents in each group was 17 people. The treatment given was acupressure at the meridian points for 5 minutes on days 2 and 3of poatpartum. Breastfeeding adequacy was assessed before treatment and on day 4 postpartum. Data were analyzed by Mc Nemar analysis. The results showed that there was a significant difference in breast milk production before and after acupressure with p value of 0.000 (p value <0.05). There was a significant difference in breast milk production in the control group before and after being given the breastfeeding technique leaflet p value 0.001 (p value <0.05). Conclusion: There is no significant difference in breast milk production after being given acupressure and after being given leaflets about breastfeeding techniques at PKU Muhamadiyah Bantul p value 0.100 (p value> 0.05).  Keywords: acupressure, breast milk production, postpartum COLOSTRUM : Jurnal Kebidanan ◽ 2021 ◽ Vol 2 (1) ◽ pp. 70-77 Author(s): Fadilah Widyaningsih ◽ Tri Marini SN Keyword(s): Breast Milk ◽ Milk Production ◽ Post Partum ◽ Rank Test ◽ Control Group ◽ P Value ◽ Food Factor ◽ Breast Milk Production ◽ Postpartum Mothers Banana blossom is a food that has many benefits and is easily obtained by the community because it can be easily planted in the yard of the house. Banana blossom processing in the community can be done by boiling and steaming. Banana blossom also contains protein, minerals (especially phosphorus, calcium and iron) and a number of vitamins A, B1 and C. There are many reasons for mothers do not give exclusive breastfeeding to their babies. However, what is most often found is insufficient breast milk. There are several factors that influence milk production, one of which is the food factor. One of the efforts to increase breastmilk production that will be carried out is by giving Kepok Banana blossom Simplicia. This study aims to determine the effect of Kepok banana blossom simplicia on increasing breast milk production in postpartum mothers at Nurhayati and Nining Pelawati Midwifery Pratama Clinic, Lubuk Pakam sub district in 2019. This type of research was a Quasi Experiment with a Non Equivalent Control Group design and purposive sampling. Samples were normal postpartum mothers from day 7 to day 21 with parity of 1-2 and were willing to take part in the study through informed consent. Based on the results of the Wilcoxon Rank test and the Mann-Whitney test, it was obtained a p-value of 0.000 (α <0.05), which means that there was a significant effect on the simplicia of Kepok banana blossom on increasing milk production in postpartum mothers and it is expected that it can be used as an effort to increase milk production in postpartum mothers and to meet the needs of babies without having formula milk. Journal of Health Sciences ◽ 10.33086/jhs.v12i1.553 ◽ 2019 ◽ Vol 12 (1) ◽ pp. 60-66 Author(s): Kusumastuti , ◽ Umi Laelatul Qomar ◽ Siti Mutoharoh Keyword(s): Breast Milk ◽ Milk Production ◽ Intervention Group ◽ Control Group ◽ P Value ◽ Chi Square ◽ Control Group Design ◽ Breast Milk Production ◽ Equivalent Control Abstract: Lactation is the whole breastfeeding process from breastmilk produced to the baby's process of sucking and swallowing milk. The process of lactation is strongly influenced by the hormone prolactin and oxytocin. Expulsion of prolactin and oxytocin hormones can be stimulated by massage one of them with woolwich massage and oxytocin massage. Determine the effect of combination of Woolwich and oxytocin massages on breast milk production of postpartum mother. Thepresent study used quasi non-equivalent control group design experiment with data analysis using chi-square. The study revealed the differences of breast milk production of the Woolwich massage group and the oxytocin massage compared to the control group. The subjects were0-7 days post-partum mothers who breastfed their baby exclusively, amounting to 22 respondents in each group. Variables of breast milk production, the combination of Woolwich and oxytocin massage have p-value of  higher than 0.05 so it is not statistically significant but clinically it showed that the breast milk production of intervention group had 17% higher than the control group. Although there was no significant effect of the combination of Woolwich and oxytocin massage on breast milk production but there was clinically differenceof outcomes that is visible in intervention groups. pp. 94-98 Author(s): Sultina Sarita ◽ Syahrianti Syahrianti ◽ Keyword(s): Breast Milk ◽ Milk Production ◽ Community Health Centers ◽ Sampling Technique ◽ Control Group ◽ Control Group Design ◽ Breast Milk Production ◽ Pocket Book ◽ Significant Difference ◽ Postpartum Mothers Background: Normal Childbirth Care or APN is a standard or a reference for childbirth assistance and a breast crawl, or called IMD, in Indonesia; however, there is lack of its implementation in the midwifery practice. Breast crawl is considered effective in increasing breast milk production among postpartum mothers.Objective: The aim of the study was to determine the effectiveness of breast crawl by midwives to increase the breast milk production in postpartum mothers in Kendari, Indonesia.Methods: True experimental study with posttest only control group design. A total of 30 midwives participated using a total sampling technique. Respondents were given intervention regarding the implementation of breast crawl using the breast crawl pocket book as a research instrument. Data were analyzed using Mann-Whitney test to determine the differences in respondents who were given IMD intervention using the breast crawl pocket book and the control group who did not use the breast crawl pocket book.Result: There was a significant difference between respondents who were given intervention using pocket books and the control group who did not use pocket books with p = 0.039 (0.05).Conclusion: There was a significant difference between the breast milk production of postpartum mothers who were given breast crawl intervention and those who were not given breast crawl intervention by midwives. This intervention is therefore recommended as one of midwifery cares in the community health centers. Author(s): Sumirah Budi Pertami ◽ Budiono Budiono ◽ Ira Rahmawati Keyword(s): Breast Milk ◽ Milk Production ◽ Control Group ◽ Postpartum Women ◽ Control Group Design ◽ Breast Milk Production ◽ Post Test ◽ Quasi Experimental ◽ Postpartum Mothers ◽ And Control Background: Exclusive breastfeeding is the gold standard of infant feeding in the first six months of their life. In Indonesia, breastfeeding practice has become a social norm among women. However, some women have experienced inadequate breast milk production especially in the first and second days after birth. This study aimed to investigate the effect of endorphin and oxytocin massage on breast milk production among primiparous postpartum women and their baby weight. Methods: a quasi-experimental design was utilized with pre-test and post-test control group design.  The population in this study were 175 postpartum mothers in  Lawang Primary Health Care Indonesia. Forty  postpartum mothers were selected and allocated to the intervention and control groups. Ten women were received endorphin massage, ten were given oxytocin massage, ten mothers were received the combination of endorphin and oxytocin massage and the remaining ten were allocated into the control group. The dependent variables were breast milk production and infant body weight, whereases the independent variables were endorphin massage, oxytocin massage and combination of endorphin and oxytocin massages. Data were analyzed by using t-test followed by One Way ANOVA Test with α = 0.05 Results: The results indicated differences between the four group regarding the breast milk production and an increase in baby weight with p = 0, 00<α = 0, 05 and p = 0.046 <α = 0.05 respectively. Conclusion: According to the results, the combination of endorphin and oxytocin massage is the most effective technique to increase breast milk production among postpartum women and therefore, facilitating the baby weight gain in the first six months of their life. 10.36089/nu.v1i2.56 ◽ 2019 ◽ Vol 1 (2) ◽ pp. 46-56 Author(s): Hamimatus Zainiyah Keyword(s): Breast Milk ◽ Milk Production ◽ Post Partum ◽ Sampling Technique ◽ Control Group ◽ P Value ◽ Control Group Design ◽ Breast Milk Production ◽ Equivalent Control ◽ Breast Care During pregnancy, the prolactin hormone from placenta increases but breast milk usually does has come out because it is still inhibited by high estrogen levels. On the second or third day of postpartum, estrogen and progesterone levels drop significantly, so the effect of prolactin is more dominant and secretion of breast milk begins. By breastfeeding earlier the secretion of breast milk gets smoother. Based on a preliminary study, 46.0% of post partum mothers experienced non-breast milk. This study aims to determine the differences in the production of breast milk in post partum mothers who performed acupressure techniques for lactation and breast care techniques. This research is a kind of research of quasy exsperimental design with non equivalent control group design. The samples were 7 post partum mothers that used acupressure techniques for lactation and 7 post partum mothers used breast care techniques from in February to March 2017. The sampling technique used was non probability sampling (consecutive sampling). From the statistic test of Independent T-Test indicates that α = 0.05 and P value 0.03 thus P Value<alpha (0.03 <0.05) so that H0 is rejected and Ha accepted there was a difference in breast milk production in post partum mothers performed acupressure and breast care techniques in the BPM Sri Wahyuni area, S.ST Surabaya. Midwives are able to socialize this acupressure technique to all post partum mothers, so that midwives can help secretion of breast milk production for post partum mothers with acupressure techniques with relatively cheaper costs Pijat oksitosin oleh suami meningkatkan produksi ASI ibu primipara Jurnal Keperawatan ◽ 10.32583/keperawatan.v11i3.580 ◽ 2019 ◽ Vol 11 (3) ◽ pp. 231-236 Author(s): Machmudah Machmudah ◽ Aam Zakiah Adawiah Keyword(s): Breast Milk ◽ Milk Production ◽ Post Partum ◽ Wilcoxon Test ◽ Control Group ◽ Control Group Design ◽ Breast Milk Production ◽ Bowel Movements ◽ Defecation Frequency ◽ Postpartum Mothers Dukungan orang terdekat khususnya suami sangat dibutuhkan dalam mendukung ibu selama memberikan ASI-nya kepada ibu. Tujuan dalam penelitian ini adalah membuktikan pengaruh pijat oksitosin oleh suami terhadap produksi ASI pada ibu post partum normal. Jenis penelitian quasi esperiment dengan rancangan pretest-posttest with control group. Jumlah sampel 26 ibu post partum normal yang menyusui. Dilakukan pengukuran produksi ASI menggunakan lembar observasi dengan parameter BB bayi, frekuensi BAK bayi, Frekuensi BAB bayi, frekuensi menyusu bayi. Analisis data menggubakan uji wilcoxon. Ada pengaruh pijat oksitosin oleh suami terhadap Produksi ASI dibuktikan dengan meningkatnya frekuensi BAK, Frekuensi BAB, Frekuensi menyusu pada responden. Pemberian pijat oksitosin oleh suami berpengaruh terhadap produksi ASI dengan parameter frekuensi BAK bayi (p<0,05), frekuensi BAB bayi (p<0,05), frekuensi menyusui bayi (p<0,05). Pemberian pijat oksitosin oleh suami dapat meningkatkan produksi ASI pada ibu yang dilihat dari frekuensi BAK bayi, frekuensi BAB bayi dan frekuensi menyusu bayi.   Kata kunci: pijat oksitosin oleh suami, produksi ASI MASSAGE OF OXYTOSIN BY HUSBAND TO INCREASE PRODUCTION OF PRIMIPARA MOTHER'S BREAST MILK   ABSTRACT The support of the closest person, especially the husband, is needed in supporting the mother while giving her breast milk  to the mother. The objective was to prove the effect of oxytocin massage by the husband in producing breast milk of normal postpartum. This research used a quasi-experiment with a pretest-posttest with control group design. The sample was 26 normal postpartum mothers who breastfeed. Breastmilk production was measured using an observation sheet with parameters for baby's defecation, baby's urination frequency, baby's defecation frequency, and baby's breastfeeding frequency. The data analysis using the Wilcoxon-test. There was the influence of oxytocin massage by the husband in producing breast milk as evidenced by the increasing frequency of urinating, frequency of bowel movements, frequency of breastfeeding in respondents. In giving oxytocin massage by the husband influence the production of breast milk with the parameters of baby's urination frequency  (p <0.05), baby's defecation frequency (p <0.05), baby's breastfeeding frequency (p <0.05). The Oxytocin massage by the husband can increase breast milk production in the mother as seen from baby's urination frequency, baby's defecation frequency, and the baby's breastfeeding frequency.   Keywords: oxytocin massage by husband, breast milk production Jurnal Kesehatan ◽ 10.35730/jk.v11i2.532 ◽ 2020 ◽ Vol 11 (2) ◽ pp. 93 Author(s): Kadek Yuke Widyantari Keyword(s): Breast Milk ◽ Music Therapy ◽ Milk Production ◽ Scoping Review ◽ Control Group ◽ Relaxation Therapy ◽ Breast Milk Production ◽ Increase Milk Production ◽ Therapy Interventions ◽ Postpartum Mothers Author(s): Keyword(s): Breast Milk ◽ Milk Production ◽

https://www.sciencegate.app/document/10.33546/bnj.188

Spain: bodies of undocumented immigrants found by entry point | Statista This statistic presents the number recovered bodies of dead irregular migrants in Spain in 2019, by route of entry. Society› Demographics Premium Premium statistics Industry-specific and extensively researched technical data (partially from exclusive partnerships). A paid subscription is required for full access. Read more Bodies of irregular migrants in Spain 2019, by point of entry Published byTeresa Romero,Jul 5, 2021 This statistic presents the number recovered bodies of dead irregular migrants in Spain in 2019, by route of entry. During that year, the number of bodies recovered in Canary Islands was around 79. 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Applied Sciences | Free Full-Text | In Vitro Analysis of Organic Ester Functional Groups in Carious Dentine Background: With the implementation of minimally invasive selective caries removal protocols to treat cavitated, deep carious dentine lesions, there is a need to investigate specific biochemical moiety distributions to help characterise and distinguish between infected (contaminated) and affected (demineralised) zones within the dentine lesion. The present in vitro investigation aimed to compare the distribution of ester functional groups (1740 cm−1) within carious dentine tissue (infected and affected dentine). The null hypothesis stipulated that there are no differences in ester function intensity/distribution within carious dentine lesions. Materials and Methods: From a total of five extracted human molar teeth with carious dentine lesions, 246 points from 10 sections of carious dentine were examined using high-resolution Raman spectroscopy and characterised into infected, affected and sound dentine. The peak intensity of the characteristic vibration mode of the ester function was calculated from sample scans. Results: Analyses indicated a statistically significant difference in the spectroscopic vibration bands of esters between the infected and affected dentine zones. Conclusion: The ester functional group is higher in intensity in the caries-infected dentine zone compared to the affected tissue. This finding could be used to develop an objective indicator for the selective operative management of carious dentine. In Vitro Analysis of Organic Ester Functional Groups in Carious Dentine by Mohammed Alturki 1,2 , Ulrica Almhöjd 1,3 , Garrit Koller 4,5 , Fiona Warburton 6 and Centre of Oral Clinical Translational Sciences, Faculty of Dentistry, Oral & Craniofacial Sciences, King’s College London, London SE1 9RT, UK 2 Department of Restorative Dental Sciences, College of Dentistry, King Saud University, Riyadh 11451, Saudi Arabia 3 Department of Cariology, Institute of Odontology, The Sahlgrenska Academy, University of Gothenburg, Box 450, SE-405 30 Gothenburg, Sweden 4 Centre for Host Microbiome Interactions, Faculty of Dentistry, Oral & Craniofacial Sciences, King’s College London, London SE1 9RT, UK 5 LCN—London Centre for Nanotechnology, 19 Gordon St, Bloomsbury, London WC1H 0AH, UK 6 Faculty of Dentistry, Oral & Craniofacial Sciences, King’s College London, London SE1 9RT, UK * Author to whom correspondence should be addressed. Appl. Sci. 2022 , 12 (3), 1088; https://doi.org/10.3390/app12031088 Received: 30 November 2021 / Revised: 7 January 2022 / Accepted: 18 January 2022 / Published: 20 January 2022 (This article belongs to the Topic Advances in Dental Biomaterials and Oral Tissues Engineering ) Abstract : −1 ) within carious dentine tissue (infected and affected dentine). The null hypothesis stipulated that there are no differences in ester function intensity/distribution within carious dentine lesions. Materials and Methods: From a total of five extracted human molar teeth with carious dentine lesions, 246 points from 10 sections of carious dentine were examined using high-resolution Raman spectroscopy and characterised into infected, affected and sound dentine. The peak intensity of the characteristic vibration mode of the ester function was calculated from sample scans. Results: Analyses indicated a statistically significant difference in the spectroscopic vibration bands of esters between the infected and affected dentine zones. Conclusion: The ester functional group is higher in intensity in the caries-infected dentine zone compared to the affected tissue. This finding could be used to develop an objective indicator for the selective operative management of carious dentine. Keywords: caries-infected ; caries-affected ; dentine ; Raman spectroscopy ; lucifer yellow ; amide I ; ester ; minimally invasive dentistry ; caries 1. Introduction Characterising and discriminating between denatured, necrotic, bacterially contaminated and demineralised tissue in a cavitated carious lesion is of clinical relevance when performing minimally invasive operative interventions [ 1 , 2 ]. Carious dentine consists of two main histological zones [ 3 , 4 ]. The outer, superficial, highly bacterially contaminated and denatured zone of tissue is clinically wet, soft and sticky—the caries-infected (bacterially contaminated) dentine. The remaining, deeper caries-affected (demineralised) dentine can be healed and repaired by the dentine–pulp complex and, therefore, can be retained and sealed off using biointeractive restorative materials [ 1 , 5 , 6 , 7 ]. This selective, minimally invasive operative approach preserves more tissue and has been shown to improve the long-term survival of the dentine–pulp complex [ 1 , 5 , 7 , 8 ]. The histological transition between these two zones of tissue, however, is diffuse and difficult to identify clinically as well as in the laboratory, which leads to subjectivity and variations between operators/researchers and, therefore, often unnecessary excessive removal of tooth tissue by clinicians. Therefore, there is an unmet need to objectively delineate between caries-infected and affected dentine, marking the clinical endpoint of carious dentine excavation in both laboratory investigations and the clinic [ 5 , 9 , 10 ]. In vitro studies have predominantly characterised carious dentine tissues mechanically. Ogawa and co-workers used microhardness to divide carious dentine into outer (superficial) and inner lesion layers [ 11 ]. Banerjee et al. (2010) further classified the carious dentine lesion according to its Knoop hardness value and its intrinsic autofluorescent characteristics, with caries-infected dentine (CID) <25 KHN and caries-affected dentine (CAD) between 25 and 40 KHN. However, the hardness measurement only indirectly represents the basic mechanical mineral integrity of the tissue, without a direct correlation with the biochemical characteristics of the lesion [ 12 , 13 ], the latter having a role in tissue repair and biomaterial adhesion. Biochemically, carious dentine contains high levels of organic compounds, proportionate with a relatively lower inorganic content, compared to sound dentine [ 4 , 14 , 15 ]. Alturki et al. (2021) characterised the carious dentine lesion using the Raman spectroscopic protein/mineral ratio (amide I/phosphate) according to the tissue microhardness. That study concluded the amide I/phosphate Raman spectral ratio could be used as a biochemical method to characterise carious dentine with a direct association with lesion histology. In addition to amide I (-CO-NH-), which represents the main organic component in carious dentine, Almhöjd et al. (2014) showed that the protein ester functional group (-CO-OR-) can be considered a unique functional group in carious dentine, as it is not observed in sound dentine. This is due to the fact that esters are formed in an environment that contains less water and a low pH compared to that found in carious dentine [ 16 ]. However, its distribution has yet to be mapped through the histological zones of infected and affected dentine. Determining any difference in intensity, and therefore distribution, of the ester functional groups between infected and affected zones of carious dentine could help develop clinical indicators to aid practitioners with selective, minimally invasive operative carious dentine removal technologies and develop research towards developing novel chemical adhesion mechanisms for biointeractive restorative materials and tissue repair mechanisms. Therefore, this in vitro study aimed to compare the distribution of ester functional groups within carious dentine tissue (infected and affected dentine). The null hypothesis was that there were no differences in the Raman spectroscopic peak vibrational intensities of ester functional groups between infected and affected carious dentine tissues. In addition, lucifer yellow (LY) was used to subjectively visualise any colour changes in the carious dentine zones, as it has the chemical ability to react with such ester functional groups, thus acting as a biochemical label [ 14 ]. 2. Materials and Methods 2.1. Ethics and Sample Collection Extracted human teeth with cavitated carious dentine lesions were collected under an ethics protocol reviewed and approved by NHS Health Research Authority (16/SW/0220). 2.2. Study Design Five extracted human molar teeth with carious dentine lesions having an international caries detection and assessment system (ICDAS) lesion score >4 [ 17 ] were collected (ethics protocol reviewed and approved by NHS Health Research Authority (16/SW/0220)) and stored for no longer than three months in distilled water in a cold cabinet (+4 °C) before sectioning. Samples were sectioned longitudinally through the lesions using a slow-speed (200 rpm) water-cooled diamond blade (diamond wafering blade XL 12205, Benetec Ltd., London, UK), and digital images of the cut surfaces were captured (lens: Nikkor AF-S Micro 60mm f/2.8G ED, D800E, Nikon, Tokyo, Japan). To obtain a flat surface for accurate measurements, the cut surfaces were polished (MetaServ 3000, Buehler, Lake Bluff, IL, USA) sequentially with silicon waterproof abrasive papers (P1200 for 10 s, P2500 for 10 s and P4000 for 4 min). The samples were first analysed using Raman spectroscopy, and then lucifer yellow was added as a biochemical ester functional group label. 2.3. Raman Spectroscopy A high-resolution Raman spectroscope (inVia, Renishaw Plc, Wotton-under-Edge, Gloucestershire, UK) running in Streamline scanning mode was used to scan the flat tooth surfaces, and a 5/0.12 NA air objective was used. A total of 246 point scans were captured from 10 carious dentine sections. In each section, an average of 18 point scans were captured along 3 line scans starting from the outermost part of the lesion in the direction from the enamel–dentine junction towards the pulp. A further 6 line point scans were captured in clinically sound dentine in the same section. The vibrational intensity of ester functional groups was calculated for each point scan (measured at 1740 cm −1 ). Spectrum acquisition was conducted using a 785 nm laser (10 mW for mapping mode and 0.5 mW for point scanning) using a 600 line/mm grate across the spectral bandwidth of 4000–400 cm −1 for each sample. Baseline correction was performed by Raman processing software (WiRe, Renishaw, UK). 2.4. Molecular Label Preparation Lucifer yellow (carbazide; LY) was used to verify the Raman results in the carious dentine zones (infected and affected dentine), as it reacts specifically with ester groups in carious dentine tissues [ 18 ]. All tooth sections were initially exposed to 0.5 M NaBH 4 for one hour in ethanol to remove any unwanted reactions between the lucifer yellow and any ketones or aldehydes present in the tissues. This step left only the organic ester functional groups able to react with lucifer yellow [ 14 ]. Subsequently, all sections were then immersed in a 15 mM ethanol solution of lucifer yellow for 24 h. To reduce the risk of any unwanted electrostatic or hydrogen bonding, sections were placed in a saline solution (NaCl, 1 M) for another 24 h. In order to ensure that only covalently bound complexes were retained within the carious tissue, a final step was introduced by immersing the specimens in alkaline solution (0.5 M NaOH) for a final 24 h period. 2.5. Data Collection and Analysis Statistical analysis and spectral correlation processing. The ratio of Raman vibrational intensity peaks at 1650 cm −1 /960 cm −1 (amide I/phosphate) has been reported as a biochemical method to distinguish between the histological carious dentine zones [ 19 ]. The peak ratio at 1740 cm −1 /960 cm −1 was calculated in each point scan to characterise the ester (1740 cm −1 ) group distribution in infected, affected and sound dentine [ 19 ]. All peaks were normalised to the phosphate peak (960 cm −1 ), as this is the most intense and consistent Raman peak in dental hard tissues [ 20 ]. A descriptive analysis was performed. All analyses were carried out using IBM SPSS, version 25.0. 3. Results Two hundred and forty-six examined points from 10 sections of carious dentine were analysed using Raman spectroscopy and characterised into infected, affected and sound dentine zones. The results showed a significant difference in the ester functional group intensities between the three histological dentine zones ( Table 1 , Figure 1 ). The descriptive analysis showed that the mean intensity of ester functional groups in the caries-infected dentine was almost three times higher than that in the caries-affected dentine (mean intensities: 3.3, 0.8 and 0.04 in infected, affected and sound, respectively). Lucifer yellow (LY) was observed on the whole tooth section surface after the initial 24 h immersion in LY solution, with more noticeable discolouration found within the carious dentine lesion ( Figure 2 ). However, after immersion in the salt and alkaline solutions, the LY staining was observed only in the carious dentine lesion (covalently bound). 4. Discussion Contemporary carious dentine excavation should be selective, preserving the remineralisable carious tissues and removing the infected and necrotic tissue only [ 1 , 6 ]. Thus, clinicians and researchers are continually trying to develop and optimise technologies that can objectively discriminate between these histological carious dentine zones. Biochemical alterations have been shown to play a significant role in carious dentine, specifically the organic components, with regard to tissue repair [ 14 , 21 , 22 ]. In vitro studies have used the organic components to distinguish carious dentine, as they become more exposed after demineralisation [ 14 , 20 , 23 , 24 ]. Specifically, the ester functional group has been isolated in carious dentine but not in sound dentine [ 18 , 25 ]. However, its distribution has not been characterised histologically to differentiate between carious dentine zones. Esters are naturally formed from reactions between structures with alcohol functions, such as carbohydrates and carboxylic acids, from protonated proteins and are catalysed by acidic agents, described as Fischer esterification. This process is influenced by water levels and changes in acidity, which means that it increases in an environment that contains low water content and low pH [ 16 ]. The present study demonstrated that the increased relative intensity of ester functional groups was higher in caries-infected dentine compared to the adjacent caries-affected dentine. This finding is consistent with the established knowledge that there are more proteins in infected dentine compared to sound tissue [ 14 , 21 ]. In addition, it is in agreement with the nature of ester formation, as it forms in a more acidic environment, such as that found in caries-infected dentine. However, in the present study, there was a trace amount of ester function found in sound dentine ( Table 1 ). This could be explained by the fact that the spectral data of the vibration bands of ester functional groups do not differentiate between types of esters, which also include glycerol-based phospholipids such as cholesterol- and phosphatidyl esters, a normal component of cell membranes [ 26 , 27 ]. Not all types of esters in carious dentine zones have been characterised. Only carbonyl ester functions (COOR) have been identified in carious tissues (Almhöjd et al., 2014, 2017) [ 14 , 18 ]. Besides these carbonyl ester functions (COOR), there might also be phosphor esters from lipids [ 28 , 29 ], trans- or diesters and from sugar acetals [ 30 ] or cyclic esters that can arise from lactones (lactic acid esters) originating from cariogenic bacteria [ 31 , 32 ]. In the present study, Raman microscopy detected all vibrational bands of ester functional groups. Therefore, a detailed analysis of ester functional groups in each carious dentine zone could be investigated further in the future. LY was used in this study because it can react covalently with all ester groups [ 18 ]. In the present study, a change in the colour of the carious dentine was subjectively observed, which is explained by the fact that significant amounts of ester functional groups in infected and affected dentine visibly reacted with LY, but no changes in sound tissues were perceived. To the authors’ knowledge, the present study is the first to objectively show the concentration and distribution of ester functional groups within carious dentine tissues, highlighting this important biochemical alteration and correlating it to the histological zones of caries-infected and caries-affected dentine. This, in turn, could pave the way for the investigation of objective clinical markers to delineate these zones of carious dentine to aid the clinician in their selective removal as part of the minimally invasive operative approach to deep cavitated lesions. New selective operative technologies might use the ester functional group distribution to target caries-infected dentine for selective removal, and innovations in biointeractive restorative material development might explore the further use of organic components for tissue repair and adhesion. Further analysis of the biochemical nature of ester functional groups in each of the carious dentine zones would contribute to further characterisation of the tissues in this regard. 5. Conclusions This in vitro study concluded that ester functional groups are distributed differently within carious dentine lesions, with a higher concentration in the caries-infected zone compared to the adjacent caries-affected dentine, which is detectable using Raman spectroscopy. Thus, the null hypothesis was rejected. Author Contributions M.A., G.K., U.A. and A.B. contributed to the conception, design, data acquisition, analysis and interpretation, as well as drafting and critical revision the manuscript. F.W. advised on the statistical analysis, interpretation of results and critical revision of the manuscript. All authors have read and agreed to the published version of the manuscript. Funding This research was funded by College of Dentistry, King Saud University, Riyadh, Saudi Arabia, for grant sponsorship of the first author (grant no. 4/52/141590). The funder had no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript. Institutional Review Board Statement The study was conducted in accordance with the Declaration of Helsinki, and approved by the Health Research Authority (16/SW/0220). Informed Consent Statement Not applicable. Data Availability Statement The data presented in this study are stored in a digital secured place of King’s College London and available upon reasonable request. Acknowledgments The authors gratefully acknowledge the assistance provided by Peter Pilecki and the College of Dentistry, King Saud University, Riyadh, Saudi Arabia, for grant sponsorship of the first author (grant no. 4/52/141590). The funder had no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript. Conflicts of Interest The authors declare no conflict of interest in relation to this study. References Banerjee, A. Selective Removal of Carious Dentin. In Management of Deep Carious Lesions ; Schwendicke, F., Ed.; Springer International Publishing: Cham, Switzerland, 2018; pp. 55–70. ISBN 978-3-319-61370-3. [ Google Scholar ] Slimani, A.; Terrer, E.; Manton, D.J.; Tassery, H. Carious Lesion Detection Technologies: Factual Clinical Approaches. Br. Dent. J. 2020 , 229 , 432–442. [ Google Scholar ] [ CrossRef ] Ogushi, K.; Fusayama, T. Electron Microscopic Structure of the Two Layers of Carious Dentin. J. Dent. Res. 1975 , 54 , 1019–1026. [ Google Scholar ] [ CrossRef ] Fusayama, T. Two Layers of Carious Dentin; Diagnosis and Treatment. Oper Dent. 1979 , 4 , 63–70. [ Google Scholar ] Kidd, E.A.M. How “clean” Must a Cavity Be before Restoration? Caries Res. 2004 , 38 , 305–313. [ Google Scholar ] [ CrossRef ] [ PubMed ] Schwendicke, F.; Dörfer, C.E.; Paris, S. Incomplete Caries Removal: A Systematic Review and Meta-Analysis. J. Dent. Res. 2013 , 92 , 306–314. [ Google Scholar ] [ CrossRef ] [ PubMed ] Innes, N.P.T.; Frencken, J.E.; Bjørndal, L.; Maltz, M.; Manton, D.J.; Ricketts, D.; Van Landuyt, K.; Banerjee, A.; Campus, G.; Doméjean, S.; et al. Managing Carious Lesions: Consensus Recommendations on Terminology. Adv. Dent. Res. 2016 , 28 , 49–57. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Schwendicke, F. Removing Carious Tissue: Why and How? Monogr. Oral. Sci. 2018 , 27 , 56–67. [ Google Scholar ] [ CrossRef ] [ PubMed ] Banerjee, A.; Kidd, E.A.; Watson, T.F. In Vitro Evaluation of Five Alternative Methods of Carious Dentine Excavation. Caries Res. 2000 , 34 , 144–150. [ Google Scholar ] [ CrossRef ] [ PubMed ] Isolan, C.P.; Sarkis-Onofre, R.; Lima, G.S.; Moraes, R.R. Bonding to Sound and Caries-Affected Dentin: A Systematic Review and Meta-Analysis. J. Adhes. Dent. 2018 , 20 , 7–18. [ Google Scholar ] [ CrossRef ] Ogawa, K.; Yamashita, Y.; Ichijo, T.; Fusayama, T. The Ultrastructure and Hardness of the Transparent Layer of Human Carious Dentin. J. Dent. Res. 1983 , 62 , 7–10. [ Google Scholar ] [ CrossRef ] Banerjee, A.; Cook, R.; Kellow, S.; Shah, K.; Festy, F.; Sherriff, M.; Watson, T. A Confocal Micro-Endoscopic Investigation of the Relationship between the Microhardness of Carious Dentine and Its Autofluorescence. Eur. J. Oral Sci. 2010 , 118 , 75–79. [ Google Scholar ] [ CrossRef ] [ PubMed ] Lippert, F.; Lynch, R.J.M. Comparison of Knoop and Vickers Surface Microhardness and Transverse Microradiography for the Study of Early Caries Lesion Formation in Human and Bovine Enamel. Arch. Oral Biol. 2014 , 59 , 704–710. [ Google Scholar ] [ CrossRef ][ Green Version ] Almhöjd, U.S.; Norén, J.G.; Arvidsson, A.; Nilsson, Å.; Lingström, P. Analysis of Carious Dentine Using FTIR and ToF-SIMS. Oral. Health Dent. Manag. 2014 , 13 , 735–744. [ Google Scholar ] Alturki, M.; Koller, G.; Almhöjd, U.; Banerjee, A. Chemo-Mechanical Characterization of Carious Dentine Using Raman Microscopy and Knoop Microhardness. R. Soc. Open Sci. 2020 , 7 , 200404. [ Google Scholar ] [ CrossRef ] [ PubMed ] Carey, F.A.; Sundberg, R.J. Advanced Organic Chemistry , 5th ed.; Springer: New York, NY, USA, 2007; ISBN 978-0-387-44897-8. [ Google Scholar ] Pitts, N.B.; Ekstrand, K.R. International Caries Detection and Assessment System (ICDAS) and Its International Caries Classification and Management System (ICCMS)—Methods for Staging of the Caries Process and Enabling Dentists to Manage Caries. Community Dent. Oral Epidemiol. 2013 , 41 , e41–e52. [ Google Scholar ] [ CrossRef ] Almhöjd, U.S.; Lingström, P.; Nilsson, Å.; Norén, J.G.; Siljeström, S.; Östlund, Å.; Bernin, D. Molecular Insights into Covalently Stained Carious Dentine Using Solid-State NMR and ToF-SIMS. Caries Res. 2017 , 51 , 255–263. [ Google Scholar ] [ CrossRef ] [ PubMed ] Alturki, M.; Koller, G.; Warburton, F.; Almhöjd, U.; Banerjee, A. Biochemical Characterisation of Carious Dentine Zones Using Raman Spectroscopy. J. Dent. 2021 , 105 , 103558. [ Google Scholar ] [ CrossRef ] Wang, Y.; Spencer, P.; Walker, M.P. Chemical Profile of Adhesive/Caries-Affected Dentin Interfaces Using Raman Microspectroscopy. J. Biomed. Mater. Res. A 2007 , 81 , 279–286. [ Google Scholar ] [ CrossRef ][ Green Version ] Liu, Y.; Yao, X.; Liu, Y.W.; Wang, Y. A Fourier Transform Infrared Spectroscopy Analysis of Carious Dentin from Transparent Zone to Normal Zone. Caries Res. 2014 , 48 , 320–329. [ Google Scholar ] [ CrossRef ][ Green Version ] Seredin, P.; Goloshchapov, D.; Prutskij, T.; Ippolitov, Y. Phase Transformations in a Human Tooth Tissue at the Initial Stage of Caries. PLoS ONE 2015 , 10 , e0124008. [ Google Scholar ] [ CrossRef ] Maske, T.T.; Isolan, C.P.; van de Sande, F.H.; Peixoto, A.C.; Faria-E-Silva, A.L.; Cenci, M.S.; Moraes, R.R. A Biofilm Cariogenic Challenge Model for Dentin Demineralization and Dentin Bonding Analysis. Clin. Oral. Investig. 2015 , 19 , 1047–1053. [ Google Scholar ] [ CrossRef ] Lopes, C.D.C.A.; Limirio, P.H.J.O.; Novais, V.R.; Dechichi, P. Fourier Transform Infrared Spectroscopy (FTIR) Application Chemical Characterization of Enamel, Dentin and Bone. Appl. Spectrosc. Rev. 2018 , 53 , 747–769. [ Google Scholar ] [ CrossRef ] Almhöjd, U.S.; Lingström, P.; Melin, L.; Nilsson, Å.; Norén, J.G. Staining of Carious Dentine Using Dyes with Covalent and Electrostatic Binding Properties—An in-Vitro Study. Oral. Health Dent. Manag. 2015 , 14 , 7. [ Google Scholar ] Rostand, K.S.; Esko, J.D. Cholesterol and Cholesterol Esters: Host Receptors for Pseudomonas Aeruginosa Adherence. J. Biol. Chem. 1993 , 268 , 24053–24059. [ Google Scholar ] [ CrossRef ] Voet, D.; Voet, J.G. Biochemistry (Second Edition). Biochem. Educ. 1995 , 23 , 104–105. [ Google Scholar ] [ CrossRef ][ Green Version ] Gotliv, B.-A.; Veis, A. Peritubular Dentin, a Vertebrate Apatitic Mineralized Tissue without Collagen: Role of a Phospholipid-Proteolipid Complex. Calcif. Tissue Int. 2007 , 81 , 191–205. [ Google Scholar ] [ CrossRef ] [ PubMed ] Gotliv, B.A.; Veis, A. The Composition of Bovine Peritubular Dentin: Matching TOF-SIMS, Scanning Electron Microscopy and Biochemical Component Distributions. New Light on Peritubular Dentin Function. Cells Tissues Organs 2009 , 189 , 12–19. [ Google Scholar ] [ CrossRef ] Tipson, R.S.S.; Horton, D. Advances in Carbohydrate Chemistry and Biochemistry ; Academic Press: San Diego, CA, USA, 1995; Volume 51, ISBN 978-0-12-007251-4. [ Google Scholar ] Goh, S.Y.; Tan, W.-S.; Khan, S.A.; Chew, H.P.; Kasim, N.H.A.; Yin, W.-F.; Chan, K.-G. Unusual Multiple Production of N-Acylhomoserine Lactones a by Burkholderia Sp. Strain C10B Isolated from Dentine Caries. Sensors 2014 , 14 , 8940–8949. [ Google Scholar ] [ CrossRef ] Muras, A.; Mayer, C.; Otero-Casal, P.; Exterkate, R.A.M.; Brandt, B.W.; Crielaard, W.; Otero, A.; Krom, B.P. Short-Chain N-Acylhomoserine Lactone Quorum-Sensing Molecules Promote Periodontal Pathogens in In Vitro Oral Biofilms. Appl. Environ. Microbiol. 2020 , 86 , e01941-19. [ Google Scholar ] [ CrossRef ] Figure 1. One of the sections showing three Raman spectra: infected, affected and sound dentine after normalising the intensities to the phosphate peak, showing differences in the ester functional group peak ratio (1740/960 cm −1 ) between the three zones (indicated by the arrows). Figure 1. One of the sections showing three Raman spectra: infected, affected and sound dentine after normalising the intensities to the phosphate peak, showing differences in the ester functional group peak ratio (1740/960 cm −1 ) between the three zones (indicated by the arrows). Figure 2. (A) A photomicrograph of one of the samples after sectioning and (B) the direction line of the Raman point scans that were taken to calculate the intensity of ester functional groups. Figure 2. (A) A photomicrograph of one of the samples after sectioning and (B) the direction line of the Raman point scans that were taken to calculate the intensity of ester functional groups. Table 1. A descriptive analysis was used to measure the differences in ester functional group intensities at 1740 cm −1 between infected, affected and sound zones (characterised according to [ 19 ]), which shows a significant difference between the three zones as the confidence intervals do not overlap between the zones. Table 1. A descriptive analysis was used to measure the differences in ester functional group intensities at 1740 cm −1 between infected, affected and sound zones (characterised according to [ 19 ]), which shows a significant difference between the three zones as the confidence intervals do not overlap between the zones. Infected Dentine ** Mean 3.39 * 95% Confidence Interval for Mean Lower 2.95 Upper 3.84 Std. Deviation 2.11 Affected Dentine ** Mean 0.87 95% Confidence Interval for Mean Lower 0.76 Upper 0.98 Std. Deviation 0.58 Sound Dentine ** Mean 0.04 95% Confidence Interval for Mean Lower 0.02 Upper 0.06 Std. Deviation 0.07 * Peak ratio (ester/phosphate). ** These zones were characterised by using the amide I/phosphate ratio: infected > 1.76, affected < 1.76 [ 19 ]. The sound dentine readings were taken from pure clinically dentine sound areas. Alturki, Mohammed, Ulrica Almhöjd, Garrit Koller, Fiona Warburton, and Avijit Banerjee. 2022. "In Vitro Analysis of Organic Ester Functional Groups in Carious Dentine" Applied Sciences12, no. 3: 1088. https://doi.org/10.3390/app12031088 MDPI and ACS Style Alturki, M.; Almhöjd, U.; Koller, G.; Warburton, F.; Banerjee, A. In Vitro Analysis of Organic Ester Functional Groups in Carious Dentine. Appl. Sci. 2022, 12, 1088. https://doi.org/10.3390/app12031088 AMA Style Alturki M, Almhöjd U, Koller G, Warburton F, Banerjee A. In Vitro Analysis of Organic Ester Functional Groups in Carious Dentine. Applied Sciences. 2022; 12(3):1088. https://doi.org/10.3390/app12031088 Chicago/Turabian Style Alturki, Mohammed, Ulrica Almhöjd, Garrit Koller, Fiona Warburton, and Avijit Banerjee. 2022. "In Vitro Analysis of Organic Ester Functional Groups in Carious Dentine" Applied Sciences12, no. 3: 1088. https://doi.org/10.3390/app12031088

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Sustainability | Free Full-Text | Optimal Design of Subway Train Cross-Line Operation Scheme Based on Passenger Smart Card Data The network operation of the subway can reduce the number of passenger transfers and improve subway operation efficiency. Based on the subway smart card data, this paper proposes an optimal design method for the cross-line operation scheme of subway trains. The method firstly calculates the OD matrix between subway stations, the passenger section flow, and the transfer flow according to the passenger smart card data. It then optimizes the design of the subway train cross-line operation plan, including determining the routing type of cross-line operation and the number of trains running. Finally, for the lines with cross-line operation conditions in the urban subway system, we design the cross-line operation schemes under all possible combinations. According to the volume of the cross-line passenger flow that the cross-line trains can carry, the top-ranked operation plans are preferably recommended. Taking the Chengdu subway network as an example, the research results show that, in the Chengdu subway network, the North Railway Station, Xibo City Station, and South Railway Station bear the highest transfer demand. The transfer demands that can be undertaken by cross-line trains are 23,934, 16,710, and 13,024 trips per hour, respectively. This shows that the proposed design method can accurately and reasonably screen out the transfer stations and lines with an urgent need for cross-line trains. Optimal Design of Subway Train Cross-Line Operation Scheme Based on Passenger Smart Card Data School of traffic and transportation engineering, Central South University, Changsha 410075, China Author to whom correspondence should be addressed. Sustainability 2022 , 14 (11), 6420; https://doi.org/10.3390/su14116420 Received: 20 April 2022 / Revised: 19 May 2022 / Accepted: 23 May 2022 / Published: 24 May 2022 (This article belongs to the Topic Sustainable Transportation ) Abstract : The network operation of the subway can reduce the number of passenger transfers and improve subway operation efficiency. Based on the subway smart card data, this paper proposes an optimal design method for the cross-line operation scheme of subway trains. The method firstly calculates the OD matrix between subway stations, the passenger section flow, and the transfer flow according to the passenger smart card data. It then optimizes the design of the subway train cross-line operation plan, including determining the routing type of cross-line operation and the number of trains running. Finally, for the lines with cross-line operation conditions in the urban subway system, we design the cross-line operation schemes under all possible combinations. According to the volume of the cross-line passenger flow that the cross-line trains can carry, the top-ranked operation plans are preferably recommended. Taking the Chengdu subway network as an example, the research results show that, in the Chengdu subway network, the North Railway Station, Xibo City Station, and South Railway Station bear the highest transfer demand. The transfer demands that can be undertaken by cross-line trains are 23,934, 16,710, and 13,024 trips per hour, respectively. This shows that the proposed design method can accurately and reasonably screen out the transfer stations and lines with an urgent need for cross-line trains. Keywords: passenger smart card data ; metro train operation scheme ; OD demand ; cross-line train 1. Introduction Rail transit construction has an important impact on urban development [ 1 , 2 ]. In recent years, with the gradual network construction of urban subways in China, the network operation of urban subways has become an urgent requirement. Subway network operation can reduce the number of passenger transfers and total travel time. Subway network operation requires trains to run across different subway lines, and cross-line operation is one of the train crossing schemes. However, the cross-line operation of the subway involves many conditions such as lines, vehicles, power supply, and signals [ 3 , 4 , 5 ], and the line reconstruction is difficult and costly in the operation stage. In order to ensure that the huge investment cost can effectively improve the operation efficiency of the subway network, it is particularly important to design a scientific and reasonable plan for the cross-line operation of subway trains. 1.1. Literature Review Existing research on the optimization of subway train operation mostly focuses on a single subway line [ 6 , 7 ], including the optimization of departure frequency and running intervals, optimization of the train stop plan, optimization of train routing plan, etc. For the optimization of the departure frequency and running interval, Niu and Zhou proposed a genetic algorithm to solve the integer programming models to optimize the timetables of metro lines [ 8 ]. Sun et al. proposed three models to design capacitated demand-sensitive timetables for metro services considering capacity constraints [ 9 ]. Zhou and Oldache used the simulated annealing algorithm to optimize the timetables for metro services [ 10 ]. For the optimization of train stop plans, Gao et al. proposed a mixed-integer linear programming model to design a skip-stop pattern for metro lines in over-crowded situations after disruptions [ 11 ]. Jamili et al. proposed a robust mathematical model to design skip-stop patterns for metro lines [ 12 ]. Lee et al. designed a skip-stop pattern considering four types of train choice behavior based on passenger smart card data [ 13 ]. For the optimization of the train routing plan, Zwaneveld et al. described an algorithm based on valid inequalities and the branch-and-cut approach to solve the problem of routing trains through stations [ 14 ]. Liu et al. developed an MINLP model to solve the real-time track reallocation problem [ 15 ]. Xu et al. constructed a multi-objective model to determine the receiving routes and departure routes of trains [ 16 ]. For the cross-line operation of trains, valuable theoretical and practical experience has been accumulated worldwide. Many scholars have conducted in-depth research on the management and coordination mechanism of rail transit cross-lines based on the rail transit network in the Tokyo metropolitan area [ 17 , 18 , 19 ]. Ming and Ye studied the traffic demand and engineering technical conditions of cross-line operations in Tokyo [ 20 ]. Yang et al. optimized the frequencies, stopping patterns, and operation zones of cross-line trains [ 21 ]. In China, Chongqing Rail Transit Line 4 and Circle Line have implemented cross-line operation, and scholars have studied their reasonable distribution layout for cross-line operation through passenger flow analysis [ 22 ]. Guangzhou Metro Line 14 and Line 21 have implemented cross-line operations as well, and some scholars have studied the design scheme of their line signal systems [ 23 ]. 1.2. The Main Contributions of This Paper With the help of passenger smart card data, it is possible to analyze the travel demand of the subway network in Beijing [ 24 ], Shanghai [ 26 ], etc. However, there are few studies exploring the proper metro stations and lines for cross-line operation based on passenger smart card data. Therefore, this paper proposes an optimization design method for subway train cross-line operation schemes based on passenger smart card data. This method can accurately and reasonably screen out the transfer stations and lines with an urgent need for cross-line trains. The method firstly calculates the OD demand matrix between stations of the subway network, the passenger section flow, and the transfer flow in all directions of the transfer station based on the data from the passenger smart card. Passenger smart card data contain a passenger’s entry time, entry station name, exit time, and exit station name. Secondly, for two subway lines, according to the property that the transfer flow in a certain direction of the transfer station is smaller than the passenger section flow of the corresponding adjacent section of the transfer station, the method optimizes the design of the subway train cross-line operation plan, including determining the routing type of train cross-line operation and the number of trains running. This plan can help cross-line trains meet the transfer demand as much as possible and fully consider the crossover conditions of the subway lines around the transfer station. Finally, for the lines with cross-line operation conditions in the subway network, the cross-line operation schemes are designed under all possible combinations. According to the volume of cross-line passenger flow that the cross-line trains can carry, the top-ranked cross-line operation plans are preferably recommended. Taking the Chengdu subway network as an example, the design process of the subway train cross-line operation scheme is expounded. The next section will introduce the research methodology, which contains the operation scheme design of the independent subway line, the design of the subway cross-line operation scheme between two subway lines, and the design of the cross-line operation scheme of the subway network. Section 3 shows the rationality of the proposed method through examples, and finally explains the conclusions of this paper and the problems to be further studied. 2. Research Methodology In this section, G N , A denotes the subway network; N denotes the subway station set; n denotes the subway station, n ∈ N ; A denotes the section set of subway lines, and a denotes the section of the subway line, a ∈ A . The target of this research is choosing proper subway lines and stations for the implementation of cross-line operation, reducing the transfer cost, and then reducing the total travel cost, that is: min Z = ∑ f r s v f r s C f r s (1) In the formula, f r s denotes the path between the OD pair r s , v f r s denotes the traffic flow assigned to the path f r s . C f r s denotes the travel time cost of the path f r s . C f r s can be divided into two types according to whether they involve subway transfers, as shown in Formula (2): C f r s = ∑ a δ f r s a t a a ∈ A f r s d o e s n o t i n v o l v e s s u b w a y t r a n s f e r s ∑ a δ f r s a t a + m t * a ∈ A f r s i n v o l v e s s u b w a y t r a n s f e r s (2) In the formula, δ f r s a is a 0–1 variable. If the path f r s contains section a, then δ f r s a is equal to 1; otherwise, δ f r s a is equal to 0. t a denotes the travel time cost of section a . t * denotes the transfer cost and m denotes the number of transfers. For some passengers, operating cross-line trains can reduce their transfer cost, that is m t * . The following part of this paper will study how to choose subway lines and stations to implement cross-line operation to reduce the transfer costs of as many passengers as possible. 2.1. Design of Train Operation Scheme of the Independent Subway Line For the operation scheme of the independent subway line, the maximum passenger section flow of the line sections shall be calculated through the OD matrix between subway stations, and the number of trains shall be determined in combination with the train operation routing and the stop scheme. The inter-station OD matrix data are the original data for calculating the maximum passenger section flow. According to the inter-station OD matrix data, the number of passengers getting on and off in the upward and downward directions of each station can be calculated, and then the passenger section flow can be calculated. Finally, the maximum passenger section flow of the whole line can be obtained. The calculation formula of passenger section flow is: P i + 1 = P i − P X i + P S i (3) In the formula, P i + 1 represents the passenger section flow in section i +1, P i represents the passenger section flow in section i , P X i represents the number of people getting off at station i , and P S i represents the number of people getting on at station i . The number of trains can be calculated according to the maximum passenger section flow of the line and the seating capacity of the train, in combination with the train routing and the stop scheme [ 27 ]. Considering the limitation of the minimum departure interval, the calculation formula of the number of trains can be expressed as follows: n t = min P m a x β P n u m , 3600 T m i n (4) In the formula, n t refers to the number of trains during different time periods throughout the day; P m a x refers to the maximum passenger section flow of the line; P n u m refers to the seating capacity of the train; β represents the full-load coefficient of the line, and the recommended value is 0.9; T m i n represents the minimum departure interval, which is determined by the technical conditions of the line, and the value in this paper is 120 s. 2.2. Design of Cross-Line Operation Scheme between Two Subway Lines 2.2.1. Calculation of Potential Traffic Demand for Cross-Line Operation When a subway line operates independently, the passenger section flow can be calculated according to Formula (3), and then the maximum passenger section flow can be found. For two independent subway lines with one transfer station, the passenger section flow can also be calculated by Formula (3), but the difference from the independent operation of one subway line is that the number of passengers getting on at a station includes not only the number of passengers going to other stations of the line but also the number of passengers who transfer to other stations of another line. For example, the number of passengers getting on at station 8 on line 2 in the downward direction includes not only the number of passengers going to stations 9, 10, 7, 11, 12, and 13 on line 2 but also the number of passengers transferring to stations 1, 2, 3, 4, 5, 6, and 7 on line 1. When the trains of the two subway lines run independently, the passenger transfer flow in the eight directions of the transfer station constitutes the potential transfer demand of the cross-line trains. According to the crossover conditions between subway lines, the potential transfer demand of cross-line trains can be divided into conditional and achievable potential transfer demand and unconditional and unrealizable potential transfer demand. As shown in Figure 1 , the two lines have a total of 13 stations, one of which is a transfer station, and there are four crossover lines. According to the conditions of crossover lines between subway lines, the potential transfer demands A, B, C, and D of cross-line trains are conditional and achievable potential transfer demands; the potential transfer demands E, F, G, and H of cross-line trains are unconditional and unrealizable potential transfer demands. Since the paths of all transfer OD must pass through the section adjacent to the transfer station, property 1 is satisfied. Property 1: The potential transfer demand in all directions of subway cross-line trains must be less than the maximum section flow of any line of two subway lines. Take the simple subway network in Figure 1 as an example to illustrate how to calculate the potential transfer demand of cross-line trains. The subway network consists of two lines, each line contains seven stations, and there is one transfer station between the two lines, as shown in Figure 1 . The inter-station OD demand matrix corresponding to the subway network is a matrix of 13 × 13, as shown in Figure 2 . According to the inter-station OD matrix, the number of people getting on and off in the upward and downward direction at each station can be obtained as shown in Table 1 . Then, according to Formula (3), the passenger section flow can be calculated as shown in Table 2 . Figure 2 not only shows the inter-station OD demand matrix corresponding to the subway network but also uses black blocks to indicate the transfer OD demand that may be undertaken by the cross-line operation. The transfer OD demands represented by each part of the black block correspond to the running direction of the cross-line trains shown in Figure 1 . The potential transfer demand in each cross-line running direction equals the sum of the corresponding OD demand data blocks. For example, the potential transfer demand in direction A in Figure 1 is equal to the sum of the OD demand represented by the black block A in Figure 2 . As shown in Table 3 , the potential transfer demand in direction A is equal to 990 trips per hour, that is, the sum of the OD demand, represented by black block A in Figure 2 , is 990 trips per hour. 2.2.2. Routing Scheme Design of Cross-Line Operation For the independently operated Metro line 1 and Metro line 2 with one transfer station, there are 5 possible subway cross-line routings, as shown in Figure 3 . Figure 3 a shows the routings of Metro Line 1 and Metro Line 2; Figure 3 b–f shows the five types of cross-line routings. It is worth noting that due to the limitations of the construction conditions of the crossover, the cross-line routing can only operate in one direction under normal operation. The cross-line routing shown in Figure 3 b can undertake the transfer flow of four directions—Direction A, B, C, and D, but it involves four crossover constructions. The cross-line routings shown in Figure 3 c–f can undertake the transfer flow of two directions. Figure 3 c can undertake the transfer flow of Direction A, B; Figure 3 d can undertake the transfer flow of Direction B, C; Figure 3 e can undertake the transfer flow of Direction C, D; Figure 3 f can undertake the transfer flow of Direction A, D. In addition, they only involve 2 crossover constructions. In engineering practice, it is necessary to select the appropriate type of cross-line routing considering the difficulty of crossover constructions near the transfer station and passenger transfer flow demands. 2.2.3. Calculate the Number of Trains for Different Routings After determining the subway cross-line routing, the OD demand data of the potential transfer flow undertaken by the cross-line train is also determined. When operating the subway cross-line trains, for cross-line routing 1 shown in Figure 3 b, the OD data blocks undertaken by the cross-line train are A, B, C, D; for cross-line routing 2 shown in Figure 3 c, the OD data blocks undertaken by the cross-line train are A, B; for cross-line routing 3 shown in Figure 3 d, the OD data blocks undertaken by the cross-line train are B, C; for cross-line routing 4 shown in Figure 3 e, the OD data blocks undertaken by the cross-line train are C, D; for cross-line routing 5 shown in Figure 3 f, the OD data blocks undertaken by the cross-line train are A, D. After determining the potential transfer OD demand undertaken by the cross-line train, the sum value of each transfer OD data block can be calculated separately, that is, the crossover section flow of all directions. The maximum value of the crossover section flow is the maximum section flow of the cross-line operation. Then, according to Formula (4), the number of trains required for the subway cross-line operation can be obtained. Based on the number of trains required for the subway cross-line operation and the number of trains for the independent operation of the subway line, one can calculate the number of trains required for the operation of the original subway line after running the subway cross-line train. This value is equal to the number of trains for the independent operation of the subway line minus the number of trains required for the subway cross-line operation. 2.3. Design of Cross-Line Operation Scheme of Subway Network For the subway network, since there may be multiple transfer stations on a subway line that intersects with multiple subway lines, it is difficult to directly calculate the passenger section flow based on Formula (3). However, the passenger section flow of the subway network can be obtained by using the method of network traffic flow assignment. The method process is as follows: Firstly, according to the data of passenger smart cards, we calculate the OD demand between stations of the subway network. We find the k-shortest paths between any OD pairs in the subway network. Secondly, we use the multinomial logit model to calculate the probability of each path being selected [ 28 ] and assign the travel demand to each path according to the probability. The calculation formula is as follows: v f r s = v r s P f r s (5) P f r s = e − C f r s ∑ f r s e − C f r s f r s ∈ F r s (6) In the above formula, P f r s denotes the probability of selecting the path f r s for a certain OD pair r s , f r s ∈ F r s . C f r s denotes the travel time cost of the path f r s . v f r s denotes the traffic flow assigned to the path f r s . v r s denotes the traffic demand of the OD pair r s . Finally, via the statistical summary of the traffic flow of all paths, the passenger section flow of the subway network and the passenger transfer flow of each transfer station can be obtained. The calculation formula of passenger section flow is as follows: x a = ∑ f r s v f r s δ f r s a (7) In the above formula, x a denotes the section flow of section a of the subway line. δ f r s a is a 0–1 variable. If the path f r s contains section a, then δ f r s a is equal to 1; otherwise, δ f r s a is equal to 0. For the subway network, the design method process of the cross-line train operation plan is shown in Figure 4 . 3. Case Analysis This section takes the Chengdu Metro Network as an example to show the process of the design method for subway train cross-line operation schemes proposed by this paper. Chengdu Metro Network refers to urban rail transit serving Chengdu, Sichuan Province, China. Its first line, Chengdu Metro Line 1, was officially opened on 27 September 2010, making Chengdu the twelfth city in mainland China to open rail transit. Based on the subway smart card data of the Chengdu Metro Network on 23 April 2021, this section designs the operation scheme of cross-line trains. Taking the combination of Chengdu Metro Lines 1 and 2 and the transfer station Tianfu Square Station as an example, the design process of the cross-line operation scheme proposed in this paper is comprehensively displayed. According to the smart card data of t Chengdu Metro Network, the OD demand data between Chengdu Metro Lines 1 and 2 and the transfer demand data in all directions undertaken by the transfer station Tianfu Square Station are calculated. On this basis, considering the infrastructure conditions of the transfer station Tianfu Square Station, the type of cross-line routing is determined to be cross-line routing 5, and the estimated number of trains for subway cross-line operation is 5 trains per hour. Through the analysis of the entire Chengdu Metro Network, it is concluded that transfer stations with the most urgent need for the implementation of subway cross-line operation in the Chengdu subway network are the North Railway Station, Xibo City, and South Railway Station. The number of trains required for operating the cross-line trains is greater than or equal to 10 trains per hour. 3.1. Schematic Design of Cross-Line Operation of Chengdu Metro Lines 1 and 2 3.1.1. Traffic Demand Analysis There are smart card data of 3.036 million Chengdu subway passengers every day, and each data must contain four basic fields: Entry time, entry station name, exit time, and exit station name, as shown in Table 4 . The Chengdu subway network consists of 12 lines in total (excluding the tram line Rong 2), and the 12 lines have a total of 46 transfer stations, as shown in Figure 5 . Based on the smart card data of Chengdu subway passengers, the traffic situation of the Chengdu subway network on 23 April 2021 was analyzed. The OD matrix between stations can be easily obtained through statistical analysis of smart card data of Chengdu subway passengers. Through the inter-station OD matrix, based on Formula (5), the passenger section flow of Chengdu Metro Lines 1 and 2 in the morning and evening rush hours can be obtained, as shown in Figure 6 . Figure 6 illustrates that the section flow of the Chengdu subway during the morning rush hour is slightly larger than that during the evening rush hour. Therefore, this paper uses the data of the morning rush hour for the design of the cross-line operation scheme. The section flow values during the morning rush hour of Metro Line 1 are shown in Figure 7 , and the section flow values during the morning rush hour of Metro Line 2 are shown in Figure 8 . The section flow during the morning rush hour of Metro Line 1 in the upward direction is mainly concentrated between People North Road Station and Tianfu Third Street Station, and in the downward direction is mainly concentrated between Four Rivers Station and Financial City Station; the section flow during the morning rush hour of Metro Line 2 in the upward direction is mainly concentrated between Xipu Station and Dongmen Bridge Station, and in the downward direction is mainly concentrated between Chengdu Administration College Station and Tianfu Square Station. From Figure 7 and Figure 8 , it can be seen that, during the morning rush hour, the maximum passenger section flow of Metro Line 1 is 34,629 trips per hour and that of Metro Line 2 is 22,640 trips per hour. By consulting the relevant information, it is concluded that the seating capacity of trains running on Chengdu Metro Lines 1 and 2 is 1468. If the implementation of cross-line trains is not considered, based on Formula (4), the number of trains running on Metro Line 1 should be 26 trains per hour, and the number of trains running on Metro Line 2 should be 17 trains per hour. Based on the OD demand data of the Chengdu subway network, this paper calculates the transfer passenger flow of Tianfu Square Station in eight directions, that is from Xipu direction to Weijianian direction, from Xipu direction to Science City direction, from Longquanyi direction to Science City direction, from Longquanyi direction to Weijianian direction, from Weijianian direction to Xipu direction, from Weijianian direction to Longquanyi direction, from Science City direction to Xipu direction, and from Science City direction to Longquanyi direction, as shown in Figure 9 . Figure 9 shows that during the morning rush hour, the transfer flows from the Longquanyi direction to the Science City direction and from the Xipu direction to the Science City direction are larger; during the evening rush hour, the transfer flows from the Science City direction to the Longquanyi direction and from the Science City direction to the Xipu direction are larger. This phenomenon is due to the fact that most people live in the Longquanyi direction and Xipu direction at night and need to work in the Science City direction during the day. Therefore, in the morning, they transfer from the Xipu direction and Longquanyi direction to the Science City direction, and in the evening, they will transfer from the Science City direction to the Xipu direction and Longquanyi direction. 3.1.2. Subway Cross-Line Operation Design Based on the main distribution of passenger transfer flow, for Metro Lines 1 and 2 and the transfer station Tianfu Square Station, if the subway cross-line operation is implemented, cross-line routing 5 should be selected. In order to determine the maximum passenger section flow, this paper lists the passenger transfer flow in all directions during the morning and evening rush hour, as shown in Table 5 . It can be seen from Table 5 that although the maximum value of transfer flow occurs in the morning rush hour, it is not considered the potential maximum passenger section flow of the subway cross-line operation, because the transfer direction is inconsistent with the one-way direction of the cross-line train. The value of the maximum passenger section flow should be the transfer flow from Science City direction to Longquanyi direction during the evening rush hour, that is, 6171 trips per hour. After the implementation of the cross-line train, based on Formula (4), it can be obtained that the number of trains required for cross-line operation is five trains per hour. After obtaining the number of trains required for the subway cross-line operation and the number of trains required for the independent operation of the subway line, it can be obtained that after operating the subway cross-line train, the numbers of trains required to operate the Metro Lines 1 and 2 are 21 trains per hour and 12 trains per hour, respectively. Table 6 shows the comparison of the number of trains running at each routing before and after running the cross-line train. 3.2. Analysis of Cross-Line Operation Scheme of Chengdu Subway Network 3.2.1. Build the Subway Network In order to implement traffic assignment, the subway network must be built. Network construction can generally be composed of four parts: A simulation module, result module, network module, and new facility module [ 29 ]. For the design method of the cross-line operation scheme of the subway network, simulation modules can be provided by traffic planning software, such as TransCAD. Based on the OD demand data, it enables the traffic assignment. The result module can also be provided by traffic planning software, which enables one to analyze the passenger section flow and the transfer demand of each transfer station. The network module is the vital module for the design method of the cross-line operation scheme, and it must be based on network topology information and geographic information. The construction process of the network module will be described in detail later in this paper. The new facility Module enables one to add new subway stations and lines to the subway network, and the construction process is similar to the network module. Subway station information, subway line information, correspondence between subway stations and lines, and the travel time of sections are fundamental information to build the Network Module of the subway network. The part data of the fundamental information about the Chengdu subway network are shown in Table 7 , Table 8 , Table 9 and Table 10 , respectively. The complete data of the fundamental information about the Chengdu subway network can be obtained on Github [ 30 ]. Table 7 and Table 8 show the name and geographical location of subway lines and stations. Based on these data, using the GeoPandas module of Python and the traffic planning software, TransCAD can easily draw the map of the subway network. Table 9 shows the stations that the subway line passes through in sequence, including beginning stations and end stations. Table 10 shows the travel time of sections of the subway network. This paper assumes subway trains will stop at all stations of each routing and the transfer cost is set as 3.5 min. Based on the information from Table 9 and Table 10 , the network module of the subway network can be obtained. 3.2.2. Subway Cross-Line Operation Design Through the inter-station OD matrix, based on the subway network and Formula (5), the passenger section flow of the Chengdu subway network in the morning and evening rush hour can be obtained as shown in Figure 10 . For all combinations, which can be formed by 46 transfer stations and 12 subway lines in the Chengdu subway network, according to the above-mentioned design process of the cross-line operation plan of Chengdu Metro Lines 1 and 2, we calculated the number of trains for cross-line operation and the maximum passenger section flow. This paper lists the top 24 cross-line operation plans with the largest passenger section flow, as shown in Table 11 . Table 11 shows that, in the Chengdu subway network, the transfer stations with the most urgent need for the implementation of the subway cross-line operation are the North Railway Station, Xibo City Station, and South Railway Station. The transfer lines corresponding to the North Railway Station are Metro Line 1 and Metro Line 7, and the number of trains required for running cross-line trains is 18 trains per hour. The best transfer lines corresponding to the Xibo City Station are Metro Line 1 and Metro Line 6, and the number of trains required for running cross-line trains is 13 trains per hour. The transfer lines corresponding to the South Railway Station are Metro Line 1 and Metro Line 7, and the number of trains required for running cross-line trains is 10 trains per hour. After the implementation of the subway cross-line operation at the three transfer stations, 23,934, 16,710, and 13,024 passengers save on transfer time during rush hour, and the traffic efficiency of the subway network can be improved. In order to intuitively show the superiority of the method proposed in this paper, Figure 11 shows the number of people entering and exiting the subway station during rush hour. During rush hour, the top three transfer stations with the largest number of people entering and exiting are Shiji City Station, Fuhuayuan Station, and Chunxi Road Station. This is not consistent with the results of the largest passenger section flow, as shown in Table 12 . It shows that, compared to the method proposed by this paper, if we use the number of people entering and exiting the subway station to determine which station implements the cross-line operation, there will be 38,451 fewer passengers who can save on transfer time during rush hour. 4. Discussion (1) Taking the cross-line operation scheme design of Chengdu Metro Line 1 and Line 2 as an example comprehensively demonstrates the design process of the subway cross-line operation scheme based on passenger smart card data proposed in this paper; that is, through the analysis of passenger travel demands, determining the subway cross-line routing, the number of trains required for running cross-line trains, etc. The research results show that if Tianfu Square Station is used as the transfer station, and Metro Line 1 and Metro Line 2 are used as transfer lines, the number of trains required to implement cross-line operation is five trains per hour. (2) The analysis of the travel demand of the entire subway network in Chengdu shows that the transfer stations with the most urgent need for the implementation of the subway cross-line operation are the North Railway Station, Xibo City Station, and South Railway Station, and the numbers of trains required for implementing cross-line operation are all greater than or equal to 10 trains per hour. After the implementation of subway cross-line operation, these operation schemes can help more than 13,000 passengers save on transfer time during rush hour and improve traffic efficiency. (3) In the design process of the cross-line operation scheme of the subway network, this paper ignores the impact of the cost of subway line reconstruction. In future research directions, we aim to consider the cost of subway line reconstruction and explore more path selection models to ensure the path selection results are more in line with the actual situation. Author Contributions Conceptualization, M.L. and H.L.; methodology, M.L.; formal analysis, M.L.; resources, H.L.; data curation, H.L.; writing—original draft preparation, H.L.; writing—review and editing, H.L. All authors have read and agreed to the published version of the manuscript. Funding This research was funded by Fundamental Research Funds for the Central Universities of Central South University, grant number 1053320200043. Data Availability Statement Publicly available datasets were analyzed in this study. This data can be found here: https://github.com/muxian123/The-four-fundamental-pieces-of-information-about-the-Chengdu-subway-network (accessed on 19 April 2022). Conflicts of Interest The authors declare no conflict of interest. References Li, J.; Qian, Y.; Zeng, J.; Yin, F.; Zhu, L.; Guang, X. Research on the Influence of a High-Speed Railway on the Spatial Structure of the Western Urban Agglomeration Based on Fractal Theory—Taking the Chengdu–Chongqing Urban Agglomeration as an Example. Sustainability 2020 , 12 , 7550. [ Google Scholar ] [ CrossRef ] Gong, D.; Li, G. Research on Multi-objective Optimized Target Speed Curve of Subway Operation Based on ATO System. Int. Core J. Eng. 2020 , 6 , 133–137. [ Google Scholar ] Griffin, T. Light rail transit sharing the railtrack system. Proc. Inst. Civ. Eng. Transp. 1996 , 117 , 98–103. [ Google Scholar ] [ CrossRef ] Drechsler, G. Light railway on conventional railway tracks in karlsruhe, germany. Proc. Inst. Civ. Eng. Transp. 1996 , 117 , 81–87. [ Google Scholar ] [ CrossRef ] Novales, M.; Orro, A.; Bugarin, M.R. The tram-train: State of the art. Proc. Inst. Mech. Eng. 2002 , 216 , 1–13. [ Google Scholar ] [ CrossRef ] Zhou, S.; Liu, H.; Wang, B.; Chen, B.; Zhou, Y.; Chang, W. Public Norms in the Operation Scheme of Urban Rail Transit Express Trains: The Case of the Beijing Changping Line. Sustainability 2021 , 13 , 7187. [ Google Scholar ] [ CrossRef ] Qian, Y.; Da, C.; Zeng, J.; Wang, X.; Zhang, Y.; Xu, D. A bidirectional quasi-moving block cellular automaton model for single-track railways. Phys. A Stat. Mech. Its Appl. 2022 , 598 , 127327. [ Google Scholar ] [ CrossRef ] Niu, H.; Zhou, X. Optimizing urban rail timetable under time-dependent demand and oversaturated conditions. Transp. Res. Part C Emerg. Technol. 2013 , 36 , 212–230. [ Google Scholar ] [ CrossRef ] Sun, L.; Jin, J.G.; Lee, D.H.; Axhausen, K.W.; Erath, A. Demand-driven timetable design for metro services. Transp. Res. Part C Emerg. Technol. 2014 , 46 , 284–299. [ Google Scholar ] [ CrossRef ] Zhou, W.; Oldache, M. Integrated optimization of line planning, timetabling and rolling stock allocation for urban railway lines. Sustainability 2021 , 13 , 13059. [ Google Scholar ] [ CrossRef ] Gao, Y.; Kroon, L.; Schmidt, M.; Yang, L. Rescheduling a metro line in an over-crowded situation after disruptions. Transp. Res. Part B Methodol. 2016 , 93 , 425–449. [ Google Scholar ] [ CrossRef ] Jamili, A.; Aghaee, M.P. Robust stop-skipping patterns in urban railway operations under traffic alteration situation. Transp. Res. Part C Emerg. Technol. 2015 , 61 , 63–74. [ Google Scholar ] [ CrossRef ] Lee, E.H.; Lee, I.; Cho, S.-H.; Kho, S.-Y.; Kim, D.-K. A travel behavior-based skip-stop strategy considering train choice behaviors based on smartcard data. Sustainability 2019 , 11 , 2791. [ Google Scholar ] [ CrossRef ][ Green Version ] Zwaneveld, P.J.; Kroon, L.G.; Van Hoesel, S.P.M. Routing trains through a railway station based on a node packing model. Eur. J. Oper. Res. 2001 , 128 , 14–33. [ Google Scholar ] [ CrossRef ][ Green Version ] Liu, W.; Zhu, X.; Kang, L. Real-time track reallocation for emergency incidents at large railway stations. Math. Probl. Eng. 2015 , 296394. [ Google Scholar ] [ CrossRef ][ Green Version ] Xu, J.; Wang, Z.; Yao, S.; Xue, J. Train operations organization in high-speed railway station considering variable configuration. Sustainability 2022 , 14 , 2365. [ Google Scholar ] [ CrossRef ] Cao, Z.; Asakura, Y.; Tan, Z. Coordination between node, place, and ridership: Comparing three transit operators in Tokyo. Transp. Res. Part D Transp. Environ. 2020 , 87 , 102518. [ Google Scholar ] [ CrossRef ] Kurosaki, F. A comparative study of passenger through train operation between Japan and Europe. J. East. Asia Soc. Transp. Stud. 2017 , 12 , 316–330. [ Google Scholar ] [ CrossRef ] Kurosaki, F.; Higashino, S. A study of IC card systems within Japanese urban railway lines: Considering the Integration of Transportation Services. J. East. Asia Soc. Transp. Stud. 2019 , 13 , 397–413. [ Google Scholar ] [ CrossRef ] Ming, R.; Ye, X. Through operations between subways and suburban lines in Tokyo. Urban Mass Transit 2009 , 12 , 26–30. [ Google Scholar ] Yang, A.; Wang, B.; Huang, J.; Li, C. Service replanning in urban rail transit networks: Cross-line express trains for reducing the number of passenger transfers and travel time. Transp. Res. Part C Emerg. Technol. 2020 , 115 , 102629. [ Google Scholar ] [ CrossRef ] Le, M.; Wen, C.; Wang, N.; Yang, J.; Jiang, Y. Distribution design of cross line operation between Metro Line 4 and the Circle Line in Chongqing city. Urban Mass Transit 2020 , 23 , 14–17. [ Google Scholar ] Wang, C. Signal system for the cross-line operation of Guangzhou Metro Line 14 and Line 21. Urban Mass Transit 2017 , 20 , 36–38. [ Google Scholar ] Yang, C.; Yan, F.; Ukkusuri, S.V. Unraveling traveler mobility patterns and predicting user behavior in the Shenzhen metro system. Transp. A Transp. Sci. 2018 , 14 , 576–597. [ Google Scholar ] [ CrossRef ] Yong, N.; Ni, S.; Shen, S.; Chen, P.; Ji, X. Uncovering stable and occasional human mobility patterns: A case study of the Beijing subway. Phys. A Stat. Mech. Its Appl. 2018 , 492 , 28–38. [ Google Scholar ] [ CrossRef ] Kim, K. Exploring the difference between ridership patterns of subway and taxi: Case study in Seoul. J. Transp. Geogr. 2018 , 66 , 213–223. [ Google Scholar ] [ CrossRef ] He, B. Discussion upon the Calculation of Passenger Train Quantity Launched in Passenger Dedicated Lines. Railw. Transp. Econ. 2005 , 2 , 61–62+65. [ Google Scholar ] Matejka, F.; Mckay, A. Rational Inattention to Discrete Choices: A New Foundation for the Multinomial Logit Model. Am. Econ. Rev. 2015 , 105 , 272–298. [ Google Scholar ] [ CrossRef ][ Green Version ] Kostrzewski, M. Implementation of distribution model of an international company with use of simulation method. Procedia Eng. 2017 , 192 , 445–450. [ Google Scholar ] [ CrossRef ] Available online: https://github.com/muxian123/The-four-fundamental-pieces-of-information-about-the-Chengdu-subway-network (accessed on 17 May 2022). Figure 1. Schematic diagram of subway network. Figure 1. Schematic diagram of subway network. Figure 2. Schematic diagram of OD matrix. Figure 2. Schematic diagram of OD matrix. Figure 3. Schematic diagram of subway cross-line operation. ( a ) Metro Line routings; ( b ) Cross-line routing 1; ( c ) Cross-line routing 2; ( e ) Cross-line routing 4; ( f ) Cross-line routing 5. Figure 3. Schematic diagram of subway cross-line operation. ( a ) Metro Line routings; ( b ) Cross-line routing 1; ( c ) Cross-line routing 2; ( e ) Cross-line routing 4; ( f ) Cross-line routing 5. Figure 4. Design method process of the cross-line train operation plan. Figure 4. Design method process of the cross-line train operation plan. Figure 5. Schematic diagram of Chengdu subway network. Figure 5. Schematic diagram of Chengdu subway network. Figure 6. Passenger section flow of Chengdu Metro Lines 1 and 2 in the morning and evening rush hour. ( a ) Morning rush hour; ( b ) Evening rush hour. Figure 6. Passenger section flow of Chengdu Metro Lines 1 and 2 in the morning and evening rush hour. ( a ) Morning rush hour; ( b ) Evening rush hour. Figure 7. Passenger section flow of Metro Line 1 during morning rush hour. Figure 7. Passenger section flow of Metro Line 1 during morning rush hour. Figure 8. Passenger section flow of Metro Line 2 during morning rush hour. Figure 8. Passenger section flow of Metro Line 2 during morning rush hour. Figure 9. Passenger transfer flows in eight directions during different time periods. Figure 9. Passenger transfer flows in eight directions during different time periods. Figure 10. Passenger section flow of Chengdu subway network in the morning and evening rush hour. ( a ) Morning rush hour; ( b ) Evening rush hour. Figure 10. Passenger section flow of Chengdu subway network in the morning and evening rush hour. ( a ) Morning rush hour; ( b ) Evening rush hour. Figure 11. The number of people entering and exiting the subway station during rush hour. ( a ) Morning rush hour; ( b ) Evening rush hour. Figure 11. The number of people entering and exiting the subway station during rush hour. ( a ) Morning rush hour; ( b ) Evening rush hour. Table 1. Number of people getting on and off at each station (trips per hour). Table 1. Number of people getting on and off at each station (trips per hour). Metro Line 1 Metro Line 2 Upward Direction Downward Direction Upward Direction Downward Direction Station Number of Passengers Getting on Number of Passengers Getting Off Number of Passengers Getting On Number of Passengers Getting Off Station Number of Passengers Getting On Number of Passengers Getting Off Number of Passengers Getting On Number of Passengers Getting Off 1 1230 0 0 1290 8 0 1260 1290 0 2 1130 100 100 1190 9 100 1160 1190 100 3 1030 200 200 1090 10 200 1060 1090 200 7 2460 2190 2370 2280 7 2280 2460 2190 2370 4 200 1120 1060 200 11 1120 200 200 1030 5 100 1220 1160 100 12 1220 100 100 1130 6 0 1320 1260 0 13 1320 0 0 1230 Table 2. Passenger section flow. Table 2. Passenger section flow. Metro Line 1 Section Section Flow of Upward Direction (Trips per Hour) Section Flow of Downward Direction (Trips per Hour) Metro Line 2 Section Section Flow of Upward Direction (Trips per Hour) Section Flow of Downward Direction (Trips per Hour) 1–2 1230 1290 8–9 1290 1260 2–3 2260 2380 9–10 2380 2320 3–7 3090 3270 10–7 3270 3180 7–4 3360 3180 7–11 3090 3360 4–5 2440 2320 11–12 2260 2440 5–6 1320 1260 12–13 1230 1320 Table 3. Potential transfer demand. Table 3. Potential transfer demand. Potential Transfer Demand of Possible Directions of Cross-Line Train Potential Transfer Demand of Impossible Directions of Cross-Line Train Direction A Direction B Direction C Direction D Direction E Direction F Direction G Direction H Potential transfer demand (trips per hour) 990 1260 1080 1170 900 900 900 900 Table 4. Part of data. Table 4. Part of data. Transaction Date Enter Time Enter Station Name Exit Time Exit Station Name 20210423 10:53:02 Xipu 11:17:08 Tianfu square 20210423 19:37:08 Xipu 20:14:03 Tianfu square 20210423 11:15:07 Tianfu square 11:22:21 Xipu 20210423 08:15:16 Tianfu square 08:50:02 Xipu Table 5. Passenger transfer flow during morning and evening rush hour. Table 5. Passenger transfer flow during morning and evening rush hour. Transfer Flow during Morning Rush Hour (Trips per Hour) Transfer Flow during Evening Rush Hour (Trips per Hour) Xipu-Weijianian 1403 288 Xipu-Science City 5520 974 Longquanyi-Weijianian 1633 808 Longquanyi-Science City 7394 2700 Weijianian-Xipu 415 662 Weijianian-Longquanyi 1054 1031 Science City-Xipu 1440 3985 Science City-Longquanyi 2966 6171 Table 6. Changes in the number of trains running at each routing before and after running the cross-line train. Table 6. Changes in the number of trains running at each routing before and after running the cross-line train. Number of Trains (Trains per Hour) Running Time Interval (min) Metro Line 1 Before operating cross-line 26 2.3 After operating cross-line 21 2.9 Metro Line 2 Before operating cross-line 17 3.5 After operating cross-line 12 5 Cross-line routing Before operating cross-line - - After operating cross-line 5 12 Table 7. Part subway station information of the Chengdu subway network. Table 7. Part subway station information of the Chengdu subway network. Subway Station Name Subway Station Location Mian shang yin xing zhan POINT(104.538370 30.410898) San yuan wai guo yu xue xiao zhan POINT(104.153051 30.789908) Da yuan POINT(104.043775 30.552330) Mu hua lu POINT(104.008817 30.487746) Nan xun da dao POINT(103.849654 30.692276) Xiang cheng xiao xue zhan POINT(104.180097 30.805446) Guang zhou lu POINT(104.074757 30.421508) Table 8. Part subway line information of the Chengdu subway network. Table 8. Part subway line information of the Chengdu subway network. Subway Line Direction Subway Line location Metro Line 10 0 LINESTRING(104.019205 30.627921,104.018534 30.627220,...) Metro Line 10 1 LINESTRING(103.779287 30.411013,103.781375 30.413648,...) Metro Line 17 0 LINESTRING(103.976043 30.647566,103.974564 30.647177,...) Metro Line 17 1 LINESTRING(103.791832 30.758752,103.792310 30.758067,...) Metro Line 18 1 LINESTRING(104.065209 30.606333,104.065229 30.605643,...) Metro Line 18 0 LINESTRING(104.452430 30.336950,104.451925 30.335890,...) Table 9. Part correspondence information between subway stations and lines of the Chengdu subway network. Table 9. Part correspondence information between subway stations and lines of the Chengdu subway network. Stations of Chengdu Metro Line 2 Subway Station Name ID Subway Station Name ID Xi pu 1 Dong men da qiao 17 Tian he lu 2 Niu wang miao 18 Bai cao lu 3 Niu shi kou 19 Jin zhou lu 4 Dong da lu 20 Jin ke bei lu 5 Ta zi shan gong yuan 21 Ying bin da dao 6 Cheng dong dong ke zhan 22 Cha dian zi ke yun zhan 7 Cheng yu li jiao 23 Yang xi li jiao 8 Hui wang ling 24 Yi pin tian xia 9 Hong he 25 Shu han lu dong 10 Cheng dong xing zheng xue yuan 26 Bai guo lin 11 Da mian pu 27 Zhong yi da sheng yi yuan 12 Lian shan po 28 Tong hui men 13 Jie pai 29 Ren min gong yuan 14 Shu fang 30 Tian fu guang chang 15 Long ping lu 31 Chun xi lu 16 Long quan yi 32 Table 10. Travel time of part sections of the Chengdu subway network. Table 10. Travel time of part sections of the Chengdu subway network. Section Name (Downward Direction) Travel Time (min) Section Name (Upward Direction) Travel Time (min) Xi pu—Tian he lu 3 Tian he lu—Xi pu 3 Tian he lu—Bai cao lu 3 Bai cao lu—Tian he lu 3 Bai cao lu—Jin zhou lu 3 Jin zhou lu—Bai cao lu 3 Jin zhou lu—Jin ke bei lu 2 Jin ke bei lu—Jin zhou lu 2 Table 11. The top 24 cross-line operation plans with the largest passenger section flow. Table 11. The top 24 cross-line operation plans with the largest passenger section flow. Transfer Station Transfer Line 1 Transfer Line 2 Maximum Section Flow (Trips per Hour) Number of Trains for Cross-Line Operation (Trains per Hour) Number of Trains for Transfer Line 1 after Running Cross-Line Train (Trains per Hour) Number of Trains for Transfer Line 2 after Running Cross-Line Train (Trains per Hour) North railway station Metro line 1 Metro line 7 23,934 18 8 1 Xibo city Metro line 1 Metro line 6 16,710 13 13 1 Xibo city Metro line 1 Metro line 18 14,902 11 15 1 South railway station Metro line 1 Metro line 7 13,024 10 16 9 South railway station Metro line 1 Metro line 18 11,727 9 17 3 Chengdu east railway station Metro line 2 Metro line 7 10,277 8 9 11 Haichang road Metro line 1 Metro line 18 10,253 8 18 4 Yipintianxia Metro line 2 Metro line 7 10,088 8 9 11 Peopel north road Metro line 1 Metro line 6 9948 8 18 6 Sima bridge Metro line 3 Metro line 7 9855 7 11 12 Luoma city Metro line 1 Metro line 4 9384 7 19 4 Kuishudian Metro line 4 Metro line 7 8994 7 4 12 North station west second road Metro line 5 Metro line 7 8463 6 9 13 Taiping garden Metro line 3 Metro line 7 8160 6 12 13 Xipu Metro line 2 Metro line 6 7585 6 11 8 Incubation park Metro line 1 Metro line 18 7451 6 20 6 Niuwang temple Metro line 2 Metro line 6 7065 5 12 9 Huilong Metro line 5 Metro line 6 6693 5 10 9 Palace of culture Metro line 4 Metro line 7 6442 5 6 14 Provincial gymnasium Metro line 1 Metro line 3 6248 5 21 13 Tianfu square Metro line 2 Metro line 1 6171 5 12 21 Tcm provincial hospita Metro line 2 Metro line 4 5510 4 13 7 South railway station Metro line 18 Metro line 7 5476 4 8 15 Southwest jiaotong university Metro line 6 Metro line 7 5346 4 10 15 Table 12. The results of different methods. Table 12. The results of different methods. The Top 3 Transfer Stations with the Largest Number of People Entering and Exiting Maximum Section Flow (Trips per Hour) The Top 3 Transfer Stations with the Largest Passenger Section Flow Maximum Section Flow (Trips per Hour) North railway station 23,934 Shiji city station 5113 Xibo city 16,710 Fuhuayuan station 7451 South railway station 13,024 Chunxi road station 2653 Li M, Li H. Optimal Design of Subway Train Cross-Line Operation Scheme Based on Passenger Smart Card Data. Sustainability. 2022; 14(11):6420. https://doi.org/10.3390/su14116420 Chicago/Turabian Style Li, Maosheng, and Hangcong Li. 2022. "Optimal Design of Subway Train Cross-Line Operation Scheme Based on Passenger Smart Card Data" Sustainability14, no. 11: 6420. https://doi.org/10.3390/su14116420 Article Metrics MDPI and ACS Style Li, M.; Li, H. Optimal Design of Subway Train Cross-Line Operation Scheme Based on Passenger Smart Card Data. Sustainability 2022, 14, 6420. https://doi.org/10.3390/su14116420 AMA Style Li M, Li H. Optimal Design of Subway Train Cross-Line Operation Scheme Based on Passenger Smart Card Data. Sustainability. 2022; 14(11):6420. https://doi.org/10.3390/su14116420 Chicago/Turabian Style Li, Maosheng, and Hangcong Li. 2022. "Optimal Design of Subway Train Cross-Line Operation Scheme Based on Passenger Smart Card Data" Sustainability14, no. 11: 6420. https://doi.org/10.3390/su14116420

https://www.mdpi.com/2071-1050/14/11/6420

(PDF) Impaired clustered protocadherin-α (cPcdh-α) leads to aggregated retinogeniculate terminals and impaired visual acuity in mice PDF | Clustered protocadherins (cPcdhs) comprising cPcdh-α, -β, and -γ, encode a large family of cadherin-like cell-adhesion molecules specific to... | Find, read and cite all the research you need on ResearchGate Impaired clustered protocadherin-α (cPcdh-α) leads to aggregated retinogeniculate terminals and impaired visual acuity in mice February 2015 Journal of Neurochemistry 133(1) DOI: 10.1111/jnc.13053 Abstract and Figures Clustered protocadherins (cPcdhs) comprising cPcdh-α, -β, and -γ, encode a large family of cadherin-like cell-adhesion molecules specific to neurons. Impairment of cPcdh-α results in abnormal neuronal projection patterns in specific brain areas. To elucidate the role of cPcdh-α in retinogeniculate projections, we investigated the morphological patterns of retinogeniculate terminals in the lateral geniculate nucleus of mice with impaired cPcdh-α. We found huge aggregated retinogeniculate terminals in the dorsal lateral geniculate nucleus, whereas no such aggregated terminals derived from the retina were observed in the olivary pretectal nucleus and the ventral lateral geniculate nucleus. These aggregated terminals appeared between P10 and P14, just before eye opening and at the beginning of the refinement stage of the retinogeniculate projections. Reduced visual acuity was observed in adult mice with impaired cPcdh-α, whereas the orientation selectivity and direction selectivity of neurons in the primary visual cortex were apparently normal. These findings suggest that cPcdh-α is required for adequate spacing of retinogeniculate projections, which may be essential for normal development of visual acuity.This article is protected by copyright. All rights reserved. Retinogeniculate axon terminals in cPcdh-α KO mice. (a and b) Gross distribution patterns of cholera toxin β subunit (CTb)-positive brown terminals derived from the contralateral eye in olivary pretectal nucleus (OPT), dorsal LG (LGd), intergeniculate leaflet (IGL; arrow head), and ventral LG (LGv) of a WT (a) and a cPcdh-α KO (b) mouse, aged 8–10 weeks. Sections were processed by Nissl staining. (c and d) Higher magnification of LGd shown in a (c) and b (d). Many aggregated terminals (arrows) were scattered in the LGd of cPcdh-α KO mice (d). (e and f) Other images of CTb-positive terminals at a higher magnification in a WT (e) and a cPcdh-α (f) mouse. These sections were not counter-stained. (g and h) Corticogeniculate terminals in LGd (arrowheads) labeled after biotinylated dextran amine (BDA) injection into the primary visual cortex (V1) of a WT (g) and a cPcdh-α KO (h) mouse. No aggregated terminals were found. Scale bars, 1 mm in (a and b), 20 μm in (c and d), and 10 μm in (e–h). … Quantitative analysis of isolated retinogeniculate terminals. (a and b) Typical schematic distribution of retinal terminals at the center level of dorsal LG (LGd), including the area that receives inputs from the ipsilateral eye. Aggregated terminals were arbitrarily categorized into large (≥ 100 μm2), medium (< 100 and > 40 μm2), and small (≤ 40 μm2) types. Only the large and medium types are shown. The areas surrounded by a dotted line received inputs from the ipsilateral eye. (c) Cumulative distribution of area sizes. The isolated aggregated terminal size measurement was taken on the binarized digital images of cholera toxin β subunit (CTb)-stained sections. (d) Sizes of isolated retinogeniculate terminals. (e) Number of fine type isolated retinogeniculate terminals (< 13.1 μm2) per mm2 in LGd sections. In the analyses shown in (c–e), the areas that received inputs from the ipsilateral eye were omitted. … Ontogenetic observation of retinogeniculate terminals. (a, c, and e) Retinogeniculate terminals at P10 (a), P14 (c), and P19 (e) in WT mice. (b, d, and f) Retinogeniculate terminals at P10 (b), P14 (d), and P19 (f) in cPcdh-α KO mice. Scale bars, 20 μm in (a–f). … Reduced visual acuity in cPcdh-α KO mice. (a) Sample traces of neuronal calcium response to moving grating patterns (from 0° to 315° in 45° steps) in a WT mouse (left) and a cPcdh-α KO mouse (right). (b) Cumulative distributions of the orientation selectivity index (OSI) of 481 neurons in WT mice and 423 neurons in cPcdh-α KO mice (left). The distribution of the direction selectivity index (DSI) of 197 neurons in WT mice and 164 neurons in cPcdh-α KO mice is also shown (right). DSI was estimated in neurons with an OSI > 0.4. (c) Schematic drawing of the Prusky water maze used to estimate visual acuity. (d) Examples of the results obtained for a WT and a cPcdh-α KO mouse. (e) Visual acuity in 10 WT and nine cPcdh-α KO mice. … Figures - uploaded by Hiroaki Tsukano University, Niigata, Japan (CREST), Osaka University, Suita, Japan Abstract and - c , encode a large family of cadherin-like cell-adhesion molecules speci fi c to neurons. Impairment of cPcdh- a results in abnormal neuronal projection patterns in speci fi c brain areas. To elucidate the role of cPcdh- a in retinogeniculate projections, we investigated the morphological patterns of retinogeniculate terminals in the lateral geniculate (LG) nucleus of mice with impaired cPcdh- a . We found huge aggregated retinogeniculate terminals in the dorsal LG nucleus, whereas no such aggre- gated terminals derived from the retina were observed in the olivary pretectal nucleus and the ventral LG nucleus. These aggregated terminals appeared between P10 and P14, just before eye opening and at the beginning of the re fi nement stage of the retinogeniculate projections. Reduced visual acuity was observed in adult mice with impaired cPcdh- a , whereas the orientation selectivity and direction selectivity of neurons in the primary visual cortex were apparently normal. These fi ndings suggest that cPcdh- a is required for adequate spacing of retinogeniculate projections, which may be essential for normal development of visual acuity. Keywords: aggregated terminals, lateral geniculate nucleus, protocadherin, retinal terminals, visual acuity. J. Neurochem. (2015) 133 ,6 6 – 72. the cadherin superfamily of neuronal adhesion molecules (Kohmura et al. 1998; Junghans et al. 2005; Shapiro et al. 2007; Yagi 2012). Mammalian cPcdh genes consist of three families, cPcdh- a ,- b , and - c , each of which has a clustered structure in a small genome locus on a single chromosome (Wu 2005). In normal mice, olfactory sensory neurons expressing the same odorant receptor project their axons into a speci fi c glomerulus in the main olfactory bulb (Mori and Sakano 2011). However, these projection patterns are abnormal in cPcdh- a knockout (KO) mice, in which the constant region of cPcdh- a is deleted (Hasegawa et al. 2008). Furthermore, abnormal aggregation of serotonergic axons is observed in cPcdh- a KO mice (Katori et al. 2009), suggesting that cPcdh- a is important for the normal development of axonal projection patterns derived from speci fi c types of neurons. In the visual system of mice, cPcdh- a is expressed in the optic nerve fi bers, in the lateral geniculate nucleus (LG), and in the primary visual cortex (V1) for 2 weeks after birth (Morishita et al. 2004). This period before eye opening is very important for eye-speci fi c segregation of the retinogen- iculate fi bers (Jaubert-Miazza et al. 2005; Guido 2008), the Received November 22, 2014; revised manuscript received January 16, 2015; accepted January 26, 2015. Correspondence and present address: Reiko Meguro, School of Health Sciences, Faculty of Medicine, Niigata University, 2-746 Asahi-machi, Chuo-ku, Niigata 951-8518, Japan. E-mail: [email protected] Abbreviations used : BDA, biotinylated dextran amine; cPcdh, clus- tered protocadherin; CTb, cholera toxin b subunit; DAB, diaminobenzi- dine; IGL, intergeniculate lea fl et; LG, lateral geniculate nucleus; LGd, dorsal LG; LGv, ventral LG; OPT, olivary pretectal nucleus; PBS, phosphate-buffered saline; V1, primary visual cortex. 66 © 2015 International Society for Neurochemistry, J. Neurochem. (2015) 133 , 66- -72 JOURNAL OF NEUROCHEMISTRY | 2015 | 133 | 66–72 doi: 10.1111/jnc.13053 re fi nement of retinogeniculate synapses (Chen and Regehr 2000; Hooks and Chen 2006, 2008; Liu and Chen 2008), and the maturation of inhibitory neurons (Golding et al. 2014) in the dorsal LG (LGd). In the present study, we thus investigated the morphology of retinogeniculate terminals in cPcdh- a KO mice and found abnormally aggregated terminals mainly in LGd. These aggregated terminals found in adults were also observed at P14 but not until P10. The abnormal aggregation can result in impaired transmission of visual information through LGd. As expected, we found signi fi cantly deteriorated visual acuity in cPcdh- a KO mice, which was determined using a behavioral test (Prusky et al. 2000), whereas the orientation tuning of V1 neurons was apparently normal in cPcdh- a KO mice. Materials and methods Animal experiments We used male and female mice of cPcdh- a -KO (Hasegawa et al. 2008) and C57BL/6J strain obtained from Charles River Japan (Tokyo, Japan). All experiments were performed in accordance with the guide for the Care and Use of Laboratory Animals of the Science Council of Japan, and were approved by the Animal Experiment Committee of Niigata University. Neural tracing experiments Five wild type (WT) mice and fi ve cPcdh- a KO mice of aged 8 – 10 weeks were anesthetized intraperitoneally with a mixture of ketamine (300 mg/kg; Daiichi-Sankyo, Tokyo, Japan) and xylazine (30 mg/mL; Bayer, Leverkusen, Germany). The cholera toxin b subunit (CTb, 5 ng in 1 l L of saline; List Biological Laboratories, Campbell, CA, USA), an anterograde neural tracer, was injected into the left eye using a glass pipette. Five days after the injection, the mice were anesthetized with an overdose of pentobarbital (100 mg/kg, i.p.) and perfused with phosphate-buffered saline, followed by 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.2). For the developmental study, two WT and two cPcdh- a KO mice at each of P10, P14, and P19 were tested in a similar manner with a minimum survival time of 48 h for CTb transportation (Wu et al. 2000). The transported CTb in LG was visualized as described previously (Horie et al. 2013). Brie fl y, 50 l m coronal sections, prepared using a cryostat, were incubated with a goat polyclonal anti-CTb antibody (1 : 8000 dilution; List Biological Laboratories) overnight, followed by incubation with a biotinylated anti-goat IgG antibody (1 : 200 dilution; Jackson, West Grove, PA, USA) for 2 h. The sections were further incubated with an avidin – biotin complex (1 : 100 dilution; Vector Laboratories, Burlingame, CA, USA) and the CTb immunoreactivity was visualized with oxidized diaminobenzidine. The reacted sections were mounted on gelatin-coated glass slides and air-dried. Some serial CTb-labeled sections were counter-stained with cresyl violet. Sections were observed using a microscope (BX-50; Olympus, Tokyo, Japan) and images were acquired using a CCD camera (CS-600; Olympus). The terminal size measurement was taken using a program based on Matlab (Mathworks, Natick, MA, USA). To visualize the corticogeniculate fi bers in two of the each WT and cPcdh- a mice, biotinylated dextran amine (BDA-3000; Invitrogen, Carlsbad, CA, USA) was injected into V1, as described previously (Horie et al. 2013; Tohmi et al. 2014). Two-photon calcium imaging of V1 neurons Orientation selectivity and direction selectivity in V1 neurons were tested using two-photon calcium imaging in fi ve WT and fi ve cPcdh- a KO mice. The two-photon calcium imaging was performed as described previously (Honma et al. 2013; Yoshitake et al. 2013; Tohmi et al. 2014). Brie fl y, V1 was identi fi ed using fl avoprotein fl uorescence imaging in mice anesthetized with urethane (1.75 g/kg, i.p.). After removing the skull above V1, a glass pipette fi lled with the cocktail of fura-2 AM (Invitrogen) and sulforhodamine 101 (SR- 101; Invitrogen) was inserted at the center of V1 and was advanced into the supragranular layers, down to a depth of 200 – 300 l m from the surface. The cocktail was injected using a pressure of 4 – 15 kPa for 5 – 10 min, so that the cells in the area located 200 – 300 mm from the tip of the pipette were labeled with fura-2. Astrocytes labeled with SR-101 were excluded from further analysis. After the mice were set under a two-photon microscope (TCS SP5 MP; Leica Microsystems, Wetzlar, Germany), grating patterns moving in eight directions (from 0 ° to 315 ° in 45 ° steps) were administered to the mice and V1 neuronal responses were analyzed, as described previously (Honma et al. 2013; Yoshitake et al. 2013). Behavioral assessment of visual acuity Visual acuity of mice was measured via a visual water maze task developed by Prusky and coworkers (Prusky et al. 2000; Prusky and Douglas 2004). Ten WT and nine cPcdh- a KO mice (aged 7 – 12 weeks) were trained and tested in the visual water maze task in 32 trials every day to assess their visual acuity. During the training phase, they were trained to choose a sine-wave vertical grating pattern at a low spatial frequency (0.14 cycles/degree) from a homogeneous gray pattern with the same averaged luminous intensity. In the subsequent testing phase, the limit of their discriminatory ability was assessed at spatial frequencies higher than 0.14 cycles/degree. A preliminary grating threshold was established at which the mice achieved 70% correct choices. The spatial frequencies around the threshold were retested and the average performance was calculated to plot a frequency-of-seeing curve. The point at 70% accuracy was determined from the curve and was de fi ned as the visual acuity. Statistical tests The Kormogorv – Smirnov test was used to evaluate the differences in the cumulative distributions of terminal area sizes between WT and cPcdh- a mice. For group comparisons, the Mann – Whitney U test was used. Results Abnormal aggregation of retinogeniculate terminals in cPcdh- a KO mice The gross anatomical features of the thalamus and the tectum were checked in WT (Fig. 1a) and cPcdh- a KO (Fig. 1b) mice aged 8 – 10 weeks. The distribution of CTb-positive terminals originating from the contralateral eye was apparently similar between the two types of mice. At a higher magni fi cation, © 2015 International Society for Neurochemistry, J. Neurochem. (2015) 133 , 66- -72 Aggregated LGd terminals in cPcdh- a KO mice 67 however, many aggregated terminals were found in LGd of cPcdh- a KO mice (Fig. 1d and f) but not in WT mice (Fig. 1c and e). Some aggregated terminals (arrows in Fig. 1d) were comparable in size to the soma of Nissl-stained LGd neurons. The abnormally aggregated terminals were intermingled with apparently normal terminals in the LGd of cPcdh- a KO mice. Such abnormal terminals were also observed in the ipsilateral retinogeniculate projection, within the other side of LGd. Some aggregated terminals were also found in the intergen- iculate lea fl et, whereas few were observed in the olivary pretectal nucleus and in the ventral LG (LGv) in cPcdh- a KO and WT mice (data not shown). As a control, we tested the distribution pattern of corticogeniculate terminals, which represent another main input into the LGd. BDA-positive terminals in LGd, labeled after BDA injection into V1, were not aggregated, and were apparently similar between a WT (Fig. 1g) and a cPcdh- a (Fig. 1h) mouse. Quantitative analysis of isolated retinogeniculate terminals in cPcdh- a -KO mice We arbitrarily categorized the aggregated retinogeniculate terminals into three types on the basis of their approximate sizes: large ( ≥ 100 l m 2 ), medium ( < 100 and > 40 l m 2 ), and small ( ≤ 40 l m 2 ) types. The large type was distributed all over the LGd in cPcdh- a KO mice, except the area receiving CTb-negative terminals from the ipsilateral eye (Fig. 2b). In WT mice, only a few terminals of the large type appeared near the lateral surface of the LGd (Fig. 2a). The medium terminals were observed throughout the LGd in both types of mice (Fig. 2a and b), and the small terminals were as well (data not shown). A quantitative analysis of size distribution in the isolated terminals with or without aggregation revealed a signi fi cant difference between WT and cPcdh- a KO mice ( p < 0.001, Fig. 2c). The mean size of the isolated terminals was 13.1 l m 2 in WT mice and 27.7 l m 2 in cPcdh- a KO mice. This difference was statis- tically signi fi cant ( p < 0.005, Fig. 2d). Fine terminals ( < 13.1 l m 2 ) occupied 71% of isolated terminals in WT mice and 56% of those in cPcdh- a KO mice. The density of fi ne terminals in cPcdh- a KO mice was 62.5% of that observed in WT mice; this difference was also statistically signi fi cant ( p < 0.005, Fig. 2e). Appearance of the aggregated retinogeniculate terminals at P14 in cPcdh- a KO mice We examined the post-natal development of retinogeniculate terminals in cPcdh- a KO mice. Gross distribution patterns were similar between WT and cPcdh- a KO mice at all stages after P10, including the normal completion of eye-speci fi c segregation patterns up to P10 (data not shown). In WT mice, the retinogeniculate terminals did not change much in shape and size between P10 and P19, whereas their densities were reduced between P10 and P19 (Fig. 3a, c, and e). In cPcdh- a mice, the aggregated retinogeniculate terminals, which were not present at P10, were observed at P14, at the day of eye opening (Fig. 3b and d). Dark rearing had no effect on these results (data not shown), suggesting that visual experience is not required for aggregation. At P19, the aggregated terminals increased in size and were as large as those (a) (b) (c) (d) (e) (f) (g) (h) Fig. 1 Retinogeniculate axon terminals in cPcdh- a KO mice. (a and b) Gross distribution patterns of cholera toxin b subunit (CTb)-positive brown terminals derived from the contralateral eye in olivary pretectal nucleus (OPT), dorsal LG (LGd), intergeniculate lea fl et (IGL; arrow head), and ventral LG (LGv) of a WT (a) and a cPcdh- a KO (b) mouse, aged 8 – 10 weeks. Sections were processed by Nissl staining. (c and d) Higher magni fi cation of LGd shown in a (c) and b (d). Many aggregated terminals (arrows) were scattered in the LGd of cPcdh- a KO mice (d). (e and f) Other images of CTb-positive terminals at a higher magni fi cation in a WT (e) and a cPcdh- a (f) mouse. These sections were not counter-stained. (g and h) Corticogeniculate termi- nals in LGd (arrowheads) labeled after biotinylated dextran amine (BDA) injection into the primary visual cortex (V1) of a WT (g) and a cPcdh- a KO (h) mouse. No aggregated terminals were found. Scale bars, 1 mm in (a and b), 20 l m in (c and d), and 10 l mi n( e – h). © 2015 International Society for Neurochemistry, J. Neurochem. (2015) 133 , 66- -72 68 R. Meguro et al. observed in adult mice aged 8 – 10 weeks (Fig. 3f). The mean size of the isolated terminals were 11.72, 12.58, 14.63 l m 2 in two WT mice and 16.57, 20.63, 25.39 l m 2 in two cPcdh- a KO mice, at P10, P14, and P19, respectively. Deteriorated visual acuity in cPcdh- a KO mice The presence of the aggregated retinogeniculate terminals suggests abnormal visual functions in cPcdh- a KO mice. Therefore, we performed two-photon calcium imaging of the V1 neurons. However, the orientation selectivity and direction selectivity were apparently similar between WT and cPcdh- a KO mice (Fig. 4a and b), as reported previously (Yoshitake et al. 2013). We further tested the visual acuity of mice using a behavioral test (Prusky et al. 2000), and compared the results between WT and cPcdh- a KO mice (Fig. 4c). WT and cPcdh- a KO mice similarly performed low-frequency tasks at 0.14 cycles/degree during the training phase. The visual acuity of WT mice [0.56  0.01 cycles/degree (mean  SEM), n = 10] was comparable to the values reported previously (Prusky et al. 2000; Prusky and Douglas 2004). However, the visual acuity of cPcdh- a KO mice (0.41  0.01 cycles/ degree, n = 9) was signi fi cantly deteriorated compared with that observed in WT mice ( p < 0.0003; Fig. 4d and e). Discussion Mechanisms underlying terminal aggregation in cPcdh- a KO mice In the present study, we found abnormal terminal aggrega- tion in the LGd of cPcdh- a KO mice. Impairment of cPcdh- a results in abnormal projection patterns from olfactory sensory neurons expressing a given odorant receptor onto a (c) (a) (cPcdh- From ipsilateral eye Size of t erminals (µ m 2 ) 0 10 20 30 WT n = 5 cPcdh KO n = 5 - p < 0.005 p < 0.005 Number o f fine terminals/mm 2 (d) (e) (b) Large Medium LGd LGd IGL IGL (WT) 100 m 0 500 1000 WT n = 5 cPcdh KO n = 5 - m 2 Results obtained from 5 m ice each WT cPcdh- KO Medium Large 0 20 40 60 80 100 % 0 50 100 150 200 250 KO) Fig. 2 Quantitative analysis of isolated retinogenicul ate terminals. (a and b) Typical schematic distribution of retinal terminals at the center level of dorsal LG (LGd), including the area that receives inputs from the ipsilateral eye. Aggregated terminals were arbitrarily categorized into large ( ≥ 100 l m 2 ), medium ( < 100 and > 40 l m 2 ), and small 2 ) types. Only the large and medium types are shown. The areas surrounded by a dotted line received inputs from the ipsilateral eye. (c) Cumulative distribution of area sizes. The isolated aggregated terminal size measurement was taken on the binarized digital images of cholera toxin b subunit (CTb)-stained sections. (d) Sizes of isolated retinogeniculate terminals. (e) Number of fi ne type isolated retinogen- iculate terminals ( < 13.1 l m 2 ) per mm 2 in LGd sections. In the analyses shown in (c – e), the areas that received inputs from the ipsilateral eye were omitted. (a) (b) (c) (d) (e) (f) Fig. 3 Ontogenetic observation of retinogenicul ate terminals. (a, c, and e) Retinogeniculate terminals at P10 (a), P14 (c), and P19 (e) in WT mice. (b, d, and f) Retinogeniculate terminals at P10 (b), P14 (d), and P19 (f) in cPcdh- a KO mice. Scale bars, 20 l mi n( a – f). © 2015 International Society for Neurochemistry, J. Neurochem. (2015) 133 , 66- -72 single glomerulus in the olfactory bulb (Hasegawa et al. 2008). Abnormal aggregation of serotonergic fi bers is also found in cPcdh- a mice (Katori et al. 2009). The corticocor- tical pathways connecting the primary somatosensory cor- tices in both hemispheres are also impaired in cPcdh- a KO mice (Yamashita et al. 2012). These fi ndings indicate that cPcdh- a is required for normal development of axonal projection patterns derived from speci fi c types of neurons. Although cPcdh- a is expressed in most neurons, impairment of cPcdh- a has dominant effects exclusively in speci fi c types of neurons or synapses. The retinal fi ber innervating areas, such as LGd, LGv, and the superior colliculus, also express cPcdh- a , and almost all the optic tract fi bers contain cPcdh- a protein during the early post-natal stage, with abnormality appearing in the LGv and the superior colliculus. These fi ndings suggest that other cPcdhs or cell-adhesion molecules might replace cPcdh- a in other neurons or synapses. Distinct subsets of the cPcdh cluster are expressed differentially in individual neurons, and enormous cell surface diversity may result from their combinatorial expression (Kohmura et al. 1998; Wang et al. 2002a; Esumi et al. 2005; Kaneko et al. 2006). The cell surface diversity between neurons expressing similar sets of cPcdhs (Shapiro and Colman 1999; Phillips et al. 2003; Yagi 2012). Alter- natively, cPcdh- c supposedly plays a role in self-avoidance between neurites that express the same sets of cPcdh- c (Lefebvre et al. 2012; Zipursky and Grueber 2013). The abnormal terminal aggregation observed in the LGd of cPcdh- a KO mice can be explained by either of the following mechanisms: it may be attributed to non-speci fi c synapse formation between a single LGd neuron and many retinal ganglion cells, or the failure of avoidance between retino- geniculate terminals, which avoid one another in WT mice. In cPcdh- a KO mice, terminal aggregation was not observed at P10, and appeared just before normal eye opening at P14; thus, cPcdh- a is likely to play an important role in the re fi nement process of retinogeniculate terminals between P10 and P14 (Chen and Regehr 2000; Hooks and Chen 2006, 2008; Liu and Chen 2008). The slight mean size difference of retinogeniculate terminals already appeared at P10, the starting point of the re fi nement process with lots of excess terminals to be eliminated. cPcdh- a might have some effects on the size of immature terminals. In contrast, cPcdh- c is required for neuronal survival and for preparing cord (Wang et al. 2002b; Prasad et al. 2008; Prasad and Weiner 2011; Chen et al. 2012). Moreover, cPcdh- c -deleted mutant mice lack voluntary movements and re fl exes and die shortly after birth (Wang et al. 2002b; Chen et al. 2012). Therefore, cPcdh- a and cPcdh- c may play distinct roles in the neural circuit formation at different developmental stages. Relationship between terminal aggregation and impaired visual acuity in cPcdh- a KO mice In the present study, we observed aggregation of retinogen- iculate terminals and impaired visual acuity in cPcdh- a KO mice. However, the relationship between the two phenomena remains unclear, because there could be other abnormalities in retinal circuits, in which cPcdh- c plays an important role (Lefebvre et al. 2012), or in circuits higher than LGd, such 30 70 100 0.3 0.4 0.5 0.6 WT Cycles/degree % Correct 0.5 0.6 WT cPcdh -KO ( n = 10) ( n = 9) p < 0.0003 (a) (b) (c) Platform Wate r Wate r Platform (e) (d) 1.0 0 100 0 OSI DSI WT WT cPcdh KO - 10 s 10% F/F 0 50 50 0.5 0.5 cPcdh KO - cPcdh KO - Cumulative distribution C ycles/degree Fig. 4 Reduced visual acuity in cPcdh- a KO mice. (a) Samp le traces of neuronal calcium response to moving grating patterns (from 0 ° to 315 ° in 45 ° steps) in a WT mouse (left) and a cPcdh- a KO mouse (right). (b) Cumulative distributions of the orientation selectivity index (OSI) of 481 neurons in WT mice and 423 neurons in cPcdh- a KO mice (left). The distribution of the direction selectivity index (DSI) of 197 neurons in WT mice and 164 neurons in cPcdh- a KO mice is also Schematic drawing of the Prusky water maze used to estimate visual acuity. (d) Examples of the results obtained for a WT and a cPcdh- a KO mouse. (e) Visual acuity in 10 WT and nine cPcdh- a KO mice. © 2015 International Society for Neurochemistry, J. Neurochem. (2015) 133 , 66- -72 70 R. Meguro et al. as geniculocortical projections, intracortical circuits in V1 or higher visual areas. To date, orientation selectivity, direction selectivity, and ocular dominance plasticity in V1 are apparently normal in cPcdh- a KO mice (Fig. 4 and Yoshi- take et al. 2013). The estimation of visual acuity required visual discrimination learning, in which no apparent differ- ence was found between WT and cPcdh- a KO mice when a grating pattern of 0.14 cycles/degree was used. However, V1 plasticity induced by spatial mismatches between whisker and visual inputs is impaired in cPcdh- a KO mice, suggest- ing the presence of some abnormalities in higher areas of these mice (Yoshitake et al. 2013). The possibility of retinal circuit and geniculocortical circuit abnormality also needs to be considered. However, an important fi nding in the present study is that the magnitude of impaired visual acuity in cPcdh- a KO mice was comparable to that of the aggregated retinothalamic terminals. We investigated visual acuity in the present study, since the aggregated retinogeniculate terminals had suggested impaired visual acuity in cPcdh- a KO mice. However, we cannot exclude the possibility that other parameters such as the size of receptive fi eld are abnormal in cPcdh- a KO mice. Normal retinogeniculate synapses are highly ef fi cient in signal transmission (Budisantoso et al. 2012), and aggrega- tion of terminals is unlikely to further assist signal transmis- sion. Instead, non-speci fi c mixing of the information conveyed by each retinal ganglion axon at the aggregated terminals probably impairs signal transmission. Therefore, the fi ne spatial information conveyed through LGd may be mediated mainly via fi ne terminals ( < 13.1 l m 2 ). The density of fi ne terminals in LGd was reduced to 62.5% in cPcdh- a KO mice compared with WT mice. Assuming that retinogeniculate terminals with spatial information were distributed in the grid, the visual acuity discriminating two vertical (or horizontal) lines are inversely proportional to the grid intervals, or proportional to the square root of the terminal density. When the visual acuity determined using grating patterns would be proportional to the square root of the fi ne-terminal density, it would deteriorate to 79% in cPcdh- a KO mice. Because the visual acuity observed in WT mice was 0.56 cycles/degree, the expected visual acuity in cPcdh- a KO mice could be 0.44 cycles/degree. This expected value is very close to 0.41 cycles/degree, which was the value obtained in behavioral experiments. This simpli fi ed argument suggests that the aggregation of retinogeniculate terminals in cPcdh- a KO mice explains a considerable part of the deteriorated visual acuity in cPcdh- a KO mice, although the possibility that other abnormalities may contribute to the observed visual impairment cannot be excluded (Katori et al. 2009; Lefebvre et al. 2012; Yoshitake et al. 2013). There- fore, morphological and functional studies on the retinogen- iculate synapses in mice with genetically manipulated cPcdhs may be useful for elucidating the functions of cPcdhs in the future. Acknowledgments and conflict of interest disclosure This work was supported by JSPS KAKENHI Grant Number 23659091 and JST-CREST. The authors declare no competing fi nancial interests. All experiments were conducted in compliance with the ARRIVE guidelines. References Budisantoso T., Matsui K., Kamasawa N., Fukazawa Y. and Shigemoto R. (2012) Mechanisms underlying signal fi ltering at a multisynapse contact. J. Neurosci. 32 , 2357 – 2376. Chen C. and Regehr W. G. (2000) Developmental remodeling of the retinogeniculate synapse. Neuron 28 , 955 – 966. Chen W. V., Alvarez F. J., Lefebvre J. L. et al. (2012) Functional signi fi cance of isoform diversi fi cation in the protocadherin gamma gene cluster. Neuron 75 , 402 – 409. Esumi S., Kakazu N., Taguchi Y., Hirayama T., Sasaki A., Hirabayashi T., Koide T., Kitsukawa T., Hamada S. and Yagi T. (2005) Monoallelic yet combinatorial expression of variable exons of the protocadherin-alpha gene cluster in single neurons. Nat. Genet. 37 , 171 – 176. Golding B., Pouchelon G., Bellone C. et al. (2014) Retinal input directs the recruitment of inhibitory interneurons into thalamic visual circuits. Neuron 81 , 1057 – 1069. Guido W. (2008) Re fi nement of the retinogeniculate pathway. J. Physiol. 586 , 4357 – 4362. Hasegawa S., Hamada S., Kumode Y., Esumi S., Katori S., Fukuda E., Uchiyama Y., Hirabayashi T., Mombaerts P. and Yagi T. (2008) The protocadherin-alpha family is involved in axonal coalescence of olfactory sensory neurons into glomeruli of the olfactory bulb in after crossing nerve transfer. PLoS ONE 7 , e35676. Yoshitake K., Tsukano H., Tohmi M., Komagata S., Hishida R., Yagi T. and Shibuki K. (2013) Visual map shifts based on whisker-guided cues in the young mouse visual cortex. Cell Rep. 5 , 1365 – 1374. Zipursky S. L. and Grueber W. B. (2013) The molecular basis of self- avoidance. Annu. Rev. Neurosci. 36 , 547 – 568. © 2015 International Society for Neurochemistry, J. Neurochem. (2015) 133 , 66- -72 72 R. Meguro et al. ... As mentioned in the Introduction section, there are many studies showing that refinement of one of the key elements in the visual system, the retinogeniculate projection, depends on immune molecules during development. Visual information relaying from the retina to the visual thalamus has also been shown to control visual acuity (Meguro et al., 2015) . Therefore, we tested if MIA affects retinogeniculate refinement in the offspring mice. ... ... In the current study, we found that the retinogeniculate synapse maturation is hindered by prenatal immune activation. In addition to optical clarity and photoreceptor arrangement at the fovea, visual information relay from the retina to the visual thalamus has been shown to play an important role in determining visual acuity (Meguro et al., 2015) . Refined retinogeniculate synapses have a significantly stronger synaptic strength than unrefined and resolving synapses and are highly efficient in relaying visual information (Budisantoso et al., 2012). ... Maternal immune activation alters visual acuity and retinogeniculate axon pruning in offspring mice Yixiu Yan Shan Cheng Matthew P Anderson Individuals with autism spectrum disorder (ASD) have been found to have a variety of sensory processing deficits. Here we report that maternal immune activation, a known factor for ASD, impairs visual acuity in the offspring mice. By intraperitoneally injecting polyinosinic-polycytidylic acid (polyI:C) to induce maternal immune activation during embryonic days 10 to 14, we found that polyI:C treatment impairs visual acuity in young adult offspring mice as examined by their optomotor responses. Concurrently, polyI:C treatment suppresses retinogeniculate axon elimination, resulting in a high fraction of weak optical fibers innervating the relay neurons in the visual thalamus. The results link in-utero maternal inflammation to defective optical fiber pruning and arrested developmental strengthening of single optic fibers which may underlie impaired visual acuity. ... Besides the role of Pcdhα in olfactory system, the optic nerve fiber projections in the dorsal lateral geniculate nucleus (LGd) and in the primary visual cortex aggregate in Pcdhα hypomorphic mutants (Meguro et al 2015) . This phenotype is reminiscent of the axonal arborization defects of serotonergic neurons (Chen et al 2017), although it is not known whether this is due to defective tilling. ... ... This phenotype is reminiscent of the axonal arborization defects of serotonergic neurons (Chen et al 2017), although it is not known whether this is due to defective tilling. Perhaps not surprisingly, these mutant mice display defects in visual discrimination behaviors that assess visual acuity (Meguro et al 2015) . Pcdhα genes have also been implicated in dendritic development and spine morphogenesis (Suo et al 2012), and more recently in cortical neuron migration (Fan et al 2018). ... Writing, Reading, and Translating the Clustered Protocadherin Cell Surface Recognition Code for Neural Circuit Assembly Chiamaka L. Nwakeze The ability of neurites of individual neurons to distinguish between themselves and neurites from other neurons and to avoid self (self-avoidance) plays a key role in neural circuit assembly in both invertebrates and vertebrates. Similarly, when individual neurons of the same type project into receptive fields of the brain, they must avoid each other to maximize target coverage (tiling). Counterintuitively, these processes are driven by highly specific homophilic interactions between cell surface proteins that lead to neurite repulsion rather than adhesion. Among these proteins in vertebrates are the clustered protocadherins (Pcdhs), and key to their function is the generation of enormous cell surface structural diversity. Here we review recent advances in understanding how a Pcdh cell surface code is generated by stochastic promoter choice; how this code is amplified and read by homophilic interactions between Pcdh complexes at the surface of neurons; and, finally, how the Pcdh code is translated to cellular function, which mediates self-avoidance and tiling and thus plays a central role in the development of complex neural circuits. Not surprisingly, Pcdh mutations that diminish homophilic interactions lead to wiring defects and abnormal behavior in mice, and sequence variants in the Pcdh gene cluster are associated with autism spectrum disorders in family-based genetic studies in humans. ... Serotonergic fibers gather around the dorsal lateral geniculate nucleus and the medial geniculate nucleus but are scarce in the central regions of these nuclei (Table SⅢ) (39). In PCDHA knockout mice, huge aggregates, formed by the terminals of retinal ganglion cells projecting to the dorsal lateral geniculate nucleus, contribute to vision loss (40) . ... Novel variants identified in a three‑generation family with concomitant exotropia Xuefeng Shi Concomitant exotropia is a condition where there is a misalignment between both eyes, which is more prevalent in Asians than in Caucasians. It is an eye disease related to the neural development of binocular vision and eye movement control. Studies have indicated that genetic factors contribute to the development of concomitant exotropia; however, the underlying mutations have not been thoroughly investigated to date. In the present study, whole-exome sequencing was performed in a three-generation family with concomitant exotropia. In the proband and the proband's father, bioinformatics analyses identified a duplication of the genomic region spanning genes PCDHA1-7 and a heterozygous mutation c.3775G>A (p.A1259T) of the COL3A1 gene, which is located in the conserved COLFI domain and leads to decreased stability of the encoded protein product. Furthermore, a deletion of amino acid S165 in the gene NCOA7 was discovered in the family members, including the proband, the proband's mother and maternal grandfather. S165 was predicted to be a conserved phosphokinase site of CK1/VRK and CK1/CK1. The genes in which these variants reside are all involved in cortical neuronal development. The present study reveals novel variants of concomitant exotropia and suggests that aberrant cortical neuronal development may contribute to the origin of concomitant strabismus. ... 56 In the mice study, an impaired Pcdha cluster resulted in reduced visual acuity. 57 Moreover, the deletion of Pcdha and Pcdhg clusters led to more dramatic defects in the survival of inner retinal neurons and dendritic self-avoidance of starburst amacrine cells in mice retinas than Pcdhg deletion alone. 58 Other protocadherin genes besides the α cluster were also analyzed in myopia studies. ... Decreased Levels of DNA Methylation in the PCDHA Gene Cluster as a Risk Factor for Early-Onset High Myopia in Young Children Justyna A Karolak Marzena Gajecka Purpose: High myopia (HM), an eye disorder with at least -6.0 diopters refractive error, has a complex etiology with environmental, genetic, and likely epigenetic factors involved. To complement the DNA methylation assessment in children with HM, we analyzed genes that had significantly lower DNA methylation levels. Methods: The DNA methylation pattern was studied based on the genome-wide methylation data of 18 Polish children with HM paired with 18 controls. Genes overlapping CG dinucleotides with decreased methylation level in HM cases were assessed by enrichment analyses. From those, genes with CG dinucleotides in promoter regions were further evaluated based on exome sequencing (ES) data of 16 patients with HM from unrelated Polish families, Sanger sequencing data of the studied children, and the RNA sequencing data of human retinal ARPE-19 cells. Results: The CG dinucleotide with the most decreased methylation level in cases was identified in a promoter region of PCDHA10 that overlaps intronic regions of PCDHA1-9 of the PCDHA gene cluster in myopia 5q31 locus. Also, two single nucleotide variants, rs200661444, detected in our ES, and rs246073, previously found as associated with a refractive error in a genome-wide association study, were revealed within this gene cluster. Additionally, genes previously linked to ocular phenotypes, myopia-related traits, or loci, including ADAM20, ZFAND6, ETS1, ABHD13, SBSPON, SORBS2, LMOD3, ATXN1, and FARP2, were found to have decreased methylation. Conclusions: Alterations in the methylation pattern of specific CG dinucleotides may be associated with early-onset HM, so this could be used to develop noninvasive biomarkers of HM in children and adolescents. ... Unlike classical cadherins which typically mediate strong cell adhesion, the impacts of Pcdhs on adhesion remains unclear. For example, Pcdhs promote moderate association with cell membrane (Schreiner and Weiner, 2010;Molumby et al., 2016;Tarusawa et al., 2016), but may also promote anti-adhesive functions to avoid interaction with membranes (Lefebvre et al., 2012;Kostadinov and Sanes, 2015; Meguro et al., 2015) . This combination of pro-adhesion and antiadhesion likely specifies synaptic connections which are fundamental for proper functioning of neural circuit. ... Long-Term Enhancement of NMDA Receptor Function in Inhibitory Neurons Preferentially Modulates Potassium Channels and Cell Adhesion Molecules Qiang Zhou Effectively enhancing the activity of inhibitory neurons has great therapeutic potentials since their reduced function/activity has significant contributions to pathology in various brain diseases. We showed previously that NMDAR positive allosteric modulator GNE-8324 and M-8324 selectively increase NMDAR activity on the inhibitory neurons and elevates their activity in vitro and in vivo. Here we examined the impact of long-term administering M-8324 on the functions and transcriptional profiling of parvalbumin-containing neurons in two representative brain regions, primary auditory cortex (Au1) and prelimbic prefrontal cortex (PrL-PFC). We found small changes in key electrophysiological parameters and RNA levels of neurotransmitter receptors, Na ⁺ and Ca ²⁺ channels. In contrast, large differences in cell adhesion molecules and K ⁺ channels were found between Au1 and PrL-PFC in drug-naïve mice, and differences in cell adhesion molecules became much smaller after M-8324 treatment. There was also minor impact of M-8324 on cell cycle and apoptosis, suggesting a fine safety profile. ... Deficits in serotonergic wiring have also been reported in the -KO and c1/2-KO mice (Katori S et al. 2009;Chen WV et al. 2017) and alterations in serotonergic tone could also affect overall interneuron activity. Interestingly, deficits in visual acuity and aggregation of retinogeniculate terminals have been reported to appear between P10 and P14 in the -KO (Meguro R et al. 2015) , coinciding with the emergence of cIN phenotype we observed at P13. ... Loss of Clustered Protocadherin Diversity Alters the Spatial Distribution of Cortical Interneurons in Mice Full-text available Nov 2020 Cortical interneurons (cINs) are locally-projecting inhibitory neurons that are distributed throughout the cortex. Due to their relatively limited range of influence, their arrangement in the cortex is critical to their function. cINs achieve this arrangement through a process of tangential and radial migration, and apoptosis during development. In this study, we investigated the role of clustered protocadherins (cPcdhs) in establishing the spatial patterning of cINs through the use of genetic cPcdh knockout mice. cPcdhs are expressed in cINs, and are known to play key functions in cell spacing and cell survival, but their role in cINs is poorly understood. Using spatial statistical analysis, we found that the two main subclasses of cINs, parvalbumin-expressing (PV) and somatostatin-expressing (SST) cINs, are non-randomly spaced within subclass, but randomly with respect to each other. We also found that the relative laminar distribution of each subclass was distinctly altered in whole α- or β-cluster mutants. Examination of perinatal timepoints revealed that the mutant phenotypes emerged relatively late, suggesting that cPcdhs may be acting during cIN morphological elaboration and synaptogenesis. We then analyzed an isoform-specific knockout for pcdh-αc2 and found that it recapitulated the α-cluster knockout, but only in SST cells, suggesting that subtype-specific expression of cPcdh isoforms may help govern subtype-specific spatial distribution. ... Deficits in serotonergic wiring have also been reported in the a-KO and ac1/2-KO mice (Katori S et al. 2009;Chen WV et al. 2017) and alterations in serotonergic tone could also affect overall interneuron activity. Interestingly, deficits in visual acuity and aggregation of retinogeniculate terminals have been reported to appear between P10 and P14 in the a-KO (Meguro R et al. 2015) , coinciding with the emergence of cIN phenotype we observed at P13. This could alter the activity state of the visual cortex, where we observe the most robust mutant phenotype. ... Loss of clustered protocadherin diversity alters the spatial distribution of cortical interneurons Cortical interneurons (cINs) are locally-projecting inhibitory neurons that are distributed throughout the cortex. Due to their relatively limited range of influence, their arrangement in the cortex is critical to their function. cINs achieve this arrangement through a process of tangential and radial migration, and apoptosis during development. In this study, we investigated the role of clustered protocadherins (cPcdhs) in establishing the spatial patterning of cINs. cPcdhs are expressed in cINs, and are known to play key functions in cell spacing and cell survival, but their role in cINs is poorly understood. Using spatial statistical analysis, we found that the two main subclasses of cINs, parvalbumin-expressing (PV) and somatostatin-expressing (SST) cINs, are non-randomly spaced within subclass, but randomly with respect to each other. We also found that the relative laminar distribution of each subclass was distinctly altered in whole α- or β-cluster mutants. Examination of perinatal timepoints revealed that the mutant phenotypes emerged relatively late, suggesting that cPcdhs may be acting during cIN morphological elaboration and synaptogenesis. We then analyzed an isoform-specific knockout for pcdh-αc2 and found that it recapitulated the α-cluster knockout, but only in SST cells, suggesting that subtype-specific expression of cPcdh isoforms may help govern subtype-specific spatial distribution. ... Third, clustered Pcdhs, like Dscam1 isoforms, are expressed in a combinatorial and stochastic fashion in at least some populations (Esumi et al., 2005;Kaneko et al., 2006;Toyoda et al., 2014;Mountoufaris et al., 2018). Fourth, clustered Pcdhs mediate self-avoidance in several neuronal populations, including dendrites of retinal starburst amacrine cells and cerebellar Purkinje cells and axons of olfactory sensory and serotonergic neurons; they also mediate self-/non-self-discrimination in at least starburst amacrine cells Meguro et al., 2015; Mountoufaris et al., 2017;Chen et al., 2017;Katori et al., 2017;Ing-Esteves et al., 2018). Although specific isoforms play unique roles in some cases Katori et al., 2017), in general, as with Dscam1, the specific isoforms expressed in a neuron are unimportant, it is only important that they are different from one another. ... Synaptic Specificity, Recognition Molecules, and Assembly of Neural Circuits Developing neurons connect in specific and stereotyped ways to form the complex circuits that underlie brain function. By comparison to earlier steps in neural development, progress has been slow in identifying the cell surface recognition molecules that mediate these synaptic choices, but new high-throughput imaging, genetic, and molecular methods are accelerating progress. Over the past decade, numerous large and small gene families have been implicated in target recognition, including members of the immunoglobulin, cadherin, and leucine-rich repeat superfamilies. We review these advances and propose ways in which combinatorial use of multifunctional recognition molecules enables the complex neuron-neuron interactions that underlie synaptic specificity. Self-avoidance alone does not explain the function of Dscam1 in mushroom body axonal wiring Alternative splicing of Drosophila Dscam1 into 38,016 isoforms provides neurons with a unique molecular code for self-recognition and self-avoidance. A canonical model suggests that the homophilic binding of identical Dscam1 isoforms on the sister branches of mushroom body (MB) axons supports segregation with high fidelity, even when only a single isoform is expressed. Here, we generated a series of mutant flies with a single exon 4, 6, or 9 variant, encoding 1,584, 396, or 576 potential isoforms, respectively. Surprisingly, most of the mutants in the latter two groups exhibited obvious defects in the growth, branching, and segregation of MB axonal sister branches. This demonstrates that the repertoires of 396 and 576 Dscam1 isoforms were not sufficient for the normal patterning of axonal branches. Moreover, reducing Dscam1 levels largely reversed the defects caused by reduced isoform diversity, suggesting a functional link between Dscam1 expression levels and isoform diversity. Taken together, these results indicate that canonical self-avoidance alone does not explain the function of Dscam1 in MB axonal wiring. CRISPR/Cas9 interrogation of the mouse Pcdhg gene cluster reveals a crucial isoform-specific role for Pcdhgc4

https://www.researchgate.net/publication/272029469_Impaired_clustered_protocadherin-a_cPcdh-a_leads_to_aggregated_retinogeniculate_terminals_and_impaired_visual_acuity_in_mice

Aggressive stem cells might improve transplant outcome | EurekAlert! Science News Investigators at St. Jude Children's Research Hospital have demonstrated in mice a way that might reduce the time it takes for a bone marrow transplant to rebuild a child's immune system, and so reduce the risk of potentially fatal virus infections that can occur during this time. Aggressive stem cells might improve transplant outcome St. Jude researchers find that harvesting aggressive stem cells from donated bone marrow could speed rebuilding of the immune system, reduce risk of infection IMAGE:Raymond Barfield, M.D., Ph.D., Division of Bone Marrow Transplantation at St. Jude Children's Research Hospitalview more Credit: Biomedical Communications, St. Jude Children's Research Hospital Investigators at St. Jude Children's Research Hospital have demonstrated in mice a way that might reduce the time it takes for a bone marrow transplant to rebuild a child's immune system, and so reduce the risk of potentially fatal virus infections that can occur during this time. The St. Jude team showed that the current way of harvesting specific stem cells from donated bone marrow to capture many of the stem cells called CD34+ cells fails to capture many of the cells that might be more vigorous in reproducing and rebuilding the immune system. CD34+ cells give rise to a variety of blood cells including T lymphocytes, a critical part of the immune system that orchestrates the attack on invading microorganisms. The researchers showed in mice that most of the CD34+ stem cells left behind by the current harvesting method are especially vigorous in multiplying and producing T lymphocytes. In humans, the loss of many of these vigorous stem cells from the transplant might contribute to the delay in rebuilding the immune system--a time lag that leaves children exposed to serious infections. The St. Jude finding is important because children whose own bone marrow stem cells have been destroyed by chemotherapy or radiation treatments for cancer routinely undergo bone marrow transplantation. Therefore, reducing the time it takes for the immune system to regenerate T lymphocytes would be an important step in reducing infection risk in these children and improving their long-term outcomes. A report on this work appears in the current online issue of the journal Stem Cell. Stem cells that give rise to T lymphocytes--whether they are a person's own stem cells or donated cells--normally gather in the thymus gland in the chest. There the stem cells receive their "marching orders" as new T lymphocyte warriors against invading bacteria, viruses and other potentially dangerous targets including cancer cells, and they develop a specialized homing mechanism called a T cell receptor that lets them find the specific target they are genetically programmed to attack. The key to the new finding is the discovery by the St. Jude team that the antibody currently used to seek out and "capture" CD34+ stem cells from the donated bone marrow actually identifies only a certain group of them, according to Raymond Barfield, M.D., Ph.D., assistant member of the Bone Marrow Transplantation division at St. Jude. The antibody usually used to capture CD34+ cells, called a class II antibody, recognizes a particular target on the CD34+ cells, Barfield said. But this antibody does not as easily recognize more vigorous CD34+ cells that mature and reproduce faster. These more aggressive stem cells have a different target on their surface that is best recognized by another type of antibody, called class III. The St. Jude investigators first demonstrated that class II and III antibodies bind to different populations of CD34+ cells by adding both of these antibodies to pieces of human thymus glands. The pieces of thymus glands used in the study had been removed from children during operations unconnected to this research and would otherwise have been discarded. Using the different types of antibodies, the scientists determined which one produced T lymphocytes faster by measuring the amount of rings of DNA called sjTRECs the cells released. T lymphocytes discard sjTRECs as they develop from stem cells in the thymus and "learn" to recognize a specific target. (St. Jude researchers previously showed that by measuring sjTRECs clinicians could predict whether a bone marrow stem cell transplant would successfully restore the recipient's T lymphocyte population: [http://www.stjude.org/media/0,2561,453_5489_17063,00.html]). The investigators found that the lymphocytes captured by class III antibodies produced more sjTRECs than stem cells captured by class II antibodies; that is, the class III antibody stem cells matured faster. The researchers then infused the cells captured by class II and III antibodies into mice that had been specially bred to lack an immune system. The cells traveled to the thymus, where they were trained to produce different families of T lymphocytes, each of which was genetically programmed to attack a specific target. Half of the mice that received cells selected with class III antibodies produced sjTRECs compared with 14 percent of mice transplanted with cells selected using the class II antibody. "These findings show that the thymus of mice that got class III antibody stem cells were able to produce T lymphocytes quicker than mice receiving class II antibody stem cells," Barfield noted. "This suggests if we harvest CD34+ cells using class III as well as class II antibodies we are likely to get better results when we transplant them into children." The results also suggest that the time from transplantation to a fully restored immune system might be reduced using this strategy. The St. Jude team said that more studies are needed to confirm this initial finding and to translate it into better treatment for children. "Such information will help clinicians rethink how they can do bone marrow stem cell transplantation so that the risk of infection is lowered in their patients," Barfield said. ### Other authors of this study include Mario Otto (now at Akron Children's Hospital, Ohio); Xiaohua Chen, William J. Martin, Wing Leung, James Knowles, Marti Holladay and Jim Houston (all of St. Jude); and Rupert Handgretinger (University of Tübingen, Germany). This work was supported in part by a Cancer Center Support grant, ALSAC and the Assisi Foundation of Memphis. St. Jude Children's Research Hospital St. Jude Children's Research Hospital is internationally recognized for its pioneering work in finding cures and saving children with cancer and other catastrophic diseases. Founded by late entertainer Danny Thomas and based in Memphis, Tenn., St. Jude freely shares its discoveries with scientific and medical communities around the world. No family ever pays for treatments not covered by insurance, and families without insurance are never asked to pay. St. Jude is financially supported by ALSAC, its fund-raising organization. For more information, please visitwww.stjude.org. Stem Cell Cancer Center Support, American Lebanese Syrian Associated Charities, Assisi Foundation of Memphis Keywords

https://archive.eurekalert.org/pub_releases/2006-12/sjcr-asc120706.php

Diseases | Free Full-Text | Update Treatment for HBV Infection and Persistent Risk for Hepatocellular Carcinoma: Prospect for an HBV Cure Since the discovery of the hepatitis B virus (HBV) by Blumberg et al. in 1965, its genome, sequence, epidemiology, and hepatocarcinogenesis have been elucidated. Globally, hepatitis B virus (HBV) is still responsible for the majority of hepatocellular carcinoma (HCC). HCC is the sixth-most common cancer in the world and the second-most common cancer death. The ultimate goal of treating HBV infection is the prevention of HCC. Fortunately, anti-HBV treatment with nucleos(t)ide analogues (NAs), which began with lamivudine in 1998, has resulted in remarkable improvements in the survival of patients with chronic hepatitis B and a reduced incidence of HCC. These results were documented with lamivudine, entecavir, and tenofovir. Nonetheless, as the duration of antiviral treatment increases, the risk for HCC still remains despite undetectable HBV DNA in serum, as reported by different investigators with observation up to 4–5 years. In our own experience, we are witnessing the development of HCC in patients who have received antiviral treatment. Some have enjoyed negative serum HBV DNA for over 12 years before developing HCC. Current treatment with NAs can effectively suppress the replication of the virus but cannot eradicate the covalently closed circular DNA (cccDNA) that is within the nucleus of hepatocytes. There still remains a great need for a cure for HBV. Fortunately, several compounds have been identified that have the potential to eradicate HBV, and there are ongoing clinical trials in progress in their early stages. Update Treatment for HBV Infection and Persistent Risk for Hepatocellular Carcinoma: Prospect for an HBV Cure by Joseph Yoo 1 , Hie-Won Hann 1,2,* , Robert Coben 1 , Mitchell Conn 1 and Anthony J. DiMarino 1 Liver Disease Prevention Center Department of Medicine, Thomas Jefferson University Hospital, Philadelphia, PA 19107, USA * Author to whom correspondence should be addressed. Diseases 2018 , 6 (2), 27; https://doi.org/10.3390/diseases6020027 Received: 20 March 2018 / Revised: 13 April 2018 / Accepted: 16 April 2018 / Published: 20 April 2018 Abstract : Since the discovery of the hepatitis B virus (HBV) by Blumberg et al. in 1965, its genome, sequence, epidemiology, and hepatocarcinogenesis have been elucidated. Globally, hepatitis B virus (HBV) is still responsible for the majority of hepatocellular carcinoma (HCC). HCC is the sixth-most common cancer in the world and the second-most common cancer death. The ultimate goal of treating HBV infection is the prevention of HCC. Fortunately, anti-HBV treatment with nucleos(t)ide analogues (NAs), which began with lamivudine in 1998, has resulted in remarkable improvements in the survival of patients with chronic hepatitis B and a reduced incidence of HCC. These results were documented with lamivudine, entecavir, and tenofovir. Nonetheless, as the duration of antiviral treatment increases, the risk for HCC still remains despite undetectable HBV DNA in serum, as reported by different investigators with observation up to 4–5 years. In our own experience, we are witnessing the development of HCC in patients who have received antiviral treatment. Some have enjoyed negative serum HBV DNA for over 12 years before developing HCC. Current treatment with NAs can effectively suppress the replication of the virus but cannot eradicate the covalently closed circular DNA (cccDNA) that is within the nucleus of hepatocytes. There still remains a great need for a cure for HBV. Fortunately, several compounds have been identified that have the potential to eradicate HBV, and there are ongoing clinical trials in progress in their early stages. Keywords: HBV ; HCC ; anti-HBV therapy ; HBV cure risk for HCC 1. Introduction ], much has been elucidated regarding its genome, sequence, epidemiology, and hepatocarcinogenesis. However, despite all that is known about the virus, and global vaccination programs, HBV infection continues to be a significant public health problem worldwide. It is estimated that there are currently greater than 240–300 million people chronically infected with HBV, and 75% of these individuals live in the Asia-Pacific region. HBV is responsible for 30% of all cases of cirrhosis and over 50% of hepatocellular carcinoma (HCC) with an estimated 50 million new cases diagnosed annually in endemic countries [ 2 , 3 ]. Once infected with HBV, one of the main risks is the development of cirrhosis, hepatic decompensation, and ultimately HCC. Without treatment, 15–40% of these individuals will develop these sequelae during their lifetime [ 4 , 5 ]. Though the exact mechanism is not yet completely understood, the current thought is that the replication of the virus itself is not directly cytotoxic to cells, but rather, it is the response of the host immune system towards infected hepatocytes that ultimately leads to immune mediated liver damage [ 5 , 6 ]. HBV is indeed the most significant hepatocarcinogen responsible for the development of HCC, which currently ranks as the sixth-most common cancer worldwide and the second-most common cause of cancer death [ 7 , 8 , 9 , 10 ]. Since the first report by Beasley et al. [ 4 ], the connection between HBV and HCC has been well established. Subsequent reports have found that individuals with chronic hepatitis B (CHB) have up to a 30-fold increased risk of HCC, with approximately 25% of individuals infected with CHB developing HCC during their lifetime [ 5 , 11 , 12 , 13 , 14 , 15 ]. Although most patients with HCC had evidence of cirrhosis [ 16 ], HBV has been shown to cause HCC even in the absence of cirrhosis, and inactive carriers of HBV (HBeAg negative, HBV DNA <10,000 copies/mL, normal liver enzymes, no cirrhosis) continue to carry a five-fold increased risk of HCC compared to HBsAg-negative controls [ 17 ]. 2. Antiviral Therapy and Incidence of HCC Currently-available therapies have shown the ability to delay disease progression and reduce the incidence of HCC with lamivudine, entecavir, and tenofovir disoproxil fumarate [ 18 , 19 , 20 ]. Furthermore, in patients who have already developed HCC, concomitant antiviral therapy after initial tumor ablation resulted in not only improved survival, but also reduced recurrence of tumor [ 21 , 22 , 23 , 24 , 25 , 26 ]. However, as the antiviral treatment period is prolonged, the risk for HCC persists in CHB patients even after more than a decade of successful viral suppression as reported earlier [ 27 ]. In our experience at the Liver Disease Prevention Center, Division of Gastroenterology and Hepatology of Thomas Jefferson University Hospital, over two thousand patients have been treated for chronic HBV infection over the last twenty-five years. Underlining the risk of HCC, between July 2007 and May 2015, there were 12 patients who developed HCC after having been on anti-viral treatment for over 18 years. The details are shown in Table 1 below. After the detection of the small tumor, all patients underwent local tumor ablation with continued antiviral therapy and were closely monitored. Five patients developed new or recurrent tumor and eventually died. On the other hand, 7/12 have remained tumor free (Patient 8 underwent liver transplantation) and have continued antiviral therapy. Our experience is perhaps the longest with oral antiviral therapy since lamivudine, the first nucleoside analogue, was approved in 1998. Given the persistent risk of HCC even after two decades of antiviral (nucleos(t)ides) therapy, there is a desperate need to develop new drugs to cure HBV. In addition to the persistent risk for an initial diagnosis of HCC, we have also observed the development of de novo or recurrent HCC appearing at 10, 11, and 15 years after the successful ablation of the initial HCC despite continued HBV suppression with antiviral treatment (personal communication). With the recent development of a cure for the hepatitis C virus, the time now seems ripe to unpack the remaining mysteries of HBV and develop a cure for a virus which continues to cause significant morbidity and mortality worldwide. 3. Understanding Hepatitis B Virus Much of the genome and replication cycle of HBV has been elucidated since its original discovery in 1965 [ 1 ], and that understanding has yielded various targeted approaches in the effort to discover a cure. HBV is composed of an outer envelope and a viral nucleocapsid within the envelope. The outer envelope contains three primary envelope glycoproteins, or surface antigens, which are central in the diagnosis of HBV infection [ 28 ]. The viral nucleocapsid, or core, contains a DNA polymerase-reverse transcriptase as well as the HBV viral genome, a relaxed partially double-stranded circular DNA molecule (rcDNA) with a length of about 3.2 kb [ 29 , 30 ]. The viral replication cycle begins when HBV recognizes highly-sulfated heparin sulfate proteoglycans (HSPGs) on the surface of liver cells, and binds sodium taurocholate co-transporting polypeptide (NTCP or SLC10A1) to gain entry into the host cell [ 31 , 32 ]. Upon entry into a host cell, the partially double-stranded viral DNA is converted into a covalently closed circular DNA (cccDNA) within the nucleus of the host cell. This stable cccDNA molecule remains within the nucleus of the infected host cell, and is used as the template for the transcription of four viral mRNA intermediates. The mRNA intermediates are then transported to the cytoplasm and translated for the production of seven HBV proteins, including viral surface, core, polymerase, and X proteins. The HBV X protein has been of particular interest as it is felt to play a critical role in the development of HCC, as will be discussed in more detail later. The largest of these mRNA intermediates, the pregenomic RNA (pgRNA) is critical for viral replication as it encodes the viral core, polymerase, and also serves as a template for viral DNA synthesis [ 5 , 33 , 34 ]. 4. Hepatocarcinogenesis As previously stated, infection with HBV has a well-established connection with the development of HCC [ 10 , 11 , 12 , 13 , 14 , 15 ], and HBV DNA sequences have been found to be integrated into the cellular DNA of HCC patients since the early 1980s [ 35 ]. Currently, the connection between HBV and HCC has been so firmly established, that HBV is considered a group 1 human carcinogen by the World Health Organization. Nevertheless, the entire carcinogenic process of HBV-associated HCC is not fully elucidated, and studies have suggested that the mechanism is most likely multifactorial. In addition to the integration of HBV DNA into host chromosomes, there could be the activation of cellular oncogenes, inactivation of tumor suppressor genes, chronic liver injury, inflammation and regeneration (cellular DNA synthesis, impaired cellular repair etc.), activation of cellular proto-oncogenes or suppression of growth regulating genes, and an increase of the HBx protein, which plays an important role in the transcription of HBV DNA as well as in hepatocarcinogenesis. As suggested earlier, the pathogenesis of hepatocellular injury and ultimately cirrhosis seems to result from the host immune response against infected hepatocytes, as opposed to being a result of viral replication itself. Similarly, the carcinogenesis of HBV infection results from creating an environment of chronic inflammation, as well as interactions the virus has with signaling pathways in both the innate and adaptive immune system of the infected host. For example, NF-kB, a signaling molecule involved in the pro-inflammatory response of the innate immune system, and the Tim-3/galectin-9 signaling pathway, associated with the HBV-mediated T cell dysfunction within the adaptive immune system, have both been shown to play a role in the development of HBV-associated HCC [ 36 ]. One of the more important factors in the hepatocarcinogenesis of HBV is likely to be the integration of the virus into the host genome. By integrating into the host genome, endogenous genes can be affected and, subsequently, their expression can become dysregulated. One such example is the TERT gene, which encodes telomerase reverse transcriptase, and is often found to be overexpressed in cancer cells [ 36 ]. In 2012, Sung et al. [ 37 ] published the results of sequencing 81 HBV-positive and seven HBV-negative HCC genomes, along with adjacent normal tissues. The results showed that HBV integration was observed more frequently in the tumors (86.4%) compared to adjacent liver tissues (30.7%). Furthermore, recurrent integration events (≥4) were found at known cancer-related genes, including TERT, resulting in overexpression of these genes in tumor versus normal tissue. In addition to causing altered expression of endogenous genes by integration into the host genome, the HBV genome itself, and the viral proteins it encodes for, carry properties that increase the risk for HCC. One such property is the relatively high rate of mutation of the HBV genome, and recent work by Park et al. has now identified eight specific mutations within the HBV genome that indicate an increased risk for the development of HCC [ 36 , 38 ]. HBV X (HBx), as previously stated, is a viral protein that is also felt to play an important role in the pathogenesis and hepatocarcinogenesis of HBV. Specifically, it has been shown to interact with Bcl-2, a well-known protein involved in regulating cell death and apoptosis [ 39 ]. The connection of HBx protein and HCC is further supported by the fact that the HBx protein is found in most patients with HBV-related HCC, and that the HBx gene is the most frequently integrated viral sequence in HCC [ 36 ]. 5. Current Therapies Current treatments for hepatitis B include interferon and the nuceleos(t)ide analogs lamivudine, adefovir, entecavir, telbivudine, tenofovir disoproxil fumarate and, recently FDA-approved tenofovir alafenamide. Interferon works primarily through immune modulation and has only a weak antiviral effect, while the nucleos(t)ide analogs suppress the replication of the viral genome by selectively targeting the viral reverse transcriptase [ 5 , 40 ]. Ultimately, the goal of treatment with either interferon or a nucleos(t)ide analog is to achieve a functional cure by preventing the development of cirrhosis and HCC. Indicators of successful therapy (functional cure) are the loss of HBsAg and/or seroconversion to anti-HBs, and undetectable HBV DNA in serum. However, without eradicating cccDNA, HBV continues to carry the risk of development of HCC even if a functional cure with antiviral treatment is achieved, especially in patients who have already developed cirrhosis [ 27 ]. 6. Emerging Therapies Multiple emerging drug therapies are currently in the early stages of development as part of the growing effort to find a true cure for HBV [ 41 ]. Generally speaking, the desired emerging therapies would include: (1) prevention of the entry of the whole virion into an uninfected hepatocyte by blocking the liver specific receptor (sodium taurocholate co-transporting polypeptide (NTCP) [ 32 , 42 ]; (2) prevention of the virion release into circulation; (3) directly targeting cccDNA, such as the formation of, or destabilization and elimination of HBV cccDNA. It is the stable HBV cccDNA, found within the nucleus of infected cells, that stands as the biggest obstacle in finding a cure, as it allows HBV to persist and integrate into the host genome [ 34 ]; (4) RNA interference during the transcription of RNA including pregenomic RNA; (5) direct acting antivirals (DAAs) that target the HBV capsid; (6) the conversion of rcDNA to cccDNA; (7) Immune modulation of innate and adaptive immune responses, (such as toll-like receptor agonists, HBV specific T cell vaccine) ; and (8) Inhibition of the viral DNA polymerase (currently done with nucleos(t)ide analogues). Levrero et al. has introduced the recent development of potential cure drugs with detailed updates [ 43 ]. 7. Conclusions In light of the growing understanding of HBV and the focus on the need for continued HBV basic science research, there still remain several unanswered questions. There are obstacles to solving the remaining questions, such as the current lack of a robust tissue culture system for in vitro studies and the lack of practical animal models for in vivo studies. However, the future is promising, and each remaining unanswered question about HBV represents an unexplored opportunity to discover a new potential strategy in the effort to cure HBV. Author Contributions H.H., and J.Y. conceived and wrote the paper, H.H., R.C., M.C., and A.J.D. contributed and analyzed the long-term follow-up data. Conflicts of Interest The authors declare no conflict of interest. References Blumberg, B.S.; Alter, H.J.; Visnich, S. A New Antigen in Leukemia Sera. JAMA 1984 , 252 , 252–257. [ Google Scholar ] [ CrossRef ] [ PubMed ] Schweitzer, A.; Horn, J.; Mikolajczyk, R.T.; Krause, G.; Ott, J.J. Estimations of worldwide prevalence of chronic hepatitis B virus infection: A systematic review of data published between 1965 and 2013. Lancet 2015 , 386 , 1546–1555. [ Google Scholar ] [ CrossRef ] MacLachlan, J.H.; Cowie, B.C. Hepatitis B virus epidemiology. Cold Spring Harb. Perspect. Med. 2015 , 5 , a021410. [ Google Scholar ] [ CrossRef ] [ PubMed ] Beasley, R.P.; Lin, C.C.; Hwang, L.Y.; Chien, C.S. Hepatocellular carcinoma and hepatitis B virus: A prospective study of 22 707 men in Taiwan. Lancet 1981 , 318 , 1129–1133. [ Google Scholar ] [ CrossRef ] Ganem, D.; Prince, A.M. Hepatitis B virus infection—Natural history and clinical consequences. N. Engl. J. Med. 2004 , 350 , 1118–1129. [ Google Scholar ] [ CrossRef ] [ PubMed ] Chisari, F.V.; Ferrari, C. Hepatitis B virus immunopathogenesis. Ann. Rev. Immunol. 1995 , 13 , 29–60. [ Google Scholar ] [ CrossRef ] [ PubMed ] Ferlay, J.; Shin, H.R.; Bray, F.; Forman, D.; Mathers, C.; Parkin, D.M. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int. J. Cancer 2010 , 127 , 2893–2917. [ Google Scholar ] [ CrossRef ] [ PubMed ] Sanyal, A.J.; Yoon, S.K.; Lencioni, R. The etiology of hepatocellular carcinoma and consequences for treatment. Oncologist 2010 , 15 (Suppl. 4), 14–22. [ Google Scholar ] [ CrossRef ] [ PubMed ] Szabó, E.; Páska, C.; Novák, P.K.; Schaff, Z.; Kiss, A. Similarities and differences in hepatitis B and C virus induced hepatocarcinogenesis. Pathol. Oncol. Res. 2004 , 10 , 5. [ Google Scholar ] [ CrossRef ] [ PubMed ] Gurtsevitch, V.E. Human oncogenic viruses: Hepatitis B and hepatitis C viruses and their role in hepatocarcinogenesis. Biochemistry (Moscow) 2008 , 73 , 504–513. [ Google Scholar ] [ CrossRef ] [ PubMed ] Franceschi, S.; Montella, M.; Polesel, J.; La Vecchia, C.; Crispo, A.; Dal Maso, L.; Casarin, P.; Izzo, F.; TommasI, L.G.; Chemin, I.; et al. Hepatitis viruses, alcohol, and tobacco in the etiology of hepatocellular carcinoma in Italy. Cancer Epidemiol. Biomark. Prev. 2006 , 15 , 683–689. [ Google Scholar ] [ CrossRef ] [ PubMed ] Matsubara, K.; Tokino, T. Integration of hepatitis B virus DNA and its implications for hepatocarcinogenesis. Mol. Biol. Med. 1990 , 7 , 243–260. [ Google Scholar ] [ PubMed ] Fourel, G.; Trepo, C.; Bougueleret, L.; Henglein, B.; Ponzett, A.; Tiollais, P.; Buendia, M.A. Frequent activation of N-myc genes by hepadnavirus insertion in woodchuck liver tumours. Nature 1990 , 347 , 294. [ Google Scholar ] [ CrossRef ] [ PubMed ] Popper, H.; Roth, L.; Purcell, R.H.; Tennant, B.C.; Gerin, J.L. Hepatocarcinogenicity of the woodchuck hepatitis virus. Proc. Natl. Acad. Sci. USA 1987 , 84 , 866–870. [ Google Scholar ] [ CrossRef ] [ PubMed ] Kim, C.M.; Koike, K.; Saito, I.; Miyamura, T.; Jay, G. HBx gene of hepatitis B virus induces liver cancer in transgenic mice. Nature 1991 , 351 , 317. [ Google Scholar ] [ CrossRef ] [ PubMed ] Yang, J.D.; Kim, W.R.; Coelho, R.; Mettler, T.A.; Benson, J.T.; Sanderson, S.O.; Therneau, T.M.; Kim, B.; Roberts, L.R. Cirrhosis is present in most patients with hepatitis B and hepatocellular carcinoma. Clin. Gastroenterol. Hepatol. 2011 , 9 , 64–70. [ Google Scholar ] [ CrossRef ] [ PubMed ] Chen, J.D.; Yang, H.I.; Iloeje, U.H.; You, S.L.; Lu, S.N.; Wang, L.Y.; Su, J.; Sun, C.A.; Liaw, Y.F.; Chen, C.J. Carriers of inactive hepatitis B virus are still at risk for hepatocellular carcinoma and liver-related death. Gastroenterology 2010 , 138 , 1747–1754. [ Google Scholar ] [ CrossRef ] [ PubMed ] Liaw, Y.F.; Sung, J.J.; Chow, W.C.; Farrell, G.; Lee, C.Z.; Yuen, H.; Tanwandee, T.; Tao, Q.M.; Shue, K.; Keene, O.N.; et al. Lamivudine for patients with chronic hepatitis B and advanced liver disease. N. Engl. J. Med. 2004 , 351 , 1521–1531. [ Google Scholar ] [ CrossRef ] [ PubMed ] Hosaka, T.; Suzuki, F.; Kobayashi, M.; Seko, Y.; Kawamura, Y.; Sezaki, H.; Akuta, N.; Suzuki, Y.; Saitoh, S.; Arase, Y.; et al. Long-term entecavir treatment reduces hepatocellular carcinoma incidence in patients with hepatitis B virus infection. Hepatology 2013 , 58 , 98–107. [ Google Scholar ] [ CrossRef ] [ PubMed ] Kim, W.; Loomba, R.; Berg, T.; Aguilar Schall, R.E.; Yee, L.J.; Dinh, P.V.; Flaherty, J.F.; Martins, E.B.; Therneau, T.M.; Jacobson, I.; et al. Impact of long-term tenofovir disoproxil fumarate on incidence of hepatocellular carcinoma in patients with chronic hepatitis B. Cancer 2015 , 121 , 3631–3638. [ Google Scholar ] [ CrossRef ] [ PubMed ] Kubo, S.; Tanaka, H.; Takemura, S.; Yamamoto, S.; Hai, S.; Ichikawa, T.; Kodai, S.; Shinkawa, H.; Sakaguchi, H.; Tamori, A.; et al. Effects of lamivudine on outcome after liver resection for hepatocellular carcinoma in patients with active replication of hepatitis B virus. Hepatol. Res. 2007 , 37 , 94–100. [ Google Scholar ] [ CrossRef ] [ PubMed ] Huang, G.; Lau, W.Y.; Wang, Z.G.; Pan, Z.Y.; Yuan, S.X.; Shen, F.; Zhou, W.P.; Wu, M.C. Antiviral therapy improves postoperative survival in patients with hepatocellular carcinoma: A randomized controlled trial. Ann. Surg. 2015 , 261 , 56–66. [ Google Scholar ] [ CrossRef ] [ PubMed ] Yin, J.; Li, N.; Han, Y.; Xue, J.; Deng, Y.; Shi, J.; Guo, W.; Zhang, H.; Wang, H.; Cheng, S.; et al. Effect of antiviral treatment with nucleotide/nucleoside analogs on postoperative prognosis of hepatitis B virus-related hepatocellular carcinoma: A two-stage longitudinal clinical study. J. Clin. Oncol. 2013 , 31 , 3647–3655. [ Google Scholar ] [ CrossRef ] [ PubMed ] Chuma, M.; Hige, S.; Kamiyama, T.; Meguro, T.; Nagasaka, A.; Nakanishi, K.; Yamamoto, Y.; Nakanishi, M.; Kohara, T.; Sho, T.; et al. The influence of hepatitis B DNA level and antiviral therapy on recurrence after initial curative treatment in patients with hepatocellular carcinoma. J. Gastroenterol. 2009 , 44 , 991–999. [ Google Scholar ] [ CrossRef ] [ PubMed ] Wu, C.Y.; Chen, Y.J.; Ho, H.J.; Hsu, Y.C.; Kuo, K.N.; Wu, M.S.; Lin, J.T. Association between nucleoside analogues and risk of hepatitis B virus–related hepatocellular carcinoma recurrence following liver resection. JAMA 2012 , 308 , 1906–1913. [ Google Scholar ] [ CrossRef ] [ PubMed ] Hann, H.W.; Coben, R.; Brown, D.; Needleman, L.; Rosato, E.; Min, A.; Hann, R.S.; Park, K.B.; Dunn, S.; DiMarino, A.J. A long-term study of the effects of antiviral therapy on survival of patients with HBV-associated hepatocellular carcinoma (HCC) following local tumor ablation. Cancer Med. 2014 , 3 , 390–396. [ Google Scholar ] [ CrossRef ] [ PubMed ] Dargan, A.; Wong, S.Y.; Coben, R.; Conn, M.; Dimarino, A.J.; Hann, H.W. Persistent risk for hepatocellular carcinoma after more than a decade of successful hepatitis B virus suppression. Miner. Gastroenterol. E Dietol. 2017 , 63 , 74. [ Google Scholar ] Ganem, D. Assembly of hepadnaviral virions and subviral particles. In Hepadnaviruses ; Springer: Heidelberg, Berlin, Germany, 1991; pp. 61–83. [ Google Scholar ] Robinson, W.S.; Lutwick, L.I. The virus of hepatitis, type, B. N. Engl. J. Med. 1976 , 295 , 1168–1175. [ Google Scholar ] [ CrossRef ] [ PubMed ] Summers, J.; O’Connell, A.; Millman, I. Genome of hepatitis B virus: Restriction enzyme cleavage and structure of DNA extracted from Dane particles. Proc. Natl. Acad. Sci. USA 1975 , 72 , 4597–4601. [ Google Scholar ] [ CrossRef ] [ PubMed ] Schulze, A.; Gripon, P.; Urban, S. Hepatitis B virus infection initiates with a large surface protein–dependent binding to heparan sulfate proteoglycans. Hepatology. 2007 , 46 , 1759–1768. [ Google Scholar ] [ CrossRef ] [ PubMed ] Summers, J.; Mason, W.S. Replication of the genome of a hepatitis B-like virus by reverse transcription of an RNA intermediate. Cell 1982 , 29 , 403–415. [ Google Scholar ] [ CrossRef ] Halegoua-De Marzio, D.; Hann, H.W. Then and now: The progress in hepatitis B treatment over the past 20 years. World J. Gastroenterol. 2014 , 20 , 401. [ Google Scholar ] [ CrossRef ] [ PubMed ] Zeisel, M.B.; Lucifora, J.; Mason, W.S.; Sureau, C.; Beck, J.; Levrero, M.; Kann, M.; Knolle, P.A.; Benkirane, M.; Durantel, D.; et al. Towards an HBV cure: State-of-the-art and unresolved questions—Report of the ANRS workshop on HBV cure. Gut 2015 , 64 , 1314–1326. [ Google Scholar ] [ CrossRef ] [ PubMed ] Shafritz, D.A.; Shouval, D.; Sherman, H.I.; Hadziyannis, S.J.; Kew, M.C. Integration of hepatitabletis B virus DNA into the genome of liver cells in chronic liver disease and hepatocellular carcinoma: Studies in percutaneous liver biopsies and post-mortem tissue specimens. N. Engl. J. Med. 1981 , 305 , 1067–1073. [ Google Scholar ] [ CrossRef ] [ PubMed ] Xu, H.Z.; Liu, Y.P.; Guleng, B.; Ren, J.L. Hepatitis b virus-related hepatocellular carcinoma: Pathogenic mechanisms and novel therapeutic interventions. Gastrointest. Tumors. 2014 , 1 , 135–145. [ Google Scholar ] [ CrossRef ] [ PubMed ] Sung, W.K.; Zheng, H.; Li, S.; Chen, R.; Liu, X.; Li, Y.; Lee, N.P.; Lee, W.H.; Ariyaratne, P.N.; Tennakoon, C.; et al. Genome-wide survey of recurrent HBV integration in hepatocellular carcinoma. Nat. Genet. 2012 , 44 , 765. [ Google Scholar ] [ CrossRef ] [ PubMed ] Park, Y.M.; Jang, J.W.; Yoo, S.H.; Kim, S.H.; Oh, I.M.; Park, S.J.; Jang, Y.S.; Lee, S.J. Combinations of eight key mutations in the X/preC region and genomic activity of hepatitis B virus are associated with hepatocellular carcinoma. J. Viral Hepat. 2014 , 21 , 171–177. [ Google Scholar ] [ CrossRef ] [ PubMed ] Geng, X.; Huang, C.; Qin, Y.; McCombs, J.E.; Yuan, Q.; Harry, B.L.; Palmer, A.E.; Xia, N.S.; Xue, D. Hepatitis B virus X protein targets Bcl-2 proteins to increase intracellular calcium, required for virus replication and cell death induction. Proc. Natl. Acad. Sci. USA 2012 , 109 , 18471–18476. [ Google Scholar ] [ CrossRef ] [ PubMed ] Dianzani, F. Biological basis for the clinical use of interferon. Gut 1993 , 34 , S74–S76. [ Google Scholar ] [ CrossRef ] [ PubMed ] Wang, X.Y.; Chen, H.S. Emerging antivirals for the treatment of hepatitis, B. World J. Gastroenterol. 2014 , 20 , 7707. [ Google Scholar ] [ CrossRef ] [ PubMed ] Ni, Y.; Lempp, F.A.; Mehrle, S.; Nkongolo, S.; Kaufman, C.; Fälth, M.; Stindt, J.; Königer, C.; Nassal, M.; Kubitz, R.; et al. Hepatitis B and D viruses exploit sodium taurocholate co-transporting polypeptide for species-specific entry into hepatocytes. Gastroenterology 2014 , 146 , 1070–1083. [ Google Scholar ] [ CrossRef ] [ PubMed ] Levrero, M.; Testoni, B.; Zoulim, F. HBV Cure: Why, how, when? Curr. Opin. Virol. 2016 , 18 , 135–143. [ Google Scholar ] [ CrossRef ] [ PubMed ] Table 1. Development of HCC in patients with cirrhosis on long-term antiviral therapy. Table 1. Development of HCC in patients with cirrhosis on long-term antiviral therapy. Pt Date startTx Chang in Child Class On Tx Date HCC Dx Yrs on anti-HBV Tx at HCC DX Yrs with HBV DNA (-) Age (yr) at HCC Dx Size (cm) and Site of HCC HBVDNA at HCC Dx Anti-HBV Tx Status 1 4/1998 B→A 7/2007 9.3 3.4* 53 1.1 Junction UD # LAM + TDF alive 2 6/2002 A→A 8/2007 5.2 4.7 70 1.0 Rt UD LAM + TDF alive 3 1/1998 B→A 3/2008 10.2 8.2 68 2.8 × 2.5 UD LAM + TDF dead 4 5/1998 A→A 2/2008 9.8 6.7 76 1.8 × 0.9 Lt UD LAM + TDF alive 5 7/2004 B→B 9/2009 5.2 4.7 52 3.9 Rt UD LAM + TDF alive 6 7/2001 B→B 9/2010 9.2 4.1 54 2.8 Rt UD LAM + TDF dead 7 2/2004 A→A 6/2013 9. 3 7.7 57 2.5 Lt med UD TDF dead 8 2/1996 A→A 7/2013 17.4 9.7 73 1.6 × 1.4 Rt UD TDF dead 9 8/1997 A→A 6/2014 16.8 5.9 54 2.2 × 1.9 Lt lat UD ETV alive 10 5/1996 A→A 10/2014 18.4 10.4 74 3.4 Rt UD LAM + TDF dead 11 2/2000 A→A 4/2015 15.2 12.4 62 3.4 × 3.4 Rt UD TDF alive 12 2/2000 B→A 5/2015 15.3 12.2 65 3.8 Rt UD TDF alive MDPI and ACS Style Yoo, J.; Hann, H.-W.; Coben, R.; Conn, M.; DiMarino, A.J. Update Treatment for HBV Infection and Persistent Risk for Hepatocellular Carcinoma: Prospect for an HBV Cure. Diseases 2018, 6, 27. https://doi.org/10.3390/diseases6020027 AMA Style Yoo J, Hann H-W, Coben R, Conn M, DiMarino AJ. Update Treatment for HBV Infection and Persistent Risk for Hepatocellular Carcinoma: Prospect for an HBV Cure. Diseases. 2018; 6(2):27. https://doi.org/10.3390/diseases6020027 Chicago/Turabian Style Yoo, Joseph, Hie-Won Hann, Robert Coben, Mitchell Conn, and Anthony J. DiMarino. 2018. "Update Treatment for HBV Infection and Persistent Risk for Hepatocellular Carcinoma: Prospect for an HBV Cure" Diseases6, no. 2: 27. https://doi.org/10.3390/diseases6020027

https://www.mdpi.com/2079-9721/6/2/27/htm

OUTDOOR MEIDA PROPERTIES, INC. vs. PAWLISHECK, MISC 20-000166 OUTDOOR MEDIA PROPERTIES, INC., Plaintiff, v. SUSAN PAWLISHECK, LINDA TREMBLAY, KEVIN HENDERSON, DEREK DEPETRILLO, PAUL DESCOTEAUX, DREW DANA, and JOHN SCHILLIZZI, in their capacities as members of the TOWN OF SALISBURY ZONING BOARD OF APPEALS, SCOTT VANDEWALLE, in his capacity as the TOWN OF SALISBURY BUILDING INSPECTOR AND ZONING ENFORCEMENT OFFICER, and the TOWN OF SALISBURY, Defendants MISC 20-000166 APRIL 29, 2021 ESSEX, ss. ROBERTS, J. MEMORANDUM OF DECISION DENYING PLAINTIFF'S MOTION FOR SUMMARY JUDGMENT AND ENTERING SUMMARY JUDGMENT FOR DEFENDANTS INTRODUCTION Plaintiff Outdoor Media Properties, Inc. ("OMP") commenced this action on April 9, 2020 with the filing of a complaint challenging the decision of the defendant town of Salisbury's ("the Town") zoning board of appeals ("the ZBA") to uphold the decision of defendant Scott Vandewalle, the Town's building inspector and zoning enforcement officer ("the Building Inspector"), to deny two applications for the construction of a two-sided freestanding electronic billboard ("the Project") at 109 Rabbit Road, Salisbury ("109 Rabbit Road"), one pursuant to the State Building Code ("the Building Code Application") and the other pursuant to the Town's Sign Bylaw, Chapter 214 of the Town's bylaws ("SBL" and "the SBL Application") (collectively, the Building Code Application and the SBL Application are referred to herein as "the Applications"). In its complaint, OMP sought review pursuant to G. L. c. 40A, § 17, of the denial of the Applications (Count I), a determination pursuant to G. L. c. 240, § 14A, of the validity of the SBL and the Town's zoning bylaw ("ZBL") as applied to OMP's proposed construction of the Project (Count II), and a declaration pursuant to G. L. c. 231A regarding ten issues between the parties bearing on whether the Building Inspector properly denied the Applications (Count III). At an initial case management conference on June 9, 2020, the parties reported their intent to promptly file cross-motions for summary judgment, being of the view that no discovery was required. As a result, a briefing schedule was established and a hearing scheduled for October 6, 2020. Instead of cross-motions for summary judgment, OMP filed a motion for partial summary judgment on July 21, 2020 ("the Motion") and the municipal defendants filed an opposition thereto but no cross-motion of their own. At the October 6, 2020 hearing, the court raised several issues of both fact and law that resulted in some discovery regarding the Town's past practices with respect to other applications for sign permits and some further legal research. Supplemental briefs were filed by both sides and further hearings were held on February 10 and February 17, 2021. Thereafter, further supplemental briefs were filed by both sides. For the reasons stated on the record on February 17, 2021 and further elaborated below, Plaintiff's Motion For Partial Summary Judgment is DENIED and summary judgment will enter instead for defendants pursuant to Mass. R. Civ. P. 56 (c). UNDISPUTED FACTS[Note 1] The following facts established in the record and pertinent to the Motion are undisputed or are deemed admitted. The Parties 1. OMP is a Massachusetts corporation, is the lessee of 109 Rabbit Road, and was the applicant in the Applications at issue here. Motion at II. Undisputed Facts ("UF") ¶ 1. 2. The Town is an incorporated town located in Essex County. UF ¶ 2. 3. The ZBA is the permit granting authority in the Town and has five members and two associate members, all of whom serving during times material hereto are named as defendants herein. See UF ¶ 3. 4. The Building Inspector held his office within the Town at all times material hereto. UF ¶ 4. The Property 5. 109 Rabbit Road is a 13.62 acre parcel fronting on Rabbit Road in the Town and abutting Interstate Highway 95 northbound at its rear. UF ¶ 5. 6. 109 Rabbit Road is located partially in the Commercial II (C-2) zoning district and partially in the Limited Industrial (L-I) district as described in Sections 300-6 and 300-7 of the ZBL, Appendix ("App.") Ex. 2, and the Town Zoning Map, App. Ex. 3. UF ¶ 6. Relevant ZBL Provisions 7. Prior to May 2019, the ZBL contained the following definitions relevant to the regulation of signs:[Note 2] SIGN Any permanent or temporary structure, device, letter, word, model, banner, pennant, insignia, trade flag, or representation used as, or which is in the nature of, an advertisement, announcement or direction. SIGN, OSCILLATING OR FLASHING Any permanent or temporary structure which is in the nature of an advertisement, announcement, or direction and which is designed to attract the eye by intermittent or repeated motion or illumination. [Amended 10- 23-2006 ATM by Art. 5] STRUCTURE A combination of materials assembled for occupancy or use, such as a building, bridge, trestle, tower, framework, retaining wall, tank, tunnel, tent, stadium reviewing stand, platform, shelters, piers, wharves, bin, fence, sign, or the like. 8. Signs are not identified in the ZBL's table of uses as primary or accessory uses/structures regulated thereby. UF ¶ 10; App. Ex. 2 (ZBL 300 Attachment 1). 9. The ZBL does not permit use variances. Relevant SBL Provisions 10. The SBL was enacted in May 1996 as a general bylaw passed under the Town's Home Rule powers and without compliance with the requirements imposed on the enactment of zoning bylaws under G. L. c. 40A. UF ¶ 12; App. Ex. 4. 11. Section 214-1 of the SBL provides that: The purpose of this bylaw is to regulate, restrict and place such limitations on the size, location, type, illumination and other characteristics of signs to assure that they will be appropriate to the land, building or use to which they are appurtenant; be uniform within zoning districts; be protective of property values and the safety of the public; and serve the informational purposes for which signs are intended while not detracting from the aesthetic qualities and characteristics of the Town of Salisbury. Terms used in this bylaw that are not defined herein shall have the meanings given in the Salisbury Zoning Bylaw. 12. Pursuant to Section 214-2 of the SBL, "[n]o sign shall be erected, altered or relocated without a permit issued by the Building Inspector, except as otherwise permitted herein." UF ¶ 14; App. Ex. 4. 13. The SBL contains regulations regarding the dimensions, construction, content, location and number of signs that may be permitted. UF ¶ 15. 14. Pursuant to Section 214-12(A) of the SBL, "[a]ny person aggrieved by his or her inability to obtain or retain a sign permit may file for a special permit in writing within 30 days of such decision to the Zoning Board of Appeals." UF ¶ 16; App. Ex. 4. 15. Pursuant to Section 214-12(B) of the SBL, an applicant requesting "the relaxation of these bylaws and the issuance of a special permit" must prove five things: (1) "[t]here is a hardship of the condition of the land or preexisting building location which necessitates the granting of the special permit[;] (2) [t]he granting of the special permit will not have a negative effect on the surrounding property values[;] (3) [s]ign scale is appropriate in relation to development scale, viewer distance and travel speed and sign sizes on adjacent properties[;] (4) [s]ign materials, colors, lettering style and form are compatible with building design and use[;] [and] (5) [t]he issuance of the special permit is not contrary to the purpose of this bylaw." UF ¶ 19; App. Ex. 4. OMP's Special Permit Applications Pursuant To The SBL 16. In 2016, OMP applied to the ZBA for a special permit to authorize construction of the Project as then located at 109 Rabbit Road. UF ¶ 22; App. Ex. 5. 17. At the September 13, 2016 public hearing with respect to OMP's application, the Chair of the ZBA announced the hearing as follows: "To act on a petition for relief-- a Special Permit--with respect to the Salisbury Zoning Bylaw, Section 300 [and] M.G.L. Chapter 40A. ... [OMP is] seeking a Special Permit under Section 214-12 of the Salisbury Zoning Bylaws [sic] regarding the allowance of a free-standing billboard." UF ¶ 23; App. Ex. 5. 18. Following public comments and a discussion on the application, the ZBA voted 4-1 to approve the application. UF ¶ 24; App. Ex. 5. 19. The ZBA made none of the findings set forth in Section 214-12(B) of the SBL. App. Ex. 5. 20. After the ZBA's vote, the Chair of the ZBA explained the procedural steps to be followed and concluded by stating that "once the decision is filed [in the Essex County (South) Registry of Deeds ("the Registry")], you must present proof of [such] filing with the Building Inspector's office. At that time, you will be issued your building permit pursuant to 780 CMR - the Mass. State Building Code." UF ¶ 25; App. Ex. 5. 21. The ZBA's decision memorializing its vote to approve this application was issued on September 19, 2016 ("the 2016 Special Permit"), provides for "the allowance of a free standing billboard," and notes that this relief was granted by a supermajority vote of 4-1. UF ¶ 26; App. Ex. 6. 22. The 2016 Special Permit further states that it was issued by the ZBA "[b]y virtue of its authority under M.G.L. Ch. 40A and the Salisbury Zoning Bylaw." UF ¶ 27; App. Ex. 6. 23. The 2016 Special Permit additionally provides that "[a]ppeals of this decision shall be made in accordance with the M.G.L. Ch. 40A, § 17 and shall be filed within twenty (20) days of the date of filing of this decision in the Office of the Salisbury Town Clerk." UF ¶ 28; App. Ex. 6. 24. The 2016 Special Permit additionally provides that: By law, this decision shall not take effect until a copy of such notice has been certified by the Town Clerk that; [sic] Twenty (20) days has elapsed since the decision was filed and no appeal has been filed. The Petitioner shall present and record the certified decision at the Essex Registry of Deeds in Salem MA. The cost of the recording shall be borne by the Petitioner. A registered copy must then be presented to the Building Inspector in order to apply for an appropriate permit. This Decision will lapse if Substantial use of the Petitioned Relief is not made within two (2) years for a Special Permit and one (1) year for a Variance. UF ¶ 28; App. Ex. 6. 25. The 2016 Special Permit was filed with the Salisbury Town Clerk ("Town Clerk") on September 19, 2016. UF ¶ 29; App. Ex. 6. 26. On October 11, 2016, the Town Clerk certified that no appeal of the 2016 Special Permit had been filed. UF ¶ 30; App. Ex. 6. 27. The 2016 Special Permit was recorded in the Registry on October 12, 2016 at Book 35345, Page 501. UF ¶ 31; App. Ex. 6. 28. In 2018, OMP appeared before the ZBA again to request a new special permit for the Project, which would now be relocated on the lot at 106 Rabbit Road to an area 200 feet north of the location approved in the 2016 Special Permit and which would specifically be a double-sided electronic billboard. UF ¶ 34; App. Ex. 7. 29. At the February 27, 2018 public hearing on this request, the Chair of the ZBA announced the hearing as follows: "[t]o act on a petition for relief--a Special Permit--with respect to the Salisbury Zoning Bylaws, Section 300 and M.G.L. Chapter 40A. At 109 Rabbit Road, Map 18, Lot 3. Applicant: Outdoor Media Properties is seeking a Special Permit under Section 300-214-8(E) of the Salisbury Zoning Bylaws [sic] regarding the relocation and modification of a free-standing static billboard to a double-sided digital billboard." App. Ex. 7. 30. After discussion, the ZBA voted 5-0 to approve the application. UF ¶ 35; App. Ex. 7. 31. After the vote, and as was the case with respect to the hearing on the 2016 Special Permit, the Chair of the ZBA explained the procedural steps to follow and concluded by stating that "once the decision is filed [in the Registry], you must present proof of the filing to the building inspector's office. At that time, you will be issued your building permit pursuant to 780 CMR - the Mass. State Building Code." UF ¶ 36; App. Ex. 7. 32. The ZBA's decision memorializing its vote to approve this application was issued on March 2, 2018 ("the 2018 Special Permit"), provides that the relief granted was to "[r]elocate and modify a free-standing static billboard to a double-sided digital billboard," and notes that this relief was granted by a unanimous vote of 5-0. UF ¶ 37; App. Ex. 8. 33. The 2018 Special Permit, like the 2016 Special Permit, also states that it was issued by the ZBA "[b]y virtue of its authority under M.G.L. Ch. 40A and the Salisbury Zoning Bylaw." UF ¶ 38;[Note 3]App. Ex. 8. 34. The 2018 Special Permit, like the 2016 Special Permit, additionally provides that "[a]ppeals of this decision shall be made in accordance with the M.G.L. Ch. 40A, § 17 and shall be filed within twenty (20) days of the date of filing of this decision in the Office of the Salisbury Town Clerk." UF ¶ 38; App. Ex. 8. 35. The 2018 Special Permit additionally provides that: By law, this decision shall not take effect until a copy of such notice has been certified by the Town Clerk that; [sic] Twenty (20) days has elapsed since the decision was filed and no appeal has been filed. The Petitioner shall obtain the original certified decision from the office of the Building Inspector and record that document at the Essex Registry of Deeds in Salem MA. The cost of the recording shall be borne by the Petitioner. A registered copy must then be presented to the Building Inspector in order to apply for an appropriate permit. This Decision will lapse if Substantial use of the Petitioned Relief is not made within two (2) years for a Special Permit and one (1) year for a Variance. App. Ex. 8. 36. The 2018 Special Permit was filed with the Town Clerk on March 2, 2018. UF ¶ 39; App. Ex. 8. 37. On March 23, 2018, the Town Clerk certified that no appeal of the 2018 Special Permit had been filed. UF ¶ 40; App. Ex. 8. 38. The 2018 Special Permit was recorded in the Registry on April 4, 2018 at Book 36626, Page 323. UF ¶ 41; App. Ex. 8. 39. Following the issuance of the 2018 Special Permit and through August 2019, OMP was engaged in ongoing and diligent actions in furtherance of the 2018 Special Permit, including site work, securing a development agreement for the Project, permitting before the Commonwealth's Office of Outdoor Advertising ("OOA"), and local permitting before the Town's Conservation Commission, all of which were needed to obtain building permits and/or a sign permit to proceed with the construction of the Project. UF ¶ 42. The May 2019 Annual Town Meeting 40. On May 3, 2019, the warrants for the Town's annual town meeting were posted. App. Ex. 11 (May 13, 2019 Report to the Boards by Neil J. Harrington, Town Manager). 41. Warrant Article Eight stated: To see if the Town will vote to amend the [SBL], by inserting the following new definition after the definition of "Electronic Message Boards' [sic] I the list contained in §214-3: ELECTRONIC OUTDOOR ADVERTISING A use whereby an outdoor sign or billboard, whether double-faced, back-to back, or V-shaped, with a screen(s) that is 200 square feet or larger, serves to advertise, direct or call attention to any business, article, substance, or service, or anything that is digitally or electronically projected, on or by a structure of any kind on real property or upon the ground itself, and that advertises services, products or commodities that are not available on said real property or parcel. Electronic Outdoor Advertising is a use to be regulated by the Zoning Bylaw and not the [SBL]. or to take any other action relative thereto. ON PETITION OF THE TOWN MANAGER App. Ex. 10. 42. Warrant Article Twelve stated: To see if the Town will vote to amend the Zoning Bylaws of the Town, Chapter 300, by amending the list of definitions contained in Article 1, §300-5 (Definitions) by inserting the text shown in bold and deleting the text shown in strike-through, as shown on the amended List of Definitions, a copy of which is on file with the Town Clerk; or to take any other action relative thereto. ON PETITION OF THE TOWN MANAGER App. Ex. 10. 43. In relevant part, the text referenced in Warrant Article Twelve added to the ZBL text defining "Electronic Outdoor Advertising" as follows: A use whereby an outdoor sign or billboard, whether double-faced, back-to-back, or V-shaped, with a screen(s) that is 200 square feet or larger, serves to advertise, direct or call attention to any business, article, substance, or service, or anything that is digitally or electronically projected, on or by a structure of any kind on real property or upon the ground itself, and that advertises services, products or commodities that are not available on said real property or parcel. UF ¶ 45; App. Ex. 9. 44. The Town and its respective departments, boards, divisions and personnel did not comply with the notice and hearing requirements set forth in G. L. c. 40A, § 5, with respect to Warrant Article 8. 45. Both Warrant Article 8 and Warrant Article 12 ("the 2019 Amendments") were approved by voice vote at the May 20, 2019 Annual Town Meeting. UF ¶ 50. 46. During the discuss of Warrant Article 8 at the Annual Town Meeting, the Town Manager made the following statement: I'm sure everyone is aware of the controversy that arose relative to the proposed billboard overlooking the highway that is now currently pending before the State Outdoor Advertising Board [i.e., the Project]. Due to such a large groundswell of opposition, the Board of Selectmen directed the Planning Director and myself to put together a warrant article that would ban these types of billboards in the future, and that's what the intent of this article is. It defines "Electronic Outdoor Advertising" as a use. It's regulated by the Zoning Board, and in each of the various zones of the Town, it's listed whether or not something is allowed as a use, and in this particular case it's not allowed as a use in any zone. And, since we do not have use variances, someone could not appeal a decision to deny a permit for an electronic billboard to the Zoning Board because we have no use variances. The purpose of this article is to prohibit going forward the electronic billboards in Town. App. Ex. 13. 47. Warrant Article 8 and Warrant Article 12 were thereafter approved by the office of the Massachusetts Attorney General. UF ¶ 52; App. Ex. 14. The Applications and the Actions Thereon by the Building Inspector and the ZBA 48. The Applications were filed with the Building Inspector on September 25, 2019. UF ¶ 53; App. Ex. 15. 49. Between the approval of the 2018 Special Permit in April 2018 and the filing of the Applications in September 2019, OMP engaged in (1) negotiations to acquire from a third party its OOA approval for an electronic billboard, (2) clearing the Project site, and (3) filing a Notice of Intent ("NOI") with the Town's Conservation Commission. App. Ex. 1 (Petition For Relief by Administrative Appeal ("the Petition"), Ex. 9 (November 5, 2019 Statement of Elissa Albertelli-Campbell, President, OMP ("Campbell Statement"))). 50. At the time of the May 2019 annual Town meeting, hearings were still in process before the Conservation Commission on OMP's NOI (an Order of Conditions was not issued by the Conservation Commission until August 8, 2019 and did not become final until August 22, 2019), and the transfer of the OOA permit to OMP from the third party had not yet been approved by OOA (the application to the OOA for the transfer of the permit was granted on June 11, 2019). App. Ex. 1 (Ex. 9, Campbell Statement). 51. On October 8, 2019, the Building Inspector issued two letters denying the Applications. UF ¶ 55. 52. The letter addressed to the SBL Application states: The Permit Application to Erect a Sign dated 9/25/19 is hereby DENIED in accordance with §214-3 of the Salisbury General Bylaws. The proposed Digital Sign meets the Salisbury Sign Bylaw definition of ELECTORNIC OUTDOOR ADVERTISING 'A use whereby an outdoor sign or billboard, whether double-faced, back-to-back, or Vshaped, with a screen(s) that is 200 square feet or larger, serves to advertise, direct or call attention to any business, article, substance, or service, or anything that is digitally or electronically projected, on or by a structure of any kind on real property or upon the ground itself, and that advertises services, products or commodities that are not available on said real property or parcel. Electronic Outdoor Advertising is a use to be regulated by the Zoning Bylaw and not the General Bylaw'. Salisbury Zoning Bylaw §300-27 states that 'It shall be unlawful for any owner or person to erect, construct, reconstruct, or alter a structure or change the use or lot coverage, increase the intensity of use, or extend or displace the use of any building or other structure on a lot without applying for and receiving from the Building Inspector the required building permit therefor'. App. Ex. 16. 53. The letter addressed to the Building Code Application states: The Application for a Building Permit dated 9/25/2019 for Construct Digital Sign per Plans is hereby DENIED in accordance with §300-27 of the Salisbury Zoning Bylaw; 'It shall be unlawful for any owner or person to erect, construct, reconstruct, or alter a structure or change the use of or [sic] lot coverage, increase the intensity of use, or extend or displace the use of any building or other structure on a lot without applying for and receiving from the Building Inspector the required building permit therefor'. §300-5 defines a Structure as 'A combination of materials assembled for occupancy or use, such as a building, bridge, trestle, tower, framework, retaining wall, tank, tunnel, tent, stadium reviewing stand, platform, shelters, piers, wharves, bin, fence, sign, or the like'. The proposed Digital Sign meets the Salisbury Sign Bylaw definition of Electronic Outdoor Advertising 'A use whereby an outdoor sign, or billboard, whether double-faced, back-to-back, or Vshaped, with a screen(s) that is 200 square feet or larger, serves to advertise, direct or call attention to any business, article, substance, or service, or anything that is digitally or electronically projected, on or by a structure of any kind on real property or upon the ground itself, and that advertises services, products or commodities that are not available on said real property or parcel. Electronic Outdoor Advertising is a use to be regulated by the Zoning Bylaw and not the General Bylaw'. Salisbury Zoning Bylaw §300-27 states that 'It shall be unlawful for any owner or person to erect, construct, reconstruct, or alter a structure or change the use or lot coverage, increase the intensity of use, or extend or displace the use of any building or other structure on a lot without applying for and receiving from the Building Inspector the required building permit therefor'. App. Ex. 17. 54. Both letters provide for an appeal within thirty days, in the case of the denial of the SBL Application, in accordance with Chapter 214, § 214-12 of the SBL by filing with the Town Clerk and, in the case of the denial of the Building Code Application, in accordance with Chapter 300 of the ZBL and G. L. c. 40A, § 15 by filing with the Town Clerk. App. Exs. 16, 17. 55. On November 5, 2019, within the thirty-day appeal period, OMP filed its Petition pursuant to G. L. c. 40A, §§ 8 and 15. UF ¶ 58. 56. By decision dated March 4, 2020, the ZBA denied OMP's Petition by a vote of 3-1 without providing any statement of reasons for the denial. UF ¶ 59; App. Ex. 18. Other Freestanding Electronic Billboard Applications 57. In addition to this Project, three other sites in the Town have been the subject of applications for freestanding electronic billboards: 100 Elm Street; 75 Main Street and 74 Main Street. See Plaintiff's Supplemental Memorandum In Further Support Of Its Motion For Partial Summary Judgment ("Supp. Mem."), Exs. 2, 3. 58. A special permit was granted for the 100 Elm Street site in 2013, on which was proposed a thirty-foot tall freestanding electronic billboard. Supp. Mem. Ex. 2. 60. A special permit was granted for the 75 Main Street site in 2014, on which was proposed an eighty-nine foot freestanding electronic billboard. Supp. Mem. Ex. 3. 61. The 75 Main Street site is located in the Town's commercial district. Supp. Mem. Ex. 3. 62. At the same time, a special permit for the 74 Main Street site on which was proposed a fifty-five foot freestanding electronic billboard was denied. Supp. Mem. Ex. 3. 63. The 74 Main Street site was located in the Town's commercial district. Supp. Mem. Ex. 3. 64. The actions regarding 75 Main Street and 74 Main Street were the subject of litigation, resulting in the Appeals Court's decision in Clear Channel Outdoor, Inc. v. Zoning Bd. of Appeals of Salisbury,94 Mass. App. Ct. 594(2018) annulling the ZBA's actions with respect to both applications and returning both matters to the ZBA for further consideration. 65. According to OMP's president, the holder of the OOA permit for 75 Main Street agreed to transfer its permit to OMP on January 15, 2019, within a month of the Appeals Court's Clear Channel decision, App. Ex. 1 (Ex. 9, Campbell Statement at ¶ 9), and the transfer was approved by OOA on June 11, 2019. App. Ex. 1 (Ex. 9, Campbell Statement ¶ 12). 66. The 74 Main Street application, having been returned to the ZBA for further consideration, was then denied by a unanimous vote of 5-0 "because of the effect on the surrounding community and negative property values." Supp. Mem. Ex. 3. 67. Counsel for the parties agreed at the February 10, 2021 hearing on this matter that no building permit had been sought for either the 75 Main Street or the 74 Main Street project. STANDARD OF REVIEW Generally, summary judgment may be entered if the "pleadings, depositions, answers to interrogatories, and responses to requests for admission ... together with the affidavits ... show that there is no genuine issue as to any material fact and that the moving party is entitled to a judgment as a matter of law." Mass. R. Civ. P. 56 (c). In viewing the factual record presented as part of the motion, the court draws "all logically permissible inferences" from the facts in favor of the non-moving party. Willitts v. Roman Catholic Archbishop of Boston,411 Mass. 202, 203 (1991). "Summary judgment is appropriate when, 'viewing the evidence in the light most favorable to the nonmoving party, all material facts have been established and the moving party is entitled to a judgment as a matter of law.'" Regis College v. Town of Weston,462 Mass. 280, 284 (2012), quoting Augat, Inc. v. Liberty Mut. Ins. Co.,410 Mass. 117, 120 (1991). In addition, "[p]ursuant to rule 56 (c), 'summary judgment, when appropriate, may be rendered against the moving party.' See, e.g., Perseus of N.E., MA, Inc. v. Commonwealth,429 Mass. 163, 168, 706 N.E.2d 681 (1999)." Locator Servs. Group, Ltd. v. Treasurer & Receiver Gen.,443 Mass. 837, 851 (2005). DISCUSSION In its Motion, OMP seeks partial judgment in its favor on Count II, brought pursuant to G.L. c. 240, § 14A, and Count III, brought pursuant to G. L. c. 231A, § 8, declaring the following: (1) that the SBL, being for all intents a zoning regulation that was not passed in conformance with G. L. c. 40A, is invalid; (2) that the definition of "Electronic Outdoor Advertising" inserted in the SBL at the May 2019 annual town meeting is also invalid for the same reason; (3) that the definition of "Electronic Outdoor Advertising" inserted in the SBL at the May 2019 annual town meeting is further invalid because, by its express terms, it provides that "Electronic Outdoor Advertising is a use to be regulated . . . not [by] the [SBL];" and (4) that, even if the SBL is not invalidated, the 2018 Special Permit was in full force and effect and authorized the issuance of all building permits or sign permits for the Project. Motion at 3. As the municipal defendants point out, G. L. c. 240, § 14A provides that "[t]he owner of a freehold estate in possession in land" may bring a petition in the land court, and OMP is not the owner of 106 Rabbit Road, but a lessee. App. Ex. 1 (Ex. 3, Notice Of Lease).[Note 4]Accordingly, relief under that statute is not available to OMP here and its requests will be considered solely under G. L. c. 231A. Sorting through what can best be described as a mash-up of zoning and general bylaw provisions in the Town makes settling on an analytical framework for this case more than a little difficult. By way of example, the ZBL has, since its inception, included signs as structures and currently contains provisions regulating some signs.[Note 5]The SBL, adopted in 1996, incorporates definitions from the ZBL insofar as they are not defined in the SBL, makes the Building Inspector the administrator of the SBL, and provides for appeals to the ZBA by any person aggrieved by his inability to obtain a sign permit. App. Ex. 4. The hearings for both the 2016 Special Permit and the 2018 Special Permit were introduced by the Chair of the ZBA as petitions "with respect to the Salisbury Zoning Bylaw, Section 300" and G. L. c. 40A. App. Exs. 5, 7. The decisions issued on both the 2016 Special Permit and the 2018 Special Permit stated that they were issued by the ZBA "[b]y virtue of its authority under M.G.L. Ch. 40A and the Salisbury Zoning Bylaw," and stated that "[a]ppeals of this decision shall be made in accordance with the M.G.L. Ch. 40A, § 17." App. Exs. 6, 8. Matters were not helped by the 2019 Amendments, where, under the amended SBL, electronic outdoor advertising signs were defined as "a use to be regulated by the [ZBL] and not the [SBL]," and the ZBL was amended to define electronic outdoor advertising signs as a "use," even though neither those nor any other signs appear in the ZBL's Table Of Use Regulations. App. Ex. 2. Turning first to OMP's argument that, regardless of the applicability of the SBL and its amendment to the Project, the 2018 Special Permit was in full force and effect and authorized the issuance of all building permits or sign permits for the Project, this court disagrees. The decision in Albahari v. Zoning Bd. of Appeals,76 Mass. App. Ct. 245(2010) ("Albahari II"), is dispositive on the effect of the bylaw amendments on the Applications, standing alone. There, the defendant Lucille Owocki ("Ms. Owocki"), the owner of two contiguous parcels of land in Brewster, applied for a permit to build a single-family home thereon. The town building commissioner denied the application but, following litigation, ultimately issued the permit on September 22, 2005. Id. By then, however, the town zoning bylaw had been amended in a way that made it impossible for Ms. Owocki to build in accordance with her plans (the issue was whether Ms. Owocki's property was in a wetlands conservancy district based on soil type, and the bylaw was amended to add a soil type as a conservancy soil, similar if not identical to that already listed in the bylaw, that existed on Ms. Owocki's property). In the prior litigation, Albahari v. Board of Appeals of Brewster, 2004 Mass. App. Lexis 516, 61 Mass. App. Ct. 1104 (2004) (Rule 23.0 Decision), the Appeals Court affirmed the decision of the trial court that the bylaw in effect at the time of the zoning board of appeal's decision was controlling, but declined to decide which version of the bylaw would apply to the actual issuance of the permit. 76 Mass. App. Ct. at 247. Back before the municipal officials, the building commissioner granted Ms. Owocki a building permit and, on an appeal by the neighbors, the local board upheld him, finding that deficiencies in Ms. Owocki's application were either rectified before the first publication of the notice of the proposed bylaw change, or were routine, ministerial details that were easily corrected, such that the prior bylaw applied to her application. Id. at 247-248. On appeal, a judge of this court reversed, finding that the remaining deficiencies in Ms. Owocki's application were more than routine, such that the amended bylaw applied. On further appeal to the Appeals Court, that court stated that: [t]he permit in this case did not actually issue before the first publication of notice of a hearing on the proposed by-law change. Were that all there were to it, Owocki would not be entitled to the permit she now seeks, for she acquired no rights simply by filing the permit application. Id. at 249 (citations omitted). However, quoting its earlier opinion, the Albahari II court described the issue now before it as "whether the permit in question should be deemed to have been 'issued' for the purpose of G. L. c. 40A, § 6 ... or whether the circumstances are such as to have created in defendant Owocki a 'vested' right to the permit." Id. The court then turned to Framingham Clinic, Inc. v. Zoning Bd. of Appeals of Framingham,382 Mass. 283, 297 (1981) for the proposition that "[t]he burden is on the applicant to demonstrate in the Superior Court an entitlement to a building permit. ... Upon such a showing, the issuance of a permit is a matter of duty, not discretion, and relief in the form of an order that a permit issue is appropriate." According to the Albahari II court, [a]lthough the Framingham Clinic case did not involve a by-law change, the case is relevant because it stands for the proposition that the right to a permit vests at the time the applicant is unconditionally entitled to its issuance. Until that time, ...the applicant is subject to any changes in zoning provisions that are properly noticed before the permit issues. 76 Mass. App. Ct. at 250-251. In applying its analysis to the case before it, the Albahari II court stated: The question, then, is not whether Owocki's permit issued before publication of the first notice of a hearing on the proposed by-law, but whether Owocki was unconditionally entitled to a permit at any time before the first notice, for if she was, then issuance became a duty and created rights she could enforce. OMP in this case has even less of a claim to a building permit than Ms. Owocki in Albahari II. Unlike Ms. Owocki, OMP did not even file its application for a building permit until September 25, 2019, months after the 2019 Amendments were adopted, apparently because OMP required the OOA permit from the Commonwealth and the Order of Conditions from the Town's Conservation Commission before doing so. OMP not having a vested right to a building permit when the 2019 Amendments were enacted, the Applications are subject to those amendments, unless the prior grant of the 2018 Special Permit changes that result. This court is of the view that it does not. And, while OMP argues that the SBL and its amendment are invalid, this court is of the further view that the same result obtains whether the 2018 Special Permit is treated as issued under the SBL or the ZBL (and, if not void, it has to have been issued under one or the other), so it need not reach the issue of the SBL's validity. As the municipal defendants argue, relying on Spenlinhauer v. Town of Barnstable,80 Mass. App. Ct. 134, 137-138 (2011), "changes in zoning ordinances protect some prior existing uses, see G. L. c. 40A, § 6, but general ordinances typically do not." So, assuming the SBL is valid and the 2018 Special Permit validly issued thereunder, it would not be protected from the subsequently enacted 2019 Amendments. Alternatively, if the SBL is invalid,[Note 6]then it follows that the 2018 Special Permit issued pursuant to it is a nullity, doubtless not the result sought by OMP here, or that the 2018 Special Permit, while valid, is only protected from zoning amendments for twelve months pursuant to G. L. c. 40A, § 6. Regarding the latter option, § 6 provides in pertinent part: A zoning ordinance or by-law shall provide that construction or operations under a building or special permit shall conform to any subsequent amendment of the ordinance or by-law unless the use or construction is commenced within a period of not more than 12 months after the issuance of the permit and in cases involving construction, unless such construction is continued through to completion as continuously and expeditiously as is reasonable. This zoning freeze, which "is an issue separate and distinct from the construction necessary to prevent a permit from 'lapsing'" under G. L. c. 40A, § 9, Martin R. Healy, et al., Massachusetts Zoning Manual, § 7.2.2(b) (MCLE, Inc. 6th ed. 2017, Supp. 2019), requires that "[w]ork or operations authorized under a building permit or special permit must be commenced within a period of not more than twelve months in order to be within the protection of the statute." Id. Furthermore, preliminary site work, such as demolition and debris removal preparatory to construction, are not sufficient to constitute "construction" under the statute. See Alexander v. Building Inspector of Provincetown,350 Mass. 370, 374-375 (1966) ("preparation for new construction by removal of standing buildings and leveling of consequent debris" would not support a finding that construction had lawfully begun) (as applied to predecessor to G. L. c. 40A, § 6); Murphy v. Board of Selectmen,1 Mass. App. Ct. 407, 409 (1973) ("site preparation activity would not qualify as construction as that word is used in § 11") (as applied to predecessor to G. L. c. 40A, § 6). Here, the ZBA's decision granting the 2018 Special Permit was dated March 1, 2018, filed with the Town Clerk on March 2, 2018, and recorded at the Registry on April 4, 2018. App. Ex. 8. The Applications were filed with the Building Inspector on September 25, 2019, some seventeen months later. At that point, work at 106 Rabbit Road consisted of "excavation, removal of overburden, material hauling, and leveling the pad area for the proposed Sign," which was accomplished in January 2019. App. Ex. 1 (Ex. 9, Campbell Statement ¶ 12). While OMP appears to have been diligently pursuing the other permits required for the Project, it did not commence use or construction within twelve months of the issuance of the 2018 Special Permit. As a result, the 2018 Special Permit is subject to the 2019 ZBL amendment. While OMP sought only a partial summary judgment on the issues addressed above, this decision has broader ramifications for OMP's building permit application. Under other circumstances, the court would be inclined to remand this matter in order to allow the ZBA to reconsider the building inspector's denial of the Applications and, this time, to provide a rationale for its decision, whatever it might be. It is clear, though, that if, as the court has determined, the 2018 Special Permit is subject to the 2019 ZBL amendment, then the ZBA has no option but to uphold the building inspector's decision to deny the Building Code Application based on the fact that the ZBL does not permit electronic outdoor advertising as a use and does not allow for use variances.[Note 7]See G. L. c. 40A, § 10 ("Except where local ordinances or by-laws shall expressly permit variances for use, no variance may authorize a use or activity not otherwise permitted in the district in which the land or structure is located"). CONCLUSION For the reasons set forth above, Plaintiff's Motion For Partial Summary Judgment is DENIED. Instead, judgment will enter pursuant to Mass. R. Civ. P. 56 (c) on Count I upholding the decision of the ZBA to affirm the Building Inspector's denial of the Building Code Application, and on Count III declaring (1) that OMP was not entitled to the issuance of a building permit, its right to such a permit not having vested at the time of the bylaw amendments at issue here and (2) that, whether considered under the SBL or the ZBL, the Applications were subject to the 2019 Amendments. SO ORDERED FOOTNOTES [Note 1]These are set forth in numbered paragraphs in OMP's Motion, Sec. II Undisputed Facts, and admitted by the municipal defendants in Defendants' Responses To Plaintiff's Statement Of Undisputed Material Facts ("Response") docketed on August 28, 2020. [Note 2]The municipal defendants disputed this statement in their Response because the copy of the ZBL contained in the Appendix as Ex. 2 is not certified as the version of the ZBL in effect prior to May 2019. However, at the hearing on October 6, 2020, counsel for the defendants agreed that Ex. 2 could be relied on as accurately reflecting the ZBL as amended on various dates stated therein, including prior to May 2019. [Note 3]In the Response, due to a clerical error, paragraph 38 of the UF is renumbered as paragraph 39, and the numbering of all subsequent paragraphs in the Response is adjusted accordingly. [Note 4]The court notes that the Notice Of Lease appears to contain an incorrect deed reference. The deed in to 109 Rabbit Road's current owner, KLS Management Group, LLC ("KLS"), is dated May 25, 2016 and recorded in the Registry at Book 34950, Page 304. The Notice Of Lease references a deed in to KLS dated January 20, 2011 recorded in the Registry at Book 30495, Page 332. [Note 5]See App. Ex. 2, § 300-96(H) (imposing regulations in addition to those set forth in the SBL regarding signs for "adult use"); § 300-125(E) (imposing regulations in addition to those set forth in the SBL regarding signs for wireless communication facilities); § 300-141.7(C) (imposing regulations in addition to those set forth in the SBL regarding signs for wind energy facilities). [Note 6]There is much to commend this argument, considering the purpose and effect of the SBL, the procedural mechanisms adopted by it, and the practice of local officials, at least in this case, to treat an appeal from the building inspector's denial of a sign permit as an appeal pursuant to the ZBL and G. L. c. 40A. See Rayco Inv. Corp. v. Selectmen of Raynham,368 Mass. 385, 386-387 (1975) (where town had previously regulated trailer parks through its zoning bylaw, general ordinance regulating the number of trailer park licenses in the town not adopted in compliance with G. L. c. 40A was invalid); Spenlinhauer, supra (invalidating "Comprehensive Occupancy" general ordinance not adopted in compliance with G. L. c. 40A insofar as it purported to regulate off-street parking where the town's zoning bylaw contained a section devoted to "Off street Parking Regulations" and where the town had historically regulated street parking through its zoning bylaw, not its general ordinances). Given the decision here, however, the court need not reach the issue. [Note 7]As argued by the municipal defendants, the Project also violates the height limit in its zoning district and, on that additional ground, the ZBL Application would be denied in the absence of a variance.

http://masscases.com/cases/land/2021/2021-20-000166-MEMORANDUM.html

Importation of hybrid human-associated Trypanosoma cruzi strains of Southern South American origin, Colombia. - Free Online Library Free Online Library: Importation of hybrid human-associated Trypanosoma cruzi strains of Southern South American origin, Colombia.(DISPATCHES, Report) by " Emerging Infectious Diseases" ; Health, general Biodiversity Biological diversity Chagas disease Causes of Distribution Research Chagas' disease Infection Physiological aspects Importation of hybrid human-associated Trypanosoma cruzi strains of Southern South American origin, Colombia. Chicago style: The Free Library . S.v. Importation of hybrid human-associated Trypanosoma cruzi strains of Southern South American origin, Colombia.." Retrieved May 31 2023 from https://www.thefreelibrary.com/Importation+of+hybrid+human-associated+Trypanosoma+cruzi+strains+of...-a0461529940 APA style: Importation of hybrid human-associated Trypanosoma cruzi strains of Southern South American origin, Colombia.. (n.d.) >The Free Library. (2014). Retrieved May 31 2023 from https://www.thefreelibrary.com/Importation+of+hybrid+human-associated+Trypanosoma+cruzi+strains+of...-a0461529940 Chagas disease is the most common parasitic infection in Latin America, annually affecting [approximately equal to] 5-6 million persons and putting another 70 million at risk (1). The etiologic agent, Trypanosoma cruzi, displays remarkable genetic diversity, which is widely thought to contribute to the considerable biologic, epidemiologic, and clinical variation observed in regions where the disease is endemic (2). Seven discrete typing units (DTUs) are currently recognized (TcI-TcVI and TcBat) (2); TcV and TcVI are natural interlineage hybrids of TcII and TcIII (3). It is unknown whether these hybrids arose from multiple independent recombination events (3) or a single incidence of hybridization followed [down arrow]by clonal divergence (4). Molecular dating indicates these lineages evolved recently (<1 million years ago) (3,4), suggesting that genetic exchange may still be driving the emergence of novel recombinants (3,4). For analysis, we assembled a panel of 57 T. cruzi biologic clones from a range of representative hosts/vectors across South America: 24 uncharacterized clones from Colombia and 33 reference clones (Figure 1; online Technical Appendix 1 Table 1, http://wwwnc.cdc.gov/EID/article/22/8/15-0786-Techapp1.pdf). From 2002-2010, we isolated the uncharacterized clones from humans; triatomine vectors (Panstrongylus geniculatus, Rhodnius prolixus, and Triatoma venosa insects); and sylvatic mammalian hosts (Dasypus spp. armadillos) in 3 T. cruzi-endemic departments in northern Colombia. We genotyped all isolates using nuclear housekeeping genes GPX, GTP, Met-II, TcAPX, and TcMPX (6,8) (online Technical Appendix 1 Table 2); 25 microsatellite loci (online Technical Appendix 1 Table 3) (9); and 10 mitochondrial gene fragments (10). Diploid multilocus sequence typing (MLST) data were analyzed by locus and concatenated according to their relative chromosomal positions in MLSTest (11); heterozygous variable sites were handled as average states. Gene haplotypes were inferred using PHASE version 2.1 (12). PCR products were cloned and sequenced to confirm ambiguous gene phases. We constructed maximum-likelihood and Bayesian phylogenies for nuclear haplotypic and concatenated mitochondrial data (13). For microsatellite loci, we defined sample clustering using a neighbor-joining tree based on pairwise distances between multilocus genotypes (Figure 2) (13). We calculated DTU-level heterozygosity (Bonferroni-corrected) and evaluated genetic diversity using sample size-corrected allelic richness and private allele frequency per locus (Table). To examine TcV/TcVI allele inheritance, we classified genotypes at each locus as hybrid (TcII/ TcIII) or nonhybrid (TcII/TcII or TcIII/TcIII) based on the presence or absence of specific parental alleles (online Technical Appendix 2, http://wwwnc.cdc.gov/EID/ article/22/8/15-0786-Techapp2.xlsx). All putative hybrids from Colombia were highly heterozygous and minimally diverse. They possessed TcII and TcIII alleles at an approximate 1:1 ratio and, compared with parental DTUs, they displayed fewer private alleles or single-nucleotide polymorphisms; these strains fulfilled all the expectations of progeny from a recent Mendelian hybridization event(s) (Table). Based on nuclear MLST and microsatellite data, all hybrids from Colombia were classified as TcVI, not novel recombinants. Examination of TcII and TcIII alleles across 5 nuclear loci showed that hybrid haplotypes from Colombia were shared among other TcVI strains from the Southern Cone region of South America and showed negligible affinities to parental alleles from Colombia (online Technical Appendix 1 Figures 1, 2). Microsatellite profiles also supported this allopatric inheritance: only a minority of private parental alleles from Colombia were common to local TcVI hybrids. At mitochondrial loci, TcVI clones from Colombia were noticeably divergent from local TcIII maxicircle haplotypes and those observed in reference TcVI strains (Figure 2). Of note, 1 hybrid from Colombia (AACf2 cl11), which was unequivocally classified as TcVI by both types of nuclear loci, possessed a TcV-type mitochondria. All isolates in this study were biologic clones, ruling out mixed infections as a potential confounder. Overall, our data support the hypothesis that 2 separate recombination events led to the formation of TcV and TcVI. These interlineage hybrids have distinct nuclear and mitochondrial MLST genotypes and related but independent microsatellite profiles, and most alleles that distinguish between hybrid DTUs (70.4% [38/54 alleles]) were also present in their corresponding parental strains. Inter-lineage differences (fixed at 84% [21/25 of loci]) between TcV and TcVI are not consistent with allelic sequence divergence (Meselson effect); for such divergence, a much higher frequency of private alleles, compared with parental genotypes, would be expected at rapidly evolving micro-satellite loci. TcVI clones from Colombia had more private micro-satellite alleles per locus (0.86) than their southern counterparts (0.43), despite their unequivocal origin in the Southern Cone. This phenomenon could be attributable to de novo mutations or a founder effect with respect to the northerly introduction of TcVI. Support for the latter cause is evidenced by an overall reduction in genetic diversity among hybrids from Colombia compared with TcVI strains from the Southern Cone (allelic richness 1.87 vs. 2.46, respectively). However, we cannot discount some sampling bias because reference Southern Cone strains represented a much wider geographic range. A novel observation among TcVI strains from Colombia was the presence of an anomalous TcV maxicircle. This pattern of inheritance could reflect 1) recent mitochondrial introgression from TcV into TcVI, leaving undetectable signatures of nuclear hybridization by our markers or, possibly, none at all (10,14), or 2) potential backcrossing of TcVI into TcIII. Genetic exchange has not been described among hybrid DTUs, but it might be expected to be more permissive between closely related strains (14). We also isolated hybrid AACf2 cl11 from a dog. T. cruzi hybridization has been proposed to arise within mammalian cells (14), and mixed infections in such hosts are common. Alternatively, TcV and TcVI may have evolved from the beneficiaries of different alleles during a single hybridization event between heterozygous parents with mixed TcIII-type mitochondrial complements; although, to date, reported levels of mitochondrial heteroplasmy in T. cruzi are low (10). Conclusions Our understanding of the geographic and ecologic distribution of T. cruzi DTUs is changing because of parallel improvements in sampling strategies and genotyping techniques. Human Chagas disease in Colombia is currently associated with DTUs TcI, TcII (to a lesser extent), and oral outbreaks of TcIV (5). In this study, we isolated T. cruzi hybrids from 3 domestic triatomine vectors, a reridomestic dog, and congenital infections among local patients. Given that no reservoir hosts of TcV and TcVI have been described (15), the hybrids from Colombia are more likely the result of long-range anthropogenic introduction than local sylvatic invasion, especially considering the successful establishment of these DTUs among domestic infections in the Southern Cone. Further intensive sampling efforts in northern South America are warranted to elucidate the transmission cycle ecology of TcVI and to accurately assess the epidemiologic risk of human Chagas disease associated with this low-diversity hybrid lineage. This research was supported by the Wellcome Trust and a European Commission Framework Programme project (Comparative epidemiology of genetic lineages of Trypanosoma cruzi, ChagasEpiNet; contract no. 223034). L.A.M. was funded by a BBSRC (Biotechnology and Biological Sciences Research Council) doctoral training grant. Dr. Messenger is postdoctoral researcher at the London School of Hygiene and Tropical Medicine. Her research interests include population genetics, molecular epidemiology, clinical parasitology, and disease control. References (1.) World Health Organization. Chagas disease in Latin America: an epidemiological update based on 2010 estimates [cited 2015 Jun 2]. http://www.who.int/wer/2015/wer9006/en/ (2.) Messenger LA, Miles MA, Bern C. Between a bug and a hard place: Trypanosoma cruzi genetic diversity and the clinical outcomes of Chagas disease. Expert Rev Anti Infect Ther. 2015; 13:995-1029. http://dx.doi.org/10.1586/14787210.2015.1056158 (3.) Lewis MD, Llewellyn MS, Yeo M, Acosta N, Gaunt MW, Miles MA. Recent, independent and anthropogenic origins of Trypanosoma cruzi hybrids. PLoS Negl Trop Dis. 2011; 5:e1363. http://dx.doi.org/10.1371/journal.pntd.0001363 (4.) Flores-Lopez CA, Machado CA. Analyses of 32 loci clarify phylogenetic relationships among Trypanosoma cruzi lineages and support a single hybridization prior to human contact. PLoS Negl Trop Dis. 2011; 5:e1272. http://dx.doi.org/10.1371/journal.pntd.0001272 (5.) Guhl F, Ramirez JD. Retrospective molecular integrated epidemiology of Chagas disease in Colombia. Infect Genet Evol. 2013; 20:148-54. http://dx.doi.org/10.1016/j.meegid.2013.08.028 (6.) Yeo M, Mauricio IL, Messenger LA, Lewis MD, Llewellyn MS, Acosta N, et al. Multilocus sequence typing (MLST) for lineage assignment and high resolution diversity studies in Trypanosoma cruzi. PLoS Negl Trop Dis. 2011; 5:e1049. http://dx.doi.org/10.1371/journal.pntd.0001049 (7.) Venegas J, Rojas T, Diaz F, Miranda S, Jercic MI, Gonzalez C, et al. Geographical structuring of Trypanosoma cruzi populations from Chilean Triatoma infestans triatomines and their genetic relationship with other Latino American counterparts. Ann Trop Med Parasitol. 2011; 105:625-46. http://dx.doi.org/10.1179/204777 3211Y.0000000002 (8.) Lauthier JJ, Tomasini N, Barnabe C, Rumi MM, D'Amato AM, Ragone PG, et al. Candidate targets for multilocus sequence typing of Trypanosoma cruzi: validation using parasite stocks from the Chaco region and a set of reference strains. Infect Genet Evol. 2012; 12:350-8. http://dx.doi.org/10.1016/j.meegid.2011.12.008 (9.) Llewellyn MS, Miles MA, Carrasco HJ, Lewis MD, Yeo M, Vargas J, et al. Genome-scale multilocus microsatellite typing of Trypanosoma cruzi discrete typing unit I reveals phylogeograhic structure and specific genotypes linked to human infection. PLoS Pathog. 2009; 5:e1000410. http://dx.doi.org/10.1371/journal. ppat.1000410 (11.) Tomasini N, Lauthier JJ, Llewellyn MS, Diosque P. MLSTest: novel software for multi-locus sequence data analysis in eukaryotic organisms. Infect Genet Evol. 2013; 20:188-96. http://dx.doi.org/10.1016/j.meegid.2013.08.029 (12.) Stephens M, Smith N, Donnelly P. A new statistical method for haplotype reconstruction from population data. Am J Hum Genet. 2001; 68:978-89. http://dx.doi.org/10.1086/319501 (13.) Messenger LA, Garcia L, Vanhove M, Huaranca C, Bustamante M, Torrico M, et al. Ecological host fitting of Tcl in Bolivia: mosaic population structure, hybridization and a role for humans in Andean parasite dispersal. Mol Ecol. 2015; 24:2406-22. http://dx.doi.org/10.1111/mec.13186 (14.) Messenger LA, Miles MA. Evidence and importance of genetic exchange among field populations of Trypanosoma cruzi. Acta Trop. 2015; 151:150-5. http://dx.doi.org/10.1016/j.actatropica.2015.05.007 (15.) dos Santos Lima V, Xavier SC, Maldonado IF, Roque AL, Vicente AC, Jansen AM. Expanding the knowledge of the geographic distribution of Trypanosoma cruzi TcII and TcV/TcVI genotypes in the Brazilian Amazon. PLoS ONE. 2014; 9:e116137. http://dx.doi.org/10.1371/journal.pone.0116137 Author affiliations: London School of Hygiene and Tropical Medicine, London, UK (L.A. Messenger, M.S. Llewellyn, M.A. Miles); Universidad del Rosario, Bogota, Colombia (J.D. Ramirez); Universidad de Los Andes, Bogota (F. Guhl) DOI: http://dx.doi.org/10.3201/eid2208.150786 Address for correspondence: Louisa A. Messenger, Department of Pathogen Molecular Biology, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London WC1E 7HT, UK; email: [email protected] Table. Population genetic parameters for Trypanosoma cruzi discrete typing units, South America, 2002-2010 * No. multilocus Proportion No. Discrete genotypes/no. shared polymorphic typing isolates alleles [+ or -] SD loci unit TcII 14/15 (5/6) 0.44 [+ or -] 0.23 24 (15) (0.062 [+ or -] 0.053) TcIII 13/13 (4/4) 0.48 [+ or -] 0.15 22 (21) (0.30 [+ or -] 0.16) TcV 8/8 0.15 [+ or -] 0.092 22 TcVI 21/21 (14/14) 0.24 [+ or -] 0.87 21 (20) (0.22 [+ or -] 0.103) Mean no. Discrete private alleles typing per locus [+ or -] SE Mean [A.sub.r] [+ or -] SE unit ([dagger]) TcII 1.76 [+ or -] 0.20 3.94 [+ or -] 0.29 (0.68 [+ or -] 0.14) (1.65 [+ or -] 0.12) TcIII 2.35 [+ or -] 0.48 4.26 [+ or -] 0.43 (1.76 [+ or -] 0.27) (2.35 [+ or -] 0.18) TcV 0.16 [+ or -] 0.07 2.38 [+ or -] 0.20 TcVI 0.43 [+ or -] 0.12 2.46 [+ or -] 0.21 (0.86 [+ or -] 0.20) (1.87 [+ or -] 0.11) Mean % Loci with Discrete expected/observed deficit/excess typing heterozygosity heterozygosity unit ([dagger]) ([double dagger]) TcII 0.58/0.65 29.2/20.8 (0.91/0.58) (40.0/0) TcIII 0.45/0.70 4.5/27.3 (0.46/0.69) (9.5/38.1) TcV 0.85/0.58 54.6/4.5 TcVI 0.60/0.49 41.7/16.7 (0.71/0.54) (40.0/15.0) * Values represent findings for reference clones derived from a range of representative hosts and vectors across South America and, in parentheses, clones isolated from humans, triatomine vectors, and sylvatic mammalian hosts in northern Colombia. Values were calculated from microsatellite data for 25 analyzed loci. Ar, allelic richness. ([dagger]) Across all loci. ([double dagger]) After sequential Bonferroni correction. Article Details Title Annotation: DISPATCHES Author: Messenger, Louisa A.; Ramirez, Juan David; Llewellyn, Martin S.; Guhl, Felipe; Miles, Michael A. Publication: Report Geographic Code: 4EUUK Date: Aug 1, 2016 Words: 2362

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Biomarkers of Antidepressant Response and Suicidal Events in Depressed Youth - Full Text View - ClinicalTrials.gov Biomarkers of Antidepressant Response and Suicidal Events in Depressed Youth The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been evaluated by the U.S. Federal Government. Read our disclaimer for details. ClinicalTrials.gov Identifier: NCT03547219 Recruitment Status : Completed First Posted : June 6, 2018 Last Update Posted : April 4, 2019 Sponsor: National Research Foundation of Korea Information provided by (Responsible Party): Jae-Won Kim, Seoul National University Study Details Study Description Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Brief Summary: The aims of this study are to (1) identify inflammatory/neurotrophic and neurodevelopmental markers that predict antidepressant response and suicidal risk in depressed youth and (2) predict their treatment response and the occurrence of suicidal events at the individual level using machine learning approach. Condition or disease Intervention/treatment Phase Depression Adolescent Drug: Escitalopram Not Applicable Study Design Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Layout table for study information Study Type : Interventional  (Clinical Trial) Actual Enrollment : 152 participants Allocation: N/A Intervention Model: Single Group Assignment Masking: None (Open Label) Primary Purpose: Basic Science Official Title: Biomarkers of Antidepressant Response and Suicidal Events in Depressed Youth Actual Study Start Date : August 5, 2015 Actual Primary Completion Date : August 2, 2018 Actual Study Completion Date : August 2, 2018 MedlinePlus Genetics related topics: Depression MedlinePlus related topics: Antidepressants Drug Information available for: Citalopram hydrobromide Citalopram Escitalopram Escitalopram oxalate U.S. FDA Resources Arms and Interventions Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Arm Intervention/treatment Experimental: Escitalopram Participants with depression were treated with escitalopram(ranging from 5mg to 30mg) for 8 weeks. Escitalopram was initiated at 5mg for 1 week, followed by an increase to 10mg at week 2. After week 2, doses of escitalopram were titrated according to symptoms and adverse effects. Specific, indicated psychotherapy for depression was not allowed during the study. Drug: Escitalopram Other Name: Lexapro Outcome Measures Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Primary Outcome Measures : the change from baseline in CDRS-R at week 8 or upon termination [ Time Frame: baseline and 8 weeks or upon termination ] The Children's Depression Rating Scale-Revised (CDRS-R) is a clinician-rated scale to measure the severity of depression of children and adolescents. The CDRS-R consists of 17 items. Depression symptoms are rated on a 5-point scale from 1 to 5 for sleep, appetite, and tempo of speech items and a 7-point scale from 1 to 7 for remaining 14 items. The total score ranges form 17 (normal) to 113 (severe depression). Change from baseline CDRS-R at 8 weeks or upon termination was calculated as baseline minus 8 weeks in the adjusted CDRS-R total score (CDRS-R total minus 17, the minimum possible total score). Secondary Outcome Measures : CGI-I at 8 weeks or upon termination [ Time Frame: baseline and 8 weeks or upon termination ] The Clinical Global Impression - Improvement scale (CGI-I) is a 7 point scale that requires the clinician to assess how much the patient's illness has improved or worsened relative to a baseline state at the beginning of the intervention. and rated as: 1=very much improved; 2= much improved; 3=minimally improved; 4=no change from baseline; 5=minimally worse; 6=much worse; 7=very much worse since the initiation of treatment. CGI-I was measured at 8 weeks or upon termination Eligibility Criteria Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the contacts provided below. For general information, Learn About Clinical Studies. Layout table for eligibility information Ages Eligible for Study: 12 Years to 17 Years   (Child) Sexes Eligible for Study: All Accepts Healthy Volunteers: No Criteria Inclusion Criteria: clinical diagnosis of major depressive disorder(MDD) according to criteria of Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) current episode at least 4 weeks in duration at baseline with a score of at least 40 on the Children's Depression Rating Scale-Revised (CDRS-R), and CGI-Severity ≥4 at baseline Exclusion Criteria: intelligence quotient (IQ) lower than 70 psychotic features or first-degree relatives with a history of bipolar I disorder alcohol or substance abuse within the past 6 months history of schizophrenia, bipolar disorder, eating disorder, or autism history of neurological diseases including convulsive disorders or brain damage concurrent medications with psychotropic effects (other than stimulants for ADHD) chronic medical conditions (e.g., asthma, inflammatory bowel disease, diabetes) and/or chronic medication with psychotropic effects (e.g., anticonvulsants) or chronic use of NSAIDS or other drugs with known impact on inflammatory pathways. Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Information from the National Library of Medicine To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor. Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT03547219 Locations Layout table for location information Korea, Republic of Seoul National University Hospital Seoul, N/A = Not Applicable, Korea, Republic of, 03080 Sponsors and Collaborators Seoul National University National Research Foundation of Korea Investigators Layout table for investigator information Principal Investigator: Jae-Won Kim, MD, PhD Seoul National University and Seoul National University Hospital Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Publications automatically indexed to this study by ClinicalTrials.gov Identifier (NCT Number): Choi CH, Lee J, Lee KH, Hong SB, Kim SH, Han JY, Kim JW, Cho SC, Kim JW. Effects of Antidepressant Treatment on Symptom Measures of Attention in Adolescents with Depression: A Preliminary Open-Label Study. J Child Adolesc Psychopharmacol. 2021 May;31(4):288-293. doi: 10.1089/cap.2020.0101. Epub 2021 Jan 8. Layout table for additonal information Responsible Party: Jae-Won Kim, Professor, Seoul National University ClinicalTrials.gov Identifier: NCT03547219 History of Changes Other Study ID Numbers: 2015R1A2A2A01004501 First Posted: June 6, 2018 Key Record Dates Last Update Posted: April 4, 2019 Last Verified: April 2019 Layout table for additional information Studies a U.S. FDA-regulated Drug Product: No Studies a U.S. FDA-regulated Device Product: No Keywords provided by Jae-Won Kim, Seoul National University: Depression Adolescent Biomarker Inflammation Brain-derived neurotrophic factor (BDNF) Functional MRI Antidepressant Machine learning Additional relevant MeSH terms: Layout table for MeSH terms Depression Behavioral Symptoms Escitalopram Selective Serotonin Reuptake Inhibitors Neurotransmitter Uptake Inhibitors Membrane Transport Modulators Molecular Mechanisms of Pharmacological Action Neurotransmitter Agents Serotonin Agents Physiological Effects of Drugs

https://clinicaltrials.gov/ct2/show/NCT03547219

Network Working Group B. Joshi Request for Comments: 5240 Infosys Technologies Ltd. Category: Standards Track R. Bijlani June 2008 Protocol Independent Multicast (PIM) Bootstrap Router MIB Status of This Memo This document specifies an Internet standards track protocol for the Internet community, and requests discussion and suggestions for improvements. Please refer to the current edition of the "Internet Official Protocol Standards" (STD 1) for the standardization state and status of this protocol. Distribution of this memo is unlimited. Abstract This document defines a portion of the Management Information Base (MIB) for use with network management protocols in the Internet community. In particular, it describes managed objects used for managing the Bootstrap Router (BSR) mechanism for PIM (Protocol Independent Multicast). Table of Contents 1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . 2 2. The Internet-Standard Management Framework . . . . . . . . . . 2 3. Conventions . . . . . . . . . . . . . . . . . . . . . . . . . 2 4. Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 5. Definitions . . . . . . . . . . . . . . . . . . . . . . . . . 3 6. Security Considerations . . . . . . . . . . . . . . . . . . . 17 7. IANA Considerations . . . . . . . . . . . . . . . . . . . . . 19 8. Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . 19 9. References . . . . . . . . . . . . . . . . . . . . . . . . . . 20 9.1. Normative References . . . . . . . . . . . . . . . . . . . 20 9.2. Informative References . . . . . . . . . . . . . . . . . . 20 Joshi & Bijlani Standards Track [Page 1] RFC 5240 PIM BSR MIB June 2008 1. Introduction This memo defines a portion of the Management Information Base (MIB) for use with network management protocols in the Internet community. In particular, it describes managed objects used for managing the Bootstrap Router (BSR) mechanism for PIM [RFC4601], [RFC5059]. This document was created by moving some of the PIM BSR-specific MIB tables from one of the earlier versions of PIM MIB [RFC5060]. 2. The Internet-Standard Management Framework For a detailed overview of the documents that describe the current Internet-Standard Management Framework, please refer to section 7 of RFC 3410 [RFC3410]. Managed objects are accessed via a virtual information store, termed the Management Information Base or MIB. MIB objects are generally accessed through the Simple Network Management Protocol (SNMP). Objects in the MIB are defined using the mechanisms defined in the Structure of Management Information (SMI). This memo specifies a MIB module that is compliant to the SMIv2, which is described in STD 58, RFC 2578 [RFC2578], STD 58, RFC 2579 [RFC2579] and STD 58, RFC 2580 [RFC2580]. 3. Conventions The key words "MUST", "MUST NOT", "REQUIRED", "SHALL", "SHALL NOT", "SHOULD", "SHOULD NOT", "RECOMMENDED", "MAY", and "OPTIONAL" in this document are to be interpreted as described in RFC 2119 [RFC2119]. 4. Overview This MIB module contains four tables. The tables are: 1. The Candidate-RP Table, which contains one row for each multicast group address prefix for which the local router is configured to advertise itself as a Candidate-RP (C-RP). This table exists on routers that are configured as Candidate-RP. 2. The Elected BSR RP-Set Table, which contains one row for each Group-to-RP mapping that was received in C-RP advertisements. This table exists on a router that is an elected BSR (E-BSR). 3. The Candidate-BSR Table, which contains one row for each Candidate-BSR configuration for the local router. This table exists on routers that are configured as Candidate-BSR. Joshi & Bijlani Standards Track [Page 2] RFC 5240 PIM BSR MIB June 2008 4. The Elected-BSR Table, which contains one row for each elected BSR. This table exists on a router that is an elected BSR. This MIB module uses textual conventions defined in the INET-ADDRESS- MIB [RFC4001]. 5. Definitions PIM-BSR-MIB DEFINITIONS ::= BEGIN IMPORTS MODULE-IDENTITY, OBJECT-TYPE, NOTIFICATION-TYPE, mib-2, Unsigned32, TimeTicks FROM SNMPv2-SMI RowStatus, TruthValue, StorageType FROM SNMPv2-TC MODULE-COMPLIANCE, OBJECT-GROUP, NOTIFICATION-GROUP FROM SNMPv2-CONF InetAddressType, InetAddressPrefixLength, InetAddress, InetZoneIndex FROM INET-ADDRESS-MIB; pimBsrMIB MODULE-IDENTITY LAST-UPDATED "200805280000Z" -- 28 May 2008 ORGANIZATION "IETF Protocol Independent Multicast (PIM) Working Group" CONTACT-INFO "Email: [email protected] WG charter: http://www.ietf.org/html.charters/pim-charter.html" DESCRIPTION "The MIB module for management of the Bootstrap Router (BSR) mechanism for PIM routers. Copyright (C) The IETF Trust (2008). This version of this MIB module is part of RFC 5240; see the RFC itself for full legal notices." REVISION "200805280000Z" -- 28 May 2008 DESCRIPTION "Initial version, published as RFC 5240." ::= { mib-2 172 } -- -- Top-level structure -- pimBsrNotifications OBJECT IDENTIFIER ::= { pimBsrMIB 0 } pimBsrObjects OBJECT IDENTIFIER ::= { pimBsrMIB 1 } Joshi & Bijlani Standards Track [Page 3] RFC 5240 PIM BSR MIB June 2008 -- -- Conformance Information -- pimBsrConformance OBJECT IDENTIFIER ::= { pimBsrMIB 2 } pimBsrCompliances OBJECT IDENTIFIER ::= { pimBsrConformance 1 } pimBsrGroups OBJECT IDENTIFIER ::= { pimBsrConformance 2 } -- -- The BSR Candidate-RP Table -- pimBsrCandidateRPTable OBJECT-TYPE SYNTAX SEQUENCE OF PimBsrCandidateRPEntry MAX-ACCESS not-accessible STATUS current DESCRIPTION "The (conceptual) table listing the IP multicast group prefixes for which the local router is to advertise itself as a Candidate-RP." ::= { pimBsrObjects 1 } pimBsrCandidateRPEntry OBJECT-TYPE SYNTAX PimBsrCandidateRPEntry MAX-ACCESS not-accessible STATUS current DESCRIPTION "An entry (conceptual row) in the pimBsrCandidateRPTable." INDEX { pimBsrCandidateRPAddressType, pimBsrCandidateRPAddress, pimBsrCandidateRPGroupAddress, pimBsrCandidateRPGroupPrefixLength } ::= { pimBsrCandidateRPTable 1 } PimBsrCandidateRPEntry ::= SEQUENCE { pimBsrCandidateRPAddressType InetAddressType, pimBsrCandidateRPAddress InetAddress, pimBsrCandidateRPGroupAddress InetAddress, pimBsrCandidateRPGroupPrefixLength InetAddressPrefixLength, pimBsrCandidateRPBidir TruthValue, pimBsrCandidateRPAdvTimer TimeTicks, pimBsrCandidateRPPriority Unsigned32, pimBsrCandidateRPAdvInterval Unsigned32, pimBsrCandidateRPHoldtime Unsigned32, pimBsrCandidateRPStatus RowStatus, pimBsrCandidateRPStorageType StorageType } Joshi & Bijlani Standards Track [Page 4] RFC 5240 PIM BSR MIB June 2008 pimBsrCandidateRPAddressType OBJECT-TYPE SYNTAX InetAddressType MAX-ACCESS not-accessible STATUS current DESCRIPTION "The Inet address type of the Candidate-RP." ::= { pimBsrCandidateRPEntry 1 } pimBsrCandidateRPAddress OBJECT-TYPE SYNTAX InetAddress (SIZE (4|8|16|20)) MAX-ACCESS not-accessible STATUS current DESCRIPTION "The (unicast) address that will be advertised as a Candidate-RP. The InetAddressType is given by the pimBsrCandidateRPAddressType object." ::= { pimBsrCandidateRPEntry 2 } pimBsrCandidateRPGroupAddress OBJECT-TYPE SYNTAX InetAddress (SIZE (4|8|16|20)) MAX-ACCESS not-accessible STATUS current DESCRIPTION "The IP multicast group address that, when combined with the corresponding value of pimBsrCandidateRPGroupPrefixLength, identifies a group prefix for which the local router will advertise itself as a Candidate-RP. The InetAddressType is given by the pimBsrCandidateRPAddressType object. This address object is only significant up to pimBsrCandidateRPGroupPrefixLength bits. The remainder of the address bits are zero. This is especially important for this field, which is part of the index of this entry. Any non-zero bits would signify an entirely different entry." ::= { pimBsrCandidateRPEntry 3 } pimBsrCandidateRPGroupPrefixLength OBJECT-TYPE SYNTAX InetAddressPrefixLength (4..128) MAX-ACCESS not-accessible STATUS current DESCRIPTION "The multicast group address mask that, when combined with the corresponding value of pimBsrCandidateRPGroupAddress, identifies a group prefix for which the local router will advertise itself as a Candidate-RP. The InetAddressType is given by the Joshi & Bijlani Standards Track [Page 5] RFC 5240 PIM BSR MIB June 2008 pimBsrCandidateRPAddressType object." ::= { pimBsrCandidateRPEntry 4 } pimBsrCandidateRPBidir OBJECT-TYPE SYNTAX TruthValue MAX-ACCESS read-create STATUS current DESCRIPTION "If this object is set to TRUE, this group range is advertised with this RP as a BIDIR-PIM group range. If it is set to FALSE, it is advertised as a PIM-SM group range." DEFVAL { false } ::= { pimBsrCandidateRPEntry 5 } pimBsrCandidateRPAdvTimer OBJECT-TYPE SYNTAX TimeTicks MAX-ACCESS read-only STATUS current DESCRIPTION "The time remaining before the local router next sends a Candidate-RP-Advertisement to the elected BSR for this zone." ::= { pimBsrCandidateRPEntry 6 } pimBsrCandidateRPPriority OBJECT-TYPE SYNTAX Unsigned32 (0..255) MAX-ACCESS read-create STATUS current DESCRIPTION "The priority for this Candidate-RP advertised in Candidate-RP-Advertisements." REFERENCE "RFC 5059, section 3.2" DEFVAL { 192 } ::= { pimBsrCandidateRPEntry 7 } pimBsrCandidateRPAdvInterval OBJECT-TYPE SYNTAX Unsigned32 (1..26214) UNITS "seconds" MAX-ACCESS read-create STATUS current DESCRIPTION "A Candidate-RP generates Candidate-RP-Advertisements periodically. This object represents the time interval in seconds between two consecutive advertisements." REFERENCE "RFC 5059, sections 3.2 and 5" DEFVAL { 60 } Joshi & Bijlani Standards Track [Page 6] RFC 5240 PIM BSR MIB June 2008 ::= { pimBsrCandidateRPEntry 8 } pimBsrCandidateRPHoldtime OBJECT-TYPE SYNTAX Unsigned32 (0..65535) UNITS "seconds" MAX-ACCESS read-create STATUS current DESCRIPTION "Holdtime for this Candidate-RP. The amount of time (in seconds) this Candidate-RP entry is valid. This object's value can be zero only when this C-RP is shutting down." REFERENCE "RFC 5059, section 4.2" DEFVAL { 150 } ::= { pimBsrCandidateRPEntry 9 } pimBsrCandidateRPStatus OBJECT-TYPE SYNTAX RowStatus MAX-ACCESS read-create STATUS current DESCRIPTION "The status of this row, by which new entries may be created, or old entries deleted from this table. This status object can be set to active(1) without setting any other columnar objects in this entry. All writable objects in this entry can be modified when the status of this entry is active(1)." ::= { pimBsrCandidateRPEntry 10 } pimBsrCandidateRPStorageType OBJECT-TYPE SYNTAX StorageType MAX-ACCESS read-create STATUS current DESCRIPTION "The storage type for this row. Rows having the value 'permanent' need not allow write-access to any columnar objects in the row." DEFVAL { nonVolatile } ::= { pimBsrCandidateRPEntry 11 } -- Joshi & Bijlani Standards Track [Page 7] RFC 5240 PIM BSR MIB June 2008 -- The BSR Elected BSR RP-Set Table -- pimBsrElectedBSRRPSetTable OBJECT-TYPE SYNTAX SEQUENCE OF PimBsrElectedBSRRPSetEntry MAX-ACCESS not-accessible STATUS current DESCRIPTION "The (conceptual) table listing BSR-specific information about PIM group mappings learned via C-RP advertisements or created locally using configurations. This table is maintained only on the Elected BSR. An Elected BSR uses this table to create Bootstrap messages after applying a local policy to include some or all of the group mappings in this table." ::= { pimBsrObjects 2 } pimBsrElectedBSRRPSetEntry OBJECT-TYPE SYNTAX PimBsrElectedBSRRPSetEntry MAX-ACCESS not-accessible STATUS current DESCRIPTION "An entry (conceptual row) in the pimBsrElectedBSRRPSetTable." INDEX { pimBsrElectedBSRGrpMappingAddrType, pimBsrElectedBSRGrpMappingGrpAddr, pimBsrElectedBSRGrpMappingGrpPrefixLen, pimBsrElectedBSRGrpMappingRPAddr } ::= { pimBsrElectedBSRRPSetTable 1 } PimBsrElectedBSRRPSetEntry ::= SEQUENCE { pimBsrElectedBSRGrpMappingAddrType InetAddressType, pimBsrElectedBSRGrpMappingGrpAddr InetAddress, pimBsrElectedBSRGrpMappingGrpPrefixLen InetAddressPrefixLength, pimBsrElectedBSRGrpMappingRPAddr InetAddress, pimBsrElectedBSRRPSetPriority Unsigned32, pimBsrElectedBSRRPSetHoldtime Unsigned32, pimBsrElectedBSRRPSetExpiryTime TimeTicks, pimBsrElectedBSRRPSetGrpBidir TruthValue } pimBsrElectedBSRGrpMappingAddrType OBJECT-TYPE SYNTAX InetAddressType MAX-ACCESS not-accessible STATUS current DESCRIPTION Joshi & Bijlani Standards Track [Page 8] RFC 5240 PIM BSR MIB June 2008 "The Inet address type of the IP multicast group prefix." ::= { pimBsrElectedBSRRPSetEntry 2 } pimBsrElectedBSRGrpMappingGrpAddr OBJECT-TYPE SYNTAX InetAddress (SIZE (4|8|16|20)) MAX-ACCESS not-accessible STATUS current DESCRIPTION "The IP multicast group address that, when combined with pimBsrElectedBSRGrpMappingGrpPrefixLen, gives the group prefix for this mapping. The InetAddressType is given by the pimBsrElectedBSRGrpMappingAddrType object. This address object is only significant up to pimBsrElectedBSRGrpMappingGrpPrefixLen bits. The remainder of the address bits are zero. This is especially important for this field, which is part of the index of this entry. Any non-zero bits would signify an entirely different entry." ::= { pimBsrElectedBSRRPSetEntry 3 } pimBsrElectedBSRGrpMappingGrpPrefixLen OBJECT-TYPE SYNTAX InetAddressPrefixLength (4..128) MAX-ACCESS not-accessible STATUS current DESCRIPTION "The multicast group prefix length that, when combined with pimBsrElectedBSRGrpMappingGrpAddr, gives the group prefix for this mapping. The InetAddressType is given by the pimBsrElectedBSRGrpMappingAddrType object. If pimBsrElectedBSRGrpMappingAddrType is 'ipv4' or 'ipv4z', this object must be in the range 4..32. If pimBsrElectedBSRGrpMappingAddrType is 'ipv6' or 'ipv6z', this object must be in the range 8..128." ::= { pimBsrElectedBSRRPSetEntry 4 } pimBsrElectedBSRGrpMappingRPAddr OBJECT-TYPE SYNTAX InetAddress (SIZE (4|8|16|20)) MAX-ACCESS not-accessible STATUS current DESCRIPTION "The IP address of the RP to be used for groups within this group prefix. The InetAddressType is given by the pimBsrElectedBSRGrpMappingAddrType object." ::= { pimBsrElectedBSRRPSetEntry 5 } pimBsrElectedBSRRPSetPriority OBJECT-TYPE Joshi & Bijlani Standards Track [Page 9] RFC 5240 PIM BSR MIB June 2008 SYNTAX Unsigned32 (0..255) MAX-ACCESS read-only STATUS current DESCRIPTION "The priority for RP. Numerically higher values for this object indicate lower priorities, with the value zero denoting the highest priority." REFERENCE "RFC 5059, section 4.1" ::= { pimBsrElectedBSRRPSetEntry 6 } pimBsrElectedBSRRPSetHoldtime OBJECT-TYPE SYNTAX Unsigned32 (0..65535) UNITS "seconds" MAX-ACCESS read-only STATUS current DESCRIPTION "The holdtime for RP" REFERENCE "RFC 5059, section 4.1" ::= { pimBsrElectedBSRRPSetEntry 7 } pimBsrElectedBSRRPSetExpiryTime OBJECT-TYPE SYNTAX TimeTicks MAX-ACCESS read-only STATUS current DESCRIPTION "The minimum time remaining before this entry will be aged out. The value zero indicates that this entry will never be aged out." ::= { pimBsrElectedBSRRPSetEntry 8 } pimBsrElectedBSRRPSetGrpBidir OBJECT-TYPE SYNTAX TruthValue MAX-ACCESS read-only STATUS current DESCRIPTION "If this object is TRUE, this group range with this RP is a BIDIR-PIM group range. If it is set to FALSE, it is a PIM-SM group range." ::= { pimBsrElectedBSRRPSetEntry 9 } -- -- The BSR Candidate-BSR Table -- pimBsrCandidateBSRTable OBJECT-TYPE SYNTAX SEQUENCE OF PimBsrCandidateBSREntry MAX-ACCESS not-accessible STATUS current Joshi & Bijlani Standards Track [Page 10] RFC 5240 PIM BSR MIB June 2008 DESCRIPTION "The (conceptual) table containing Candidate-BSR configuration for the local router. The table contains one row for each zone for which the local router is to advertise itself as a Candidate-BSR." ::= { pimBsrObjects 3 } pimBsrCandidateBSREntry OBJECT-TYPE SYNTAX PimBsrCandidateBSREntry MAX-ACCESS not-accessible STATUS current DESCRIPTION "An entry (conceptual row) in the pimBsrCandidateBSRTable." INDEX { pimBsrCandidateBSRZoneIndex } ::= { pimBsrCandidateBSRTable 1 } PimBsrCandidateBSREntry ::= SEQUENCE { pimBsrCandidateBSRZoneIndex InetZoneIndex, pimBsrCandidateBSRAddressType InetAddressType, pimBsrCandidateBSRAddress InetAddress, pimBsrCandidateBSRPriority Unsigned32, pimBsrCandidateBSRHashMaskLength Unsigned32, pimBsrCandidateBSRElectedBSR TruthValue, pimBsrCandidateBSRBootstrapTimer TimeTicks, pimBsrCandidateBSRStatus RowStatus, pimBsrCandidateBSRStorageType StorageType } pimBsrCandidateBSRZoneIndex OBJECT-TYPE SYNTAX InetZoneIndex (1..4294967295) MAX-ACCESS not-accessible STATUS current DESCRIPTION "The zone index uniquely identifies the zone on a device to which this Candidate-BSR is attached. There is one entry for each zone in ipMcastZoneTable. Scope-level information for this zone can be extracted from ipMcastZoneTable in IP Multicast MIB [RFC5132]. Zero is a special value used to request the default zone for a given scope. Zero is not a valid value for this object." ::= { pimBsrCandidateBSREntry 1 } pimBsrCandidateBSRAddressType OBJECT-TYPE SYNTAX InetAddressType Joshi & Bijlani Standards Track [Page 11] RFC 5240 PIM BSR MIB June 2008 MAX-ACCESS read-create STATUS current DESCRIPTION "The address type of the Candidate-BSR." ::= { pimBsrCandidateBSREntry 2 } pimBsrCandidateBSRAddress OBJECT-TYPE SYNTAX InetAddress MAX-ACCESS read-create STATUS current DESCRIPTION "The (unicast) address that the local router will use to advertise itself as a Candidate-BSR. The InetAddressType is given by the pimBsrCandidateBSRAddressType object." ::= { pimBsrCandidateBSREntry 3 } pimBsrCandidateBSRPriority OBJECT-TYPE SYNTAX Unsigned32 (0..255) MAX-ACCESS read-create STATUS current DESCRIPTION "The priority value for the local router as a Candidate-BSR for this zone. Numerically higher values for this object indicate higher priorities." DEFVAL { 0 } ::= { pimBsrCandidateBSREntry 4 } pimBsrCandidateBSRHashMaskLength OBJECT-TYPE SYNTAX Unsigned32 (0..128) MAX-ACCESS read-create STATUS current DESCRIPTION "The hash mask length (used in the RP hash function) that the local router will advertise in its Bootstrap messages for this zone. This object defaults to 30 if pimBsrCandidateBSRAddressType is 'ipv4' or 'ipv4z' , and defaults to 126 if pimBsrCandidateBSRAddressType is 'ipv6' or 'ipv6z'." ::= { pimBsrCandidateBSREntry 5 } pimBsrCandidateBSRElectedBSR OBJECT-TYPE SYNTAX TruthValue MAX-ACCESS read-only STATUS current DESCRIPTION "Whether the local router is the elected BSR for this zone." Joshi & Bijlani Standards Track [Page 12] RFC 5240 PIM BSR MIB June 2008 ::= { pimBsrCandidateBSREntry 6 } pimBsrCandidateBSRBootstrapTimer OBJECT-TYPE SYNTAX TimeTicks MAX-ACCESS read-only STATUS current DESCRIPTION "The time remaining before the local router next originates a Bootstrap message for this zone. Value of this object is zero if pimBsrCandidateBSRElectedBSR is 'FALSE'." ::= { pimBsrCandidateBSREntry 7 } pimBsrCandidateBSRStatus OBJECT-TYPE SYNTAX RowStatus MAX-ACCESS read-create STATUS current DESCRIPTION "The status of this row, by which new entries may be created or old entries deleted from this table. This status object can be set to active(1) without setting any other columnar objects in this entry. All writable objects in this entry can be modified when the status of this entry is active(1)." ::= { pimBsrCandidateBSREntry 8 } pimBsrCandidateBSRStorageType OBJECT-TYPE SYNTAX StorageType MAX-ACCESS read-create STATUS current DESCRIPTION "The storage type for this row. Rows having the value 'permanent' need not allow write-access to any columnar objects in the row." DEFVAL { nonVolatile } ::= { pimBsrCandidateBSREntry 9 } -- -- The BSR Elected-BSR Table -- pimBsrElectedBSRTable OBJECT-TYPE SYNTAX SEQUENCE OF PimBsrElectedBSREntry MAX-ACCESS not-accessible STATUS current DESCRIPTION Joshi & Bijlani Standards Track [Page 13] RFC 5240 PIM BSR MIB June 2008 "The (conceptual) table containing information about elected BSRs. The table contains one row for each zone for which there is an elected BSR." ::= { pimBsrObjects 4 } pimBsrElectedBSREntry OBJECT-TYPE SYNTAX PimBsrElectedBSREntry MAX-ACCESS not-accessible STATUS current DESCRIPTION "An entry (conceptual row) in the pimBsrElectedBSRTable." INDEX { pimBsrElectedBSRZoneIndex } ::= { pimBsrElectedBSRTable 1 } PimBsrElectedBSREntry ::= SEQUENCE { pimBsrElectedBSRZoneIndex InetZoneIndex, pimBsrElectedBSRAddressType InetAddressType, pimBsrElectedBSRAddress InetAddress, pimBsrElectedBSRPriority Unsigned32, pimBsrElectedBSRHashMaskLength Unsigned32, pimBsrElectedBSRExpiryTime TimeTicks } pimBsrElectedBSRZoneIndex OBJECT-TYPE SYNTAX InetZoneIndex (1..4294967295) MAX-ACCESS not-accessible STATUS current DESCRIPTION "The zone index uniquely identifies the zone on a device to which this Elected BSR is attached. There is one entry for each zone in ipMcastZoneTable. Scope-level information for this zone can be extracted from ipMcastZoneTable in IP Multicast MIB [RFC5132]. Zero is a special value used to request the default zone for a given scope. Zero is not a valid value for this object." ::= { pimBsrElectedBSREntry 1 } pimBsrElectedBSRAddressType OBJECT-TYPE SYNTAX InetAddressType MAX-ACCESS read-only STATUS current DESCRIPTION "The address type of the elected BSR." ::= { pimBsrElectedBSREntry 2 } Joshi & Bijlani Standards Track [Page 14] RFC 5240 PIM BSR MIB June 2008 pimBsrElectedBSRAddress OBJECT-TYPE SYNTAX InetAddress (SIZE (4|8|16|20)) MAX-ACCESS read-only STATUS current DESCRIPTION "The (unicast) address of the elected BSR. The InetAddressType is given by the pimBsrElectedBSRAddressType object." ::= { pimBsrElectedBSREntry 3 } pimBsrElectedBSRPriority OBJECT-TYPE SYNTAX Unsigned32 (0..255) MAX-ACCESS read-only STATUS current DESCRIPTION "The priority value for the elected BSR for this address type. Numerically higher values for this object indicate higher priorities." ::= { pimBsrElectedBSREntry 4 } pimBsrElectedBSRHashMaskLength OBJECT-TYPE SYNTAX Unsigned32 (0..128) MAX-ACCESS read-only STATUS current DESCRIPTION "The hash mask length (used in the RP hash function) advertised by the elected BSR for this zone." ::= { pimBsrElectedBSREntry 5 } pimBsrElectedBSRExpiryTime OBJECT-TYPE SYNTAX TimeTicks MAX-ACCESS read-only STATUS current DESCRIPTION "The minimum time remaining before the elected BSR for this zone will be declared down." ::= { pimBsrElectedBSREntry 6 } -- -- PIM BSR Notifications -- pimBsrElectedBSRLostElection NOTIFICATION-TYPE OBJECTS { pimBsrElectedBSRAddressType, pimBsrElectedBSRAddress, pimBsrElectedBSRPriority } STATUS current DESCRIPTION Joshi & Bijlani Standards Track [Page 15] RFC 5240 PIM BSR MIB June 2008 "A pimBsrElectedBSRLostElection notification should be generated when current E-BSR lost election to a new Candidate-BSR. Only an E-BSR should generate this notification. This notification is generated when pimBsrCandidateBSRElectedBSR becomes FALSE." REFERENCE "RFC 5059, section 3.1" ::= { pimBsrNotifications 1 } pimBsrCandidateBSRWinElection NOTIFICATION-TYPE OBJECTS { pimBsrCandidateBSRElectedBSR } STATUS current DESCRIPTION "A pimBsrCandidateBSRWinElection notification should be generated when a C-BSR wins BSR Election. Only an E-BSR should generate this notification. This notification is generated when pimBsrCandidateBSRElectedBSR becomes TRUE." REFERENCE "RFC 5059, section 3.1" ::= { pimBsrNotifications 2 } -- -- Compliance Statements -- pimBsrCompliance MODULE-COMPLIANCE STATUS current DESCRIPTION "The compliance statement for PIM routers that implement the Bootstrap Router (BSR) mechanism." MODULE -- this module MANDATORY-GROUPS { pimBsrObjectGroup } GROUP pimBsrDiagnosticsGroup DESCRIPTION "This group is optional." ::= { pimBsrCompliances 1 } -- -- Units of Conformance -- pimBsrObjectGroup OBJECT-GROUP Joshi & Bijlani Standards Track [Page 16] RFC 5240 PIM BSR MIB June 2008 OBJECTS { pimBsrCandidateRPBidir, pimBsrCandidateRPAdvTimer, pimBsrCandidateRPPriority, pimBsrCandidateRPAdvInterval, pimBsrCandidateRPHoldtime, pimBsrCandidateRPStatus, pimBsrCandidateRPStorageType, pimBsrElectedBSRRPSetPriority, pimBsrElectedBSRRPSetHoldtime, pimBsrElectedBSRRPSetExpiryTime, pimBsrElectedBSRRPSetGrpBidir, pimBsrCandidateBSRAddress, pimBsrCandidateBSRAddressType, pimBsrCandidateBSRPriority, pimBsrCandidateBSRHashMaskLength, pimBsrCandidateBSRElectedBSR, pimBsrCandidateBSRBootstrapTimer, pimBsrCandidateBSRStatus, pimBsrCandidateBSRStorageType, pimBsrElectedBSRAddress, pimBsrElectedBSRAddressType, pimBsrElectedBSRPriority, pimBsrElectedBSRHashMaskLength, pimBsrElectedBSRExpiryTime } STATUS current DESCRIPTION "A collection of objects for managing the Bootstrap Router (BSR) mechanism for PIM routers." ::= { pimBsrGroups 1 } pimBsrDiagnosticsGroup NOTIFICATION-GROUP NOTIFICATIONS { pimBsrElectedBSRLostElection, pimBsrCandidateBSRWinElection } STATUS current DESCRIPTION "Objects providing additional diagnostics related to the Bootstrap Router (BSR) mechanism for PIM routers." ::= { pimBsrGroups 2 } END 6. Security Considerations There are a number of management objects defined in this MIB module with a MAX-ACCESS clause of read-write and/or read-create. Such objects may be considered sensitive or vulnerable in some network environments. The support for SET operations in a non-secure Joshi & Bijlani Standards Track [Page 17] RFC 5240 PIM BSR MIB June 2008 environment without proper protection can have a negative effect on network operations. These are the tables and objects and their sensitivity/vulnerability: o A new Candidate-BSR with high priority or modification of priority ( bsrCandidateBSRPriority ) of an existing Candidate-BSR can take over the functionality of an Elected BSR, which can prevent and disrupt the services. o A new Candidate-RP with lower priority or modification of priority ( bsrCandidateRPPriority ) of an existing Candidate-RP can force other routers to select itself for a particular group prefix. This can prevent and disrupt the services provided through this group prefix. The following are the read-write and read-create objects defined in this MIB module: bsrCandidateRPBidir bsrCandidateRPPriority bsrCandidateRPAdvInterval bsrCandidateRPHoldtime bsrCandidateBSRAddressType bsrCandidateBSRAddress bsrCandidateBSRPriority bsrCandidateBSRHashMaskLength Some of the readable objects in this MIB module (i.e., objects with a MAX-ACCESS other than not-accessible) may be considered sensitive or vulnerable in some network environments. It is thus important to control even GET and/or NOTIFY access to these objects and possibly to even encrypt the values of these objects when sending them over the network via SNMP. These are the tables and objects and their sensitivity/vulnerability: pimBsrCandidateRPAdvTimer pimBsrElectedBSRRPSetPriority pimBsrElectedBSRRPSetHoldtime pimBsrElectedBSRRPSetExpiryTime pimBsrElectedBSRRPSetGrpBidir pimBsrCandidateBSRElectedBSR pimBsrCandidateBSRBootstrapTimer pimBsrElectedBSRAddressType pimBsrElectedBSRAddress pimBsrElectedBSRPriority pimBsrElectedBSRHashMaskLength pimBsrElectedBSRExpiryTime Joshi & Bijlani Standards Track [Page 18] RFC 5240 PIM BSR MIB June 2008 In this MIB module, possible effects that can be induced by GET operations include: o Determination of Elected BSR, Candidate-BSRs, and Candidate-RPs in the Multicast Network topology. This information may be sensitive and may be used in preparation for Denial-of-Service (DoS) attacks including any of the attacks described above. SNMP versions prior to SNMPv3 did not include adequate security. Even if the network itself is secure (for example by using IPsec), there is still no control over whom on the secure network is allowed to access (read/change/create/delete) the objects in this MIB module. It is RECOMMENDED that implementers consider the security features as provided by the SNMPv3 framework (see [RFC3410], section 8), including full support for the SNMPv3 cryptographic mechanisms (for authentication and privacy). Further, deployment of SNMP versions prior to SNMPv3 is NOT RECOMMENDED. Instead, it is RECOMMENDED to deploy SNMPv3 and to enable cryptographic security. It is then a customer/operator responsibility to ensure that the SNMP entity giving access to an instance of this MIB module is properly configured to give access to the objects only to those principals (users) that have legitimate rights to access (read/change/create/delete) them. 7. IANA Considerations The MIB module in this document uses the following IANA-assigned OBJECT IDENTIFIER values recorded in the SMI Numbers registry: Descriptor OBJECT IDENTIFIER value ---------- ----------------------- pimBsrMIB { mib-2 172 } 8. Acknowledgments This MIB module is based on the original work in [RFC5060] by R. Sivaramu, J. Lingard, and B. Joshi. Many thanks to Bill Fenner, Stig Venaas, Nidhi Bhaskar, David Mcwalter, David Harrington, and J. W. Atwood for their feedback on this MIB module. Suggested IPv6 multicast MIBs by R. Sivaramu and R. Raghunarayan have been used for comparison while editing this MIB module. Joshi & Bijlani Standards Track [Page 19] RFC 5240 PIM BSR MIB June 2008 9. References Joshi & Bijlani Standards Track [Page 20] RFC 5240 PIM BSR MIB June 2008 9.1. Normative References [RFC2119] Bradner, S., "Key words for use in RFCs to Indicate Requirement Levels", BCP 14, RFC 2119, March 1997. [RFC2578] McCloghrie, K., Ed., Perkins, D., Ed., and J. Schoenwaelder, Ed., "Structure of Management Information Version 2 (SMIv2)", STD 58, RFC 2578, April 1999. [RFC2579] McCloghrie, K., Ed., Perkins, D., Ed., and J. Schoenwaelder, Ed., "Textual Conventions for SMIv2", STD 58, RFC 2579, April 1999. [RFC2580] McCloghrie, K., Perkins, D., and J. Schoenwaelder, "Conformance Statements for SMIv2", STD 58, RFC 2580, April 1999. [RFC4001] Daniele, M., Haberman, B., Routhier, S., and J. Schoenwaelder, "Textual Conventions for Internet Network Addresses", RFC 4001, February 2005. [RFC4601] Fenner, B., Handley, M., Holbrook, H., and I. Kouvelas, "Protocol Independent Multicast - Sparse Mode (PIM-SM): Protocol Specification (Revised)", RFC 4601, August 2006. [RFC5060] Sivaramu, R., Lingard, J., McWalter, D., Joshi, B., and A. Kessler, "Protocol Independent Multicast MIB", RFC 5060, January 2008. [RFC5059] Bhaskar, N., Gall, A., Lingard, J., and S. Venaas, "Bootstrap Router (BSR) Mechanism for Protocol Independent Multicast (PIM)", RFC 5059, January 2008. [RFC5132] McWalter, D., Thaler, D., and A. Kessler, "IP Multicast MIB", RFC 5132, December 2007. 9.2. Informative References [RFC3410] Case, J., Mundy, R., Partain, D., and B. Stewart, "Introduction and Applicability Statements for Internet- Standard Management Framework", RFC 3410, December 2002. Joshi & Bijlani Standards Track [Page 21] RFC 5240 PIM BSR MIB June 2008 Authors' Addresses Bharat Joshi Infosys Technologies Ltd. 44 Electronics City, Hosur Road Bangalore 560 100 India EMail: [email protected] URI: http://www.infosys.com/ Raina Bijlani EMail: [email protected] Joshi & Bijlani Standards Track [Page 22] RFC 5240 PIM BSR MIB June 2008 Full Copyright Statement Copyright (C) The IETF Trust (2008). This document is subject to the rights, licenses and restrictions contained in BCP 78, and except as set forth therein, the authors retain all their rights. This document and the information contained herein are provided on an "AS IS" basis and THE CONTRIBUTOR, THE ORGANIZATION HE/SHE REPRESENTS OR IS SPONSORED BY (IF ANY), THE INTERNET SOCIETY, THE IETF TRUST AND THE INTERNET ENGINEERING TASK FORCE DISCLAIM ALL WARRANTIES, EXPRESS OR IMPLIED, INCLUDING BUT NOT LIMITED TO ANY WARRANTY THAT THE USE OF THE INFORMATION HEREIN WILL NOT INFRINGE ANY RIGHTS OR ANY IMPLIED WARRANTIES OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE. Intellectual Property The IETF takes no position regarding the validity or scope of any Intellectual Property Rights or other rights that might be claimed to pertain to the implementation or use of the technology described in this document or the extent to which any license under such rights might or might not be available; nor does it represent that it has made any independent effort to identify any such rights. Information on the procedures with respect to rights in RFC documents can be found in BCP 78 and BCP 79. Copies of IPR disclosures made to the IETF Secretariat and any assurances of licenses to be made available, or the result of an attempt made to obtain a general license or permission for the use of such proprietary rights by implementers or users of this specification can be obtained from the IETF on-line IPR repository at http://www.ietf.org/ipr. The IETF invites any interested party to bring to its attention any copyrights, patents or patent applications, or other proprietary rights that may cover technology that may be required to implement this standard. Please address the information to the IETF at [email protected]. Joshi & Bijlani Standards Track [Page 23]

https://www.rfc-editor.org/in-notes/rfc5240.txt

Waddell v. Coleman, 4731 - Texas - Case Law - VLEX 890853124 0: [object Object]. 1: [object Object]. 2: [object Object]. 3: [object Object]. 4: [object Object]. 5: [object Object] Texas Waddell v. Coleman, 4731 Court Court of Appeals of Texas. Court of Civil Appeals of Texas Writing for the Court WALKER Citation 248 S.W.2d 216 Parties WADDELL et al. v. COLEMAN et al. Docket Number No. 4731,4731 Decision Date 25 October 1951 Page 216 248 S.W.2d 216 WADDELL et al. v. COLEMAN et al. No. 4731. Court of Civil Appeals of Texas, Beaumont. Oct. 25, 1951. Rehearing Denied Jan. 16, 1952. Grover C. Lowe, Woodville, A. M. Huffman, Beaumont, for appellees. WALKER, Justice. The action is in trespass to try title, to recover a tract of 100 acres of land in Hardin County, in the southeastern corner of the Frederick Helfenstein Survey. This corner is on Village Creek Page 218 and from it the boundary of the 100 acre tract runs north 850 varas along the eastern boundary of the Helfenstein Survey, and then turns and extends west a distance of 618 varas. From the point thus fixed, which is the northwestern corner of the 100 acre tract, the boundary extends south to Village Creek, and Village Creek is the southern boundary of the 100 acre tract. Under the pleadings, the appellees were cross-plaintiffs and the appellants were cross-defendants, but the appellees will be referred to as plaintiffs and their cross-action as the petition, and the appellants will be referred to as defendants and their pleading as an answer and cross-action. In addition to the formal allegations in trespass to try title and their prayer for recovery of the land, plaintiffs alleged that defendants had wrongfully cut and removed timber from the land and prayed recovery of damages. The 100 acre tract in suit was the eastern 100 acres of a 160 acre tract on which M. C. Spell (Mack Spell) and his wife, Lucy Spell, had resided at a time many years before the present controversy arose but it is often referred to in the proof as the 'Spell tract'. M. C. Spell and Lucy Spell were divorced, and Lucy Spell married one Lock. After her marriage to Lock, she and her former husband sued one Chapman in whom, apparently, the title to the land was vested, to recover some land in the Helfenstein Survey which either was or included the 160 acre tract; and by a judgment dated July 7, 1919, which was rendered by agreement, Mrs. Lock and M. C. Spell recovered an undivided 100 acres of the 160 acres, which was to be surveyed so as to include the improvements they had made on the 160 acres. The 100 acre tract was surveyed later; the field notes are dated September 29, 1919. The title of Mrs. Lock and M. C. Spell was acquired by the plaintiff Lowe and the former plaintiff Tom Coleman and Mr. Lowe and Mr. Coleman conveyed the pine timber on the 100 acre tract to the Kirby Lumber Company by a deed dated February 14, 1920. The Kirby Lumber Company cut this timber in 1920. Such title to the 100 acre tract as the defendants may have is founded upon adverse possession of a part of the 100 acre tract which, defendants say, amounted to a compliance with the 10-year statute of limitation, since the year 1920, by E. M. (or Emory) Waddell and his family. E. M. Waddell died in 1936. Of the defendants, Mrs. Emmie Waddell is the widow of E. M. Waddell; and the other defendants (except Lindsey, Diffey and Payne) are the children of E. M. Waddell and the husbands of some of these children. The rights of Lindsey, Diffey and Payne were derived from the other defendants. The defendant Mrs. Emmie Waddell was the second wife of E. M. Waddell. She and Mr. Waddell were married in 1906 and established their home on a tract of land in the Bryan Survey which is east of, and adjoins, the 100 acre tract in suit. They apparently established their residence on this tract in 1909 and they resided there until 1927, and in that year they removed from this tract in the Bryan Survey to the community of Village Mills, where they resided until Mr. Waddell's death in 1936. The western line and corners of the Waddell tract on the Bryan Survey co-incide with the eastern line and corners of the 100 acre tract in suit. Some evidence was adduced which tended to show that the eastern line of the Helfenstein Survey, in which the 100 acre tract in suit is supposedly situated, was actually separated from the western line of the Bryan Survey by a vacancy which was about 145 or 150 varas wide, and that all of the land in suit which was held in actual possession by the Waddells lay on this vacancy. However, this question was resolved in the defendants' favor by the jury. The cause was tried to a jury who returned the following answers to the Special Issues submitted to them: (1) The true eastern line of the 100 acre tract (which we construe as meaning the eastern line of the Helfenstein Survey) was the line identified in the evidence as running north 850 varas with the west line of the Bryan Survey; (2) defendants (other than Diffey and Payne) and E. M. Waddell held peaceable Page 219 and adverse possession of the 100 acres in suit, cultivating, using or enjoying, and claiming the same for a period of 10 years or more continually after the year 1920 and prior to 1948; (3) a preponderance of the evidence did not show that E. M. Waddell, within the period of 10 years found by them, acknowledged and recognized that Tom. F. Coleman and Grover C. Lowe, or either, were the owners of the land; and (4) a preponderance of the evidence did not show that E. M. Waddell at any time immediately prior to the date of the judgment in Lock v. Chapman acknowledged and recognized that Lucy Lock was the owner of the land. Subsequently, the plaintiffs filed a motion for judgment non obstante veredicto, and this motion was sustained and judgment was rendered by the trial court that plaintiffs recover the title to and the possession of the 100 acre tract in suit. The trial court also rendered judgment in behalf of plaintiffs against defendants for $3,913.49, the value of the timber cut and removed by the defendants from the land. From this judgment the defendants have appealed. Defendants have assigned nine Points of Error but these Points raise only the questions now to be discussed. (1) A deed from Kirby to Chapman is an element of plaintiffs' chain of title, and defendants say that the description of the land conveyed by this deed did not cover the land in suit. In their argument under one of their Points, they make the same criticism of Mooney's deed to Kirby, which was another element of plaintiffs' chain of title. These contentions are overruled. It is the position of all parties that the land in suit is a part of the tract known as the Mack Spell or Lucy Lock tract, or 'place', and that this Spell-Lock tract is in the Helfenstein Survey, and in Hardin County, on the north bank of Village Creek. Plaintiffs proved a patent to Frederick Helfenstein for a 1280 acre survey, and the description in Mooney's deed to Kirby and the description in Kirby's deed to Chapman obviously refer to the survey described in and conveyed by this patent. Further, the proof showed that the 1280 acre survey thus patented to Helfenstein is the particular Helfenstein Survey in which the parties have located the Spell-Lock tract. Thus Grimes' field notes of the tract awarded to Mrs. Lock and Spell in their suit against Chapman are accompanied by, and obviously refer to, some field notes which are the field notes of the Helfenstein Survey described in the patent. The patent locates the survey in 'Liberty District, Tyler County, on the north bank of Big Sandy Creek;' but Ellis, a surveyor, located the Spell-Lock tract in a Helfenstein Survey of 1280 acres which was located in both Hardin and Tyler Counties, on Village Creek, and he contended that he had found the original southeastern corner of the Helfenstein Survey which is described in the patent mentioned above, but located at a point 145 or 150 varas west of the point contended for by the defendants as the southeastern corner of the Helfenstein Survey in which the Spell-Lock tract is located. Ellis was, unquestionably, referring to the same Helfenstein Survey as is described in the patent. Hyde, another surveyor, obviously referring to the same southeastern corner, located this corner at the place for which the defendants contend as the southeastern corner of the Helfenstein Survey in which the Spell-Lock tract is located. Further, the judgment in behalf of Emory Waddell (who originated the adverse possession and claim on which defendants' claim of title by adverse possession is founded) against the Receiver of the Kirby Lumber Company not only identifies the 100 acres there awarded to Waddell with the tract recovered by Mrs. Lock and Spell from Chapman, but also describes the Helfenstein Survey in which it lies and identifies that with the survey described in the patent. As we have stated, this judgment in behalf of Mrs. Lock and Spell is an element of plaintiffs' chain of title, and defendants claim the benefit of the judgment in favor of Waddell. The deed to the defendant Lindsey from his co-defendant Mrs. Emmie Waddell, and Lindsey's later deed to Mrs. Waddell, also Page 220 use descriptions which are derived from the description in Waddell's judgment against the Receiver. Indeed, C. S. Holland's deed to E. M. Waddell, dated April 30, 1901, conveying what seems to be the Spell tract describes the Helfenstein Survey so as to identify it with that conveyed by the patent. The patentee conveyed the survey to Maes, and later deeds from the heirs of Maes to persons who are in plaintiffs' chain of title refer to the survey as being in Hardin County and identify Big Sandy Creek with Village Creek. As we read the proof, the parties are really not in disagreement about the identity of the particular Helfenstein Survey in which the Spell-Lock tract is situated. (2) In paragraph 4 of the answer which defendants filed on March 14, 1950, defendants plead certain matters as an estoppel against plaintiffs, and on June 14, 1950, plaintiffs filed a reply to this answer and defendants' accompanying cross-action) in which they excepted to the plea of estoppel. These exceptions were sustained, and Point 9 assigns error to the order sustaining the exceptions.... 3 practice notes Coleman v. Waddell, No. A-3540 United States Supreme Court of Texas June 11, 1952 ...the verdict. The Court of Civil Appeals reversed the trial court's judgment and rendered judgment for respondents on the verdict. 248 S.W.2d 216 . Its opinion may be referred to for a full statement of the case. We are turning the case on a single question, and our statement will be limited ...... Pewitt v. Renwar Oil Corp., No. 6695 United States Court of Appeals of Texas. Court of Civil Appeals of Texas June 11, 1953 ...could not render judgment for the two acres actually used by them. A reference to the opinion of the Court of Civil Appeals in this case, 248 S.W.2d 216 , will show that the Waddells were suing Coleman for the 100-acre tract and, of course, in their role as plaintiffs it became necessary for...... Powell Lumber Co. v. Dickerson, No. 14217. United States United States Courts of Appeals. United States Court of Appeals (5th Circuit) April 3, 1953 ...in connection with the improvements on that survey, these cases are without application here. Cf. Waddell v. Coleman, Tex.Civ.App., 248 S.W.2d 216 , at pages 226-227, and the thorough and informative discussion of the general question by Justice Smedley of the Texas Supreme Court in Simonds ...... 3 cases Coleman v. Waddell, No. A-3540 United States Supreme Court of Texas June 11, 1952 ...the verdict. The Court of Civil Appeals reversed the trial court's judgment and rendered judgment for respondents on the verdict. 248 S.W.2d 216 . Its opinion may be referred to for a full statement of the case. We are turning the case on a single question, and our statement will be limited ...... Pewitt v. Renwar Oil Corp., No. 6695 United States Court of Appeals of Texas. Court of Civil Appeals of Texas June 11, 1953 ...could not render judgment for the two acres actually used by them. A reference to the opinion of the Court of Civil Appeals in this case, 248 S.W.2d 216 , will show that the Waddells were suing Coleman for the 100-acre tract and, of course, in their role as plaintiffs it became necessary for...... Powell Lumber Co. v. Dickerson, No. 14217. United States United States Courts of Appeals. United States Court of Appeals (5th Circuit) April 3, 1953 ...in connection with the improvements on that survey, these cases are without application here. Cf. Waddell v. Coleman, Tex.Civ.App., 248 S.W.2d 216 , at pages 226-227, and the thorough and informative discussion of the general question by Justice Smedley of the Texas Supreme Court in Simonds ......

https://case-law.vlex.com/vid/waddell-v-coleman-no-890853124

U.S. Patent: 5136156 - Photoelectric switch - August 4, 1992 Photoelectric switch Abstract An improved photoelectric switching device for coordinate detection system comprises a first array of light emitting elements, a first array of photo-detecting elements for receiving the light from the first array of light emitting elements and producing a first block signal when the light is blocked by a stylus, such as a finger, a second array of light emitting elements, a second array of photo-detecting elements for receiving the light from the second array of light emitting elements and producing a second block signal when the light is blocked by the stylus wherein the light receiving surfaces of the second array of photo-detecting elements are aligned an angle, such as 180 degrees, with respect to those of the first array of photo-detecting elements, and a control means such as a microcomputer for driving the first and the second array of the light emitting elements and for discriminating the coordinates of the stylus on the display unit which displays a pattern having several regions allocated for certain control of optional equipment. Therefore, the stylus can be detected even if there is an inadvertent operation in either of the first and the second photo-detecting elements due to a strong incidence of ambient light in the lateral direction. <table><tbody><tr><td> Inventors:</td><td> Nounen; Michiyasu(Hyogo, JP); Taketoshi; Kouichi(Hyogo, JP); Uragata; Yoshihiro(Hyogo, JP)</td></tr><tr><td> Assignee:</td><td> Mitsubishi Denki Kabushiki Kaisha(Tokyo, JP)</td></tr><tr><td> Appl. No.:</td><td> 624464</td></tr><tr><td> Filed:</td><td> December 10, 1990</td></tr></tbody></table> Foreign Application Priority Data <table><tbody><tr><td></td><td> Nov 01, 1988[JP]</td><td> 63-277997</td></tr></tbody></table> <table><tbody><tr><td> Current U.S. Class:</td><td> 250/221; 250/222.1</td></tr><tr><td> Intern'l Class:</td><td> G01V 009/04</td></tr><tr><td> Field of Search:</td><td> 250/221,222.1 340/555,556,557,706,712 341/20,31</td></tr></tbody></table> References Cited U.S. Patent Documents <table><tbody><tr><td> 3234512</td><td> Feb., 1966</td><td> Burkhardt</td><td> 250/221.</td></tr><tr><td> 3746863</td><td> Jul., 1973</td><td> Pronovost</td><td> 250/221.</td></tr><tr><td> 3764813</td><td> Oct., 1973</td><td> Clement et al.</td><td> 250/221.</td></tr><tr><td> 4198623</td><td> Apr., 1980</td><td> Misek</td><td> 340/712.</td></tr><tr><td> 4703316</td><td> Oct., 1987</td><td> Sherbeck</td><td> 250/221.</td></tr><tr><td colspan=4> Foreign Patent Documents</td></tr><tr><td> 0330767</td><td> Sep., 1989</td><td> EP.</td><td></td></tr><tr><td> 2344066</td><td> Oct., 1977</td><td> FR.</td><td></td></tr></tbody></table> Primary Examiner: Nelms; David C. Attorney, Agent or Firm: Burns, Doane, Swecker & Mathis Parent Case Text This application is a continuation, of application Ser. No. 07/412,984, filed Sept. 26, 1989, now abandoned. Claims What is claimed is: 1. A photoelectric switching device for a vehicle comprising: a first light emitting element mounted at one side of a display unit; a first photo-detecting element mounted at the other side of the display unit for receiving the light from said light emitting element along a first light path and producing a first block signal when the first light path is blocked by an object; a second light emitting element mounted at said other side of the display unit; a second photo-detecting element mounted at said one side of the display unit for receiving the light from said second light emitting element along a second light path parallel and in a direction opposite to said first light path and producing a second block signal when the second light path is blocked by the object; and a control means for driving said first and second light emitting elements and for producing a detection signal which indicates the presence of the object when either said first or second block signal is produced by said first or second photodetecting element, whereby a plurality of regions are displayed on said display unit for indicating operable functions and in alignment with corresponding light paths. 2. A photoelectric switching device as defined in claim 1, wherein said first and second light emitting elements comprise an array of light emitting diodes, respectively and said first and second photo-detecting elements comprise an array of photo-detecting transistors, respectively. 3. A photoelectric switching device as defined in claim 1, and further including a third light emitting element the light path of which is mutually perpendicular to that of the first and second light emitting elements and a third photo-detecting element for receiving the light from said third light emitting element and producing a block signal when the light is blocked by the object. 4. A photoelectric switching device as defined in claim 2, and further including an array of light emitting diodes mutually perpendicular to said first and second array of light emitting diodes, and a third array of photo-detecting transistors mutually perpendicular to said first and second array of photo-detecting transistors, and arranged to received light from said array of light emitting diodes except when blocked by said object. 5. A photoelectric switching device as defined in claim 4, wherein the third array of light emitting diodes is mounted at the lower side of a frame and the third array of the photo-detecting transistors is mounted at the upper side of the frame. 6. A photoelectric switching device as defined in claim 2 or 4, wherein the control means is comprises an input circuit, an output circuit, a central processing unit, a random access memory, and a read only memory. 7. A photoelectric switching device as defined in claim 1, wherein a light path of the first light emitting element and a light path of the second light emitting element are arranged to pass through the same region allocated for the control of optional equipment. 8. A photoelectric switching device as defined in claim 4, wherein a light path of one light emitting diode of the first array of light emitting diodes and a light path of one light emitting diode of the second array of the light emitting diodes are arranged to pass through the same divided region or regions allocated for the control of optional equipment. 9. A photoelectric switching device as defined in claim 1 or 4, wherein the light emitting elements emit infrared light. Description BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a photoelectric switching device for a coordinate detection system and, more particularly, to such a photoelectric switching device for a coordinate data input device, a so called touch panel, to be installed in adjacent to an associated CRT (Cathode Ray Tube) display unit and the like. 2. Description of the Prior Art Japanese Laid-open Patent No. 61,224/1987 discloses an arrangement such as that shown in FIG. 1, wherein 1 denotes a liquid crystal display unit, 2a, 2a denotes two mutually perpendicular linear arrays of infrared light-emitting elements or light emitting diodes for emitting infrared light along the lateral and longitudinal light paths, respectively, 3a, 3a denotes two mutually perpendicular linear arrays of associated photo-detector elements for detecting infrared light from respective light emitting diodes, and 4 denotes a user's finger placed on the face of display unit in such a manner as to block the infrared light along the lateral and longitudinal light paths A and B being emitted from the light-emitting diodes. According to this system, each light emitting element 2a emits light one by one in a sequential manner under the control of control circuitry (not shown) and each associated photo-detector element 3a receives the light one by one in the same manner. If the light path A in the lateral direction and the light path B in the longitudinal direction are blocked respectively by a finger, for instance, the photo-detector elements 3a that correspond to the light paths A and B will produce a signal for indicating obstruction of the light paths. The control circuitry will then locate the spatial coordinates of the finger 4 by discriminating the sources of the signals produced by the photo-detector elements. Therefore, various operations such as change of display, control of optional equipment being installed, and the like are carried out by detecting the spatial coordinates of the finger 4 on a pattern (not shown) to be displayed on the face of display unit 1 for entering a switching input. Such a prior art coordinate detection system has been installed in an instrument panel of an automobile for controlling a car audio system, an air conditioner and the like. In such a prior art system, the photo-detector elements arranged in the longitudinal direction along the light path B are normally installed at the upper portion of the display unit 1 for eliminating inadvertent operations to be caused by the ambient light such as the sun light. The detectors that conforms to the lateral direction along the light path A are installed at the left side of the display unit 1 for decreasing the influence of ambient light to a greater extent in the prior art system under such condition as the coordinate detection system is installed at the central part, between a driver's seat and the passenger seat, of the instrument panel. This is because ambient light from the driver's side is usually effectively blocked by a driver since the driver always occupies driver's seat at the right side in case of the right-hand steering-wheel vehicle however, it is hard to prevent ambient light from the passenger side from reaching the right side of the display unit 1 if no one is on the passenger seat. Accordingly, in the prior art system, it has been difficult to avoid inadvertent operations merely by installing the photo-detecting elements at the left side of the display unit 1 even though ambient light from the driver's side is substantially blocked by the driver. It is, therefore, a principal object of the present invention to eliminate the aforesaid difficulties and to provide a photoelectric switching device for coordinate detection systems having less susceptibility to inadvertent operations caused by ambient light. SUMMARY OF THE INVENTION According to the present invention, a photoelectric switching device for a coordinate detection system comprises a first array of light emitting elements, a first array of photo-detecting elements for receiving the light from said first array of light emitting elements and producing a first block signal when the light is blocked by an object, a second array of light emitting elements, a second array of photo-detecting elements for receiving the light from said second array of light emitting elements and producing a second block signal when the light is blocked by the object, wherein the light receiving surfaces of the second array of photo-detecting elements are aligned in a different angle from those of the first array of photo-detecting elements, and a control means for driving said first and second arrays of light emitting elements and for producing a final detection signal indicating coordinate information of the object on the surface of the display unit when either said first or second detection signal is derived from said first and second arrays of photo-detecting elements. According to the specific embodiment of this invention, the light receiving surfaces of the first and second arrays of photo-detecting elements are aligned at a different angle with respect to each other and the control means derives a detected information signal upon receipt of either said first or second detect signal. Therefore, the presence of an object can effectively be detected by the arrangement according to the invention even if either the first or second array of photo-detecting elements is exposed to ambient light. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram of the prior art system; FIG. 2 is a block diagram of an embodiment of this invention; FIG. 3 is an enlarged showing of an object detecting unit; and FIG. 4A and FIG. 4B are flowcharts showing the operation steps performed by the microcomputer of FIG. 1. DESCRIPTION OF THE PREFERRED EMBODIMENT The present invention will now be described in more detail with reference to the accompanying drawings. Referring now to FIG. 2, there is shown a block diagram of the present invention, wherein 5 designates a display unit including a CRT tube of the type installed in an instrument panel (not shown) between the driver's seat and the passenger seat, which is the same installation as the prior art system for the right-hand steering-wheel vehicles as described in the above, 6 designates a driving circuit for the display unit 5, and 7 is a position detecting unit (which will be described with reference to FIG. 3 hereinafter) installed at the periphery of the display unit 5 in a similar manner to the prior art for detecting the spatial location of a user's finger. In addition, 8 and 9 are a radio and an air conditioner, respectively, and 10 denotes a microcomputer including a CPU 10a, RAM 10c, an input circuit 10d, and an output circuit 10e for the control of the driving circuit 6, the position detecting unit 7, the radio 8 and the air conditioner 9. FIG. 3 shows an enlarged diagram for illustrating the position detecting unit 7, in which 5a denotes a display of the display unit 5. The display 5a is divided into 12 regions Z11-Z43 provided by 3 regions in longitudinal and 4 regions in lateral directions, as shown by dotted lines in the drawing. By assigning X1 through X4 for lateral coordinates and Y1 though Y3 for longitudinal coordinates, for ease of understanding, a position on the display will be specified by coordinates (Xi, Yj) (where i=1, 2, 3, 4, and j=1, 2, 3) as shown in the drawing. D1 through D4 are light emitting elements consisting of infrared light emitting diodes mounted on the lower side of the display unit 5. They emit infrared light upwards respectively along light paths designated by P1 through P4. T1 through T4 are photo-detecting elements in the form of photo-transistors mounted on the upper side of the display unit 5 for receiving the light from the light emitting elements D1 through D4 and for generating a block signal if any one of the light paths P1 through P4 is blocked by a finger. D11, D21 and D31 constitute a first array of light emitting elements consisting of infrared light emitting diodes which are mounted on the right side (driver's seat side) of the display unit 5 for providing light paths P11, P21 and P31 by transmitting light towards the left side of the display 5a. T11, T21 and T31 constitute a first array of photo-detecting elements consisting of photo-transistors which mounted at the left side (passenger seat side) of the display 5a for receiving the light from the first array of light emitting elements and generating a first block signal when either of the light paths P11, P21 and P31 is blocked by a finger. D12, D22 and D32 constitute a second array of light emitting elements consisting of infrared light emitting diodes for providing light paths P12, P22 and P32 by transmitting the light towards the right side of the display 5a. T12, T22 and T32 constitute a second array of photo-detecting elements in the form of photo-transistors which are mounted on the right side of the display 5a for receiving the light from the second light emitting elements D12, D22 and D32 and generating a second block signal when either of the light paths P12, P22 and P32 is blocked by a finger. The light path P1 passes through the regions Z11, Z22 and Z23, while the light path P2 passes through the regions Z21, Z22 and Z23, the light path P3 through the regions Z31, Z32 and Z33, and the light path P4 through the regions Z41, Z42 and Z43. On the other hand, the light paths P11 and P12 pass through the regions Z11, Z21, Z31 and Z41 simultaneously in the opposite directions, similarly, the light paths P21 and P22 through the regions Z12, Z22, Z32 and Z42, and the light paths P31 an P32 through the regions Z13, Z23, Z33 and Z43 respectively. Further, every light emitting element (D1 and others) is driven by the microcomputer 10 through an amplifier 11 and every photo-detector (T1 and others) is connected to the microcomputer through an amplifier 12 for the transmission of each block signal as shown in FIG. 2. In accordance with the embodiment illustrated in the above, the regions Z11-Z43 in the display 5a of the display unit 5 are assigned to display specific patterns (not shown) such as channel selection buttons, volume control buttons and the like for the operation of the radio 8. With this arrangement, if a user's finger is placed on a pattern which indicates a desired operation, for instance the region Z11, the longitudinal light path P1 and the lateral light paths P11 and P12 will be blocked entirely by the finger and the photo-detector elements T1, T11 and T12 will produce block signals that correspond to the required operation. The microcomputer 10 discriminates, by receiving the derived block signals, the region Z11 where the longitudinal light path P1 and the lateral light paths P11 and P12 are blocked at the same time and then controls the radio 8, for instance, based on the operating functions assigned to the region Z11. The microcomputer 10 is capable of discriminating the regions Z11, Z21, Z31 and Z41 where the lateral light paths P11 and P12 pass through even if one of the lateral light paths P11 and P12 is not blocked with a finger and only one block signal from either the first or second photo-detecting elements T11 or T12 is produced. With this capability, even when strong ambient light falls on the display 5a from the left to the right side thereof, the first photo-detecting element T11 can produce the first block signal whilst the second photo-detecting element T12 is not capable of producing a second block signal. Accordingly, the second photo-detecting element T12 can produce the second block signal without fail when strong ambient light falls on the display 5a from the right to the left side thereof. As it is described above, in the installation of the photo-electric switching device to a vehicle, since strong ambient light such as sunlight falls generally on the display 5a from the upper side to the lower side thereof, no problems result from installing the photo-detecting elements T1 through T4 only at the upper side of the display 5a. Referring now to FIGS. 4A and 4B, there are shown flowcharts implemented by the microcomputer 10 of FIG. 1. In the main flowchart of FIG. 4A, the microcomputer performs a first step S101 of initialization upon initiation of the apparatus. A display process for delivering a signal of displaying a predetermined pattern on the display unit 5 to the drive circuit 6 is performed at step S102. The control of the position detecting unit 7 is performed at step S103 and whereby carrying out the position detecting process for discriminating a region of the display 5a touched by a finger. The detailed steps of the step S103 are shown in FIG. 4B, an initial setting of parameters "i" and "j" to "1" is performed respectively at step S201 after starting the flow. Next, step S202 outputs a signal for exciting a light emitting element Di. An output signal (which indicates the existence of a block signal) of a photo-detecting element Ti is accepted at step S203 and the status of a photo-detecting element Ti is memorized on a memory Xi at step S204 whereby, "1" is memorized if there is an output from the photo-detecting element Ti and "0" if there is not. Step S205 increments a parameter "i" to "i+1" and the program returns from S206 to S202 until the parameter becomes 4 by the repeating of steps S202-S205. With this arrangement, the status of blocking in any of the four light paths P1-P4 along the longitudinal direction of the display 5a is memorized adequately. A first light emitting element Dj1 is excited to emit light at step S207. An output (which indicates the existence of the first block signal) of a first photo-detecting element Tij is accepted at step S208 and the status of the first photo-detecting element Tij is memorized on a memory Yj1 at step S209 whereby, "1" is memorized if there is an output from the first photo-detecting element Tj1 and "0" if there is not. In a similar manner, a second light emitting element Dj2 is excited to emit light at S210, an output (which indicates the existence of a second block signal) of a photo-detecting element Tj2 is accepted at step S211 and the status of the second photo-detecting element Tj2 is memorized on a memory Yj2 at step S212 whereby, "1" is memorized if there is an output from the second photo-detecting element Tj2 and "0" if there is not. Step S213 performs disjunction of memories Yj1 and Yj2 and the resultant is memorized on a memory Yj. Step S214 increments a parameter "j" to "j+1" and the program returns from S215 to step S207 until the parameter j becomes 3 by repeating the steps of S207-S215. The discrimination of the region touched by the finger is performed at step S216 based on the contents of the memories Xi and Yj, that is, if the contents of the memories Xi and Yj are both "1" at the same time, then, the region Zij is determined to be the region touched by the finger and the program returns to the main flowchart of FIG. 4a again for carrying out the sucessive steps. S104 performs the control of the radio 8, for instance, increasing the volume thereof, if the pattern corresponding to the region being discriminated at step S216 indicated such an operation. Step S105 performs the control of the air conditioner 9, for instance, increasing a setting temperature, if the pattern denotes an operational instruction for the air conditioner 9. In this way, the steps S102-S105 are performed repeatedly. As described above, according to the embodiment, it is required to detect blockage of one light path P1 in the longitudinal direction of the display 5a for discriminating the region Z11 touched by the finger. However an object detected signal (the content of the memory Yj at step S213 corresponds to this signal in FIG. 4b) which indicates the presence of the finger in the lateral direction of the display 5a is also required provided that either one of the first block signal or the second block signal is derived from the respective photo-detectors after detecting either one of the two lateral light paths P11 and P12 being blocked. In other words, since the first and the second photo-detectors T11 and T12 being related to the light paths P11 and P12 are so aligned at a predetermined angle to each other (180 degrees for this embodiment), the partial position 2 of the finger on the display 5a can be reliably determined by performing the disjunction of the first and the second block signals even if there is a strong incidence of ambient light which results in an inadvertent operation of either one of the associated photo-detectors. This is because at least the other photo-detector operates normally since there is no incidence of ambient light thereon. It should be appreciated from the foregoing description that the present invention provides an improved switching device for detecting a partial position of finger on the display, whereby the light receiving surfaces of second photo-detecting elements are aligned in a different angle from those of the first photo-detecting elements for deriving an object detect signal from a control means if there is either one of the first and the second block signals. Therefore, this invention has a specific feature of detecting the object even if there is an inadvertent operation in either of the first and the second photo-detecting elements due to a strong incidence of ambient light. Although the present invention has been described in detail with reference to the preferred embodiment, it should be understood by those of ordinary skilled in the art that various modifications can be made without departing from the scope of this invention, for example, it is obvious that this invention may be applied to an automatic door the opening or closing of which is performed by detecting a human body with such an arrangement that at least one light emitting element is mounted for each first and second light emitting elements and at least one associated photo-detector is mounted for each first and second photo-detectors whereby detecting the human body when two light paths have blocked simultaneously. In addition to this, the light receiving surfaces of the first and the second photo-detectors are not really necessary to oppose face to face each other but it is enough if there is a definite angle therebetween. Accordingly, this invention is not to be limited except as by the appended claims.

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Parent’s Choice Rice Baby Cereal Recall | High Arsenic Level - Consumer Reports FDA says rice baby cereal sold at Walmart has been recalled because of high arsenic levels. CR experts explain how to stay safe. Rice Baby Cereal Sold at Walmart Recalled Due to High Arsenic Levels Caregivers should check for Parent’s Choice rice baby cereal sold after April 2021 <here is a image db246891de0068a4-62c0ae00f308df7a> Source: FDA Baby food manufacturer Maple Island issued anationwide recallof three lots of its 8-ounce Parent’s Choice rice baby cereal after a sample tested above the Food and Drug Administration’s guidance for inorganic arsenic, according to the FDA. The products were tested as part of a routine sampling program by the FDA. No illnesses have been reported. Parent’s Choice rice baby cereal is sold at Walmart. The retailer said in an announcement in early October that it had pulled the recalled baby food from store shelves. And on Nov. 8, Walmart said it does not plan to restock the product. While the recall notice says Maple Island’s tests of both the raw materials and the finished rice cereal product showed that they were “in compliance with the FDA’s guidelines,” the company said it issued the recall out of an abundance of caution. The recall comes after the FDA said earlier this year that it would takesteps to reduce certain heavy metals in baby food. The FDA’s announcement came on the heels of a government report that foundstartlingly high levels of these metals in some foodsmade by a variety of baby food manufacturers. Since then, the FDA sent letters to manufacturers of baby food as a reminder of their responsibility to consider the hazards oftoxic elements such as lead and arsenicin their products. This is also the second recall by a manufacturer for elevated inorganic arsenic levels in rice cereal for babies in less than six months. Beech-Nut issued a recall in June 2021 and announced it was permanently withdrawing from the infant rice cereal market after routine testing showed elevated levels of arsenic. Heavy metals in baby food have been a documented problem for several years. Tests in 2018 of 50 nationally distributed baby and toddler foods byConsumer Reports found that two-thirds had worrisome levels of at least one type of heavy metal. The problem is that the developing brains and bodies of babies and young children may be particularly vulnerable to the toxic effects of heavy metal exposure, which can lower IQ and is linked to behavior problems and ADHD, as well as an increased risk for skin and bladder cancer. (SeeCR’s advice on how to mitigate exposure.) Despite all the documented risks, the FDA has set limits only for heavy metals such as lead and arsenic in a handful of foods. “We’ve reached a critical point,” says Brian Ronholm, director of food policy at Consumer Reports. “The industry appears to be unable to control the issue, and the FDA’s go-slow regulatory approach is not serving the public interest.” “Baby food manufacturers should consider pulling infant rice cereal from the market until they are able to implement effective mitigation measures to address inorganic arsenic,” he says. “Parents and caregivers deserve better.” Maple Island did not respond to CR’s request for comment. The Details Product recalled:Three lots of 8-ounce packages of Parent’s Choice rice baby cereal sold at Walmart after April 5, 2021. The following product information can be found on the bottom left corner on the back of the package: • Lot 21083, with UPC Code #00681131082907 and a “best if used by” date of June 24, 2022 • Lot 21084, with UPC Code #00681131082907 and a “best if used by” date of June 25, 2022 • Lot 21242, with UPC Code #00681131082907 and a “best if used by” date of Nov. 30, 2022 The problem:The product tested above the guidance level for inorganic arsenic set by the FDA. The fix:Return the baby food to the Walmart store where it was purchased for a full refund or toss it into the trash. How to contact the manufacturer:Call Maple Island between 9 a.m. and 5 p.m. ET Monday through Friday at 800-369-1022 or send an email to [email protected]. Editor’s Note: This story was updated to include new information from Walmart. <here is a image 06b5313bbd6c5c80-e1e00105a735ee38>

https://www.consumerreports.org/health/food-contaminants/rice-baby-cereal-sold-at-walmart-recalled-due-to-high-arseni-a5714508306/

mittfittimikik ANNUAL REPORT OF THE TREASURER OF TIIK STATE OF GEORGIA, FOE THE FISCAL YEAE ENDING-DECEMBER 31ST, 1877. ATLANTA, GEORGIA: JAMES P. HABKISON & CO., PEINTEBS AND BINDERS. 1878. ANNUAL REPORT OF THE TREASURER OF THE STATE OF GEORGIA, FOE THE FISCAL YEAE ENDING DECEMBER 31ST, 1877. ATLANTA, GEORGIA: JAMES P. HARBISON & CO., PRINTERS AND BINDERS. 1878. MP^PHM TREASURER'S REPORT. TREASURY OF GEORGIA, ATLANTA, January 1, 1878. To His Excellency, ALFRED H. COLOUITT, Governor of Georgia : SIR--In obedience to law, I have the honor to present herewith to your Excellency the annual report of this Department, showing the financial condition of the State for and during the year 1877, in the tables and statements as follows, ro-wit: The cash balance in the Treasury, January 1, 1877 $ 564 283 33 Received in the Treasury from January 1st to December 31, 1877 bothinclusive ' ' 1)998|346 84 _ Total $2,562,630 17 Disbursements from January 1 to December 31, both inclusive, as per Executive warrants ' _ -[ 728910 80 Balance in Treasury January 1, 1878 ; % 33 719 37 Table A, beginning with the cash balance in the Treasury on the 1st of January, 1877, shows the receipts and payments in each month, beginning January 1, and ending December 31, 1877. Table B is an account current between J. W. Renfroe, Treasurer, and the State, beginning with the cash on' hand January 1, 1877, and shows the collection from the different sources of revenue, and the payments on each particular fund, for the year 1877. Table C is a concise statement of the public debt as it now appears, beginning with the bonds maturing in the present year, and shows the liability of the State on its own account for bonds running to maturity, amounting to $10,644,500. Table D shows the amount of interest to be paid quarterly and semi-annually on bonds of the State of Georgia, and the month in which it falls due, in and during the present year. 4 TREASURER'S REPORT. Table E is the usual estimate required of the Treasurer of the amounts to be paid during the ensuing year for the ordinary expenses of the government, and the usual appropriations for the support and repairs of the universi ies, asylums, and other objects of the State's care, and the probable revenue to be derived from the various sources thereof. Table /''shows the total valid bonded debt of the State on Janu- ary 1, 1878; also the amount of liabilities of the State, absolute and contingent, as endorser on railroad bonds, and the amount of property owned and possessed by the State Table G is a minute and particular descriptive list of the bonds of the State not yet due, showing their number, size, rate, date of issue and maturity; when and where payable, authority and object of issue, principal and interest due each year, the names of signers, also the size, time of payment, and signers of coupons. In my report of January 1, 1877, it will be seen that in my official transactions in respect to the bonds authorized by the act approved February 24, 1876, that there remained in the Treasury at that time $10,319.15 subject to the incoming, and payment of outstanding, coupons. Since then, I have paid Macon and Brunswick Railroad coupons, $9,030.00; North and South Railroad coupons, $78.40-total, $9,108.40, leaving a balance in the Treasury, subject to the incoming, and payment of aforesaid coupons yet outstanding, $1,210.75. RECAPITULATION. Balance in Treasury January 1st, 1877 Paid Macon and Brunswick Railroad coupons Paid North and South Railroad coupons Balance in Treasury "V"*10-819 t6 $J,0ol) uu 78 40 *"~10 7o In accordance with the provision of an act approved February 19 1877, authorizing the issuing of 6 per cent, bonds of the State of Georgia, amounting to $2,298,397.00, for the purpose of retiring by exchange or paying off the recognized bonds of the Macon and Brunswick Railroad, to the amount of $1,951,000.00, the same being first mortgage bonds, indorsed by the State, and bonds of the North and South Railroad, indorsed by the State to the amount of $240,000.00; also, bonds of the Memphis Branch Railroad, indorsed by the State to the amount of $34,000.00, with accrued interest thereon to 1st January, 1877, to the amount of $74,397.00. I have received from the Executive Department twenty-two hundred and ninety eight bonds, of $l,CO0 each, amounting to $2,298,000.00, and have paid off and exchanged said indorsed bonds, as exhibited in the following statement: VBM TREASURER S REPORT. Dr. To 2,298 State six per cent, bonds, $1,000 each Cr. $2,298,000 00 By 1,480 Bonds Macon and Brunswick Railroad, $1,000 each, paid and exchanged $1,480,000 00 By 895 Bonds Macon and Brunswick Railroad, $500 each, paid and exchanged 447,500 00 By 24 Bonds Memphis Brunch Railroad, $1,000 each, paid and exchanged 24,000 00 By 20 Bonds Memphis Branch Railroad, $500 each, paid and exchanged 10,000 00 By 238 Bonds North and South Railroad, $1,000 each, paid and exchanged 238,000 00 By coupons Macon and Brunswick Railroad, paid and exchanged 09,247 50 By coupons Memphis Branch Railroad, paid and ex- changed 4>515 00 By coupons North and South Railroad, paid and exchanged 2,531 58 By amount paid for lithographing bonds, advertising notice to holders, signing, filling out the bonds, coupons, etc By 13 State six per cent bonds on hand, $1,000 each Cash balance in the Treasury $13,000 00 7,741 07- 1,464 85 20,74107-$2,298,000 00 The foregoing tables have been carefully compiled, and, it is be- lieved, are entirely correct, and will afford all the information that is desired in regard to the financial condition of the State, her bonded debt, and her liabilities as indorser. All of which is re- spectfully submitted. J. W. RENFROE, Treasurer. TABLE A. Monthly Receipts and Disbursements from the Treasury, from January 1st to December 31st, 1877. 1877. RECEIPTS. DR. 1877. PAYMENTS. CR. w January.. . To balance in Treasury January 1st, 1877... $ 564,283 33 By payments from the Treasury January.... 223,810 43 February.... February... 108,906 34 March 68,638 07 $ 110,555 33 20,524 80 ian 289,724 73 157,792 28 April May July September.. 65,784 30 308,528 03 48,674 74' July 31,169 15 " 34,075 77 September .. 28,486 65 October 123,078 56 November... 190,799 69 51,457 76 37,898 65 w 20,688 87 ToJ 278,511 94 87,194 54 H 94,138 72 364,013 23 December... 389,623 49 December. 593,180 97 By balance in Treasury December 31st, 1877. 833,719 37 $2,562,630 17 $2,562,630 17 m i :> TABLE B. J. W. RENFROE, Treasurer of the State of Georgia, in account with the State on account of Receipts and Disbursements at the Treasury, from January 1st to December 31, 1877, both inclusive. To balance in Treasury January 1, 1877*, S 564.288 33 By civil establishment, 1876 To general tax of 1877 *..1.;...,, 815,273 79 Bv civil establishment, 1877 To general tax of 1876 378,106 91 By contingent fund, 1876 , To To general general tax tax of 1875 of previous years. ...** - 7,411 17 By contingent, fund, 1877 1,145 18 By printing fund, 1876 To temporary loan,,.,,(...:......t To rental oF^WcijtCrn arid Atlantic Railroad........ 200.1100 00 By printing fund, 1877 800,000 00 By Slate Geologist 1876 and 1877 To claimsof Western and Atlantic Railroad 152,678 94 By Agricultural Department, 1876 To earnings of Maeo^i and Brunswick Bailroad To railroad tax of 1876 and 1877 ,, 25,000 00 By Agricultural Department, 1877 38,203 90 By printing fund, Agricultural Department, 1877 To insurance tax of 1876andl877 15 223 09 By Chemical Bureau, 1876 To liquor tax of 1877 To hire of convicts of 1876 and 1877 _ a _: 25.632 05 By Chemical Bureau, 1877 11,032 07 By State School Commissioner, To showtaxof 1877 _.,, 4,076 50 By salary of chemist, 1877 To fertilizer tax of 1877 5,325 55 By overpayment of taxes To wild land tax : 8,000 00 By public buildings, 1877 To Supreme Court costs of 1877 ..: 3,021 75 By Deaf and Dumb Institute, 1876 and 1877 To rent of public buildings of 1877 2,677 50 By Executive clerks, 1876 and 1877 To insurance fees of 1877 1,045 00 By Solicitor Generals, act of 1858 To sewing machine tax of 1877 683 96 By public debt To express company tax of 1877 689 59 By land scrip fund To te egraph company tax of 1877 424 31 By temporary loan To sleeping car company tax of 1877 275 17 By legislative pay roll To office fees of 1377 402 25 By Lunatic Asylum, 1876 and 1877 To miscellaneous 2,010 96 By Lunatic Asvlum Trustees By Academy forlhe Blind, 1877 By change bills Western and Atlantic Railroad... By Siate Board of Health, 1876 By indebtedness of Western and Atlantic Railroad By school fund, 1876 and 1S77 By special appropriation., 1877 By Constitutional Convention, 1877 By books purchased for library By balance in Treasury 1st January, 1878 82,562,630 17 $ 27 254 00 72, 922 62 ,871 24 ,996 82 ,460 87 424 25 H 828 53 fa > 400 00 M 497 40 a 553 05 330 61 156 4S 445 53 750 00 ,312 21 M f, in 505 69 625 00 W H 574 21 105 00 881 14 314 14 Ow H 025 48 821 65 625 00 316 66 18, 500 00 205 30 454 52 250 00 177. 171 01 33, 851 14 740 75 240 50 719 87 J.f6?.630 17 TABLE C. Consolidated Statement of the Public Debt of the State of Georgia, (not yet due) on the 1st day of January, 1878, showing the dates of issue and maturity, and the annual interest on each issue. 1801 and 1873 1873 1866 and 1873 1872 1876 1877 Date of Issue. Amounts-- Principal and Tears to Eun. When Principal Aunual Interest Annual Interest due. due each year. on each issue. on each amount. H 70 1 month 1878 i 3 months. . 1878 100,000 $ 100,000 3,000 $ 8,000 103,000 108,000 >W icn 1879 300,000 30,000 320,000 w 1880 300,000 30,000 320,000 1881 200,000 14,000 214,000 UJ" 1883 100,000 1883 100,000 1884 100,000 8,000 8,000 8,000 108,000 108,000 ?w s 108,000 o>t 1885 100,000 1886 4,000,000 8,000 378,000 108,000 4,278,000 H 19Q 2,097,000 146,790 2,243,790 14 years. ... 1892 307,500 21,525 329,035 1896 542,000 37,940 579,940 19 years 1897 3,298,000 137,880 3,435,880 $ 10,644,500 $ 719,135 The State is endorser on the South Georgia and Florida Railroad .Bonds to the amount of $461,000.00. Endorsed in 1871. / TABLE D. Showing amount of interest to be paid quarterly and semi-annually on Bonds of the State of Georgia, and the month in which it falls due, in and during the year 1878. Classs of Bonds. Rate. Amount. January Feb'ary. April. July. ., August, October. Total Total Interest. Payments. H M Bonds issued for Stock in Atlantic >W and Gulf Railroad 6 per cent. & f900,000 $127,000 $ 24,000 $ 51,000 $ 151,000 Bonds secured by Mortgage on West ern and Atlantic Railroad 7 per cent. Bonds issued under Act of January 18, 1872 7 per cent. 3,600,000 126,000 00 307,500 10,762 50 126,000 00 10,762 50 252,000 21,525 252,000 H *,, en* 21,525 Bonds issued under Act of February 19, 1873 8 per cent. Bonds issued under Act of February 900,000 136,000 00 36,000 00 72,000 W 172,000 M D O 24, 1876 7 per cent. 542,000 18,970 00 18,970 00 37,940 37,940 3 Bonds issued to redeem M. and B., N. S. and M. Branch Bonds 6 per cent. 2,298,000 68,940 00 68,940 00 137,880 137,880 Gold Bonds, Quarterly Coupons.. 7 per cent. 2,097,000 36,097 50 Average Gold Premium 3 per cent 1,100 92 36,697 50 36,697 50 1,100 92 1,100 92 36,697 50 146,790 1,100 92 4,403 151,193 $10,644,500 $262,470 92 $127,000 $173,798 42 $262,470 92 $ 24,000 $73,798 42 $723,538 $ 923,538 *One hundred thousand dollars of this amount is the principal of the 8 per cent, bonds falling due this month (April). fOne hundred thousand dollars of this amount is the principal of the 6 per bonds falling due this month (February.) Estimate of probable and permanent receipts and disbursements at the Treasury, for the fiscal year ending December 31, 1878. RECEIPTS. From general tax of 1878 From general tax of 1877 From rent of Western and Atlantic Railroad From earnings of tne Macon and Brunswick Kailroad From insurance rax. From railroad tax From dividends From tax of former years From liquor tax From fees of inspectors of fertilizers From rents of public buildings Balance DISBURSEMENTS. $ 875,000 00 225,000 00 300,000 00 25,000 00 15,000 00 15,000 00 2.000 00 5,000 00 20,000 00 30,000 00 2,500 00 5,538 00 $1.5-30.038 00 To civil establishment To legislative pay-roll To Lunatic Asylum, support of To Lunatic Asylum, pay of chaplain and trustees To Deaf and Dumb Asylum, support of To Accademy for the Blind, support of To University of Georgia To University of Atlanta To printing fund To contingent fund To public buildings To officers executive department To inspectors of fertilizers To State agents for vaccination To educational fund ,-- To School Commissioner and clerK! To department State Geologist To department Commissioner of Agriculture To Supreme Court reports To Solicitor Generals' cost Supreme Court To public debt To special appropriations H >M $ 100, 000 100. onil 100 ,000 2,.000 w 13.,000 11 000 8. 1)1)0 1)00 w w ,000 o 12 .OIK) 6 .1)00 12, ,01111 MOO 150.,000 3. 71)11 10,000 10. ,000 3..501.) 2:,000 923..538 10. .000 $1,520,038 00 TABLE F. Showing the total valid Bonded Debt of the State of Georgia, on January 1, 1878, also the amount of liability of the State, absolute and contingent, as endorser on Railroad Bonds, and the amounOof property owned and possessed by the State. Total Bond Debt Total endorsement on (secured by first lien) bonds of South Georgia and Florida Railroad > $10,644,500 a 464,000 g CR. Western and Atlantic Railroad, 138 miles--lease 14 years to run--annual rental $300,000. Macon and Brunswick Railroad--195 miles. North and South Railroad--20 miles. Memphis Branch Railroad--5 miles. South Georgia and Florida Railroad--58 miles--(contingent)--has never defaulted in paying interest. Bonds 1st Mortgage State endorsed of Macon and Brunswick Railroad BondsOeonee Bridge Bank State of Georgia--1,833 shares Bank of Augusta-800 shares Georgia Railroad and Banking Company--180 shares Atlantic and Gulf Railroad--10,000 shares--(par $1,000,000- (no market value) Southern and Atlantic Telegraph, guaranteed by Presid' nt Union Telegraph Company--440 shares Public Buildings--not estimated ^ W o 2 H value $ u.000 value.... no vil uc- nt> value.... 1,280 i4,uuu 10,000 TABLE G. A Descriptive List of the Bonds of the State of Georgia, running to maturity, outstanding on the 31st December, 1877, in order in which they become due, showing their number, size, rate, date of issue and maturity, when and where payable, authority and object of issue, principal and interest due each year, and the names of signers; also, size, time of payment, and signers of coupons. Date of Bond. Size of No. of Rate per Bond. Issue, cent. Time to run. Payable. Month. Year, Year. February February August. February August February'.... February February August August May MOacyt.ober July July April. April April April April April April April April July May 1858 1859 1859 1860 1860 1861 1866 I860 1866 1866 1866 1866 1870 1872 1872 1873 1873 1873 1873 1873 1873 1873 1873 1873 1876 1877 500 200 6 per cent. 20 years 1 month.., 'February.. 500 300 6 per cent. 20 years 1 year February.. 500 100 6 per cent. 20 years \% years.. August 500 300 6 per cent. 20 years 2 years February.. 500 100 6 per cent. 20 years -iy% years..August 500 200 6 per cent. 20 years 3 years February.. 500 200 per cent. 20 years 8 years February.. 100 345 per cent. 20 years 8 years February.. 500 300 . per cent. 20 years 8 years .. August 100 155 6 per cent. 20 years 8 years August-- 1,0001 2,800 7 per cent. 20 years 8 years-- May 500| 1,600 7 per cent. 20 years 8 years May 1,000 2,0j9u8ul per cent. 20 years 12 years .. October ... 1,000: 207 7 per cent. 20 years 14 years... July 500i 201 7 per cent. 20 years 14 years... July 250; 400 8 per cent. 3 years 3 mouths. April 500j 200 8 per cent. 4 years 1 year April 500! 200 8 per cent. 5 years 2 years... April 500 200A 8 per cent. 6 years 3 years... April 500 200 8 per cent. 7 years 4 years... April 1.000 100 8 per cent. 8 years 5 years... April 1,000 100 8,, per cent. 9 years 6 years... April 1,000 100"8 per cent. 10 years 7 years... April 1,000 100 8 per cent. 11 years 8 years... April 1.000 542 7 per cent. 20 years 8 years... July 1,000 2,298 6 per cent. 20 years 19 years... July 1878 1879 1879 1880 1880 1881 1880 1880 1886 1880 1886 1880 1890 1892 1892 1878 1879 1880 1881 1882 1883 1884 1885 1880 1896 1897 Amount of each Issue. Annual Engraved Size of Interest, or Coupons. Printed Signers of Coupons. 100,000 150,000 50,000 150,000 50,000 100,000 100,000 34,500 150,000 15,500 2,800,000 800.000 2,097,000 207,000 100,500 100,000 100,000 100,000 100,000 100,000 100,000 100,000 100.000 100.000 542,000 2,298,000 6,000 Engraved. 9.000 Engraved. 3,000 Engraved. 9,000 Engraved. 3,000 Engraved. 6.000 Engraved.., 0,000 Engraved... 2.070 Engraved .. 9,000 Engraved... 930 Engraved... 196.000 Engraved... 56,000 Engraved... 146,790 Engraved... 14.490 Engraved... 7,035 Engraved... 8,000 Engraved... 8.000 Engraved.., 8.000 Engraved... 8,000 Engraved... 8,000 Engraved .. 8,000 Engraved... 8,000 Engraved... 8.000 Engraved... 8,000 Engraved... 37,940 Engraved... 137,880 Engraved... $ 10,644,500 % 719,135 J. B. Trippe, Treasurer J. B. Trippe, Treasurer.. .... J. B Trippe, Treasurer John Jones, Treasurer John Jones, Treasurer John Jones, Treasurer John Jones. Treasurer John Jones, Treasurer John Jones. Treasurer 00 John Jones. Treasurer., 00 W. H. Scott, for Treasurer 50 B. A. White, for Treasurer 00 Madison Bell, Comp. General.. 00N L. Angier, Treasurer 50.N, L Angier, Treasurer 00 John Jones, Treasurer (Hi John Jones, Treasurer OO! John Jones, Treasurer 00 John Jones, Treasurer 00 John Jones, Treasurer 00 John Jones, Treasurer 00 John Jones. Treasurer 00 John Jones, Treasurer 00 John Jones, Treasurer 00J W. Renf roe. Treasurer. , 00 J. W. Renfroe, Treasurer Time for Payment of Coupons. Where Payable. Authority for Issue. For What Purpose Issued. Signers of Bonds. February and August... New York and State Treasury.. lActof 27thof February, 1856 February and August... New York and State Treasury.. Act of 27th of February, 1856 Atlantic and Gulf Railroad.. Atlantic and Gulf Railroad Joseph E. Brown, Gouernor; E. P. Watkins, Secretary of State. Joseph E. Brown, Governor; E. P. WTatkins, Secretary of State. February and August... New York and State Treasury.. Act of 27th of February 1856 Atlantic and Gulf Railroad Joseph E. Brown, Governor; E. P. Watkins, Secretary of State. February and August... New York and State Treasury.. Act of 27th of February 1856 Atlantic and Gulf Railroad...... Joseph E. Brown, Governor, E. P. Watkins, Secretary of State. February and August... New York and State Treasury.. Act of 27th of February 1856 Atlantic and Gulf Railr. ad Joseph E. Brown, Governor; E. P. Watkins, Secretary of State. February and August... New York and State Treasury.. Act of 27th of February 1856 Atlantic and Gulf Railroad Joseph E. Brown, Governor; E. P. Watkins, Secretary of State. February and August... New- York and State Treasury.. Act of 27 th of February 1856 Atlantic and Gulf Railroad Charles J. Jenkins. Governor; Nathan C. Barnett, Secretary of State. February and August... New York and State Treasury.. Act of 27th of February, 1856 Atlantic and Gulf Railroad Charles J. Jenkins, Governor; Nathan C. Barnett, Secretary of State. February and August... New York and State Treasury.. Act of 27th of February 1856 Atlantic and Gulf Railroad Charles J. Jenkins, Governor; Nathan C. Barnett, Secretary of State. February and August... New York and State Treasury.. Act of 27th of February. 1856 Atlantic and Gulf Railroad Charles J. Jenkins, Governor; Nathan C. Barnett, Secretary of State. January and July New York and State Treasury.. Act of 12th of March, 1866 Renewal of old Bonds, etc Charles J. Jenkins, Governor; John T. Burns, Comptroller General. January and July New York and State Treasury. Act of 12th of March, 1866 Renewal of old Bonds, etc Charles J. Jenkins, Governor; JohnT. Burns, Comptroller General. -Jan. April, July and Oct. New York and State Treasury.. Act of 15th of Sept., 1870 Renewal of old Bonds, etc Rufus B. Bullock, Governor; David G. Cotting, Secretary of State. January and July New- York and State Treasury.. Act of 18th of January. 1873 Renewal of old Bonds, etc James M. Smith, Governor; David G. Cotting, Secretary of State. January and July New York and State Treasury., Act of 18th of January, 1878 Renewal of old Bonds, etc James M. Smith, Governor; David G. Cotting, Secretary of State. April and October New York and State Treasury. Act of 19th of February 1873 ... Renewal of old Bonds, etc James M. Smith, Governor; Nathan C. Barnett, Secretaiy of State. April and October New York and State Treasury. Act of 19th of February 1873 Renewal of old Bonds, etc James M. Smith, Governor; Nathan C. Barnett, Secretary of State. April and October New- York and State Treasury.. Act of 19th of February 1873 Renewal of old Bonds, etc James M. Smith. Governor; Nathan C. Barnett, Secretary of State. April and October April and October New York and State Treasury., New- York and State Treasury. Act of 19th of February, 1873 Act of 19th of February wrs Renewal of old Bonds, etc Renewal of old Bonds, etc James M. Smith, Governor; Nathan C. Barnett, Secretary of State. James M. Smith, Governor; Nathan C. Barnett, Secretary of State. April and October New York and State Treasury. Act of 19th of February 1873 Renewal of old Bonds, etc James M. Smith, Governor; Nathan C. Barnett, Secretary of State. April and October New York and State Treasury.. Act of 19th of February 1873.... Renewal of old Bonds, etc James M. Smith, Governor; Nathan C. Barnett, Secretary of State. April and October New York and State Treasury. Act of 19th of February 1873 Renewal of old Bonds, etc James M. Smith, Governor; Nathan C. Barnett, Secretary of State. April and October New York and State Treasury.. Act of 19th of February 1873 .... Renewal of old Bonds, etc James M. Smith, Governor; Nathan C. Barnett, Secretary of State. January and July New York and State Treasury. Act of 24th of February 1876 To pay int. on M.& B..N.& S.Bs James M. Smith, Governor; Nathan C. Barnett, Secretary of State. January and July New York and State Treasury., Act o* 19th of February 1877 To fund M. & B , N. & S., M, Alfred H. Colquitt, Governor; Nathan C. Barnett, Secretary of State. Branch Bonds The following numbers of Gold Quarterly Coupon Bonds are outlawed by the Act approved August 23. 1872. viz : 797, 798, 1,010*, 1,629, 1,6 ), l.< , 1,741, 1,771, 1,772, 1,773, 1,774, 1,791, 1,792, 1,811, 1,825, 1,826,1,840, 2,501 to 2,575, both inclusive, 2,591, 2,592, 2,593, 2,594, 2,595, 2,596, 2,597, 2,598, 2,599, and 2,600. Bond No. 1,010 has been paid, by authority of an Act approved 1877. ^MBH ^" ** m^HUHHUSp ***** m UNIVERSITY OF GEORGIA UBBARIES VHMBHBMBM

https://dlg.usg.edu/record/dlg_ggpd_y-ga-bt800-b-pa1-b1877/fulltext.text

IJMS | Free Full-Text | Nectar Chemistry or Flower Morphology—What Is More Important for the Reproductive Success of Generalist Orchid Epipactis palustris in Natural and Anthropogenic Populations? The aim of this study was to determine the level of reproductive success (RS) in natural and anthropogenic populations of generalist orchid Epipactis palustris and its dependence on flower structure and nectar composition, i.e., amino acids and sugars. We found that both pollinaria removal and female reproductive success were high and similar in all populations, despite differences in flower traits and nectar chemistry. Flower structures were weakly correlated with parameters of RS. Nectar traits were more important in shaping RS; although, we noted differentiated selection on nectar components in distinct populations. Individuals in natural populations produced nectar with a larger amount of sugars and amino acids. The sucrose to (fructose and glucose) ratio in natural populations was close to 1, while in anthropogenic ones, a clear domination of fructose and glucose was noted. Our results indicate that the flower traits and nectar composition of E. palustris reflect its generalist character and meet the requirements of a wide range of pollinators, differing according to body sizes, mouth apparatus, and dietary needs. Simultaneously, differentiation of nectar chemistry suggests a variation of pollinator assemblages in particular populations or domination of their some groups. To our knowledge, a comparison of nectar chemistry between natural and anthropogenic populations of orchids is reported for the first time in this paper. Nectar Chemistry or Flower Morphology—What Is More Important for the Reproductive Success of Generalist Orchid Epipactis palustris in Natural and Anthropogenic Populations? by Emilia Brzosko * , Andrzej Bajguz * , Justyna Burzyńska and Magdalena Chmur Faculty of Biology, University of Bialystok, Ciolkowskiego 1J, 15-245 Bialystok, Poland Authors to whom correspondence should be addressed. Int. J. Mol. Sci. 2021 , 22 (22), 12164; https://doi.org/10.3390/ijms222212164 Received: 5 October 2021 / Revised: 6 November 2021 / Accepted: 8 November 2021 / Published: 10 November 2021 (This article belongs to the Special Issue Orchid Biochemistry 2.0 ) Abstract The aim of this study was to determine the level of reproductive success (RS) in natural and anthropogenic populations of generalist orchid Epipactis palustris and its dependence on flower structure and nectar composition, i.e., amino acids and sugars. We found that both pollinaria removal and female reproductive success were high and similar in all populations, despite differences in flower traits and nectar chemistry. Flower structures were weakly correlated with parameters of RS. Nectar traits were more important in shaping RS; although, we noted differentiated selection on nectar components in distinct populations. Individuals in natural populations produced nectar with a larger amount of sugars and amino acids. The sucrose to (fructose and glucose) ratio in natural populations was close to 1, while in anthropogenic ones, a clear domination of fructose and glucose was noted. Our results indicate that the flower traits and nectar composition of E. palustris reflect its generalist character and meet the requirements of a wide range of pollinators, differing according to body sizes, mouth apparatus, and dietary needs. Simultaneously, differentiation of nectar chemistry suggests a variation of pollinator assemblages in particular populations or domination of their some groups. To our knowledge, a comparison of nectar chemistry between natural and anthropogenic populations of orchids is reported for the first time in this paper. Keywords: floral display ; fruiting ; marsh helleborine ; nectar amino acids ; nectar sugars ; pollinaria removal 1. Introduction To achieve the highest possible reproductive success, plants have evolved different strategies. In animal pollinated plants, the strategies are directed at relations with pollinators. The masters in building the most specialized interaction with their pollinating partners are representatives of Orchidaceae. The majority of them are specialists connected to only one pollinator species (67% of all orchids) or a single functional group [ 1 , 2 , 3 , 4 ]. On the opposite point of the continuum of the specialization–generalization scale are generalists, pollinated by a wide range of animals from different systematic and ecological groups. An example of the last group is the object of the present study of Epipactis palustris , which is pollinated by more than 100 species [ 5 , 6 ]. To attract pollinators, orchids adapted their flowers structurally and chemically. Many of them (30–40% species) have developed deceptive tactics (mainly food or sexual deception) [ 7 , 8 , 9 , 10 , 11 ]. The important part of Orchidaceae constitutes rewarding species, which reward pollinators through different attractants, such as nectar, fragrances, oils, resin, and wax [ 12 ]. The first of them is the most effective for pollination success in orchids [ 13 ]. Although the role of the presence of nectar for the reproductive success (RS) of orchids is unquestionable [ 9 , 11 , 13 , 14 ], its quantity and quality for pollination effectiveness are documented only for some species [ 15 , 16 , 17 , 18 , 19 ]. Most studies on nectar in orchids, although valuable, only reported about the presence of sugars without ratios between them, or even did not distinguish between the sugars in floral and extrafloral nectar [ 20 , 21 ]. Nevertheless, studies on other plants well document the great variation of nectar properties in different species, distinct populations of a given species, dependence on habitat, flower position on inflorescence, flower age, and other factors. One of the most important findings, due to an evolutionary point of view, is that nectar produced by a given plant species meets the requirements of their pollinators. Relationships between nectar properties and pollinator types confirm many studies [ 22 , 23 , 24 , 25 , 26 ]. Pollinators’ requirements of nectar properties are connected with their body size and behavior (energetic needs), the possibility for them to acquire nectar (mouth apparatus), and gustatory (taste caused by some amino acids (AAs)) [ 26 , 27 , 28 , 29 , 30 , 31 ]. Preferences of pollinators concern both nectar concentration, sugar proportion, and amino acids composition. For example, bats and hawkmoths feed on the nectar of lower concentrations of sugars, while bees prefer a higher concentration [ 22 , 32 , 33 ]. The concentration of sugars in orchid nectar sits within a wide range, from a low percentage to 90% [ 34 ]. Different pollinators also show distinct preferences to the ratio of main sugars, i.e., saccharose, glucose, and fructose. The extreme example of pollinators preferences to sugar components are some nectarivorous birds and ants, which prefer sucrose-free nectar due to their physiological constrains—the lack of invertase prevents them from attaining sucrose assimilation [ 35 ]. Pollinators also select nectar depending on its AA composition. Butterflies choose nectar with high AA concentration, while birds or flies prefer those with lower concentration [ 28 ]. Moreover, in the nectar of different species, distinct compositions of AAs were noted—with the domination of some AAs combined with lower concentrations or even absence of others [ 24 , 36 , 37 ]. The use of nectar by pollinators depends not only on its composition but also on its availability. In orchids, nectar is accumulated in shallow, cup-like structures, at the base of the labellum, in long spurs, in the base of the flower alongside the ovary, and on the side-lobes or along the central groove of the labellum [ 38 ]. Nectar located inside the corolla or in the spur is available for specific, restricted groups of pollinators with longer mouth apparatus, while exposed nectar is available for a wide range of pollinators, differing with respect to body sizes and dietary requirements. Exposed nectar is more vulnerable for evaporation and robbery than nectar located in deeper parts of flowers [ 24 , 37 , 39 ]. Moreover, nectar accumulated in open nectaries is often dominant in hexoses (e.g., fructose and glucose), while in concealed nectaries is most often sucrose dominant [ 21 , 24 , 36 , 40 ]. Flower structure plays an important role in shaping relationships between plants and their pollinators; therefore, structure shows adaptation to pollinating animals. The mutual match between pollinator and flower traits is the result of phenotypic selection [ 41 , 42 ]. Despite the fact that the general architecture of orchid flowers has the same scheme, details are differentiated in particular representatives of the family [ 12 ] that is strictly connected with the pollinator’s properties. One of the best examples of a match between flowers and pollinators are spurred orchids, for which pollinator-mediated selection on flower traits—especially nectar spur length and corolla tube width—are well documented [ 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 ]. Studies on such species show that increasing the mechanical fit between flower and pollinator increases the precision of pollen transfer, thus affecting plant fitness [ 51 ]. Pollinator-mediated selection on flower traits is also documented by studies on deceptive orchids [ 52 , 53 , 54 , 55 , 56 ]. The application of orchids’ distinct reproductive strategies translates into their level of reproductive success (RS). For example, rewarded species achieve higher RS than deceptive ones, and among rewarding species, those which produce nectar have the best effectiveness of pollination [ 7 , 11 , 13 ]. Many data document that reproductive success in orchids is strictly related to an important component of reproductive strategies—the flower’s properties [ 42 , 46 , 49 , 56 ]. Generally, orchids are known as a group with a relatively low fruit set, especially non-autogamous species, mainly due to their limited pollinators [ 11 , 13 , 57 ]. Pollinator deficiency is often noted in anthropogenic populations [ 58 ]. Under such circumstances, increasing competition for pollinators may cause intensification of selection on floral traits by increasing pollen limitation [ 51 , 59 , 60 , 61 ]. Anthropogenic habitats also offer distinct soil resources, which can shape plant traits such as their size, flower production, or nectar quantity and quality. The dependence of these traits on soil parameters is well documented in orchids [ 16 , 17 , 18 , 19 ]. Differentiation of the above-mentioned factors causes spatial and temporary variation of reproductive success [ 11 , 13 , 57 , 62 , 63 , 64 ]. Some orchids, including E. palustris , may colonize different types of secondary habitats [ 65 , 66 , 67 , 68 ]. This presents the opportunity for maintaining orchids’ diversity, since they are one of the plant groups that are the most sensitive to habitat loss and destruction due to human activity; therefore, they belong to the most endangered plant groups [ 69 , 70 ] with different threat levels of particular species [ 71 ]. The extinction risk of all known orchid species is c.a. 47% [ 72 ], and, as an example, 25% of globally extinct orchids are Australian [ 73 ]. Kull and Hutchings [ 74 ] compared changes in orchids distribution in the United Kingdom and Estonia and found that the mean decline in distribution range for 49 species in the United Kingdom was 50% and the mean decline for 33 orchid species in Estonia was 25%. Similar trends were observed by Jacquemyn, et al. [ 75 ] in Flanders and the Netherlands, where during 70 and 50 years, respectively, 81% species decreased distribution range in Flanders, and 78% species decreased distribution area in the Netherlands. Moreover, few species in each area went extinct. Reduction of distribution area and population number in Europe is noted for the object of our studies, E. palustris [ 67 ]. The threat to orchids is strengthened by the global decline of many plant pollinators, including those crucial for the pollination of orchids [ 58 , 76 ]. For example, the 25% loss of honey bee colonies in Central Europe between 1985 and 2005 has been observed [ 58 ]. The main aim of our study was to evaluate the level of pollinaria removal and fruiting and to determine the role of flower structure and nectar composition in shaping RS in natural and anthropogenic E. palustris populations. We supposed that RS should be high, due to the following traits of this orchid: (a) as a generalist, it is pollinated by a wide range of pollinators; (b) self-compatible properties enable autogamous and geitonogamous pollination; (c) the presence of nectar enhances the probability of pollination. We also hypothesized the differentiation of nectar characteristics and flower properties between populations, especially between natural and anthropogenic ones. The answer to the question “what is more important for the reproductive success of generalist orchid E. palustris in natural and anthropogenic populations—nectar components or flower morphology?” can help elucidate the evolutionary pathways of different floral traits. Moreover, although the importance of nectar for RS is unquestionable, only a few studies document the role of nectar composition for RS in orchids. Additionally, to our knowledge, this is the first paper where a comparison of nectar chemistry between natural and anthropogenic populations of orchids has been reported. 2. Results 2.1. Floral Display and Flower Structure E. palustris populations differ significantly in all floral display parameters, i.e., shoot height, inflorescence length, and flower number ( Table 1 and Table S1 , Figure S1 ). The highest shoots were observed in the natural ZAB population (62.6 ± 16.1 cm) and the lowest in the anthropogenic SIL population (42.46 ± 7.4 cm). Inflorescence length and number of flowers per shoot was the highest in the anthropogenic SOP population. In ZAB and SIL populations, all floral display traits were monotonically correlated to each other (r s = [0.38, 0.74]; i.e., r s = 0.74 for length of inflorescence vs. shoot height, r s = 0.58 for length of inflorescence vs. shoot height, respectively), while in SOP, the length of inflorescence and number of flowers depended on shoot height (r s = 0.65 and r s = 0.41, respectively). In ROS, statistically significant correlation was found between inflorescence length and number of flowers (r s = 0.53) ( Table S2 ). Table 1. Variation of floral display and flower structure in Epipactis palustris natural (Nat.) and anthropogenic (Ant.) populations. Data ( n = 30) represent the mean ( x ¯ ) ± standard error (SE), lower quartile (Q 1 ), median (Q 2 ), upper quartile (Q 3 ), and interquartile range (IQR). Different lowercase letters indicate statistically significant differences according to Tukey’s post-hoc test ( p < 0.05). Different uppercase letters indicate statistically significant differences according to the pairwise Wilcoxon Rank Sum test with Benjamini–Hochberg adjustment ( p < 0.05). Additional comparisons were shown only when populations within Nat. or Ant. (or both) do not differ significantly. All measured flower traits differed between populations ( Table 1 , Figure S1 ). The smallest flowers (both their length and width) were noted in natural ROS populations (length of flowers (FH): 10.34 ± 0.56 mm; width of flowers (FW): 20.75 ± 1.34 mm), while the largest in the SOP population (FH: 11.9 ± 1.2 mm and FW: 24.3 ± 2.0 mm). Values of other traits most often shaped according to the same pattern as FH and FW. It should be highlighted that the isthmus area (AI), on which surface nectar is secreted, was larger in anthropogenic (especially in SOP) populations than in natural populations. Spearman’s correlation analysis revealed that almost all flower traits in SOP correlated positively with each other strongly or very strongly ( Table S2 ). In the remaining 3 populations, FH was always correlated with LDS and LP (r s = [0.72, 0.96]), and AI with LI and WI (r s = [0.71, 0.85]). Furthermore, the flower structure dataset was subjected to principal component analysis (PCA) and its preliminary tests. The p -value from Bartlett’s test of sphericity was approximately equal to 0, while the calculated overall measure of sampling adequacy (MSA) from the Kaiser–Meyer–Olkin test was equal to 0.84. MSA for individual parameters ranged from 0.48 (for width of isthmus (WI)) to 0.96 for the width of flowers (FW) ( Table S3 ). Thus, according to Kaiser [ 77 ], the MSA value is high enough to perform PCA. According to Cattell’s rule, one or two components should be selected ( Figure S2 ) [ 78 ], while Kaiser’s rule indicates that three components should be retained [ 79 ]. On the basis of the first axis (Dim1), which accounts for 53.3% of the variation, a separation of all four populations is visible—the following pattern: SOP > ZAB > SIL > ROS usually occurs for all floral structure parameters. SOP and ZAB are mostly associated with positive values of Dim1 (thus higher than average values of floral parameters), while SIL and ROS– are mostly associated with negative values of Dim1. Thus, a sign-based distinction between natural and anthropogenic populations is not possible ( Figure 1 and Figure S3 ). Figure 1. Biplot of flower structure profiles for Epipactis palustris natural (Nat.) and anthropogenic (Ant.) populations, showing the first two dimensions or factors (Dim1-2) of PCA that, together, explain 63% of the variance. Biplot vectors indicate the strength and direction of factor loading for the first two factors. Individuals (populations) are color-coded by population. 2.2. Nectar Chemistry 2.2.1. Sugars Our analyses document very low E. palustris nectar amounts of three common sugars, i.e., sucrose, fructose, and glucose. We found statistically significant differences between populations in sugars quantity (sum of sugars), excluding glucose content. The total amount of sugars was significantly lower in anthropogenic than in natural populations (SIL: 34.05 mg/mL and SOP: 35.0 mg/mL vs. ZAB: 48.09 mg/mL and ROS: 40.68 mg/mL) ( Table S4 ). Participation of sucrose in nectar was also significantly lower in anthropogenic than in natural populations ( Figure 2 , Table S5 ). On the other hand, the sucrose to (fructose and glucose) ratio was more balanced in natural ZAB and ROS populations (0.93 and 0.86, respectively), while in SIL and SOP anthropogenic ones, a clear domination of fructose and glucose was found (0.57 and 0.58, respectively). No statistically significant differences in fructose to glucose ratios were found among populations ( Table S4 ). Figure 2. Boxplots of sugar amounts for Epipactis palustris natural (Nat.) and anthropogenic (Ant.) populations. Colored dots are individual samples. The crossed square shows the mean. The lower and upper hinges correspond to the lower (Q 1 ) and upper (Q 3 ) quartiles. Thus, box length shows the interquartile range (IQR). The thicker lines inside the boxes corresponds to the median. The lower whisker extends from the hinge to the smallest value at most Q 1 − 1.5 × IQR of the hinge. The upper whisker extends from the hinge to the largest value no further than Q 3 + 1.5 × IQR. Data beyond the end of the whiskers, indicated with an asterisk symbol, are outliers. Different lowercase letters indicate statistically significant differences according to Tukey’s post-hoc test ( p < 0.05). Symbol “=” means they did not differ significantly. Additional comparisons on the left or right side were shown only when the Nat. or Ant. (or both) populations did not differ significantly. 2.2.2. Amino Acids The amount of AAs in E. palustris nectar ranged from 0.39 ± 0.002 mg/mL in SOP to 0.52± 0.002 mg/mL in ZAB. Statistically significant differences between populations were noted in the sum of AAs, and the largest differences were observed between natural and anthropogenic populations ( Table 2 and Table S6 , Figure S4 ,). In total, 27 distinct AAs were detected in E. palustris nectar (20 proteogenic and 7 non-proteogenic) with their different participation in particular populations. Nevertheless, some of them dominated in all populations (glutamic acid (Glu) and glutamine (Gln)—always above 10%). Glu, tyrosine (Tyr), arginine (Arg), and β-alanine (β-Ala) had a significantly higher percentage in natural than in anthropogenic populations. On the other hand, in anthropogenic populations, participation of proline (Pro), alanine (Ala), and phenylalanine (Phe) in nectar was higher than in natural places. It should be noted that β-Ala was observed only in natural populations (ZAB and ROS), citrulline (Cit) only in one natural population (ZAB) (but only in some individuals), and Tau was absent in one anthropogenic population (SOP). It is interesting that anthropogenic populations were characterized by a higher percentage of proteogenic AAs in nectar than natural ones, while non-proteogenic AAs had higher participation in natural populations. Table 2. The concentration of amino acids (μM) and total amount of amino acids (mg/mL) in Epipactis palustris nectar. The number of classes represents the effect of amino acids on insect chemoreceptors: I—no effect; II—inhibition of chemoreceptors; III—stimulate the salt cell; IV—the ability to stimulate the sugar cell. Data ( n = 30) represent the mean ( x ¯ ) ± standard error (SE), lower quartile (Q 1 ), median (Q 2 ), upper quartile (Q 3 ), and interquartile range (IQR). Different lowercase letters indicate statistically significant differences, according to Tukey’s post-hoc test ( p < 0.05). Different uppercase letters indicate statistically significant differences according to the pairwise Wilcoxon Rank Sum test with Benjamini–Hochberg adjustment ( p < 0.05). ND—not detected. Additional comparisons were shown only when populations within Nat. or Ant. (or both) do not differ significantly. In natural populations, strong monotonic correlations were found, i.e., leucine (Leu) vs. isoleucine (Ile) (r s = {ZAB: 0.75, ROS: 0.74}), ornithine (Orn) vs. glutamine (Gln) (r s = {ZAB: −0.78, ROS: 0.60}), taurine (Tau) vs. Orn (r s = {ZAB: 0.55, ROS: 0.63}), and Tau vs. Gln (r s = {ZAB: −0.60, ROS: 0.68}). Additionally, in ZAB, a correlation between methionine (Met) and lysine (Lys) was noted (r s = −0.64), while in ROS, r s = 0.67 was reported for tryptophan (Trp) vs. threonine (Thr) and Orn vs. glycine (Gly). In anthropogenic populations, no common strong or very strong correlations were reported. However, in the case of SIL, strong monotonic correlations (r s = [0.60, 0.79]) were noted between the following: asparagine (Asn) vs. serine (Ser), histidine (His) vs Asn, Ile vs. alanine (Ala), valine (Val) vs. Leu and Trp, as well as Orn vs His. While, in case of SOP, strong monotonic correlations (r s = [0.60, 0.79]) were noted between the following: Arg vs. Ala, Trp vs. Leu and Met, as well as Orn vs. Lys. It should be also highlighted that, between natural and anthropogenic populations, no intersection of strongly or very strongly correlated AA pairs exist ( Table S7 ). Different relations between production of sugars and AAs in particular populations was noted. In both natural populations, the sum of AAs positively correlated with the sum of sugars (ZAB: r s = 0.43; p < 0.05 and ROS: r s = 0.40; p < 0.05), in ZAB with fructose and sucrose amount (r s = 0.44; p < 0.05 and r s = 0.38; p < 0.05), and in ROS with sucrose amount (r s = 0.44; p < 0.05). In anthropogenic populations, positive correlations between AAs amount and percentage of hexoses (i.e., sum of fructose and glucose) were observed (SIL: r s = 0.44; p < 0.05 and SOP: r s = 0.38; p < 0.05) and negative correlations were observed between AAs amount and percentage of sucrose (SIL: r s = −0.44; p < 0.05 and SOP: r s = 0.38; p < 0.05). We found a notable difference between natural and anthropogenic populations in participation of AA from distinct taste classes ( Figure 3 ). The percentage share of class II AAs was approximately 35–48% for natural populations and 48–56% for anthropogenic populations, while class IV was 36–42% for SOP and 42–48% for SIL. The class II of AAs had higher participation in natural populations. On the other hand, the class III group, represented in E. palustris nectar only by Pro, had about five times higher participation in anthropogenic populations than in natural populations ( Table 2 ). Figure 3. Ternary plot of amino acid classes for Epipactis palustris natural (Nat.) and anthropogenic (Ant.) populations: II (Asp, Glu, His, Arg, Lys), III (Pro), and IV (Val, Met, Trp, Phe, Ile, Leu). Blue lines show 50%, 90%, and 95% confidence intervals via the Mahalanobis Distance and use of the Log–Ratio Transformation. The first class of AAs (Asn, Gln, Ala, Cys, Gly, Ser, Thr, Tyr) does not affect the chemoreceptors of fly (data not shown). AAs’ abbreviations and full names are present in Table 2 . Moreover, AAs were subjected to principal component analysis (PCA) and its preliminary tests. The p -value from Bartlett’s test of sphericity was approximately equal to 0, while the calculated overall measure of sampling adequacy (MSA) from the Kaiser–Meyer–Olkin test was equal to 0.95 ( Table S8 ). MSA for individual AAs ranged from 0.44 (Phe was the only AA with almost no interpopulation differences and very high data deviation, Figure S2 ) to 0.98 for Tyr, Trp, and Val ( Table S8 ). Thus, according to Kaiser [ 77 ], the MSA value is high enough to perform PCA. According to Cattell’s rule, one or two components should be selected ( Figure S5 ) [ 78 ], while Kaiser’s rule indicated that three components should be retained [ 79 ]. Finally, the first two components that explain about 75.5% of the variance were preserved. PCA grouped together anthropogenic populations (SIL and SOP), as they had much higher average Pro level and lower levels of other AAs ( Figure 4 ). Differences between ZAB and ROS populations are also visible, e.g., much higher average Cit, His, β-Ala, and Ile levels, as well as lower Arg, Asn, Trp, β-aminobutyric acid (BABA), and Lys. On the basis of the first axis (Dim1), which accounts for 64.1% of the variation, there is a clear separation of the natural vs. anthropogenic populations, particularly due to differences in Pro. Based on the second axis (Dim2, 11.5%), the two natural (Nat.) populations are also separated, while the anthropogenic (Ant.) are not ( Figure 4 and Figure S6 ). Figure 4. Biplot of amino acid profiles for Epipactis palustris natural (Nat.) and anthropogenic (Ant.) populations, showing the first two dimensions or factors (Dim1-2) of PCA that, together, explain 75.52% of the variance. Biplot vectors indicate the strength and direction of factor loading for the first two factors. Individuals (populations) are color-coded by population. Ellipses around the individuals show assumed 95% multivariate normal distribution. 2.3. Reproductive Success Reproductive success in E. palustris populations was high ( Table 3 ). Female reproductive success (FRS—the proportion of developed fruits to the number of flowers on the inflorescence) shaped from 81.47 ± 4.19% in ROS to 90.60 ± 2.49% in SOP ( Table 3 ) and did not differ between populations (F = 0.862; p = 0.46). Pollinaria removal (PR) was also similar in all populations (F = 1.289; p = 0.28); although, activity of insects was about 10% higher in SOP (96.55 ± 1.61%) than in other populations ( Table 3 ). The efficiency of pollination was high—PR to FRS ratio was equal to about 1 in all populations. Although average values of indexes of reproductive success are similar in the populations studied, we observed some differences in details of the pollination process at an individual level. In ROS, SIL, and SOP populations, about one third of individuals (11, 9, and 9, respectively) had higher PR than FRS, while in ZAB only five of them had PR larger than FRS. On the other hand, we noted higher FRS than PR in ROS and SIL (7 and 9 individuals, respectively). In ZAB, only two shoots had higher fruiting than PR, while in SOP, we did not observe such cases. Table 3. Spatial and temporal variation of female reproductive success (FRS) and pollinaria removal (PR) in Epipactis palustris populations. Data ( n = 30) show the mean ± standard error. 3. Discussion In line with our expectations and the results of earlier studies [ 5 , 67 , 80 , 81 ], high levels of PR and FRS (above 80%) were found in both natural and anthropogenic E. palustris populations. Jacquemyn, et al. [ 67 ], on the basis of Claessens and Kleynen [ 5 ] data from 24 populations of this species, reported that the average fruit set shaped at 77.6% %, and Jacquemyn and Brys [ 82 ] noted 70% fruiting in Belgian populations. We also found the high RS and pollination efficiency (in all places PR to FRS ratio equaled about 1). This result contrasts with the founding of Jacquemyn and Brys [ 82 ], who noted that fruiting in E. palustris populations was higher than the level of pollinaria removal. Although pollinator deficiency is considered the main factor restricting RS in orchids [ 11 , 13 , 57 ], a high level of RS and pollination efficiency in our studies suggest that pollinators of E. palustris are abundant in all populations. Nevertheless, assemblages of insects and the dominant pollinators may differ from one part of geographical range to another [ 5 , 6 , 83 ]. High number of pollinators of this orchid (142 species [ 5 ]) and a wide range of their sizes and requirements increase the probability of pollination. Through a diversity of potential pollinators, we can also explain the lack of differences in RS between natural and anthropogenic populations. Our results indicate that in each of them, pollinators assemblages are large and diverse enough to ensure RS at the observed level. This result contrasts with the results of other studies, where fruiting was lower in anthropogenic than in natural populations. Exceptionally low levels of fruiting were observed by Jermakowicz and Brzosko [ 59 ] in anthropogenic populations of Malaxis monophyllos . On the other hand, Pellegrino and Bellusci [ 84 ] noted an almost seven times lower fruit set in anthropogenic than in natural populations of Serapias cordigera in Italy. In a population of Oncidium ascendens from rainforest from Mexico, fruit production was almost two times higher than in populations from synanthropic habitats [ 85 ]. The authors of these studies recognized that pollinator deficiency in altered habitats was the main factor, which decreased RS in these species. In our opinion, differences between species characters of orchids studied by Pellegrino and Bellusci [ 84 ] and Parra-Tabla, et al. [ 85 ] and E. palustris could also cause distinct answers for habitat types. S. cordigera is deceptive species and relies on relatively restricted groups of pollinators in comparison with E. palustris ; additionally, O. ascendens is a self-incompatible species, whose sexual reproduction depends on cross-pollination by the native bee Trigona nigra . It could be suggested that the properties of E. palustris (its generalist character, presence of nectar, and spontaneous autogamy) and pollinators behavior (penetration of many flowers on inflorescence) are advantages, which ensure effective pollination regardless of habitat. Anthropogenic habitats are generally recognized as those with poorer assemblages of pollinators, which negatively influences plant RS, but some of them seem to be suitable for plant–pollinator interactions. For example, Rewicz, et al. [ 86 ] reported higher RS in anthropogenic than in natural populations of E. helleborine , due to the larger diversity of pollinators in the first type of habitat. Although two E. palustris populations exist within the city border, in changed places, within populations’ area, and in neighboring communities, other flowering species grew, which can attract many insects, including E. palustris pollinators. Moreover, allotments are placed in the vicinity of SIL population, which may increase pollinator numbers. In the light of our results, it seems that generalists are less sensitive for pollinator deficiency, even in anthropogenic habitats. This is in accordance with findings of other authors, who state that a decrease of fruit set as a result of the reduction in insect movements is particularly strong for specialists that show a high degree of dependence on their pollinators for fruit production [ 11 , 84 , 87 ]. Higher specialization levels and anthropogenic declines in pollinator populations can also intensify selection on floral traits [ 51 ]. The generalist character of E. palustris and pollinator efficiency could explain the very weak selection on flower and floral display traits. Only ten correlations between them and parameters of reproductive success were observed (among 152 tested cases), and only four of these concerned flower structures. Five statistically significant correlations were found in the anthropogenic SOP population and all of them were negative. In this population, lower individuals with shorter inflorescences and a lower number of flowers were favored. This can reflect the behavior of pollinators in this locality. First, they may operate at the lower part of vegetation, and secondly, penetrating a given inflorescence, they are able (or need) to acquire nectar from a restricted number of flowers. On the other hand, in ZAB, fruiting was higher on longer inflorescences. In this place, E. palustris grows in tall sedges, and probably shoots should be higher than neighboring plants to be recognized by pollinators. Stronger pollinator-mediated selection on inflorescence was noted in taller than in shorter vegetation [ 46 , 88 , 89 ]. The case of ZAB population is also in accordance with the common expectation that more fruits often develop on larger inflorescences because they attract more pollinators, which visit more flowers on larger inflorescences [ 48 , 90 , 91 , 92 ]. On the other hand, the SOP population may illustrate situations, that smaller inflorescences are favored by natural selection when larger inflorescences suffer factors decreasing fitness, such as the higher probability of geitonogamy or intense herbivore activity [ 92 , 93 , 94 ]. One of the most important evolutionary mechanisms, crucial for successful pollination, is the mechanical fit between plants and their pollinators [ 2 , 7 , 8 , 41 , 95 , 96 ]. Such a match is generally stronger in specialized systems [ 51 , 97 ], which confirms, for example, the results of studies on long-spurred orchids [ 41 , 45 , 47 , 48 ]. These findings confirm the results of our studies because only four distinct flower traits influenced RS in three among four populations. These traits seem unimportant for pollinators, which may suggest that observed correlations are random or their functions are difficult to explain. The lack of strong selection pressures on these traits maintains variation in flower traits [ 98 ]. Nevertheless, it should be noted that the isthmus area was significantly larger in the anthropogenic population than in the natural population, suggesting that pollinators with distinct mouth apparatus operate in two population groups. The important point, which enables understanding the evolution of plant–pollinator interactions, is knowledge about the importance of floral rewards, including nectar, for RS [ 99 , 100 ]. Nectar properties shape the growth, survival, reproduction, and behavior of nectar-feeding animals [ 30 , 31 , 35 , 101 , 102 ]. Our results suggest stronger dependence of RS in E. palustris populations on nectar properties than flower structure. Nectar characteristics influenced mainly PR (16 statistically significant correlations observed among 21). This is in contrast to our study on another generalist orchid, Neottia ovata , where nectar properties shaped mainly FRS [ 19 ]. Similarly to Percival [ 21 ], we found three main sugars in E. palustris nectar (sucrose, fructose, and glucose) with their amount shown to be larger in natural than anthropogenic populations. Sugar components influenced RS only in the natural ROS population, where PR was positively influenced by hexose (i.e., fructose and glucose) amounts and FRS was positively influenced by fructose amount. This is interesting because in both natural populations, sucrose percentage is significantly higher than in anthropogenic ones. Positive selection on hexoses in ROS may suggest that in this population insects, which prefer nectar rich in monosaccharides are important pollinators, and the amount of hexoses in this population is not enough to provide for their needs. Similar insects could be abundant in anthropogenic populations, where hexoses were more abundant than in natural populations. Preferences for hexoses, taken up more easily than sucrose, show nonspecialized insects, i.e., syrphids, flies, and beetles [ 25 ]. Insects from these groups were noted as E. palustris pollinators [ 6 , 67 , 83 , 99 ]. Nonspecialized insects choose hexose-rich nectar (especially fructose) because it is easier absorbed due to lower viscosity [ 32 ]. Moreover, some ants (often observed by us on E. palustris shoots and noted by Jakubska-Busse and Kadej [ 103 ]) even prefer sucrose-free nectar because they are not able to assimilate this sugar due to lack of invertase [ 29 ]. The lack of selection on nectar sugars in three populations may suggest that these nectar components are not aimed at any of the pollinator group and sugar composition met the requirements of pollinated insects. Similar results were obtained for another generalist orchid, N. ovata [ 19 ]. Different sucrose to (fructose and glucose) ratios (~1 in natural and ~0.5 in anthropogenic populations) suggest distinct pollinator assemblages in these two population groups. Larger sucrose content in natural populations could indicate that such insects as honey bees and bumblebees, which prefer this sugar, are main pollinators in these places. These insects were recognized as main pollinators in some Polish E. palustris populations by Jakubska-Busse and Kadej [ 103 ]. It should be noted that different sugar ratios in natural and anthropogenic E. palustris populations only partially confirms the statement that nectar secreted in open flowers is dominated by glucose and fructose [ 36 , 40 ]. Nectar in particular populations also differed according to AAs composition. Their total amount was higher in natural populations, but proteogenic AAs have larger participation in anthropogenic ones. At the species level, we noted a high number of different AAs (27, including 20 proteogenic and 7 non-proteogenic), similarly to another generalist orchid N. ovata (28 AAs; [ 19 ]). Pais, et al. [ 40 ] found only 17 AAs in E. atropurpurea . Fewer AAs than those in our study were observed in the nectar of specialist spurred orchids [ 15 , 17 ]. Additionally, we found domination of different AAs in the two population groups—Glu, Tyr, Arg, and β-Ala were more abundant in natural populations and Pro, Ala, and Phe were more abundant in anthropogenic populations. The most common AAs in E. palustris nectar are Gln, Glu, and serine (Ser), which are always above 10%. These AAs were also found to be among the most abundant in the nectar of generalist N. ovata [ 19 ]. Gln and Glu are needed for energetically exhaustive flights, while Glu and Ser influence pollinator behavior [ 17 , 18 , 28 ]. Available studies document preferences of pollinators both to total AAs amount in nectar and to particular AAs. Although the importance of AAs for pollinator life is poorly studied, the role some of them are known. First of all, they play a nutritional function and attract or discourage pollinators. One of the most common AAs in plants [ 24 , 104 ]—important for many pollinators, especially Hymenoptera—is Pro, production of which is more expensive than other nectar components [ 105 ]. Its participation constitutes the greatest difference between natural and anthropogenic populations among measured nectar components. In anthropogenic populations, it was one of the three most abundant AAs, and its amount was about five times higher than in natural populations. Like the majority of amino acids, proline can be used in energy production [ 104 ]. This AA rewards pollinators, propels the lift phase of the flight [ 105 , 106 ], and stimulates insects’ salt receptors, which initiate feeding [ 26 , 35 , 107 ]. Carter, et al. [ 105 ] found that Pro accumulation is a plants’ answer to stress factors. Through the last function could be explained a few times, larger amounts of this AA in were found anthropogenic populations than in natural population, since changed habitats are stressful for plants. It can indicate that Pro plays an important role in the metabolism of E. palustris pollinators in anthropogenic populations. Bertazzini, et al. [ 108 ] documented a preference of honey bees for proline-enriched artificial nectar. The other AA, whose amount was larger in the anthropogenic than in the natural population, is Phe. It has a strong phagostimulatory effect on bees and its concentration is highly variable [ 102 , 109 ]. Petanidou, et al. [ 102 ] attributed the dominance of Phe in the Mediterranean to the high number of bees, especially long-tongued bees. The authors suggest that in the Mediterranean, such bees act as crucial selective factors for Phe-rich nectars. The last abundant AA in anthropogenic populations is Ala, which influences insect growth [ 17 ]. Aspartic acid (Asp), like Glu and Ser, influences pollinator behavior [ 17 , 28 ] and disgusts pollinators [ 102 ]. The second importance of Asp may explain its negative influence on PR in ROS. In the same way, a negative correlation between the percentage of Ser and FRS in SIL population could be explained. A negative response of honey bees to Ser was reported [ 108 ]. On the other hand, Kim and Smith [ 110 ] showed that Gly elicited a feeding response in honeybees. Variation in amount and participation of nectar components, and the differentiation of selection on distinct constituents in particular populations suggest, again, that different pollinators with different nutritional needs operate in distinct populations. This supposition could be strengthened by the results showing preferences of pollinators to different taste classes. The most sensitive for nectar taste were pollinators in ROS, where we found positive correlations between PR and percentage of AAs from taste class IV (stimulation the sugar receptor cell), while negative between PR and taste class II (inhibition of the three types of chemosensory cell: salt, sugar, and water). Among the remaining three populations, only in SIL AAs from taste class I positively shaped PR. This confirms the results of earlier studies [ 28 , 111 ], in which it was recognized that nectar amino acids might be detectable by pollinators and may contribute to the overall taste of nectar. We noted the importance of nectar taste for pollinators, and consequently for RS, in other orchids, both in generalist (with open nectaries) and in specialist (with the accumulation of nectar in the spur) [ 15 , 19 ]. The nectar composition can be modified by habitat properties, especially soil nutrients [ 18 , Results of our studies confirm the generalist character of E. palustris . High levels of RS in all populations indicate that both flower traits and nectar chemistry, and variation of these properties meet the needs of wide, differentiated pollinator groups. Simultaneously, the lack of selection on flower traits and stronger selection on nectar components suggest that pollinators are more sensitive to nectar properties, including taste. Moreover, selection on distinct nectar characters in particular populations may indicate that different pollinator assemblages operate within them. The most important finding of our research is documentation (to our knowledge, it is the first such report) of significant differences in nectar properties between natural and anthropogenic orchid populations. We suggest that they are caused by the differentiation of pollinators in these two habitat types or are stronger depending on soil characters (or both). However, to precisely point out the most important factor, more detailed studies should be conducted. The results of our studies importantly enrich the knowledge needed to explain mechanisms, which underlie plant–pollinator interactions. 4. Materials and Methods 4.1. Study Species E. palustris is widely distributed throughout most of Europe but is absent in the Southern Mediterranean regions and extreme north [ 112 ]. This species usually occupies calcareous, nutrient-poor, and moist–wet substrates, mainly in full sunlight. It exists in wet dune slacks, calcareous fens, and on peat, but may also occur on sandy substrates overlying heavy clay or loamy soils [ 67 ]. E. palustris is rhizomatous species. Each shoot has about six leaves in the lower part and a few more scattered ones below the inflorescence. The inflorescence produces about twelve flowers (sometimes more than 20). The flowers are usually more or less tinged with a rose, red, or brown coloration [ 67 , 81 ]. A wide spectrum of insects visits E. palustris flowers, but the main pollinators are Diptera (i.e., Empis sp., and Episyrphus sp.) and Hymenoptera (i.e., Apis mellifera , Bombus lapidarius , and Bombus lucorum ) [ 6 , 83 ]. Claessens and Kleynen [ 5 ] found 142 pollinators of this orchid in the literature. Insects are attracted by strong scent (with eugenol and vanillin, as the crucial components attracting Diptera) and nectar with attractants such as nonanal (pelargonaldehyde), decanal, eicosanol, and its derivatives [ 83 ]. The shallow nectary is located on the labellum, and nectar is secreted on the whole surface of lip callus and abaxial side of isthmus in hypochile [ 113 ]. E. palustris has the potential for spontaneous autogamy [ 67 , 80 ]. Fruiting takes place August–September, depending on location and weather conditions [ 81 ]. 4.2. Study Area This study was performed in July and August 2021 in four populations of E. palustris in northeast Poland, two of which were localized in well-preserved natural peat bogs. ZAB population is localized in the Biebrza National Park—one of the biggest areas of peatlands in Europe. ROS population is localized in Rospuda valley. It is a vast, moss-free, low, and transitional peat bog, with rich and unique flora (four plant species listed in Annex 2 of the EU Habitats Directive, fifteen plant species listed in The Polish Red Book of Plants ). The bog is also the only refuge in Poland of a rare orchid species in Europe— Herminium monorchis . Natural populations are distanced c.a. 70–100 km from Bialystok, the largest city in NE Poland, and at least a few kilometers from the nearest villages. Two other populations (SIL and SOP) exist in anthropogenic habitats at the border of Bialystok city. The SIL population exists in an abandoned gravel pit (c.a. 3 ha). The SOP population exists within the highly damaged soligenic peat bog (0.5 ha), which is under advanced secondary succession, with a lowered level of groundwater and the presence of alien species. Both anthropogenic populations are surrounded by human-changed habitats, typical for urban areas. 4.3. Fieldwork and Floral Trait Measurements In populations studies, 30–32 flowering individuals were chosen and marked. In the field, the floral display traits, i.e., the height of shoots, length of inflorescence, and the number of flowers, were quantified. The five lowest flowers from each inflorescence were collected and used for the evaluation of nectar composition and measurement of the morphological variables of flowers (full names of abbreviations are present in Table 4 ). The area of isthmus (AI), as the product of LI and WI, was amounted. The isthmus size was considered as a measure of nectar quantity. Flower traits of one individual are given as average from five measurements. Table 4. Measured flower structure properties. Samples from all populations were collected 7–12 July during the peak of flowering under sunny and hot weather (the temperature of each day was about 30 °C). The morphological measures were taken using an opto-digital microscope DSX110 (Olympus Life Science, Waltham, MA, USA) in the Laboratory of Insect Evolutionary Biology and Ecology, Faculty of Biology, University of Bialystok. To assess the level of reproductive success (RS), the shoots were marked and the number of flowers per inflorescence in full blooming were counted. During the maturation of capsules, FRS and PR were quantified. FRS was evaluated as the proportion of developed fruits to the number of flowers on the inflorescence and was given in percentages. PR was determined in percentages (PR to the total number of pollinaria for each inflorescence). The efficiencies of pollination were also evaluated, found as the ratio of PR to FRS—the higher the index, the lower the pollination efficiency within a population. 4.4. Nectar Analysis 4.4.1. Nectar Isolation Flower nectar isolation was performed using a water washing method [ 114 ]. Five flowers per sample were placed into a 2 mL Eppendorf tube, containing 1 mL of distilled water and shaken in a laboratory thermomixer (120 rpm, 21 °C, 45 min; Eppendorf Corporate, Hamburg, Germany) for the nectar efflux. Then, the flowers were removed from the tubes, and the mixture of water with nectar was evaporated to dryness by centrifugal vacuum concentrator (45 °C, Eppendorf Concentrator Plus, Eppendorf Corporate, Hamburg, Germany). The obtained pellet was dissolved in 20 µL of distilled water, then transferred into the centrifuge tube with a filter and centrifuged to remove impurities (9000× g , 5 min; MPW-55 Med. Instruments, Gliwice, Poland). The purged extract was collected in a glass vial with a 250 µL insert with polymer feet. 4.4.2. Sugar and Amino Acid Determination Determination and quantification of sugars and AAs were performed using the high-performance liquid chromatography (HPLC) method. An Agilent 1260 Infinity Series HPLC apparat (Agilent Technologies, Inc., Santa Clara, CA, USA) with quaternary pump with an in-line vacuum degasser, a thermostatted column, and a refrigerated autosampler with an autoinjector sample loop was used. For sugar analysis, a ZORBAX Carbohydrate Analysis Column (4.6 mm × 250 mm, 5 µm) (Agilent Technologies, Inc., Santa Clara, CA, USA), at a temperature of 30 °C and a refractive index detector, was applied. The mobile phase was a solution of acetonitrile and water (70:30, v / v ) at a flow rate of 1.4 mL/min. The injection volume was 10 µL. The total time of analysis was 15 min [ 15 ]. Meanwhile, for AA detection ( Table 5 ), an automatic program of derivatization was set. Thus, the o -phthalaldehyde and 9-fluorenylmethyl chloroformate reagents were used for the derivatization of primary and secondary AAs [ 15 ]. The Agilent Zorbax Eclipse Plus C 18 (4.6 × 150 mm, 5 µm) column (Agilent Technologies, Inc., Santa Clara, CA, USA), at a temperature of 40 °C, was used to separate individual AAs. Detection of primary AAs was performed by a photodiode array detector at 388 nm, while detection of secondary AAs was performed by a fluorescence detector with an excitation wavelength of 266 nm and an emission wavelength of 305 nm. The injection volume was 5 µL. The flow rate was 1 mL/min. Eluent A of the mobile phase was 40 mM NaH 2 PO 4 (pH 7.8, adjusted by 10 M NaOH solution), while eluent B was a mixture including acetonitrile, methanol, and water (45:45:10, v / v / v ). The gradient was the following: 0–5 min, 100–90% A; 5–25 min, 90–59.5% A; 25–30 min, 59.5–37% A; 30–35 min, 37–18% A; 35–37 min, 18–0% A; 37–40 min, 0% A; and 40–43 min, 100% A. Table 5. Amino acids evaluated during HPLC analysis. The analytical data were integrated using the Agilent OpenLab CDS ChemStation software (Agilent Technologies, Inc., Santa Clara, CA, USA) for liquid chromatography systems. Identification of sugars and AAs was performed by comparing retention times of individual sugars and AAs in the reference vs. test solution. The concentration of these compounds was assayed based on comparisons of peak areas obtained for the samples, investigated with those of the reference solutions. 4.5. Statistical Analysis The R programming language or statistical environment was used to perform all statistical computations and analyses, as well as to prepare graphics and transform data for tabular representation [ 115 , 116 ]. The dataset of sugars was subjected to two-way ANOVA followed by Tukey’s post-hoc test, while AAs, floral display, and flower structure datasets were supplied to either (a) two-way ANOVA followed by Tukey’s post-hoc test or (b) the Kruskal–Wallis test followed by a pairwise Wilcoxon Rank Sum test with Benjamini–Hochberg adjustment, which compared the median values of different parameters between populations, depending on ANOVA pre-conditions (verified using Shapiro–Wilk test and Bartlett’s test) ( Table S1, Table S5, Table S6, Figure S1, Figure S4 ) [ 116 , 117 , 118 , 119 , 120 ]. Furthermore, a set of descriptive statistics (mean, standard error, quartiles, and interquartile range) was calculated for AAs, sugars, floral display, and flower structure. For all tests, the significance level was α = 0.05. In order to check if a monotonic relationship exists between floral display and flower structure parameters, Spearman’s rank correlations were calculated ( Table S2 ) using the ‘rcorr’ function from the ‘Hmisc’ package. Spearman’s correlations were also calculated between AAs ( Table S7 ). Correlations were considered significant for p < 0.05. To analyze the effect of AAs on insect chemoreceptors, all identified and determined AAs were grouped into four classes [ 24 ] (full names of abbreviations are present in Table 5 ): I. Asn, Gln, Ala, Cys, Gly, Ser, Thr, and Tyr (no effect on the chemoreceptors of fly); II. Arg, Asp, Glu, His, and Lys (inhibition of fly chemoreceptors); III. Pro (stimulation of the salt cell); and IV. Ile, Leu, Met, Phe, Trp, and Val (ability to stimulate the sugar cell) and presented as a ternary plot [ 121 ]. Principal component analysis (PCA) was used to simplify the exploration of AAs. To build the PCA model, the FactoMineR package was used [ 122 ]. Two tests that indicate the suitability of the AA dataset for structure detection and reduction were performed—Bartlett’s test of sphericity [ 123 ] and the Kaiser–Meyer–Olkin test of factorial adequacy (psych package [ 124 ]). Unit variance scaling of the data was applied; thus, PCA was performed on a correlation matrix, rather than on a covariance matrix. Number of principal components to retain was selected with the help of Cattell’s and Kaiser’s rules [ 78 , 79 ]. All biplots were created using the factoextra package [ 125 ]. Moreover a PCA was also applies to flower structure dataset using an approach identical to that used for AAs dataset. Supplementary Materials The following are available online at https://www.mdpi.com/article/10.3390/ijms222212164/s1 . Author Contributions Conceptualization, E.B.; methodology, E.B., A.B. and M.C.; software, A.B.; validation, E.B., J.B. and M.C.; formal analysis, E.B. and A.B.; investigation, J.B. and M.C.; resources, J.B. and M.C.; data curation, E.B. and A.B.; writing—original draft preparation, E.B.; writing—review and editing, E.B. and A.B.; visualization, A.B.; supervision, E.B.; project administration, E.B.; funding acquisition, E.B. and A.B. All authors have read and agreed to the published version of the manuscript. Funding This work was funded by the Ministry of Education and Science as part of subsidies for maintaining research potential awarded to the Faculty of Biology of the University of Bialystok. Informed Consent Statement Not applicable. Data Availability Statement Data is contained within the current article and supplementary material . Acknowledgments We thank the Rector of University in Bialystok and Dean of Department of Biology of University in Bialystok for financial support. A.B. thanks Adam Bajguz for statistical and programming consultations. Conflicts of Interest The authors declare no conflict of interest. References Joffard, N.; Massol, F.; Grenié, M.; Montgelard, C.; Schatz, B. Effect of pollination strategy, phylogeny and distribution on pollination niches of Euro-Mediterranean orchids. J. Ecol. 2019 , 107 , 478–490. [ Google Scholar ] [ CrossRef ][ Green Version ] Ollerton, J.; Winfree, R.; Tarrant, S. How many flowering plants are pollinated by animals? Oikos 2011 , 120 , 321–326. [ Google Scholar ] [ CrossRef ] Phillips, R.D.; Reiter, N.; Peakall, R. Orchid conservation: From theory to practice. Ann. Bot. 2020 , 126 , 345–362. [ Google Scholar ] [ CrossRef ] [ PubMed ] Tremblay, R.L. Trends in the pollination ecology of the Orchidaceae: Evolution and systematics. Can. J. Bot. 1992 , 70 , 642–650. [ Google Scholar ] [ CrossRef ] Claessens, J.; Kleynen, J. The Flower of the European Orchid. Form and Function ; Claessens & Kleynen: Guelle, The Netherlands, 2011; p. 440. [ Google Scholar ] Nilsson, L.A. Pollination ecology of Epipactis palustris (L.) Crantz (Orchidaceae). Bot. Not. 1978 , 131 , 355–368. [ Google Scholar ] Ackerman, J.D. Mechanisms and evolution of food-deceptive pollination systems in orchids. Lindleyana 1986 , 1 , 108–113. [ Google Scholar ] Cozzolino, S.; Widmer, A. Orchid diversity: An evolutionary consequence of deception? Trends Ecol. Evol. 2005 , 20 , 487–494. [ Google Scholar ] [ CrossRef ] [ PubMed ] Jersáková, J.; Johnson, S.D.; Kindlmann, P. Mechanisms and evolution of deceptive pollination in orchids. Biol. Rev. 2006 , 81 , 219–235. [ Google Scholar ] [ CrossRef ] Renner, S.S. Rewardless flowers in the angiosperms and the role of insect cognition in their evolution. In Plant-Pollinator Interactions: From Specialization to generalization ; Waser, N.M., Ollerton, J., Eds.; The University of Chicago Press: Chicago, IL, USA, 2006; pp. 123–144. [ Google Scholar ] Tremblay, R.L.; Ackerman, J.D.; Zimmerman, J.K.; Calvo, R.N. Variation in sexual reproduction in orchids and its evolutionary consequences: A spasmodic journey to diversification. Biol. J. Linn. Soc. 2005 , 84 , 1–54. [ Google Scholar ] [ CrossRef ] Dressler, R. The Orchids: Natural History and Classification ; Harvard University Press: Cambridge, MA, USA, 1981; p. 356. [ Google Scholar ] [ CrossRef ] Neiland, M.R.M.; Wilcock, C.C. Fruit set, nectar reward, and rarity in the Orchidaceae. Am. J. Bot. 1998 , 85 , 1657–1671. [ Google Scholar ] [ CrossRef ] [ PubMed ] Jersáková, J.; Johnson, S.D.; Kindlmann, P.; Pupin, A.C. Effect of nectar supplementation on male and female components of pollination success in the deceptive orchid Dactylorhiza sambucina . Acta Oecol. 2008 , 33 , 300–306. [ Google Scholar ] [ CrossRef ] Brzosko, E.; Bajguz, A. Nectar composition in moth-pollinated Platanthera bifolia and P. chlorantha and its importance for reproductive success. Planta 2019 , 250 , 263–279. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Gijbels, P.; Ceulemans, T.; Van den Ende, W.; Honnay, O. Experimental fertilization increases amino acid content in floral nectar, fruit set and degree of selfing in the orchid Gymnadenia conopsea . Oecologia 2015 , 179 , 785–795. [ Google Scholar ] [ CrossRef ] Gijbels, P.; Van den Ende, W.; Honnay, O. Landscape scale variation in nectar amino acid and sugar composition in a Lepidoptera pollinated orchid species and its relation with fruit set. J. Ecol. 2014 , 102 , 136–144. [ Google Scholar ] [ CrossRef ] Gijbels, P.; Van den Ende, W.; Honnay, O. Phenotypic selection on nectar amino acid composition in the Lepidoptera pollinated orchid species Gymnadenia conopsea . Oikos 2015 , 124 , 421–427. [ Google Scholar ] [ CrossRef ] Brzosko, E.; Bajguz, A.; Chmur, M.; Burzyńska, J.; Jermakowicz, E.; Mirski, P.; Zieliński, P. How are the flower structure and nectar composition of the generalistic orchid Neottia ovata adapted to a wide range of pollinators? Int. J. Mol. Sci. 2021 , 22 , 2214. [ Google Scholar ] [ CrossRef ] [ PubMed ] Jeffrey, D.C.; Arditti, J.; Koopowitz, H. Sugar content in floral and extrafloral exudates of orchids: Pollination, myrmecology and chemotaxonomy implication. New Phytol. 1970 , 69 , 187–195. [ Google Scholar ] [ CrossRef ][ Green Version ] Percival, M.S. Types of nectar in angiosperms. New Phytol. 1961 , 60 , 235–281. [ Google Scholar ] [ CrossRef ] Baker, H.G.; Baker, I. Floral nectar sugar constituents in relation to pollinator type. In Handbook of Experimental Pollination Biology ; Jones, C.E., Little, R.J., Eds.; Van Nostrand Reinhold Company Inc.: New York, NY, USA, 1983; pp. 131–141. [ Google Scholar ] Baker, H.G.; Baker, I. The predictive value of nectar chemistry to the recognition of pollinator types. Isr. J. Bot. 1990 , 39 , 157–166. [ Google Scholar ] [ CrossRef ] Nicolson, S.W.; Thornburg, R.W. Nectar chemistry. In Nectaries and Nectar ; Nicolson, S.W., Nepi, M., Pacini, E., Eds.; Springer: Dordrecht, The Netherlands, 2007; pp. 215–264. [ Google Scholar ] [ CrossRef ] Petanidou, T. Sugars in Mediterranean floral nectars: An ecological and evolutionary approach. J. Chem. Ecol. 2005 , 31 , 1065–1088. [ Google Scholar ] [ CrossRef ] Willmer, P. Pollination by butterflies and moths. In Pollination and Floral Ecology ; Willmer, P., Ed.; Princeton University Press: Princeton, NJ, USA, 2011; pp. 322–336. [ Google Scholar ] [ CrossRef ] Adler, L.S. The ecological significance of toxic nectar. Oikos 2000 , 91 , 409–420. [ Google Scholar ] [ CrossRef ] Gardener, M.C.; Gillman, M.P. The taste of nectar—A neglected area of pollination ecology. Oikos 2002 , 98 , 552–557. [ Google Scholar ] [ CrossRef ] Heil, M. Postsecretory hydrolysis of nectar sucrose and specialization in ant/plant mutualism. Science 2005 , 308 , 560–563. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Levin, E.; McCue, M.D.; Davidowitz, G. More than just sugar: Allocation of nectar amino acids and fatty acids in a Lepidopteran. Proc. Biol. Sci. 2017 , 284 , 20162126. [ Google Scholar ] [ CrossRef ] [ PubMed ] Mevi-Schütz, J.; Erhardt, A. Amino acids in nectar enhance butterfly fecundity: A long-awaited link. Am. Nat. 2005 , 165 , 411–419. [ Google Scholar ] [ CrossRef ] [ PubMed ] Heyneman, A.J. Optimal sugar concentrations of floral nectars—Dependence on sugar intake efficiency and foraging costs. Oecologia 1983 , 60 , 198–213. [ Google Scholar ] [ CrossRef ] [ PubMed ] Pyke, G.H.; Waser, N.M. The production of dilute nectars by hummingbird and honeyeater flowers. Biotropica 1981 , 13 , 260–270. [ Google Scholar ] [ CrossRef ] Brzosko, E.; Mirski, P. Floral nectar chemistry in orchids: A short review and meta-analysis. Plants 2021 , 10 , 2315. [ Google Scholar ] [ CrossRef ] Heil, M. Nectar: Generation, regulation and ecological functions. Trends Plant Sci. 2011 , 16 , 191–200. [ Google Scholar ] [ CrossRef ] Gottsberger, G.; Schrauwen, J.; Linskens, H.F. Amino acids and sugars in nectar, and their putative evolutionary significance. Plant Syst. Evol. 1984 , 145 , 55–77. [ Google Scholar ] [ CrossRef ] Pacini, E.; Nepi, M.; Vesprini, J.L. Nectar biodiversity: A short review. Plant Syst. Evol. 2003 , 238 , 7–21. [ Google Scholar ] [ CrossRef ] Pais, M.S.; Figueiredo, A.C.S. Floral nectaries from Limodorum abortivum (L.) Sw and Epipactis atropurpurea Rafin (Orchidaceae)—Ultrastructural changes in plastids during the secretory process. Apidologie 1994 , 25 , 615–626. [ Google Scholar ] [ CrossRef ][ Green Version ] Vandelook, F.; Janssens, S.B.; Gijbels, P.; Fischer, E.; Van den Ende, W.; Honnay, O.; Abrahamczyk, S. Nectar traits differ between pollination syndromes in Balsaminaceae. Ann. Bot. 2019 , 124 , 269–279. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Pais, M.; Neves, H.; Maria, P.; Vasconcelos, A. Amino acid and sugar content of the nectar exudate from Limodorum abortivum (Orchidaceae). Comparison with Epipactis atropurpurea nectar composition. Apidologie 1986 , 17 , 125–136. [ Google Scholar ] [ CrossRef ][ Green Version ] Moré, M.; Amorim, F.W.; Benitez-Vieyra, S.; Medina, A.M.; Sazima, M.; Cocucci, A.A. Armament imbalances: Match and mismatch in plant-pollinator traits of highly specialized long-spurred orchids. PLoS ONE 2012 , 7 , e41878. [ Google Scholar ] [ CrossRef ][ Green Version ] Trunschke, J.; Sletvold, N.; Ågren, J. Manipulation of trait expression and pollination regime reveals the adaptive significance of spur length. Evolution 2020 , 74 , 597–609. [ Google Scholar ] [ CrossRef ] Alexandersson, R.; Johnson, S.D. Pollinator-mediated selection on flower-tube length in a hawkmoth-pollinated Gladiolus (Iridaceae). Proc. Biol. Sci. 2002 , 269 , 631–636. [ Google Scholar ] [ CrossRef ][ Green Version ] Anderson, B.; Alexandersson, R.; Johnson, S.D. Evolution and coexistence of pollination ecotypes in an African Gladiolus (Iridaceae). Evolution 2010 , 64 , 960–972. [ Google Scholar ] [ CrossRef ] [ PubMed ] Boberg, E.; Ägren, J. Despite their apparent integration, spur length but not perianth size affects reproductive success in the moth-pollinated orchid Platanthera bifolia . Funct. Ecol. 2009 , 23 , 1022–1028. [ Google Scholar ] [ CrossRef ] Boberg, E.; Alexandersson, R.; Jonsson, M.; Maad, J.; Ägren, J.; Nilsson, L.A. Pollinator shifts and the evolution of spur length in the moth-pollinated orchid Platanthera bifolia . Ann. Bot. 2014 , 113 , 267–275. [ Google Scholar ] [ CrossRef ][ Green Version ] Little, K.J.; Dieringer, G.; Romano, M. Pollination ecology, genetic diversity and selection on nectar spur length in Platanthera lacera (Orchidaceae). Plant Spec. Biol. 2005 , 20 , 183–190. [ Google Scholar ] [ CrossRef ] Maad, J. Phenotypic selection in hawkmoth-pollinated Platanthera bifolia : Targets and fitness surfaces. Evolution 2000 , 54 , 112–123. [ Google Scholar ] [ CrossRef ] Maad, J.; Alexandersson, R. Variable selection in Platanthera bifolia (Orchidaceae): Phenotypic selection differed between sex functions in a drought year. J. Evol. Biol. 2004 , 17 , 642–650. [ Google Scholar ] [ CrossRef ] [ PubMed ] Whittall, J.B.; Hodges, S.A. Pollinator shifts drive increasingly long nectar spurs in columbine flowers. Nature 2007 , 447 , 706–709. [ Google Scholar ] [ CrossRef ] Caruso, C.M.; Eisen, K.E.; Martin, R.A.; Sletvold, N. A meta-analysis of the agents of selection on floral traits. Evolution 2018 , 73 , 4–14. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] De Jager, M.L.; Peakall, R. Experimental examination of pollinator-mediated selection in a sexually deceptive orchid. Ann. Bot. 2019 , 123 , 347–354. [ Google Scholar ] [ CrossRef ] Paulus, H.F. Deceived males-pollination biology of the Mediterranean orchid genus Ophrys (Orchidaceae). J. Eur. Orchid. 2006 , 38 , 303–353. [ Google Scholar ] Scopece, G.; Juillet, N.; Lexer, C.; Cozzolino, S. Fluctuating selection across years and phenotypic variation in food-deceptive orchids. PeerJ 2017 , 5 , e3704. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Sletvold, N.; Ågren, J. Nonadditive effects of floral display and spur length on reproductive success in a deceptive orchid. Ecology 2011 , 92 , 2167–2174. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Sletvold, N.; Trunschke, J.; Smit, M.; Verbeek, J.; Agren, J. Strong pollinator-mediated selection for increased flower brightness and contrast in a deceptive orchid. Evolution 2016 , 70 , 716–724. [ Google Scholar ] [ CrossRef ] Bernhardt, P.; Edens-Meier, R. What we think we know vs. what we need to know about orchid pollination and conservation: Cypripedium L. as a model lineage. Bot. Rev. 2010 , 76 , 204–219. [ Google Scholar ] [ CrossRef ] Potts, S.G.; Biesmeijer, J.C.; Kremen, C.; Neumann, P.; Schweiger, O.; Kunin, W.E. Global pollinator declines: Trends, impacts and drivers. Trends Ecol. Evol. 2010 , 25 , 345–353. [ Google Scholar ] [ CrossRef ] [ PubMed ] Jermakowicz, E.; Brzosko, E. Demographic responses of boreal-montane orchid Malaxis monophyllos (L.) Sw. populations to contrasting environmental conditions. Acta Soc. Bot. Pol. 2016 , 85 , 1. [ Google Scholar ] [ CrossRef ][ Green Version ] Mitchell, R.J.; Flanagan, R.J.; Brown, B.J.; Waser, N.M.; Karron, J.D. New frontiers in competition for pollination. Ann. Bot. 2009 , 103 , 1403–1413. [ Google Scholar ] [ CrossRef ][ Green Version ] Vamosi, J.C.; Knight, T.M.; Steets, J.A.; Mazer, S.J.; Burd, M.; Ashman, T.-L. Pollination decays in biodiversity hotspots. Proc. Natl. Acad. Sci. USA 2006 , 103 , 956–961. [ Google Scholar ] [ CrossRef ][ Green Version ] Brzosko, E. The dynamics of Listera ovata populations on mineral islands in the Biebrza National Park. Acta Soc. Bot. Pol. 2002 , 71 , 243–251. [ Google Scholar ] [ CrossRef ][ Green Version ] Brzosko, E. Dynamics of island populations of Cypripedium calceolus in the Biebrza river valley (north-east Poland). Bot. J. Linn. Soc. 2002 , 139 , 67–77. [ Google Scholar ] [ CrossRef ] Brzosko, E. The dynamics of island populations of Platanthera bifolia in the Biebrza National Park (NE Poland). Ann. Bot. Fenn. 2003 , 40 , 243–253. [ Google Scholar ] Ackerman, J.D. Invasive orchids: Weeds we hate to love? Lankesteriana 2015 , 7 , 19–21. [ Google Scholar ] [ CrossRef ][ Green Version ] Adamowski, W. Expansion of native orchids in anthropogenous habitats. Pol. Bot. Stud. 2006 , 22 , 35–44. [ Google Scholar ] Jacquemyn, H.; Brys, R.; Hutchings, M.J. Biological flora of the British Isles: Epipactis palustris . J. Ecol. 2014 , 102 , 1341–1355. [ Google Scholar ] [ CrossRef ] Rewicz, A.; Bomanowska, A.; Shevera, M.; Kurowski, J.; Krasoń, K.; Zielińska, K. Cities and disturbed areas as man-made shelters for orchid communities. Not. Bot. Horti Agrobot. Cluj-Napoca 2017 , 45 , 126–139. [ Google Scholar ] [ CrossRef ][ Green Version ] The IUCN Red List of Threatened Species, Version 2021-2. Available online: https://www.iucnredlist.org/ (accessed on 27 October 2021). Fay, M.F. Orchid conservation: How can we meet the challenges in the twenty-first century? Bot. Stud. 2018 , 59 , 16. [ Google Scholar ] [ CrossRef ][ Green Version ] Kull, T.; Selgis, U.; Peciña, M.V.; Metsare, M.; Ilves, A.; Tali, K.; Sepp, K.; Kull, K.; Shefferson, R.P. Factors influencing IUCN threat levels to orchids across Europe on the basis of national red lists. Ecol. Evol. 2016 , 6 , 6245–6265. [ Google Scholar ] [ CrossRef ] Zizka, A.; Silvestro, D.; Vitt, P.; Knight, T.M. Automated conservation assessment of the orchid family with deep learning. Conserv. Biol. 2021 , 35 , 897–908. [ Google Scholar ] [ CrossRef ] Swarts, N.D.; Dixon, K.W. Terrestrial orchid conservation in the age of extinction. Ann. Bot. 2009 , 104 , 543–556. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Kull, T.; Hutchings, M.J. A comparative analysis of decline in the distribution ranges of orchid species in Estonia and the United Kingdom. Biol. Conserv. 2006 , 129 , 31–39. [ Google Scholar ] [ CrossRef ] Jacquemyn, H.; Brys, R.; Hermy, M.; Willems, J. Does nectar reward affect rarity and extinction probabilities of orchid species? An assessment using historical records from Belgium and the Netherlands. Biol. Conserv. 2005 , 121 , 257–263. [ Google Scholar ] [ CrossRef ] Goulson, D.; Lye, G.C.; Darvill, B. Decline and conservation of bumble bees. Annu. Rev. Entomol. 2008 , 53 , 191–208. [ Google Scholar ] [ CrossRef ] [ PubMed ] Kaiser, H.F. An index of factorial simplicity. Psychometrika 1974 , 39 , 31–36. [ Google Scholar ] [ CrossRef ] Cattell, R.B. The scree test for the number of factors. Multivar. Behav. Res. 1966 , 1 , 245–276. [ Google Scholar ] [ CrossRef ] [ PubMed ] Kaiser, H.F. The application of electronic computers to factor analysis. Educ. Psychol. Meas. 1960 , 20 , 141–151. [ Google Scholar ] [ CrossRef ] Tałałaj, I.; Brzosko, E. Selfing potential in Epipactis palustris , E. helleborine and E. atrorubens (Orchidaceae). Plant Syst. Evol. 2008 , 276 , 21–29. [ Google Scholar ] [ CrossRef ] Vakhrameeva, M.G.; Tatarenko, I.V.; Varlygina, T.I.; Torosyan, G.K.; Zagulski, M.N. Orchids of Russia and Adjacent Countries (within the Borders of the Former USSR) ; A.R.G. Gantner Verlag K.G.: Ruggell, Liechtenstein, 2008; p. 690. [ Google Scholar ] Jacquemyn, H.; Brys, R. Pollen limitation and the contribution of autonomous selfing to fruit and seed set in a rewarding orchid. Am. J. Bot. 2015 , 102 , 67–72. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Jakubska-Busse, A.; Kadej, M. The pollination of Epipactis Zinn, 1757 (Orchidaceae) species in Central Europe—The significance of chemical attractants, floral morphology and concomitant insects. Acta Soc. Bot. Pol. 2011 , 80 , 49–57. [ Google Scholar ] [ CrossRef ][ Green Version ] Pellegrino, G.; Bellusci, F. Effects of human disturbance on reproductive success and population viability of Serapias cordigera (Orchidaceae). Bot. J. Linn. Soc. 2014 , 176 , 408–420. [ Google Scholar ] [ CrossRef ][ Green Version ] Parra-Tabla, V.; Vargas, C.F.; Magaña-Rueda, S.; Navarro, J. Female and male pollination success of Oncidium ascendens Lindey (Orchidaceae) in two contrasting habitat patches. Biol. Conserv. 2000 , 94 , 335–340. [ Google Scholar ] [ CrossRef ] Rewicz, A.; Jaskuła, R.; Rewicz, T.; Tończyk, G. Pollinator diversity and reproductive success of Epipactis helleborine (L.) Crantz (Orchidaceae) in anthropogenic and natural habitats. PeerJ 2017 , 5 , e3159. [ Google Scholar ] [ CrossRef ][ Green Version ] Ollerton, J.; Killick, A.; Lamborn, E.; Watts, S.; Whiston, M. Multiple meanings and modes: On the many ways to be a generalist flower. Taxon 2007 , 56 , 717–728. [ Google Scholar ] [ CrossRef ] Ågren, J.; Fortunel, C.; Ehrlén, J. Selection on floral display in insect-pollinated Primula farinosa : Effects of vegetation height and litter accumulation. Oecologia 2006 , 150 , 225–232. [ Google Scholar ] [ CrossRef ] Sletvold, N.; Grindeland, J.M.; Ågren, J. Vegetation context influences the strength and targets of pollinator-mediated selection in a deceptive orchid. Ecology 2013 , 94 , 1236–1242. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Grindeland, J.M.; Sletvold, N.; Ims, R.A. Effects of floral display size and plant density on pollinator visitation rate in a natural population of Digitalis purpurea . Funct. Ecol. 2005 , 19 , 383–390. [ Google Scholar ] [ CrossRef ][ Green Version ] Kindlmann, P.; Jersáková, J. Effect of floral display on reproductive success in terrestrial orchids. Folia Geobot. 2006 , 41 , 47–60. [ Google Scholar ] [ CrossRef ] Vallius, E.; Arminen, S.; Salonen, V. Are There Fitness Advantages Associated with a Large Inflorescence in Gymnadenia conopsea ssp. conopsea ? 2006. Available online: http://www.r-b-o.eu/rbo_public?Vallius_et_al_2006.html (accessed on 29 November 2020). Calvo, R.N. Inflorescence size and fruit distribution among individuals in three orchid species. Am. J. Bot. 1990 , 77 , 1378–1381. [ Google Scholar ] [ CrossRef ] Pellegrino, G.; Bellusci, F.; Musacchio, A. The effects of inflorescence size and flower position on female reproductive success in three deceptive orchids. Bot. Stud. 2010 , 51 , 351–356. [ Google Scholar ] Van der Niet, T.; Peakall, R.; Johnson, S.D. Pollinator-driven ecological speciation in plants: New evidence and future perspectives. Ann. Bot. 2014 , 113 , 199–211. [ Google Scholar ] [ CrossRef ][ Green Version ] Vereecken, N.J.; Cozzolino, S.; Schiestl, F.P. Hybrid floral scent novelty drives pollinator shift in sexually deceptive orchids. BMC Evol. Biol. 2010 , 10 , 103. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Peralta, G.; Vázquez, D.P.; Chacoff, N.P.; Lomáscolo, S.B.; Perry, G.L.W.; Tylianakis, J.M. Trait matching and phenological overlap increase the spatio-temporal stability and functionality of plant-pollinator interactions. Ecol. Lett. 2020 , 23 , 1107–1116. [ Google Scholar ] [ CrossRef ] [ PubMed ] Jacquemyn, H.; Brys, R. Lack of strong selection pressures maintains wide variation in floral traits in a food-deceptive orchid. Ann. Bot. 2020 , 126 , 445–453. [ Google Scholar ] [ CrossRef ] [ PubMed ] Abrahamczyk, S.; Kessler, M.; Hanley, D.; Karger, D.N.; Müller, M.P.J.; Knauer, A.C.; Keller, F.; Schwerdtfeger, M.; Humphreys, A.M. Pollinator adaptation and the evolution of floral nectar sugar composition. J. Evol. Biol. 2016 , 30 , 112–127. [ Google Scholar ] [ CrossRef ] Parachnowitsch, A.L.; Manson, J.S.; Sletvold, N. Evolutionary ecology of nectar. Ann. Bot. 2019 , 123 , 247–261. [ Google Scholar ] [ CrossRef ][ Green Version ] Fowler, R.E.; Rotheray, E.L.; Goulson, D. Floral abundance and resource quality influence pollinator choice. Insect Conserv. Divers. 2016 , 9 , 481–494. [ Google Scholar ] [ CrossRef ][ Green Version ] Petanidou, T.; Van Laere, A.; Ellis, W.N.; Smets, E. What shapes amino acid and sugar composition in Mediterranean floral nectars? Oikos 2006 , 115 , 155–169. [ Google Scholar ] [ CrossRef ][ Green Version ] Jakubska-Busse, A.; Kadej, M. Pollination ecology of marsh helleborine Epipactis palustris (L.) Crantz on the Polish side of the Orlickie Mts. (Central Sudety Mts.). In Environmental Changes and Biological Assessments IV ; Kočárek, P., Plášek, V., Malachová, K., Eds.; Scripra Facultatis Rerum Naturalium Universitatis Ostraviensis: Cambridge, UK, 2008; Volume 186, pp. 247–253. [ Google Scholar ] Gardener, M.C.; Gillman, M.P. The effects of soil fertilizer on amino acids in the floral nectar of corncockle, Agrostemma githago (Caryophyllaceae). Oikos 2001 , 92 , 101–106. [ Google Scholar ] [ CrossRef ][ Green Version ] Carter, C.; Shafir, S.; Yehonatan, L.; Palmer, R.G.; Thornburg, R. A novel role for proline in plant floral nectars. Naturwissenschaften 2006 , 93 , 72–79. [ Google Scholar ] [ CrossRef ] Nepi, M.; Soligo, C.; Nocentini, D.; Abate, M.; Guarnieri, M.; Cai, G.; Bini, L.; Puglia, M.; Bianchi, L.; Pacini, E. Amino acids and protein profile in floral nectar: Much more than a simple reward. Flora 2012 , 207 , 475–481. [ Google Scholar ] [ CrossRef ] Bertazzini, M.; Medrzycki, P.; Bortolotti, L.; Maistrello, L.; Forlani, G. Amino acid content and nectar choice by forager honeybees ( Apis mellifera L.). Amino Acids 2010 , 39 , 315–318. [ Google Scholar ] [ CrossRef ][ Green Version ] Tiedge, K.; Lohaus, G. Nectar sugars and amino acids in day- and night-flowering Nicotiana species are more strongly shaped by pollinators’ preferences than organic acids and inorganic ions. PLoS ONE 2017 , 12 , e0176865. [ Google Scholar ] [ CrossRef ][ Green Version ] Kim, Y.S.; Smith, B.H. Effect of an amino acid on feeding preferences and learning behavior in the honey bee, Apis mellifera . J. Insect Physiol. 2000 , 46 , 793–801. [ Google Scholar ] [ CrossRef ] Baker, H.G.; Baker, I. Intraspecific constancy of floral nectar amino acid complements. Bot. Gaz. 1977 , 138 , 183–191. [ Google Scholar ] [ CrossRef ] Hultén, E.; Fries, M. Atlas of North European Vascular Plants: North of the Tropic of Cancer. Volumes I-III ; Koeltz Scientific Books: Königstein, Germany, 1986; p. 1172. [ Google Scholar ] Kowalkowska, A.K.; Kostelecka, J.; Bohdanowicz, J.; Kapusta, M.; Rojek, J. Studies on floral nectary, tepals’ structure, and gynostemium morphology of Epipactis palustris (L.) Crantz (Orchidaceae). Protoplasma 2015 , 252 , 321–333. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Morrant, D.S.; Schumann, R.; Petit, S. Field methods for sampling and storing nectar from flowers with low nectar volumes. Ann. Bot. 2009 , 103 , 533–542. [ Google Scholar ] [ CrossRef ][ Green Version ] Wickham, H.; Averick, M.; Bryan, J.; Chang, W.; McGowan, L.; François, R.; Grolemund, G.; Hayes, A.; Henry, L.; Hester, J.; et al. Welcome to the Tidyverse. J. Open Source Softw. 2019 , 4 , 1686. [ Google Scholar ] [ CrossRef ] R Core Team. R: A Language and Environment for Statistical Computing, R Version 4.1.1, Kick Things ; R Foundation for Statistical Computing: Vienna, Austria, 2021; Available online: https://www.R-project.org/ (accessed on 5 October 2021). Fox, J.; Weisberg, S. An R Companion to Applied Regression , 3rd ed.; SAGE Publications, Inc.: Thousand Oaks, CA, USA, 2019. [ Google Scholar ] Burda, M. Paircompviz: Multiple Comparison Test Visualization (R Package Version 1.28.0) ; 2020; Available online: https://www.bioconductor.org/packages/release/bioc/html/paircompviz.html (accessed on 5 October 2021). Graves, S.; Piepho, H.-P.; Selzer, L.; Dorai-Raj, S. multcompView: Visualizations of Paired Comparisons, R Package Version 0.1-8 ; 2019; Available online: https://cran.r-project.org/web/packages/multcompView/index.html (accessed on 5 October 2021). Mangiafico, S. Rcompanion: Functions to Support Extension Education Program Evaluation (R Package Version 2.3.26) ; 2020; Available online: https://rdrr.io/cran/rcompanion/ (accessed on 5 October 2021). Hamilton, N.E.; Ferry, M. ggtern: Ternary diagrams using ggplot2. J. Stat. Softw. 2018 , 87 , 1–17. [ Google Scholar ] [ CrossRef ][ Green Version ] Lê, S.; Josse, J.; Husson, F. FactoMineR: An R package for multivariate analysis. J. Stat. Softw. 2008 , 25 , 1–18. [ Google Scholar ] [ CrossRef ][ Green Version ] Bartlett, M.S. Tests of significance in factor analysis. Br. J. Stat. Psychol. 1950 , 3 , 77–85. [ Google Scholar ] [ CrossRef ] Revelle, W. psych: Procedures for Personality and Psychological Research, R Package Version 1.8.10 ; Northwestern University: Evanston, IL, USA, 2018. [ Google Scholar ] Kassambara, A.; Mundt, F. Factoextra: Extract and Visualize the Results of Multivariate Data Analyses, R Package Version 1.0.6 ; 2019; Available online: https://cran.r-project.org/web/packages/factoextra/index.html (accessed on 5 October 2021). Figure 1. Biplot of flower structure profiles for Epipactis palustris natural (Nat.) and anthropogenic (Ant.) populations, showing the first two dimensions or factors (Dim1-2) of PCA that, together, explain 63% of the variance. Biplot vectors indicate the strength and direction of factor loading for the first two factors. Individuals (populations) are color-coded by population. Figure 2. Boxplots of sugar amounts for Epipactis palustris natural (Nat.) and anthropogenic (Ant.) populations. Colored dots are individual samples. The crossed square shows the mean. The lower and upper hinges correspond to the lower (Q 1 ) and upper (Q 3 ) quartiles. Thus, box length shows the interquartile range (IQR). The thicker lines inside the boxes corresponds to the median. The lower whisker extends from the hinge to the smallest value at most Q 1 − 1.5 × IQR of the hinge. The upper whisker extends from the hinge to the largest value no further than Q 3 + 1.5 × IQR. Data beyond the end of the whiskers, indicated with an asterisk symbol, are outliers. Different lowercase letters indicate statistically significant differences according to Tukey’s post-hoc test ( p < 0.05). Symbol “=” means they did not differ significantly. Additional comparisons on the left or right side were shown only when the Nat. or Ant. (or both) populations did not differ significantly. Figure 3. Ternary plot of amino acid classes for Epipactis palustris natural (Nat.) and anthropogenic (Ant.) populations: II (Asp, Glu, His, Arg, Lys), III (Pro), and IV (Val, Met, Trp, Phe, Ile, Leu). Blue lines show 50%, 90%, and 95% confidence intervals via the Mahalanobis Distance and use of the Log–Ratio Transformation. The first class of AAs (Asn, Gln, Ala, Cys, Gly, Ser, Thr, Tyr) does not affect the chemoreceptors of fly (data not shown). AAs’ abbreviations and full names are present in Table 2 . Figure 4. Biplot of amino acid profiles for Epipactis palustris natural (Nat.) and anthropogenic (Ant.) populations, showing the first two dimensions or factors (Dim1-2) of PCA that, together, explain 75.52% of the variance. Biplot vectors indicate the strength and direction of factor loading for the first two factors. Individuals (populations) are color-coded by population. Ellipses around the individuals show assumed 95% multivariate normal distribution. Parameter Statistic/Comparison Natural (Nat.) Populations Anthropogenic (Ant.) Populations ZAB ROS SIL SOP Shoot height (cm) x ¯ ± SE 62.59 ± 3.05 a 51.69 ± 1.61 b 42.47 ± 1.36 c 54.62 ± 2.14 b Q 1 51.50 47.01 37.5 43.25 Q 2 (IQR) 64.75 (18.75) 53 (10) 42.75 (9) 56.75 (16.75) Q 3 70.25 57.00 46.5. 60.00 Inflorescence length (cm) x ¯ ± SE 9.43 ± 0.58 b 10.43 ± 0.53 b 10.26 ± 0.33 b 13.46 ± 0.79 a Q 1 7.25 9.00 9.00 10.62 Q 2 (IQR) 9.25 (3.75) 10 (3.00) 10.00 (3.00) 13.50 (5.38) Q 3 11.00 12.00 12.00 16.00 Nat. vs. SIL vs. SOP b b a Flower number x ¯ ± SE 9.00 ± 0.54 10.66 ± 0.46 9.53 ± 0.43 11.85 ± 0.81 Q 1 7.00 9.00 9.00 9.25 Q 2 (IQR) 8.00 (4.50) B 11.00 (3.00) A 9.00 (1.75) B 11.00 (5.75) A Q 3 11.50 12.00 10.75 15.00 Length of dorsal sepal (LDS) (mm) x ¯ ± SE 11.22 ± 0.13 a 10.01 ± 0.09 c 10.70 ± 0.10 b 11.55 ± 0.18 a Q 1 10.64 9.56 10.33 10.99 Q 2 (IQR) 11.37 (1.04) 9.99 (0.81) 10.69 (0.66) 11.34 (1.07) Q 3 11.68 10.37 10.99 12.06 Width of dorsal sepal (WDS) (mm) x ¯ ± SE 4.48 ± 0.06 a 3.83 ± 0.05 a 3.99 ± 0.04 b 4.37 ± 0.11 b Q 1 4.34 3.64 3.87 4.02 Q 2 (IQR) 4.53 (0.28) 3.84 (0.31) 4.02 (0.27) 4.34 (0.68) Q 3 4.61 3.95 4.15 4.70 ZAB vs. ROS vs. Ant. a c b Nat. vs. SIL vs. SOP ab b a Nat. vs. Ant. do not differ significantly Length of petal (LP) (mm) x ¯ ± SE 9.38 ± 0.12 a 8.18 ± 0.11 b 9.27 ± 0.10 a 9.36 ± 0.23 a Q 1 9.06 7.74 8.77 8.63 Q 2 (IQR) 9.41 (0.74) 8.03 (0.95) 9.29 (0.86) 9.31 (1.34) Q 3 9.80 8.70 9.63 9.97 ZAB vs. ROS vs. Ant. a b a Width of petal (WP) (mm) x ¯ ± SE 4.01 ± 0.05 a 3.44 ± 0.06 b 3.83 ± 0.06 a 3.78 ± 0.11 a Q 1 3.89 3.18 3.69 3.48 Q 2 (IQR) 4.06 (0.29) 3.38 (0.42) 3.86 (0.34) 3.67 (0.53) Q 3 4.17 3.60 4.03 4.01 ZAB vs. ROS vs. Ant. a c b Length of lateral sepal (LLS) (mm) x ¯ ± SE 11.34 ± 0.13 11.01 ± 0.12 11.35 ± 0.10 12.80 ± 0.19 Q 1 10.82 10.53 11.05 11.93 Q 2 (IQR) 11.35 (1.01) BC 10.87 (0.91) C 11.40 (0.65) B 12.63 (1.36) A Q 3 11.81 11.44 11.70 13.29 Nat. vs. SIL vs. SOP B B A Width of lateral sepal (WLS) (mm) x ¯ ± SE 4.17 ± 0.04 3.76 ± 0.04 3.83 ± 0.04 4.33 ± 0.10 Q 1 4.03 3.63 3.69 3.96 Q 2 (IQR) 4.15 (0.29) B 3.71 (0.34) A 3.88 (0.32) A 4.18 (0.56) B Q 3 4.32 3.96 4.01 4.52 Width of flowers (FW) (mm) x ¯ ± SE 22.70 ± 0.35 20.83 ± 0.24 22.59 ± 0.16 24.31 ± 0.38 Q 1 22.26 20.22 21.87 22.66 Q 2 (IQR) 22.94 (1.64) B 20.89 (1.43) A 22.47 (1.40) B 24.01 (2.70) C Q 3 23.90 21.65 23.26 25.37 Length of flowers (FH) (mm) x ¯ ± SE 11.47 ± 0.14 ab 10.36 ± 0.10 c 11.01 ± 0.12 b 11.94 ± 0.22 a Q 1 11.04 10.02 10.69 11.33 Q 2 (IQR) 11.45 (0.83) 10.35 (0.76) 10.97 (0.77) 11.82 (1.30) Q 3 11.86 10.78 11.46 12.63 Length of labellum (LL) (mm) x ¯ ± SE 11.91 ± 0.16 11.46 ± 0.15 11.45 ± 0.14 11.93 ± 0.26 Q 1 11.56 11.01 11.23 11.42 Q 2 (IQR) 12.06 (0.81) B 11.47 (0.89) A 11.54 (0.64) A 11.99 (1.20) B Q 3 12.37 11.88 11.87 12.62 ZAB vs. ROS vs. Ant. A B AB Width of hypochile (HW) (mm) x ¯ ± SE 5.81 ± 0.13 a 4.90 ± 0.10 b 4.84 ± 0.14 b 5.54 ± 0.12 a Q 1 5.25 4.64 4.31 5.06 Q 2 (IQR) 5.79 (1.15) 4.95 (0.61) 4.69 (0.98) 5.53 (0.83) Q 3 6.40 5.25 5.29 5.9 Length of hypochile (HL) (mm) x ¯ ± SE 5.03 ± 0.13 4.58 ± 0.07 4.44 ± 0.06 4.80 ± 0.10 Q 1 4.62 4.38 4.20 4.36 Q 2 (IQR) 4.89 (0.61) C 4.65 (0.36) AB 4.47 (0.38) A 4.76 (0.77) BC Q 3 5.23 4.74 4.58 5.12 Length of epichile (LE) (mm) x ¯ ± SE 7.11 ± 0.08 6.74 ± 0.11 7.03 ± 0.09 7.44 ± 0.11 Q 1 6.87 6.38 6.82 7.14 Q 2 (IQR) 7.10 (0.51) B 6.88 (0.74) A 7.05 (0.44) B 7.51 (0.64) C Q 3 7.38 7.12 7.26 7.78 Length of isthmus (LI) (mm) x ¯ ± SE 2.07 ± 0.04 ab 1.79 ± 0.03 ab 1.94 ± 0.03 b 2.24 ± 0.06 a Q 1 1.92 1.66 1.82 2.09 Q 2 (IQR) 2.08 (0.33) 1.79 (0.29) 1.95 (0.26) 2.31 (0.41) Q 3 2.25 1.94 2.08 2.50 Nat. vs. SIL vs. SOP b b a Width of epichile (WE) (mm) x ¯ ± SE 7.98 ± 0.11 ab 7.71 ± 0.09 ab 7.56 ± 0.09 b 8.07 ± 0.16 a Q 1 7.52 7.46 7.27 7.47 Q 2 (IQR) 7.89 (0.84) 7.66 (0.49) 7.51 (0.54) 8.11 (0.94) Q 3 8.36 7.95 7.81 8.42 Nat. vs. SIL vs. SOP ab b a Width of isthmus (WI) (mm) x ¯ ± SE 0.84 ± 0.01 0.90 ± 0.02 0.82 ± 0.01 1.02 ± 0.03 Q 1 0.82 0.81 0.75 0.94 Q 2 (IQR) 0.85 (0.06) B 0.91 (0.15) A 0.82 (0.11) B 1.01 (0.13) C Q 3 0.88 0.96 0.86 1.07 Isthmus area (AI) (mm 2 ) x ¯ ± SE 1.75 ± 0.05 b 1.60 ± 0.05 b 1.59 ± 0.05 b 2.32 ± 0.11 a Q 1 1.65 1.42 1.41 1.94 Q 2 (IQR) 1.75 (0.29) 1.58 (0.37) 1.54 (0.34) 2.28 (0.71) Q 3 1.94 1.79 1.75 2.65 Nat. vs. SIL vs. SOP b b a Table 2. The concentration of amino acids (μM) and total amount of amino acids (mg/mL) in Epipactis palustris nectar. The number of classes represents the effect of amino acids on insect chemoreceptors: I—no effect; II—inhibition of chemoreceptors; III—stimulate the salt cell; IV—the ability to stimulate the sugar cell. Data ( n = 30) represent the mean ( x ¯ ) ± standard error (SE), lower quartile (Q 1 ), median (Q 2 ), upper quartile (Q 3 ), and interquartile range (IQR). Different lowercase letters indicate statistically significant differences, according to Tukey’s post-hoc test ( p < 0.05). Different uppercase letters indicate statistically significant differences according to the pairwise Wilcoxon Rank Sum test with Benjamini–Hochberg adjustment ( p < 0.05). ND—not detected. Additional comparisons were shown only when populations within Nat. or Ant. (or both) do not differ significantly. Amino Acid Class Statistic Natural (Nat.) Populations Anthropogenic (Ant.) Populations ZAB ROS SIL SOP Proteogenic amino acids (μM) Aspartic acid (Asp) I x ¯ ± SE 357.55 ± 6.47 377.50 ± 6.73 200.25 ± 6.81 259.51 ± 5.06 Q 1 329.36 351.40 178.80 228.94 Q 2 (IQR) 359.31 (56.82) B 380.49 (50.09) A 198.98 (48.50) D 267.79 (49.53) C Q 3 386.17 401.49 227.30 278.48 Glutamic acid (Glu) I x ¯ ± SE 740.90 ± 8.71 b 884.43 ± 7.46 a 430.24 ± 6.87 d 525.11 ± 4.78 c Q 1 706.06 858.27 400.67 509.22 Q 2 (IQR) 730.92 (71.49) 885.59 (48.87) 421.47 (64.14) 522.14 (24.38) Q 3 777.55 907.13 464.81 533.60 Alanine (Ala) I x ¯ ± SE 92.59 ± 2.48 116.35 ± 1.74 84.42 ± 1.10 89.31 ± 2.40 Q 1 80.82 111.75 79.64 81.50 Q 2 (IQR) 89.79 (23.47) B 117.99 (9.50) A 84.07 (9.36) C 83.99 (12.90) C Q 3 104.29 121.25 89.01 94.40 ZAB vs. ROS vs. Ant. B A B Cysteine (Cys) I x ¯ ± SE 163.35 ± 3.07 229.32 ± 3.52 149.33 ± 3.41 93.10 ± 1.10 Q 1 152.41 217.42 136.75 89.49 Q 2 (IQR) 168.28 (20.05) B 229.17 (17.12) A 143.13 (26.49) B 92.88 (7.34) C Q 3 172.46 234.54 163.24 96.83 Glycine (Gly) I x ¯ ± SE 68.81 ± 1.92 88.60 ± 1.03 69.36 ± 1.60 57.25 ± 0.69 Q 1 61.34 83.57 63.75 54.38 Q 2 (IQR) 71.38 (16.50) B 88.39 (9.56) A 66.38 (7.20) B 56.96 (6.03) C Q 3 77.83 93.13 70.96 60.41 Serine (Ser) I x ¯ ± SE 273.61 ± 3.77 326.20 ± 2.31 225.79 ± 2.77 184.75 ± 1.71 Q 1 263.55 318.96 214.73 180.13 Q 2 (IQR) 275.68 (23.06) B 326.89 (12.68) A 227.17 (16.76) C 185.01 (9.16) D Q 3 286.61 331.64 231.50 189.29 Threonine (Thr) I x ¯ ± SE 141.01 ± 4.28 a 139.20 ± 1.34 a 123.17 ± 1.01 b 80.10 ± 0.99 c Q 1 119.21 133.71 118.35 76.39 Q 2 (IQR) 139.94 (41.64) 139.5 (9.77) 122.99 (9.15) 80.38 (7.99) Q 3 160.85 143.48 127.49 84.38 Nat. vs. SIL vs. SOP a b c Tyrosine (Tyr) I x ¯ ± SE 12.29 ± 0.43 15.35 ± 0.34 6.91 ± 0.40 6.67 ± 0.41 Q 1 10.49 13.64 6.41 5.64 Q 2 (IQR) 12.89 (2.81) B 15.39 (3.48) A 7.41 (1.91) C 7.38 (2.63) C Q 3 13.30 17.12 8.32 8.28 ZAB vs. ROS vs. Ant. B A C Arginine (Arg) II x ¯ ± SE 24.84 ± 0.79 b 34.94 ± 0.42 a 15.65 ± 0.29 c 16.15 ± 0.42 c Q 1 21.95 32.70 14.42 14.39 Q 2 (IQR) 24.59 (5.29) 34.39 (3.80) 16.13 (2.08) 16.39 (3.10) Q 3 27.25 36.50 16.49 17.49 ZAB vs. ROS vs. Ant. b a c Asparagine (Asn) II x ¯ ± SE 313.16 ± 4.99 432.35 ± 2.86 246.77 ± 3.26 237.69 ± 1.99 Q 1 302.5 421.59 232.62 228.42 Q 2 (IQR) 317.31 (25.06) B 430.94 (20.08) A 241.48 (31.30) C 238.41 (15.70) C Q 3 327.56 441.67 263.92 244.12 ZAB vs. ROS vs. Ant. B A C Glutamine (Gln) II x ¯ ± SE 529.06 ± 6.84 b 623.95 ± 5.04 a 414.67 ± 2.64 c 351.19 ± 3.72 d Q 1 509.06 600.73 406.49 332.55 Q 2 (IQR) 521.50 (30.51) 623.48 (41.83) 412.54 (15.84) 349.53 (31.96) Q 3 539.57 642.56 422.33 364.51 Histidine (His) II x ¯ ± SE 133.04 ± 3.02 a 101.81 ± 1.01 b 71.05 ± 1.10 c 68.62 ± 0.71 c Q 1 128.42 97.40 66.89 65.64 Q 2 (IQR) 132.50 (15.12) 102.48 (8.75) 69.65 (8.37) 68.49 (5.53) Q 3 143.54 106.15 75.27 71.17 ZAB vs. ROS vs. Ant. a b c Lysine (Lys) II x ¯ ± SE 167.59 ± 3.61 231.48 ± 2.75 128.46 ± 1.29 115.63 ± 0.74 Q 1 152.75 219.45 124.67 113.68 Q 2 (IQR) 162.49 (25.32) B 230.45 (18.80) A 128.49 (7.92) C 116.39 (4.53) D Q 3 178.07 238.24 132.60 118.21 Proline (Pro) III x ¯ ± SE 87.53 ± 2.92 b 82.45 ± 2.56 b 362.10 ± 7.85 a 373.20 ± 8.13 a Q 1 76.60 71.53 331.58 335.57 Q 2 (IQR) 89.14 (25.72) 83.13 (18.10) 374.09 (69.02) 380.04 (69.61) Q 3 102.32 89.62 400.60 405.18 ZAB vs. ROS vs. Ant. b b a Nat. vs. SIL vs. SOP b a a Nat. vs. Ant. differ significantly Isoleucine (Ile) IV x ¯ ± SE 162.45 ± 3.79 a 143.93 ± 1.61 b 117.30 ± 1.60 c 100.93 ± 0.62 d Q 1 148.25 138.68 111.08 98.77 Q 2 (IQR) 162.48 (30.83) 144.89 (12.13) 118.29 (10.70) 101.39 (4.52) Q 3 179.08 150.80 121.78 103.28 Leucine (Leu) IV x ¯ ± SE 333.40 ± 4.77 a 290.65 ± 3.82 b 240.59 ± 3.13 c 242.72 ± 1.78 c Q 1 312.63 278.40 224.67 237.50 Q 2 (IQR) 331.99 (31.35 291.74 (24.45) 239.15 (28.87) 243.39 (11.74) Q 3 343.97 302.85 253.54 249.23 ZAB vs. ROS vs. Ant. a b c Methionine (Met) IV x ¯ ± SE 92.10 ± 3.11 a 87.79 ± 0.79 a 67.46 ± 0.66 b 50.42 ± 0.79 c Q 1 79.01 84.48 66.03 47.66 Q 2 (IQR) 89.19 (24.54) 87.33 (5.79) 67.43 (3.47) 50.20 (5.55) Q 3 103.55 90.27 69.50 53.21 Nat. vs. SIL vs. SOP a b c Phenylalanine (Phe) IV x ¯ ± SE 44.87 ± 1.66 ab 46.20 ± 0.61 ab 48.72 ± 1.17 a 42.50 ± 0.89 b Q 1 37.49 43.47 44.35 39.93 Q 2 (IQR) 44.95 (14.70) 46.42 (4.90) 49.4 (8.94) 43.38 (6.41) Q 3 52.19 48.37 53.29 46.33 Nat. vs. SIL vs. SOP ab a b Tryptophan (Trp) IV x ¯ ± SE 189.82 ± 3.71 255.90 ± 2.97 145.15 ± 1.20 128.45 ± 1.22 Q 1 176.42 244.04 140.58 124.39 Q 2 (IQR) 188.14 (25.84) B 254.37 (22.42) A 145.04 (9.16) C 127.39 (5.84) D Q 3 202.26 266.46 149.74 130.23 Valine (Val) IV x ¯ ± SE 76.77 ± 1.51 a 69.89 ± 0.94 b 49.85 ± 0.82 c 47.51 ± 0.83 c Q 1 71.41 66.29 46.37 44.62 Q 2 (IQR) 77.69 (12.03) 68.52 (8.12) 50.05 (6.13) 47.39 (6.46) Q 3 83.44 74.40 52.50 51.08 ZAB vs. ROS vs. Ant. a b c Non-proteogenic amino acids (μM) Ornithine (Orn) x ¯ ± SE 94.95 ± 3.81 116.33 ± 2.55 67.20 ± 1.24 ND Q 1 78.61 109.38 64.80 ND Q 2 (IQR) 89.72 (37.50) B 114.95 (9.11) A 68.54 (6.44) C ND Q 3 116.11 118.50 71.24 ND Citrulline (Cit) x ¯ ± SE 4.15 ± 0.49 ND ND ND Q 1 2.48 ND ND ND Q 2 (IQR) 4.45 (3.89) ND ND ND Q 3 6.38 ND ND ND Taurine (Tau) x ¯ ± SE 16.18 ± 0.67 14.98 ± 0.51 10.32 ± 0.28 7.61 ± 0.18 Q 1 13.92 12.59 9.38 6.60 Q 2 (IQR) 15.69 (2.84) A 14.39 (3.90) A 10.18 (2.11) B 7.51 (1.90) C Q 3 16.76 16.50 11.50 8.50 α-aminobutyric acid (AABA) x ¯ ± SE 11.67 ± 0.42 a 9.34 ± 0.36 b 7.04 ± 0.30 c 10.41 ± 0.34 c Q 1 9.95 8.40 5.50 8.91 Q 2 (IQR) 11.50 (2.84) 8.57 (2.13) 7.45 (2.24) 10.02 (2.98) Q 3 12.8 10.53 7.74 11.89 ZAB vs. ROS vs. Ant. a b c Nat. vs. SIL vs. SOP a b b Nat. vs. Ant. differ significantly β-aminobutyric acid (BABA) x ¯ ± SE 14.01 ± 0.38 21.74 ± 0.64 8.55 ± 0.31 4.55 ± 0.23 Q 1 12.51 18.76 7.52 3.58 Q 2 (IQR) 13.68 (2.26) B 21.08 (4.52) A 8.54 (1.84) D 4.4 (1.92) C Q 3 14.77 23.28 9.36 5.50 ZAB vs. ROS vs. Ant. B A C γ-aminobutyric acid (GABA) x ¯ ± SE 6.84 ± 0.55 7.29 ± 0.35 3.91 ± 0.42 ND Q 1 6.54 5.50 2.52 ND Q 2 (IQR) 7.89 (2.12) A 6.96 (3.01) A 3.72 (2.98) B ND Q 3 8.66 8.50 5.5 ND ZAB vs. ROS vs. Ant. A A B Nat. vs. SIL vs. SOP A B B Nat. vs. Ant. differ significantly β-alanine (β-Ala) x ¯ ± SE 22.81 ± 1.12 16.25 ± 0.53 ND ND Q 1 18.46 14.52 ND ND Q 2 (IQR) 21.53 (8.88) A 16.04 (2.96) B ND ND Q 3 27.34 17.48 ND ND Total amount of amino acids (mg/mL) x ¯ ± SE 0.52 ± 0.003 b 0.60 ± 0.002 a 0.41 ± 0.002 c 0.39 ± 0.002 d Q 1 0.51 0.59 0.41 0.39 Q 2 (IQR) 0.52 (0.01) 0.60 (0.01) 0.42 (0.01) 0.39 (0.01) Q 3 0.53 0.60 0.42 0.40 Table 3. Spatial and temporal variation of female reproductive success (FRS) and pollinaria removal (PR) in Epipactis palustris populations. Data ( n = 30) show the mean ± standard error. Parameter Natural Populations Anthropogenic Populations ZAB ROS SIL SOP FRS (%) 94.40 ± 2.83 81.47 ± 4.19 87.99 ± 3.36 90.60 ± 2.49 PR (%) 97.03 ± 2.68 87.48 ± 3.46 85.66 ± 4.36 96.55 ± 1.61 PR/FRS 1.07 ± 0.06 1.32 ± 0.22 1.07 ± 0.10 1.08 ± 0.03 Table 4. Measured flower structure properties. Abbreviation Full Name AI isthmus area FH length of flowers FW width of flowers HL length of hypochile HW width of hypochile LDS length of dorsal sepal LE length of epichile LI length of isthmus LL length of labellum LLS length of lateral sepal LP length of petal WDS width of dorsal sepal WE width of epichile WI width of isthmus WLS width of lateral sepal WP width of petal Table 5. Amino acids evaluated during HPLC analysis. Abbreviation Full Name AABA α-aminobutyric acid Ala alanine Arg arginine Asn asparagine Asp aspartic acid BABA β-aminobutyric acid Cit citrulline Cys cysteine GABA γ-aminobutyric acid Gln glutamine Glu glutamic acid Gly glycine His histidine Ile isoleucine Leu leucine Lys lysine Met methionine Orn ornithine Phe phenylalanine Pro proline Ser serine Tau taurine Thr threonine Trp tryptophan Tyr tyrosine Val valine β-Ala β-alanine Chicago/Turabian Style Brzosko, Emilia, Andrzej Bajguz, Justyna Burzyńska, and Magdalena Chmur. 2021. "Nectar Chemistry or Flower Morphology—What Is More Important for the Reproductive Success of Generalist Orchid Epipactis palustrisin Natural and Anthropogenic Populations?" International Journal of Molecular Sciences22, no. 22: 12164. https://doi.org/10.3390/ijms222212164 Brzosko E, Bajguz A, Burzyńska J, Chmur M. Nectar Chemistry or Flower Morphology—What Is More Important for the Reproductive Success of Generalist Orchid Epipactis palustrisin Natural and Anthropogenic Populations? International Journal of Molecular Sciences. 2021; 22(22):12164. https://doi.org/10.3390/ijms222212164 Chicago/Turabian Style Brzosko, Emilia, Andrzej Bajguz, Justyna Burzyńska, and Magdalena Chmur. 2021. "Nectar Chemistry or Flower Morphology—What Is More Important for the Reproductive Success of Generalist Orchid Epipactis palustrisin Natural and Anthropogenic Populations?" International Journal of Molecular Sciences22, no. 22: 12164. https://doi.org/10.3390/ijms222212164 Int. J. Mol. Sci. , EISSN 1422-0067, Published by MDPI

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Dispersed medieval settlement remains and a Roman building immediately south west of St Cosmus and St Damian's Church, Blean - 1018785 | Historic England List entry 1018785. Grade Not applicable to this List entry. Scheduled Monument: Dispersed Medieval Settlement Remains And A Roman Building Immediately South West Of St Cosmus And St Damian's Church. May include summary, reasons for designation and history. Dispersed medieval settlement remains and a Roman building immediately south west of St Cosmus and St Damian's Church Listed on the National Heritage List for England. Search over 400,000 listed places Overview Heritage Category: Scheduled Monument List Entry Number: 1018785 Date first listed: 07-Jul-1999 Heritage Category: Scheduled Monument List Entry Number: 1018785 Date first listed: 07-Jul-1999 Location The building or site itself may lie within the boundary of more than one authority. County: Kent District: Canterbury (District Authority) Parish: Blean National Grid Reference: TR 12874 60619 Reasons for Designation Medieval rural settlements in England were marked by great regional diversity in form, size and type, and the protection of their archaeological remains needs to take these differences into account. To do this, England has been divided into three broad Provinces on the basis of each area's distinctive mixture of nucleated and dispersed settlements. These can be further divided into sub-Provinces and local regions, possessing characteristics which have gradually evolved during the last 1500 years or more. This monument lies in the Eastern Weald sub-Province of the South-eastern Province, bounded by the North and South Downs and comprising an oval arrangement of inward facing escarpments of chalk and sandstone, separated by clay vales, all ringing a higher sandstone ridge. Apart from concentrations of nucleated settlements in the Vale of Holmsdale and around Canterbury, the sub- Province is dominated by high and very high densities of dispersed settlements, giving a countryside with farmsteads and associated enclosed fields, of medieval foundation, intermixed with cottages, medieval moated sites and hamlets bearing the names `green' or `dene'. The Canterbury-Thanet local region contains the regional centre of Canterbury, on which converges routes from Dover and former ports such as Richborough and Reculver. It is a long cleared and anciently settled countryside, lacking scattered farmsteads and with villages clustered as densely as in the Midlands. It has probably been a zone of arable cultivation for several millennia.Medieval dispersed settlements, comprising hamlets of up to five dwellings or isolated farmsteads, were scattered throughout the parish or township. Often occurring in the more densely wooded, less intensively farmed areas, the form and status of dispersed settlements varied enormously. When they survive as earthworks, their most easily distinguishable features include roads and tracks, platforms on which stood houses and other buildings such as barns, and the enclosed fields or irregular field systems with which the dwellings were associated. These rural settlements can also be represented by below ground deposits. Higher status dwellings, such as moated residences or manorial complexes, may have well-defined boundaries and planned gardens. In the western and south eastern provinces of England, dispersed settlements were the most distinctive aspect of medieval life, and their archaeological remains are one of the most important sources of understanding about rural life in the five or more centuries following the Norman conquest.The medieval settlement remains at St Cosmus and St Damian in the Blean represent the predominant, dispersed form of medieval rural settlement within the Eastern Weald sub-province. The settlement remains survive well, in association with an adjacent, contemporary church and an earlier Roman building, retaining visible earthworks. Field survey indicates that the monument will contain important archaeological and environmental evidence for the development and abandonment of the medieval settlement and the earlier Roman building. Details The monument includes the remains of a dispersed medieval settlement and an earlier Roman building situated on the southern slope of a clay hill around 7km north west of Canterbury. The Roman remains are represented by below ground archaeology, and have been identified as a villa. The scatter of masonry visible in the plough soil suggests that it occupies the northern part of the monument. Analysis of associated pottery fragments indicates that the building was in use during the first to third centuries AD. The dispersed medieval settlement survives in the form of earthworks and associated buried remains. The earthworks survive to a height of up to around 0.5m and represent three adjoining north west-south east aligned rectangular enclosures identified as a manor house complex and two associated, contemporary dwellings. Part of a roughly north-south aligned track runs along the eastern side of the monument. Documentary evidence, including an entry in the Domesday Book, suggests that the settlement was in existence by the 11th century. Analysis of pottery fragments found within the settlement suggests that it had fallen into disuse by the early 15th century. Immediately beyond the monument to the north east is the associated parish church of St Cosmus and St Damian, Listed Grade II, the standing fabric of which dates mainly to the 13th century. The church and its churchyard, both of which remain in use, are not included in the scheduling. Three telegraph poles situated on the monument are excluded from the scheduling, although the ground beneath them is included.MAP EXTRACT The site of the monument is shown on the attached map extract. Legacy The contents of this record have been generated from a legacy data system. Legacy System number: 31400 Legacy System: RSM Sources Books and journalsSt Clair-Terry, S, Blean During the Middle Ages, (1986)Tatton-Brown, T, 'Medieval Village Research Group Annual Report' in Medieval Village Research Group Annual Report: Kent, , Vol. 25, (1977), 9-10 Legal This monument is scheduled under the Ancient Monuments and Archaeological Areas Act 1979 as amended as it appears to the Secretary of State to be of national importance. This entry is a copy, the original is held by the Department for Culture, Media and Sport.

https://historicengland.org.uk/listing/the-list/list-entry/1018785

Yu. L. Maistrenko's research works | National Academy of Sciences of Ukraine, Kyiv (ISP) and other places Yu. L. Maistrenko's 15 research works with 655 citations and 691 reads, including: PARTIAL SYNCHRONIZATION IN A SYSTEM OF COUPLED LOGISTIC MAPS Yu. L. Maistrenko's research while affiliated with National Academy of Sciences of Ukraine and other places Publications (15) PARTIAL SYNCHRONIZATION IN A SYSTEM OF COUPLED LOGISTIC MAPS May 2012 A. V.taborov Yu. L.maistrenko Erik Mosekilde Clustering (or partial synchronization) in a system of globally coupled chaotic oscillators is studied by means of a model of three coupled logistic maps. For this model we determine the regions in parameter space where total and partial synchronization take place, examine the bifurcations through which total synchronization (one-cluster dynamics)... On-Off Intermittency and Riddled Basins of Attraction in a Coupled Map System Article Apr 2012 Jakob Lund Laugesen Erik Mosekilde Yu. L.maistrenko Volodymyr Leonidovich Maistrenko The paper examines the appearance of on-off intermittency and riddled basins of attraction in a system of two coupled one-dimensional maps, each displaying type-III intermittency. The bifurcation curves for the transverse destablilization of low periodic orbits embeded in the synchronized chaotic state are obtained. Different types of riddling bifu... CHAOTIC SYNCHRONIZATION AND ANTISYNCHRONIZATION IN COUPLED SINE MAPS Nov 2011 Volodymyr Leonidovich Maistrenko Yu. L.maistrenko Erik Mosekilde This paper investigates different types of chaotic synchronization in a system of two coupled sine maps. Due to the bimodal nature of the individual map, there is a range of parameters in which two synchronized chaotic states coexist along the main diagonal. In certain parameter regions, various (regular or chaotic) asynchronous states coexist with... Partial synchronization in a system of coupled logistic maps Jan 2000 A V Taborov Yu. L Maistrenko Erik Mosekilde Cite Unfolding of the riddling bifurcation Article Nov 1999 Yu.L. Maistrenko Volodymyr Leonidovich Maistrenko O. Popovych Erik Mosekilde We present analytical conditions for the riddling bifurcation in coupled, identical, smooth one-dimensional maps to be soft or hard and describe a generic scenario for the transformations of the basin of attraction following a soft riddling bifurcation. Desynchronization of chaos in coupled logistic maps Full-text available Oct 1999 Yu. L. Maistrenko Volodymyr Leonidovich Maistrenko O Popovych Erik Mosekilde When identical chaotic oscillators interact, a state of complete or partial synchronization may be attained in which the motion is restricted to an invariant manifold of lower dimension than the full phase space. Riddling of the basin of attraction arises when particular orbits embedded in the synchronized chaotic state become transversely unstable... Transverse instability and riddled basins in a system of two coupled logistic maps Mar 1998 Yu. L. Maistrenko Volodymyr Leonidovich Maistrenko A. Popovich Erik Mosekilde Riddled basins denote a characteristic type of fractal domain of attraction that can arise when a chaotic motion is restricted to an invariant subspace of total phase space. An example is the synchronized motion of two identical chaotic oscillators. The paper examines the conditions for the appearance of such basins for a system of two symmetricall... Chaos Synchronization and Riddled Basins in Two Coupled One-dimensional Maps Feb 1998 Tomasz Kapitaniak Yu.L. Maistrenko The connection between chaos synchronization and phenomena of riddled basins is discussed for a family of two-dimensional piecewise linear endomorphisms which consist of two linearly coupled one-dimensional maps. Under analytically given conditions, chaotic behavior in both maps can synchronize but synchronized state is characterized by different t... On Period-adding Sequences of Attracting Cycles in Piecewise Linear Maps Feb 1998 Yu.L. Maistrenko Volodymyr Leonidovich Maistrenko Svitlana Popovych We study numerically bifurcations in a family of bimodal three-piecewise linear continuous one-dimensional maps. Attention is paid to the attracting cycles arising after the bifurcation ‘from unimodal map to bimodal map’. It is found that this type of bifurcation is accompanied by the appearance of period-adding cascades of attracting cycles γ(a11+... Riddled basins of attraction for synchronized type-I intermittency Feb 1998 M. Manscher M. Nordahn Erik Mosekilde Yu. L Maistrenko Chaotic motion restricted to an invariant subspace of total phase space may be associated with basins of attraction that are riddled with holes belonging to the basin of another limiting state. We study the emergence of such basins for a system of two coupled one-dimensional maps, each exhibiting type-I intermittency. Role of the Absorbing Area in Chaotic Synchronization Feb 1998 Yu. L. Maistrenko Volodymyr Leonidovich Maistrenko A. Popovich Erik Mosekilde When two identical chaotic oscillators interact, one or more intervals of coupling parameters generally exist in which the synchronized state is weakly stable and its basin of attraction is riddled with holes that are repelled from it. Here, we discuss the role of the absorbing area for the emergence of local vs global riddling and for controlling... On-Off Intermittency and Riddled Basins of Attraction in a Coupled Map System Jan 1998 Jakob Lund Laugesen Erik Mosekilde Yu.L. Maistrenko Volodymyr Leonidovich Maistrenko The paper examines the appearance of on-off intermitting and Existence and stability of periodic solutions for chains of connected oscillators Jul 1997 Yu. L. Maistrenko A. V. Popovich We consider a nonlinear system of difference equations. This system corresponds to chains of N symmetrically connected oscillators with sufficiently general type of connection, which includes, among others, local and global connection. We prove a theorem on the existence and stability of space-time periodic solutions of such systems for sufficientl... Cite Request full-text Bifurcations of Attracting Cycles from Time-Delayed Chua's Circuit Feb 1995 Yu. L. Maistrenko Volodymyr Leonidovich Maistrenko Svitlana Popovych Leon O. Chua We study the bifurcations of attracting cycles for a three-segment (bimodal) piecewise-linear continuous one-dimensional map. Exact formulas for the regions of periodicity of any rational rotation number (Arnold’s tongues) are obtained in the associated three-dimensional parameter space. It is shown that the destruction of any Arnold’s tongue is a... Cycles of Chaotic Intervals in a Time-Delayed Chua's Circuit Dec 1993 Yu. L. Maistrenko Volodymyr Leonidovich Maistrenko Leon O. Chua The bifurcation set in the three-dimensional parameter space of the periodically driven van der Pol oscillator is investigated by continuation of local bifurcation curves. The two regions in which the driving frequency ω is greater or less than the limit cycle frequency ω 0 of the nondriven oscillator are considered separately. For the case ω>ω 0 ,... Citations ... Kumar). also been found to exist in identical chaotic oscillators [17], pulse coupled oscillators [18], a scale free network [19], one-dimensional ring of diffusively coupled oscillators [20] and coupled logistic maps [21] . Apart from the frequency distribution and network degree distribution, these states have also been encountered with inhomogeneous coupling [22] for identical phase oscillators. ... Reference: Partially synchronized states in an ensemble of chemo-mechanical oscillators Partial synchronization in a system of coupled logistic maps Jan 2000 A. V.taborov Yu. L. Maistrenko Erik Mosekilde ... For a system of two symmetrically coupled, identical oscillators this transition typically takes place via a transverse period-doubling or a transverse pitchfork bifurcation [1] [2]. If the coupled system involves more than two coupled chaotic oscillators, a whole new range of additional phenomena can occur, including partial synchronization (or clustering [13] [14] [15] [16]) as well as various forms of wave-like dynamics. A state of partial synchronization is said to occur when the interacting oscillators synchronize with one another in different groups, but there is no synchronization among the groups. ... Reference: Partial synchronization and clustering in a system of diffusively coupled chaotic oscillators PARTIAL SYNCHRONIZATION IN A SYSTEM OF COUPLED LOGISTIC MAPS May 2012 A. V.taborov Yu. L. Maistrenko Erik Mosekilde ... The control of bistability has been reported by developing the parametrically coupled network of sine maps [40]. The simultaneous presence of chaotic synchronization and chaotic antisynchronization has been noticed in coupled sine maps and spatial synchronization with temporal periodic solution has been delineated in coupled sine circle maps [41, 42]. Through a system of coupled maps, routes to the crisis, bifurcation transitions, and dimension variability are also detailed, followed by, spatial and spatio-temporal intermittency has been revealed in [43]. ... Reference: Influence of Parametric Symmetry on the Dynamics of 3D Sinusoidal Discrete Systems CHAOTIC SYNCHRONIZATION AND ANTISYNCHRONIZATION IN COUPLED SINE MAPS Nov 2011 dx.doi.org V. L. Maistrenko Yu. L. Maistrenko Erik Mosekilde ... The skew tent map family g(y, ν, s L , s R ) = ν + s L y, y ≤ 0, ν + s R y, y ≥ 0, (4.6) can in some ways be regarded as a normal form for border-collision bifurcations [7]. As the value of ν ∈ R passes through 0, a border-collision bifurcation occurs and the resulting dynamics depends in a complicated but well understood way on the values of s L , s R ∈ R [1,15, 20] . ... Reference: Differentiable conjugacies for one-dimensional maps Cycles of Chaotic Intervals in a Time-Delayed Chua's Circuit Dec 1993 Yu. L. Maistrenko V. L. Maistrenko Leon O. Chua ... Многие задачи из механики, физики, силовой электроники, теории управления, экономики и биологии приводят к исследованию одномерных необрати-мых кусочно-гладких бимодальных отображений (рис. 1) [1] [2] [3][4][5][6]: ... Reference: Bifurcation Analysis of Piecewise Smooth Bimodal Maps Using Normal Form Bifurcations of Attracting Cycles from Time-Delayed Chua's Circuit Feb 1995 Yu. L. Maistrenko V. L. Maistrenko S.I. VIKUL Leon O. Chua ... If the trajectory of the system is bounded and y n − x n → 0 as n → ∞, where x = y is the synchronization subspace of the one-dimensional restriction of T to ∆, then the trajectory synchronizes [3]. The chaotic set in the invariant subspace attracts a set of positive Lebesgue measure points [4, 5] . ... Reference: Dynamics and Global Bifurcations in Two Symmetrically Coupled Non-Invertible Maps Role of the Absorbing Area in Chaotic Synchronization Feb 1998 Yu. L. Maistrenko V. L. Maistrenko A. Popovich Erik Mosekilde ... If the trajectory of the system is bounded and y n − x n → 0 as n → ∞, where x = y is the synchronization subspace of the one-dimensional restriction of T to ∆, then the trajectory synchronizes [3]. The chaotic set in the invariant subspace attracts a set of positive Lebesgue measure points [4, 5]. ... Reference: Dynamics and Global Bifurcations in Two Symmetrically Coupled Non-Invertible Maps Transverse instability and riddled basins in a system of two coupled logistic maps Mar 1998 Yu. L. Maistrenko V. L. Maistrenko A. Popovich Erik Mosekilde ... Globally riddled basins resemble the morphology of a fat fractal set with a maximal density concentrated around the attractor A as well as around the preimages of A. Alternatively, if there is no access for the trajectories to go to other attractors or infinity, it is typical for allmost all trajectories to return to the neighborhood of A, and consequently, by repeating such behavior, bursts away from the diagonal are produced. At the end, most of the trajectories will be eventually attracted to A. In this case, the basin of A appears to be only locally riddled (see Ashwin et al., 1996;Maistrenko et al., 1997; Maistrenko et al., 1999b) . It is filled with initial conditions that are not leading to A, but the set that consists of all of these initial conditions is characterized by having a Lebesgue measure of zero. ... Reference: Complex dynamics in a simple model of interdependent open economies Unfolding of the riddling bifurcation Nov 1999 Yu. L. Maistrenko V. L. Maistrenko O. Popovych Erik Mosekilde ... One of the important results of such configuration is the dramatic reduction in the critical value of the parameter for the onset of chaos [36]. It would give rise to the coexisting orbits, which may be periodic, chaotic, quasi-periodic, even the infinity [37, 38] . ... Reference: Codimension-one and -two bifurcation analysis of a two-dimensional coupled logistic map Chaos Synchronization and Riddled Basins in Two Coupled One-dimensional Maps Feb 1998 Tomasz Kapitaniak Yu. L. Maistrenko ... Similarly holds for the BC of a k cycle, considering the k th iterate of the map in the involved kink point (for which it corresponds to a BC of a …xed point). The result of a BC in this case can be described by using the two slopes, see [17] , the survey [31] and references therein. The skew-tent map as a normal form is also useful in a bimodal system, as shown for the bimodal piecewise linear case in [21]. ... Reference: A piecewise smooth model of evolutionary game for residential mobility and segregation On Period-adding Sequences of Attracting Cycles in Piecewise Linear Maps Feb 1998 Yu. L. Maistrenko V. L. Maistrenko S.I. VIKUL Top co-authors (11) Volodymyr Leonidovich Maistrenko

https://www.researchgate.net/scientific-contributions/Yu-L-Maistrenko-2144094324

MILLENNIUM COMFORT HOME HEALTHCARE, LLC, NPI 1023113388 - Home Health in Pharr, TX Mch Healthcare a provider in 3912 N Jackson Rd Pharr, Tx 78577. Taxonomy code 251E00000X. Accepted Insurance: Medicaid and Medicare MILLENNIUM COMFORT HOME HEALTHCARE, LLC NPI 1023113388 Home Health in Pharr, TX Table of Contents NPI Profile Information Primary Taxonomy Accepted Insurance Authorized Official About MILLENNIUM COMFORT HOME HEALTHCARE, LLC Mch Healthcareis a provider established inPharr, Texasoperating as aHome Health. The NPI number of this provider is1023113388and was assigned on September 2006. The practitioner's primary taxonomy code is251E00000X. The provider is registered as an organization and their NPI record was last updated 3 years ago. The provider's is doing business as Mch Healthcare. The authorized official of this NPI record is Michael Palomo (Ceo) NPI 1023113388 Provider Name MILLENNIUM COMFORT HOME HEALTHCARE, LLC Location Address 3912 N JACKSON RD PHARR, TX 78577 Location Phone (956) 583-1500 Mailing Address 3912 N JACKSON RD PHARR, TX 78577 NPI Entity Type Organization Is Sole Proprietor? N/A Is Organization Subpart? No Other Organization Name MCH HEALTHCARE Other Name Type Doing Business As (3) Enumeration Date 09-13-2006 Last Update Date 09-10-2020 According to the Home Health Compare program data, Mch Healthcare doesn't have a Quality of Patient Star Rating available at this time because the number of patient episodes for this measure is too small to report. To have a quality star rating computed, Home Health Agencies must have reported data for 5 of the 7 measures used in the Quality of Patient Care Star Ratings calculation. The seven measures used to calculate the quality star rating are: timely initiation of care, improvement in ambulation, bed transferring, bathing, shortness of breath, management of oral medications and relapse in acute care hospitalizations. The CLIA number of Mch Healthcare is 45D1060178 registered as a "home health agency" facility with a CLIA Certificate of Waiver. This CLIA certificate is issued to Mch Healthcare to perform only waived tests. CLIA defines waived tests as simple tests with a low risk for an incorrect result. Waived tests include certain tests listed in CLIA regulations, tests cleared by the FDA for home use and tests approved by the FDA for waived status and that meet CLIA waiver criteria. Primary Taxonomy The primary taxonomy code defines the provider type, classification, and specialization. There could be only one primary taxonomy code per NPI record. For individual NPIs the license data is associated to the taxonomy code. Taxonomy Code 251E00000X Classification Home Health Type Agencies Taxonomy Description A public agency or private organization, or a subdivision of such an agency or organization, that is primarily engaged in providing skilled nursing services and other therapeutic services, such as physical therapy, speech-language pathology services, or occupational therapy, medical social services, and home health aide services. It has policies established by a professional group associated with the agency or organization (including at least one physician and one registered nurse) to govern the services and provides for supervision of such services by a physician or a registered nurse; maintains clinical records on all patients; is licensed in accordance with State or local law or is approved by the State or local licensing agency as meeting the licensing standards, where applicable; and meets other conditions found by the Secretary of Health and Human Services to be necessary for health and safety. Accepted Insurance The NPI profile data indicates this provider might be enrolled and accepting health plans from the following insurance companies or healthcare programs: Medicaid Medicare *Please verify directly with this provider to make sure your insurance plan is currently accepted. Business Address 3912 N JACKSON RD PHARR, TX ZIP 78577 Phone: (956) 583-1500 Fax: (956) 583-8865 Mailing Address 3912 N JACKSON RD PHARR, TX ZIP 78577 Phone: (956) 583-1500 Fax: (956) 583-8865 Location Map Authorized Official The name of the person authorized to submit the NPI application or to officially change data for a health care provider. <table><tbody><tr><td> Authorized Official Name</td><td> MICHAEL PALOMO</td></tr><tr><td> Authorized Official Title</td><td> CEO</td></tr><tr><td> Authorized Official Phone</td><td> (956) 583-1500</td></tr></tbody></table> Nursing Home Compare Information The Centers for Medicare and Medicaid Services publishes Home Health Compare quality of care data to provide consumers an easy way to compare "Medicare-certified" home health agencies throughout the nation. "Medicare-certified" home health agencies are approved by Medicare and meet certain federal health and safety requirements. The Home Health Compare information helps consumers learn how well home health agencies care for their patients, how often each agency used best practices when caring for its patients and what patients said about their recent home health care experience. <table><tbody><tr><td> Quality of Patient Care Rating Quality of Patient Care Rating The quality of patient care star rating summarizes 8 of the 23 quality measures reported on Home Health Compare. It provides a single indicator of an agency's performance compared to other agencies. </td><td>- 0 out of 5 stars - The number of patient episodes for this measure is too small to report.</td></tr><tr><td> Ownership Type Ownership Type Home health agencies can be run by private for-profit corporations, non-profit corporations, religious affiliated organizations or government entities. The type of ownership may affect agency resources and how services are organized. Quality can vary in home health agencies within each of the different types of ownership. Each agency needs to be judged on its own merits. </td><td> PROPRIETARY</td></tr><tr><td> Offers Nursing Care Offers Nursing Care? The home health agency offers care given or supervised by registered nurses. Nurses provide direct care; manage, observe, and evaluate a patient’s care; and teach the patient and his or her family caregiver. Examples include: giving IV drugs, shots, or tube feedings; changing dressings; and teaching about diabetes care. Any service that could be done safely by a non-medical person (or by yourself) without the supervision of a nurse isn’t skilled nursing care. Medicare covers home health skilled nursing care that's part time and intermittent. </td><td> Yes</td></tr><tr><td> Offers Physical Therapy Offers Physical Therapy? The home health agency offers treatment of injury and disease by mechanical means, like heat, light, exercise, and massage. </td><td> Yes</td></tr><tr><td> Offers Occupational Therapy Offers Occupational Therapy? The home health agency offers services given to help you return to usual activities (like bathing, preparing meals, and housekeeping) after illness either on an inpatient or outpatient basis. </td><td> Yes</td></tr><tr><td> Offers Speech Therapy Offers Speech Therapy? The home health agency offers services to assist with problems involving speech, language, and swallowing. Communication problems can be present at birth or develop after an injury or illness, like a stroke. </td><td> Yes</td></tr><tr><td> Offers Medical Social Services Medical Social Services? The home health agency offers services to help with social and emotional concerns related to your illness. This might include counseling or help in finding resources in your community. </td><td> Yes</td></tr><tr><td> Offers Home Health Aide Offers Home Health Aide? The home health agency offers part time or intermittent services to help with daily living activities. </td><td> No</td></tr><tr><td> Medicare Certification Date</td><td> 10-15-2007</td></tr><tr><td> Number of episodes used to calculate how much Medicare spends at this agency Number of episodes used to calculate how much Medicare spends at this agency Number of episodes of care used to calculate how much Medicare spends on an episode of care at this agency, compared to Medicare spending across all agencies nationally. </td><td></td></tr></tbody></table> CLIA Information The Clinical Laboratory Improvement Amendments (CLIA) of 1988 applies to facilities or sites that test human specimens for health assessment or to diagnose, prevent, or treat disease. The CLIA Program sets standards for clinical laboratory testing and issues certificates. The NPI / CLIA crosswalk information for the NPI number 1023113388 is: <table><tbody><tr><td> CLIA Number</td><td> 45D1060178</td></tr><tr><td> Facility Type</td><td> HOME HEALTH AGENCY</td></tr><tr><td> Certificate Type</td><td> Certificate of Waiver</td></tr></tbody></table> Additional Identifiers Additional identifier(s) currently or formerly used as an identifier for the provider. The codes may include UPIN, NSC, OSCAR, DEA, Medicaid State or PIN identification numbers. <table><tr><th> Identifier</th><th> Type / Code</th><th> Identifier State</th><th> Identifier Issuer</th></tr><tbody><tr><td> 679681</td><td> OTHER (01)</td><td></td><td> MEDICARE</td></tr><tr><td> 190823801</td><td> MEDICAID (05)</td><td> TX</td><td></td></tr></tbody></table> NPI Validation Check Digit Calculation The following table explains the step by step NPI number validation process using the ISO standard Luhn algorithm. Start with the original NPI number, the last digit is the check digit and is not used in the calculation. 1 0 2 3 1 1 3 3 8 8 Step 1: Double the value of the alternate digits, beginning with the rightmost digit. 2 0 4 3 2 1 6 3 16 Step 2: Add all the doubled and unaffected individual digits from step 1 plus the constant number 24. 2 + 0 + 4 + 3 + 2 + 1 + 6 + 3 + 1 + 6 + 24= 52 Step 3: Subtract the total obtained in step 2 from the next higher number ending in zero, the result is the check digit. 60 - 52 = 8 8 The NPI number 1023113388 is valid because the calculatedcheck digit 8using the Luhn validation algorithm matches the last digit of the original NPI number. Frequently Asked Questions What is MCH HEALTHCARE NPI number? The NPI number assigned to this healthcare provider is 1023113388, registered as an "organization" on September 13, 2006 Where is MCH HEALTHCARE located? The provider is located at 3912 N Jackson Rd Pharr, Tx 78577 and the phone number is (956) 583-1500 What kind of medical provider is MCH HEALTHCARE? This medical organization specializes in Home Health What insurance does MCH HEALTHCARE accept? The provider might be accepting Medicaid and Medicare. Please consult your insurance carrier or call the provider to make sure your health plan is currently accepted. What is MCH HEALTHCARE CLIA number? The provider's CLIA number is 45D1060178 for a "home health agency" facility with a CLIA Certificate of Waiver. This CLIA certificate is issued to perform only waived tests. CLIA defines waived tests as simple tests with a low risk for an incorrect result. Waived tests include certain tests listed in CLIA regulations, tests cleared by the FDA for home use and tests approved by the FDA for waived status and that meet CLIA waiver criteria.. How do I update my NPI information? The NPI record of MCH HEALTHCARE was last updated on September 13, 2006. To officially update your NPI information contact the National Plan and Provider Enumeration System (NPPES) at 1-800-465-3203 (NPI Toll-Free) or by email [email protected]. NPI Profile data is regularly updated with the latest NPI registry information, if you would like to update or remove your NPI Profile in this website pleasecontact us.

https://npiprofile.com/npi/1023113388

Increasing Engagement With Messages Regarding CRC Screening Among Adults Aged 45-49 - Full Text View - ClinicalTrials.gov Increasing Engagement With Messages Regarding CRC Screening Among Adults Aged 45-49 (CRC45+TEXT) The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been evaluated by the U.S. Federal Government. Read our disclaimer for details. ClinicalTrials.gov Identifier: NCT05281159 Recruitment Status : Active, not recruiting First Posted : March 16, 2022 Last Update Posted : May 27, 2022 Sponsor: University of California, Los Angeles UCLA Vatche and Tamar Manoukian Division of Digestive Diseases UCLA Anderson School of Management Carnegie Mellon University UCLA Department of Medicine Information provided by (Responsible Party): Daniel M. Croymans, MD, MBA, MS, University of California, Los Angeles Study Details Study Description Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Brief Summary: In May of 2021, the United States Preventive Service Task Force (USPSTF) updated their colorectal cancer (CRC) screening guidelines by recommending screening at an earlier age for average-risk adults starting at the age of 45 years old (Grade B recommendation). This is in addition to their Grade A recommendations of continuing to screen average-risk adults ages 50-75 years old. As the investigators health system aims to screen the newly eligible population of average-risk patients between the ages of 45-49, the investigators proposed randomized controlled trial is aimed to determine the most effective patient outreach approach to increase patients' engagement with messages regarding CRC screening and screening uptake within this specific age-group. Condition or disease Intervention/treatment Phase Colorectal Cancer Colorectal Neoplasms Behavioral: Risk Information Behavioral: Risk Information + Action Not Applicable Detailed Description: Colorectal cancer (CRC) is the second leading cause of cancer-related death for both men and women in the United States. One in 17 Americans will suffer from CRC during their lifetime and early detection of cancers and polyps by screening is shown to reduce CRC mortality. In 2021, the USPSTF updated its CRC screening guidelines to start screening average-risk individuals at the age of 45 years due in part to a rising incidence of CRC and premalignant polyps in younger patients. There will be three different text messages that have been formulated utilizing behavioral science principles, and then randomize them equally among the eligible population. There will be stratified patient randomization based on arm assignment in our 'CRC45+ Choice' evaluation to prevent cross-study contamination. The arms for this investigation will be 1) Control, 2) Risk Information and 3) Risk Information + Action. In the control (standard care) arm, patients will receive the standard SMS text message. In the Risk Information arm, patients will receive a message indicating their risk for colorectal cancer, on top of the content in the Control arm. Finally, in the Risk Information + Action arm patients will receive risk information and importance of taking quick action, on top of the content in the Control arm. The investigators will also randomly assign patients into receiving our text message on one of ten workdays, stratifying by arm assignment in both the CRC45+ Choice Evaluation and CRC45+ Text Evaluation. We will send text reminders to patients two weeks after they receive the initial text message. Reminder content will be very similar to the content of initial text message, and differ by arm. For this investigation the investigators hypothesize: Hyp 1: Patients assigned to the Risk Information + Action Arm (Arm 2) will have a higher click rate than patients assigned to the Control Arm (Arm 0) Hyp 2: Patients assigned to the Risk Information + Action Arm (Arm 2) will have a higher click rate than patients assigned to the Risk Information Arm (Arm 1) Analysis: The investigators will utilize patient-level ordinary least squares (OLS) estimation, with statistical inferences based on model-robust standard errors. The primary model term will be indicator variables for arm assignment. The analysis will adjust for sex, race/ethnicity, social vulnerability index (at the ZIP code level), and MyChart logins decile. Treatment effects will be summarized using rate differences and 95% CI's. Exploratory analyses will investigate heterogenous treatment effects by demographic subgroups by splitting the sample into each demographic subgroup as well as by testing demographics and text message arm interactions. Sensitivity analyses will be performed without covariates, and using logistic regression models in place of linear regression models. Missing covariate values will be handled by including 'unknown' indicators, along with mean imputation for quantitative covariates. The investigators will additionally explore how click rates within the SMS message vary with the day of the week. Primary analyses will use intention-to-treat protocol. Study Design Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Layout table for study information Study Type : Interventional  (Clinical Trial) Estimated Enrollment : 20000 participants Allocation: Randomized Intervention Model: Parallel Assignment Intervention Model Description:Parallel Assignment Parallel-arm randomized controlled trial,Three arms Arm 0: Standard of care control (standard text messaging) Arm 1: Experimental, "Risk Information" group Arm 2: Experimental, "Risk Information + Action" group Patient-level randomization stratified by arm assignment in CRC45+ Choice Evaluation. The investigators will also randomly assign patients into receiving the text message on one of ten workdays, stratifying by arm assignment in both the CRC45+ Choice Evaluation and CRC45+ Text Evaluation Masking: None (Open Label) Primary Purpose: Prevention Official Title: Quality Improvement Study Aimed at Increasing Engagement With Messages Regarding CRC Screening Among Adults Aged 45-49 (CRC45+ TEXT). Actual Study Start Date : February 21, 2022 Estimated Primary Completion Date : August 21, 2022 Estimated Study Completion Date : November 21, 2022 Arms and Interventions Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Arm Intervention/treatment No Intervention: Standard of Care This group will receive a standard SMS text message indicating that they have a MyChart message from their provider about CRC screening and should follow a link to be screened. Experimental: "Risk Information" This group will receive an SMS text message highlighting their risk for CRC. Behavioral: Risk Information This group will receive an SMS text message highlighting their risk for CRC on top of the content in the Control arm. Experimental: "Risk Information + Action" This group will receive an SMS message highlighting their risk for CRC and the importance of taking quick action. Behavioral: Risk Information + Action This group will receive an SMS text message highlighting their risk for CRC and the importance of taking quick action, on top of the content in the Control arm. Outcome Measures Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Primary Outcome Measures : Opening link within SMS Text message [ Time Frame: 1 week after getting the initial text message ] Rate of clicking the link to MyChart within the SMS text message Secondary Outcome Measures : Opening link within SMS Text Message [ Time Frame: 1 month after getting initial text message ] Rate of clicking the link to MyChart within the SMS text message during 1 month CRC45+ Choice MyChart Message opened [ Time Frame: 1 month after getting the initial text message ] Rate of opening MyChart message Overall completion of any CRC screening test [ Time Frame: 6 months after getting the initial text message ] Rate of completion of any CRC screening (FIT Kit or Colonoscopy) within the observation window. Eligibility Criteria Information from the National Library of Medicine Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the contacts provided below. For general information, Learn About Clinical Studies. Layout table for eligibility information Ages Eligible for Study: 45 Years to 49 Years   (Adult) Sexes Eligible for Study: All Accepts Healthy Volunteers: Yes Criteria Inclusion Criteria: UCLA Health patient Age 45-49 (as of 2/1/2022) Active primary care provider Visit within the last 3 years Exclusion Criteria: Age <45 or >50 years old at time of randomization Documented family history of CRC in EMR History (personal or family) of prior adenomatous polyps in the EMR History of high-risk cancer syndromes (e.g., Lynch, FAP) Prior CRC screening with colonoscopy or FIT Inactive MyChart status at enrollment No documented SMS-capable phone number Locations Layout table for location information United States, California UCLA Health Department of Medicine, Quality Office Westwood, California, United States, 90095 UCLA Vatche and Tamar Manoukian Division of Digestive Diseases Westwood, California, United States, 90095 University of California, Los Angeles UCLA Vatche and Tamar Manoukian Division of Digestive Diseases UCLA Anderson School of Management Carnegie Mellon University UCLA Department of Medicine Layout table for additonal information Responsible Party: Daniel M. Croymans, MD, MBA, MS, Medical Director, DOM Quality, University of California, Los Angeles ClinicalTrials.gov Identifier: NCT05281159 History of Changes Other Study ID Numbers: CRC45+TEXT First Posted: March 16, 2022 Key Record Dates Last Update Posted: May 27, 2022 Last Verified: May 2022 Layout table for additional information Studies a U.S. FDA-regulated Drug Product: No Studies a U.S. FDA-regulated Device Product: No Keywords provided by Daniel M. Croymans, MD, MBA, MS, University of California, Los Angeles: Colorectal Cancer Screening Behavioral Economics Colorectal Cancer Site Map Terms and Conditions U.S. Department of Health and Human Services

https://clinicaltrials.gov/show/NCT05281159

Post-stay email marketing implications for the hotel industry: Role of email features, attitude, revisit intention and leisure involvement level | Request PDF Request PDF | Post-stay email marketing implications for the hotel industry: Role of email features, attitude, revisit intention and leisure involvement level | Despite their widespread use in the hotel industry, limited research has been conducted on post-stay emails. Therefore, this study was designed to... | Find, read and cite all the research you need on ResearchGate Post-stay email marketing implications for the hotel industry: Role of email features, attitude, revisit intention and leisure involvement level Journal of Vacation Marketing 25(3):135676671881408 DOI: 10.1177/1356766718814081 Abstract Despite their widespread use in the hotel industry, limited research has been conducted on post-stay emails. Therefore, this study was designed to examine the effects of post-stay email features on customers’ attitudes towards a brand and their revisit intention. The email features analysed in the study consisted of personalization, message copy, interactivity, financial and illustration. Further, the effectiveness of each email feature was measured based on respondents’ leisure involvement levels. Results indicated that personalization, interactivity and financial features were significant predictors of revisit intention depending on customers’ involvement levels, while attitude was a significant mediator. Overall, the findings from the study add to the knowledge of how post-stay emails can be used when formulating customer retention strategies. ... Past research suggests that personalization by name leads to customer feelings of gratefulness and positive, gratitude-based behaviors such as referral of promotional campaign (Koch and Benlian 2015). Similarly, personalization has been found to increase web-survey response behavior (Sauermann and Roach 2013), attitude toward the brand (Yang et al. 2018) ... Past research suggests that personalization by name leads to customer feelings of gratefulness and positive, gratitude-based behaviors such as referral of promotional campaign (Koch and Benlian 2015). Similarly, personalization has been found to increase web-survey response behavior (Sauermann and Roach 2013), attitude toward the brand (Yang et al. 2018), and can make customers feel unique and valued, which can help cultivate intimate relationships with customers (Yang et al. 2018) . ... A large number of service firms support email-based customer-firm interactions, yet their email-based service recovery efforts remain under-researched. In addressing this issue, we investigate the effectiveness of post-service failure apology emails by treating offending firm (i.e., sender) and aggrieved customer (i.e., recipient) personalization as varying in degree. Based on an experimental design using a hypothetical service scenario, we find that sender personalization and customer personalization in the apology email interact to affect customer-perceived distributive justice and key customer recovery outcomes. We discuss the implications of our findings for IS research and management. ... Chang et al. (2013) revealed a positive relationship between attitudes and intentions toward the sender and consumer response. In their study, Yang et al. (2019) indicated that personalization, interactivity and financial features were significant predictors of revisit intention based on customers' involvement levels, while attitude was shown to be a significant mediator. Online opt-in e-mail marketing has become a significant source of revenue for retailers (Limbu & Jensen, 2018). ... Opt-in e-mail marketing influence on consumer behaviour: A Stimuli-Organism-Response (S-O-R) theory perspective The paper examines the influence of opt-in e-mail marketing on consumer behaviour. The study attempts to extend the Stimuli-Organism-Response (S-OR) theory that has been broadly explored in consumer research. Following a critical review of the literature organisation approach, a hypothetical model has been proposed for this study, based on identified factors, such as, informational value, entertainment-based message content, layout, visual appeal, attitude toward e-mail advertising and intention towards the sender in the context of opt-in email marketing. Data were collected in South Africa through an online survey of 436 opt-in e-mail marketing subscribers. Structural equation modelling (SEM) was employed to measure the proposed hypotheses of the study. The research results suggest that even during a pandemic, e-mail marketers could employ certain features in promotional and informational e-mail marketing communication, particularly infor-mational value, entertainment-based message content, layout, visual appeal, as a means to design their e-mail marketing messages and plan e-mail advertising campaigns. The findings of the study are intended to advance the e-mail marketing knowledge base to help marketers during a pandemic, such as COVID-19. The paper provides marketers with relevant insights on how to effectively engage with e-mail subscribers. ... The use of email marketing, a versatile and affordable medium that enables direct communication with guests and agencies/operators by increasing numbers of hotel businesses, fills this gap [8]. As stated by Yang, et al. [80] , even though emails are often used in the hotel industry, little study has been done on them, adding that post-stay emails may be applied when creating customer loyalty strategies. ... Persuasive Determinants in the Hotel Industry’s Newsletter Opening Rates Email marketing plays a key role in business communications and is one of the most widely used applications by consumers. The literature review points to several determinants that, when applied, increase the open rate of newsletters. This research evaluates the impact of six determinants of persuasion on the opening rate of a newsletter in the hotel industry. The determinants are the day of sending, the time of sending, subject line personalization, scarcity appeal, curiosity appeal, and authority figure. The chosen methodology focused on real experiments, using a high-end luxury hotel, and the respective customer database. The newsletter was sent to the subscriber list, where one part received the control and the other part received a variant with the test version. Ten A/B tests were conducted for each determinant. The results obtained were not in line with what is indicated in the literature review. Although the literature review yielded results that showed that the application of determinants increased the open rate of newsletters, this study obtained findings to the opposite and did not confirm what was prescribed by the reviewed literature. The results of the A/B tests were conclusive and revealed that the determinants did not increase the open rate of newsletters. ... Furthermore, hotel marketing should adopt a digital competitive strategy (De Pelsmacker et al., 2018). Additionally, personalized needs are also an important aspect of hotel marketing concerns, especially for high-leisure sports groups (Yang et al., 2018) . ... Is traditional marketing mix still suitable for hotel banquets? An empirical study of banquet marketing in five-star hotels Are traditional marketing mix strategies still suitable for hotel banquet marketing? Using the binary logistic regression analysis method, this study used 763 banquet sales records at the Quanzhou Hilton Hotel to comprehensively test the influence of traditional marketing mix strategies on banquet marketing effects. By focusing on new marketing methods (such as video, the Internet, and WeChat marketing), this study tested the effectiveness of traditional marketing strategies in the new media era. The findings revealed that a combination of products is easier to market than a single product, whereas price is still a key factor in hotel banquet marketing. However, sales channels and personal identity have no significant effects on banquet marketing. Finally, based on the failure cases analysis, this study proposed a feasible path for promoting banquet marketing. ... generic or nonpersonalized) newsletters result in more positive evaluations of the message (Maslowska et al., 2011). Similarly, customer personalization can increase web survey response rates (Fan & Yan, 2010;Heerwegh, 2005;Sauermann & Roach, 2013), compliance (Sahni et al., 2018), and brand attitudes and can make customers feel unique and valued, facilitating more intimate customer-firm relationships (Chandra et al., 2022; Yang et al., 2019) . However, there is little discussion in the literature of appropriate forms of salutations and email signatures, or contextual cues more generally, in emails, as indicated in Table 1, which offers an overview of prior research on apology-based service recovery efforts and outcomes. ... Digital presence in service recovery: The interactive effect of customer salutations and employee photographs in email signatures Service firms can manage failure apology emails depending on how much digital presence or lack thereof they wish to maintain toward aggrieved customers. While nascent research indicates positive effects of higher levels of digital presence, the efficacy of this strategy in service recovery settings remains unclear. Drawing on the concept of digital presence and the unified theory of social relations, the authors investigate the potential detrimental effects of combining certain salutation forms with digital presence in the form of employee photographs in these emails. The data were gathered using one survey among service employees (Prestudy, N=202), and two scenario-based experiments (Study 1, N=418; Study 2, N=449). Study 1 assesses customer reactions to different apology emails and shows that personalized salutations (i.e., addressing customers by first name) and digital presence in the form of employee photographs can negatively interact to affect recovery satisfaction and repatronage intentions, with customers’ perceived rapport mediating these relationships. Moreover, the results of a second experiment (Study 2) show that compensation after a service failure can mitigate the negative interactive effects. This research thus suggests using digital presence (in the form of employee photographs) in conversations with aggrieved customers only when customer salutation personalization is absent. ... Involvement is generally understood by the degree of relevance and its importance (Sharma & Klein, 2020). It has been continuously reviewed over the years on its impact on organisational behaviour (Nnadozie et al., 2019), psychology (Brewster et al., 2020;Qian et al., 2017), and marketing (Yang et al., 2019) . Previous studies on involvement were predominantly done to explain consumers' purchase intention (McClure & Seock, 2020) and as a mediating construct (Lim et al., 2019). ... Personal Values Perceptions on Involvement and Attitude Towards Discount Sales Promotion -Individualistic and Collectivist Values Perspective This study explores the effect of personal values (individualistic and collectivist values) on consumers' involvement and attitude towards discount sales promotion. In order to create and test a more parsimonious predictive-based research model, this study used a closed-ended survey and a two-stage technique using partial least square structural equation modelling. The findings imply that individualistic and collectivist beliefs influence consumer involvement and attitude toward discount sales promotions. Furthermore, the findings of this study expand current knowledge on similar sales promotion areas where this study details the effect of personal value (from both the lens of individualistic and collectivist values perspective) on involvement and attitude towards discount sales promotion, as well as the mediating effect of involvement on personal values (individualistic ... When customers perceived these emails as meeting their personal preferences, they developed a strong relationship with the sender. In [163] , the authors demonstrated that the personalization, interactivity and price were important predictors of the possibility of revisiting the same accommodation. To the best of our knowledge, we are the first to propose a supervised machine learning approach to enable personalized email marketing in the airline travel industry. ... Semantic data driven approach for merchandizing optimization The overall objective of this PhD is to explore and propose new approaches leveraging a large volume of heterogeneous data that needs to be integrated and semantically enriched, and recent advances in machine and deep learning techniques, in order to exploit both the increased variety of offers that an airline can make to its customers as well as the knowledge it has of its customers with the ultimate goal of optimizing conversion and purchase. The overall goal of this thesis can be broken down into three main research questions: 1) What piece of content (ancillary services, third-party content) should be recommended and personalized to each traveler? 2) When should a recommendation be made and for which communication channel to optimize conversion? 3) How do we group ancillary services and third-party content and can we learn what often goes together based on purchase logs? View ... The impact of incentive framing 3037 actively using email marketing to attract new customers and to achieve other business purposes (eMarketer, 2017;Piccoli et al., 2017; Yang et al., 2018) . While hotel companies have been actively harnessing emails for disseminating promotional messages and soliciting sales from readers, surprisingly, the efficacy of email marketing has garnered limited scholarly attention. ... The impact of incentive framing format and language congruency on readers’ post-reading responses to email advertisements Purpose This paper aims to examine how incentive framing format and language congruency interactively influence readers’ post-reading responses to hotel-related email advertisements. Design/methodology/approach A 2 (incentive framing format: amount-off versus percentage-off) × 2 (language congruency: use of readers’ native language versus use of readers’ foreign language) between-subject experiment was conducted with 233 bilingual speakers from China. Findings The findings unveil that readers are more likely to be enticed to search for more information about the promoted hotel restaurant, click the call-to-action button and share the promotional message with friends and families if an email advertisement presents the incentive of a price promotion in the form of amount-off (versus percentage-off). The indirect impact of language congruency is also verified. Specifically, the impact of incentive framing format on readers’ post-reading response is more salient when information is communicated using readers’ native (versus non-native) language. Practical implications The findings provide actionable clues for hoteliers to optimize their email marketing campaigns. If hotels want to publicize a price promotion for their high-priced service (e.g. hotel restaurant dining) via email advertisements, marketers should present the incentive in the form of amount-off. Hotels should also personalize the language used in the email advertisements according to readers’ native language. Originality/value This study enriches the literature on email marketing by explicating how hoteliers can improve the efficacy of email marketing via personalizing the incentive framing format and language used in the email advertisement according to the readers’ preferences. Travel vlog ecosystem in tourism digital marketing evolution: a narrative literature review This narrative literature review advances understanding of travel vlogs as a critical component of contemporary digital tourism marketing and highlights the transformation of digital marketing over the past three decades. The paper explores the role of vlogs in modern tourism marketing practices and proposes a travel vlog ecosystem comprising three actors: the vlogger, the viewer, and the destination marketer. The paper is the first to examine the motivations and interactions of each of these actors and proposes a new model to explain these roles and relationships. The paper advances the tourism destination digital marketing literature by examining existing literature gaps and identifying future research opportunities in vlogging activities, particularly the vlogging ecosystem and the involvement of three main central actors. Given the increasing importance of social media in destination marketing strategy, this paper also provides valuable practitioners insights to inform strategic marketing activities using vlogs. E-POSTA PAZARLAMASI ALANINDA 1950-2019 YILLARI ARASINDA YAPILAN ÇALIŞMALARA YÖNELİK BİR ŞEMATİK YAZIN TARAMASI E-posta pazarlaması internetin gelişimi ile ortaya çıkan elektronik bir pazarlama aracı olarak literatürde yer almaktadır. Şematik yazın taraması (mapping review) olarak gerçekleştirilen bu çalışmanın amacı, pazarlama aracı olarak ele alınan e-posta pazarlaması alanına dair bir öngörü sağlanmasıdır. Çalışmada 1950-2019 yılları arasında uluslararası akademik dergilerde yer alan pazarlama odaklı 57 e-posta pazarlaması çalışması incelenmiştir. Çalışmaların seçiminde anahtar kelime kullanılarak filtreleme gerçekleştirilmiş, doğrudan ilgili alana ve pazarlamaya yönelik olmayan çalışmalar kapsam dışında bırakılmıştır. İncelenen çalışmalar ile e-posta pazarlamasının teorik arka planınına, odaklandığı alanlara, ele alındığı sektörlere ve çalışmaların yönelimlerine dair sonuçlar elde edilmiştir. İncelenen çalışmaların 42’si ampirik, 15’i ise kavramsaldır. En çok çalışılan alanın tüketici davranışları olduğuna, en fazla sayıda yayının “Journal of Direct, Data and Digital Marketing Practice” dergisinde yer aldığına ve 2016 yılının 11 çalışma ile ilgili literatürde en çok yayının yapıldığı yıl olduğu sonucuna ulaşılmıştır. Çalışmada elde edilen sonuçlar ile farklı bir alana yönelmek isteyen akademisyenlere niş bir alan önerisi sunularak, e-posta pazarlaması literatüründe ele alınmamış olan alanlar gösterilmektedir. Developing Relational Bonds with Luxury Hotel Guests through Personalization: A Subgroup Analysis of Generational Cohorts Empirical studies on personalization in the luxury hotel context are lacking. Personalization, financial, social, and structural bonds have been proposed together as a conceptual framework, but have not been tested empirically. Moreover, relational bond studies in the luxury hotel U.S. market are non-existent. As personalization is a growing trend and luxury hotels continue to be a high revenue generator for brands, we proposed a relational bonds model in the context of U.S. luxury hotels. Multiple regression analysis was conducted on data collected through a Qualtrics Survey Panel from 205 U.S. respondents. A subgroup analysis was performed to examine differences between Millennials and Generation Xers. The results show differences between the two generational cohorts in terms of the relational bonds regarding behavioral loyalty intentions. Konaklama İşletmelerinin Dijital Pazarlama Açısından İncelenmesi: Samsun Örneği (Examining of Accommodation Businesses in terms of Digital Marketing: Case of Samsun) Aybike Tuba Özden In the Purchase’s Essence: Sensory and symbolic elements in the descriptions of perfume sales over the Internet This study aimed to analyse whether sensory and symbolic elements used in perfume descriptions in internet sales can positively influence the level of involvement and consumer buying intention. To do so, a quasi-experimental design was adopted with the creation of three scenarios: one control group and two experimental groups. Each participant was presented to only one of the scenarios created, characterizing the model between subjects. Regarding the results, it was found that it is not possible to perceive differences between the means of the groups in relation to the dependent variables, except for a negative association between sensory aspects and the level of involvement. Therefore, it was concluded that it may not be advantageous to invest in sensory and symbolic aspects present in the product description in internet sales, it is necessary to rethink this strategy due the fact that this environment has a certain limitation with hedonics products. ICDSST 2020 is EWG-DSS’s 6th International Conference, following the five previous successful International Conferences on Decision Support System Technology (ICDSST 2019 in Madeira; ICDSST 2018 in Crete; ICDSST 2017 in Namur; ICDSST 2016 in Plymouth; and ICDSST 2015 in Belgrade) as listed below. Since 1989 when the EWG-DSS was founded, the society has organized annual events including workshops, mini-conferences and conference-streams. These events have received wide participation from researchers, academics, practitioners, developers and other professionals within the EWG-DSS network and colleagues from other international DSS communities and professional societies. Along with the fast increase of our membership and scale of our events, not only in terms of numbers but also the internationalization, in 2015 the EWG-DSS decided to re-brand its major annual event as international conference. The ICDSST conferences have the unique title of International Conference on Decision Support System Technology, but with a different theme each year following the trend and state-of-the-art in the DSS field. The name of the conference has a corporate identity with the name of the IJDSST journal, which is the official journal publication of the Euro Working Group on Decision Support System, having published several special issues from research conducted by the EWG-DSS. The ICDSST 2020 aims at the development of Decision Support Systems and Technologies (tools and knowledge) that capture the relevance and support the human factor and the cognitive orientation in decision making. E-Posta Pazarlama: Teorik Bir Bakış Email Marketing: A Theoretical Perspective Küreselleşme ve teknolojik gelişmeler, işletmeleri özellikle pazarlama ve reklam iletişiminde, müşterilere ulaşmak için yeterli etkileşime sahip yeni medya kanalları keşfetmeye zorlamaktadır. İşletmeler her zaman müşterileriyle daha kolay, daha ucuz ve daha hızlı iletişim kurmanın yollarını ararken, internet her üçünü de sunmaktadır. E-posta pazarlaması, reklam veya ticari bilgi göndermek için e-posta kullanan çevrimiçi bir pazarlama tekniğidir. E-posta pazarlama, düşük maliyeti, yüksek yatırım getirisi ve marka bağlılığı oluşturma avantajıyla da işletmeler için son derece cazip bir pazarlama aracıdır. Bu makale, güncel istatistikleri de paylaşarak e-posta pazarlamasıyla ilgili geniş bir kavramsal çerçeve oluşturmaya odaklanmaktadır. Bununla birlikte işletme yöneticilerin e-posta pazarlama kampanyalarını daha etkin bir şekilde tasarlamalarına yardımcı olmak için gereken e-posta pazarlama stratejilerini de açıklamaya çalışmaktadır. Makalede ayrıca izin pazarlaması, e-posta pazarlama metrikleri, e-posta pazarlamanın avantaj ve dezavantajları konularına da yer verilmiştir. IN THE PURCHASE'S ESSENCE: SENSORY AND SYMBOLIC ELEMENTS IN THE DESCRIPTIONS OF PERFUME SALES OVER THE INTERNET Na essência da compra: elementos sensoriais e simbólicos nas descrições de vendas de perfumes pela internet This study aimed to analyse whether sensory and symbolic elements used in perfume descriptions in internet sales can positively influence the level of involvement and consumer buying intention. To do so, a quasi-Resumo Este estudo teve o objetivo de analisar se elementos sensoriais e simbólicos utilizados nas descrições de perfumes em vendas pela internet podem influenciar positivamente o nível de envolvimento e intenção de compra Paula et al. (2020) 214 Consumer Behavior Review, 4(3) 199-228 experimental design was adopted with the creation of three scenarios: one control group and two experimental groups. Each participant was presented to only one of the scenarios created, characterizing the model between subjects. Regarding the results, it was found that it is not possible to perceive differences between the means of the groups in relation to the dependent variables, except for a negative association between sensory aspects and the level of involvement. Therefore, it was concluded that it may not be advantageous to invest in sensory and symbolic aspects present in the product description in internet sales, it is necessary to rethink this strategy due the fact that this environment has a certain limitation with hedonics products.

https://www.researchgate.net/publication/329278839_Post-stay_email_marketing_implications_for_the_hotel_industry_Role_of_email_features_attitude_revisit_intention_and_leisure_involvement_level

eli 8' dinghy? | Boat Design Net Has anyone ever heard of this type of boat? It is an 8' sailing dinghy built in the 70's and 80's by eli laminates co. of NY. When I contacted the... eli 8' dinghy? Converting Eli to sail Mine was just a rowing dingy and I want to convert it to sail. Any of you guys help me with some measurements to do the conversion? I need help with the center board and rudder (traces here would be ideal) and the placement on the keel. Hi, I have an 8' Eli dinghy which I bought 8 years ago that came with an unused sail kit. I have had it out a couple of times but find the seating impossible. The straight tiller and non kick up rudder is a another problem. I would like to sell it to someone who understands it. This is it's Serial number ELX20062A990 which make me think it was from Sept.1990? John I picked one up this year with a sailing rig and boards that I am planning on restoring this winter to use as a tender. I did a bit of digging and was able to find a copy of the original brochure. If you have any specific questions, I might be able to help. Cheers, Kris Looks like I might be the last one to the party but I thought I'd try anyhow. I have an 8' Eli Tri Hull rowing dingy. I have only owned it since the mid 70's. It was basically rated for a 2 hp outboard but because I got such a deal on a Johnson 2 1/2 hp outboard I had to get it. It served my family well, going to and from our mooring in Holland and then in Muskegon, Michigan. My wife and I used it to shuttle our 2 girls and 2 springers and a weekend of gear from shore to the mooring. It's been towed behind or stowed on deck of our 5 sailboats, and left at the mooring over the years. It was unsinkable, even when it rolled with the Johnson on it and a dog under it. Both survived the ordeal and we still have the functioning Johnson. It was ok for a couple of adults and the dogs but we started making 2 trips after the girls arrived. Since that time a 4 hp Yamaha became available and it also went on the transom of our little tri hull. Just last year three rather tall adults borrowed it. We have a picture of them and their trip to get ice with about 2 inches of freeboard. I think the capacity plate is just a suggestion, not really a limit???? Tongue in cheek, that's a joke son, so don't write and criticize. We just purchased a RIB so our little tri hull will be hitting the want ads soon. I think I can sell it but it's more than family. Oh, I never would've thought it had a sailing rig either. How do they sail? Summer might get here if Covid 19 ever leaves. Still waiting for a launch date. Stay safe. Looks like I might be the last one to the party but I thought I'd try anyhow. I have an 8' Eli Tri Hull rowing dingy. I have only owned it since the mid 70's. It was basically rated for a 2 hp outboard but because I got such a deal on a Johnson 2 1/2 hp outboard I had to get it. It served my family well, going to and from our mooring in Holland and then in Muskegon, Michigan. My wife and I used it to shuttle our 2 girls and 2 springers and a weekend of gear from shore to the mooring. It's been towed behind or stowed on deck of our 5 sailboats, and left at the mooring over the years. It was unsinkable, even when it rolled with the Johnson on it and a dog under it. Both survived the ordeal and we still have the functioning Johnson. It was ok for a couple of adults and the dogs but we started making 2 trips after the girls arrived. Since that time a 4 hp Yamaha became available and it also went on the transom of our little tri hull. Just last year three rather tall adults borrowed it. We have a picture of them and their trip to get ice with about 2 inches of freeboard. I think the capacity plate is just a suggestion, not really a limit???? Tongue in cheek, that's a joke son, so don't write and criticize. We just purchased a RIB so our little tri hull will be hitting the want ads soon. I think I can sell it but it's more than family. Oh, I never would've thought it had a sailing rig either. How do they sail? Summer might get here if Covid 19 ever leaves. Still waiting for a launch date. Stay safe. MIDad, can you post a photo of your 8' Eli Tri Hull rowing dingy? I have a year 1986 7ft model, attached is a photo, suppose to be the same boat as yours, only 1 ft shorter. File size: 286.2 KB

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Global environmental drivers of influenza. - PDF Download Free In temperate countries, influenza outbreaks are well correlated to seasonal changes in temperature and absolute humidity... Global environmental drivers of influenza Ethan R. Deylea, M. Cyrus Maherb,c, Ryan D. Hernandezd, Sanjay Basue, and George Sugiharaa,1 a Scripps Institution of Oceanography, University of California, San Diego, La Jolla, CA 92093; bHuman Longevity Inc., Mountain View, CA 94041; cDepartment of Epidemiology and Translational Science, University of California, San Francisco, CA 94158; dDepartment of Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA 94158; and eStanford Prevention Research Center, Stanford University School of Medicine, Stanford, CA 94305 diverse group of drivers and mechanisms has been put forward to explain the wintertime occurrence of seasonal influenza outbreaks. Laboratory experiments show that relative humidity controls droplet size and aerosol transmission rates (1). Experiments with mammalian models showed that viral shedding by hosts increases at low temperature (2). Strong laboratory evidence has emerged that absolute humidity has a controlling effect on airborne influenza transmission (3). Nevertheless, questions remain as to how these potential causal agents are expressed at the population level as epidemic control variables. At the population level, environmental factors covary, multiple mechanisms can coact, and infection dynamics are influenced by many other important processes (4), such as human crowding, rapid viral evolution, and international travel patterns. Perhaps not surprisingly, statistical analyses of population level data have produced contradictory results. Although correlations between influenza incidence and both temperature and absolute humidity are easy to find in temperate countries (5) and individual US states (6), such associations are weak or altogether absent in data from tropical countries (5). Here we use new methods appropriate for disease dynamics to identify the causal drivers of influenza acting at the population level. Using time series data across countries and latitudes, we find that absolute humidity drives influenza across latitudes, and that this effect is modulated by temperature. At low temperatures, absolute humidity negatively affects influenza incidence (drier conditions improve survival of the influenza virus when it is cold), but at high temperatures, absolute humidity positively affects influenza (wetter conditions improve survival of the influenza virus when it is warm). This population-level finding supports the conclusions of theoretical work on viral envelope stability (7) and sorts out disagreements in laboratory studies (8). Our analysis is divided into three parts. First, we show how dynamic resonance can explain the success of linear statistical methods in detecting causal effects in temperate latitudes and how www.pnas.org/cgi/doi/10.1073/pnas.1607747113 mirage correlations are symptoms of their failure in the tropics. This motivates taking an empirical dynamic modeling (EDM) approach (9). Second, we use the EDM method of detecting causality, convergent cross-mapping (CCM), to show how absolute humidity and temperature combine to produce a unified explanation for influenza outbreaks that applies across latitudes. Finally, we probe the mechanistic effects of each variable, showing that absolute humidity has the most direct effect on influenza, and that this effect is modulated nonlinearly by temperature. Results Correlation and Seasonality. It is well known that correlative ap- proaches can fail to provide an accurate picture of cause and effect in a dynamic system, and this is especially true in nonlinear systems where interdependence between variables is complex. Such systems are known to produce mirage correlations that appear, disappear, and even reverse sign over time (9). Persistent correlations tend to only occur in specific circumstances; most notably, when there is synchrony between driver and response variables (the effect of the driving variable is strong enough that the response becomes enslaved to the driver). This is key, because basic host–pathogen dynamics are known to exhibit dynamical resonance when strongly forced by periodic drivers (10). Dynamical resonance causes the intrinsic nonlinear epidemiological dynamics to become synchronized (phase-locked) to the simple cyclic motion of the environmental driver. However, when drivers do not induce synchrony, the underlying nonlinear dynamics can cause the statistical relationship between driver and response to become very complex. Indeed, the same simple epidemiological SIRS model of ref. 10 (a basic epidemiological model consisting of “susceptibleinfected-recovered-susceptible”) illustrates how the identical Significance Patterns of influenza outbreak are different in the tropics than in temperate regions. Although considerable experimental progress has been made in identifying climate-related drivers of influenza, the apparent latitudinal differences in outbreak patterns raise basic questions as to how potential environmental variables combine and act across the globe. Adopting an empirical dynamic modeling framework, we clarify that absolute humidity drives influenza outbreaks across latitudes, find that the effect of absolute humidity on influenza is U-shaped, and show that this U-shaped pattern is mediated by temperature. These findings offer a unifying synthesis that explains why experiments and analyses disagree on this relationship. Author contributions: E.R.D., M.C.M., R.D.H., S.B., and G.S. designed research; E.R.D. and G.S. performed research; E.R.D. and M.C.M. analyzed data; M.C.M., R.D.H., and S.B. contributed to interpretation of results; and E.R.D. and G.S. wrote the paper. The authors declare no conflict of interest. This article is a PNAS Direct Submission. Freely available online through the PNAS open access option. See Commentary on page 12899. 1 PNAS | November 15, 2016 | vol. 113 | no. 46 | 13081–13086 ECOLOGY In temperate countries, influenza outbreaks are well correlated to seasonal changes in temperature and absolute humidity. However, tropical countries have much weaker annual climate cycles, and outbreaks show less seasonality and are more difficult to explain with environmental correlations. Here, we use convergent cross mapping, a robust test for causality that does not require correlation, to test alternative hypotheses about the global environmental drivers of influenza outbreaks from country-level epidemic time series. By moving beyond correlation, we show that despite the apparent differences in outbreak patterns between temperate and tropical countries, absolute humidity and, to a lesser extent, temperature drive influenza outbreaks globally. We also find a hypothesized U-shaped relationship between absolute humidity and influenza that is predicted by theory and experiment, but hitherto has not been documented at the population level. The balance between positive and negative effects of absolute humidity appears to be mediated by temperature, and the analysis reveals a key threshold around 75 °F. The results indicate a unified explanation for environmental drivers of influenza that applies globally. mechanistic effect of a driver can produce very different behavior when only the periodicity is changed (Fig. 1). Simulating the model with a strongly seasonal environmental driver produces consistent seasonal outbreaks that correlate very strongly to the driver (Fig. 1A, where seasonality is quantified by the Pearson’s correlation between the raw values and day of year mean). However, this changes when the driver is replaced with another signal that has the same magnitude, but with much weaker seasonality (low correlation between raw values and day of year mean). The weakened seasonality results not only in much less seasonality in influenza but also much lower correlation between influenza infections and climate (Fig. 1B). The mechanisms have not changed, only the spectrum (periodicity) of the driver. Moreover, not all regions of the world have strong climate seasonality. Although the seasonal cycle in a temperate country such as Germany can explain more than 90% of the variance in absolute humidity across multiple decades, in tropical countries such as Singapore, it explains less than 30%. Indeed, there is a strong correspondence between seasonality of climate and seasonality of influenza. Fig. 2 shows the seasonality in absolute humidity (Fig. 2A) and influenza (Fig. 2B) across countries. The countries with the least seasonality in environment (yellow shades) also have the least seasonal influenza (Spearman’s ρ = 0.73). This is most pronounced in the tropics, where it is important to note that correlations between influenza and environment are also most elusive. A 4 2 0 2004 −2 −4 Environmental Driver 4000 2000 0 # Infected 6000 Strongly Seasonal Driver 2006 2008 2010 2012 2014 Year B 4 2 0 2004 Environmental Driver 4000 2000 0 # Infected 6000 Weakly Seasonal Driver 2006 2008 2010 2012 2014 Year Fig. 1. Stochastic SIRS model with strongly and weakly seasonal drivers. (A) A strongly periodic environment induces synchrony through dynamical resonance, causing peaks in infection to correlate with seasonal lows in the seasonal environment. (B) If the same SIRS model is driven by a seasonal signal with the same variance, but much weaker seasonality, there is no dynamic resonance, infection peaks show much weaker seasonality, and correlation between infection and environment is much lower. 13082 | www.pnas.org/cgi/doi/10.1073/pnas.1607747113 Fig. 2. Correspondence between seasonality of environment and seasonality of influenza infection. Countries are colored from the least seasonal to most seasonal for absolute humidity (A) and influenza infection (B). The Spearman correlation between the two is high: ρ = 0.73. These results illustrate a crucial point: lack of seasonality in influenza and lack of correlation between influenza and climate do not indicate a lack of environmental forcing. Indeed, a poor correlation between climate and influenza is to be expected with a nonlinear relationship where seasonality is weak; however, where seasonality is strong, dynamical resonance can emerge to produce the appearance of a linear relationship (correlation). To examine these ideas critically and to develop a more unified understanding of how climate affects influenza globally requires an approach that can cope with the nonlinear interactions inherent in the system. Cross-mapping. Convergent cross-mapping is a method for detecting causality in nonlinear dynamic systems (see the following playlist of three brief animations (adapted from the supplement of ref. 12): https://www.youtube.com/watch?v=fevurdpiRYg&list=PLSSmlAMhY3bnogGTe2tf7hpWpl508pZZ).It looks for signatures of the cause in the time series of the affected variable. The idea is that if a variable, X, can predict the current or previous state of another variable, Y, then Y causally influenced X. For example, if sardine abundance can be used to recover sea surface temperature, then temperature had a causal effect on sardines. We examine cross-map prediction between influenza and the purported environmental drivers: absolute humidity, temperature, relative humidity, and precipitation. The mutual seasonality of influenza and these environmental variables makes it especially important to distinguish causal interactions from spurious correlation. Thus, we compare the cross-map prediction measured for the observational environmental time series with the null expectation obtained from cross-mapping with surrogate time series having the same seasonal cycle as the actual driver, but with randomized Deyle et al. Absolute Humidity Temperature Relative Humidity Precipitation distance from equator Malaysia Colombia Peru Philippines Senegal Thailand Madagascar New Caledonia Paraguay South Africa Chile Japan Portugal Spain Romania France Slovenia Switzerland Germany Poland United Kingdom Denmark Latvia Sweden Norway Finland 0 0.5 1 0 0.5 1 0 0.5 1 Fig. 3. Detecting cross-map causality beyond shared seasonality of environmental drivers on influenza. Red circles show the unlagged cross-map skill (ρCCM) for observed influenza predicting purported seasonal drivers: absolute humidity, temperature, relative humidity, and precipitation. Together with this, box-and-whisker plots show the null distributions for ρCCM expected from random surrogate time series that shares the same seasonality as the true environmental driver. Countries are ordered according to distance from the equator (absolute latitude). Filled circles indicate that the measured ρCCM is significantly better than the null expectation (P ≤ 0.05). Deyle et al. Multivariate EDM. To examine whether AH or T has a stronger causal effect on influenza according to previous work (12), we use a multivariate EDM approach that looks for improvement in forecasting when a suspected causal variable is included. In brief, if a multivariate empirical dynamic model containing a potential driving variable Y produces better forecasts of X than without, then Y causally influenced X. The results are summarized in Fig. 4. The results show strong evidence that AH and T are drivers of influenza, as including either variable leads to improved forecast skill (P < 0.001 globally). However, in the tropics (where AH and T generally have weaker correlation), we find that including AH and T together as embedding coordinates leads to slightly more improvement over either one alone. In temperate countries, where correlations between AH and T are extremely high (generally > 0.9), these variables contain almost identical information, and hence there is less difference in forecast skill on average between embeddings with AH, T, or both. Overall, multivariate forecast improvement suggests the possibility that AH and T taken together have a nonlinear effect on influenza that is global, but that this effect is concealed in the temperate region by their strong correlation to each other. To explore the mechanism for this interaction further, we predict the hypothetical change in influenza incidence, denoted Δflu, at historical points that would occur from small increases and decreases in the environmental driver [EDM scenario exploration (13)]. This simple device allows us not only to directly quantify the magnitude and direction of effect but also to keep track of how the effect changes through time and with varying environmental conditions. Fig. 5A shows country by country the magnitude of the effect of absolute humidity on influenza, Δflu/ ΔAH. Countries at high latitudes generally show a negative effect of absolute humidity on influenza, whereas low-latitude countries generally show a positive effect. This is consistent with the hypothesized U-shaped response of influenza to AH suggested by certain experiments (8). We can examine this effect further by plotting the scenario exploration results for all the countries together. Fig. 5B shows that the effect of AH on influenza is negative at low AH (Δflu/ ΔAH < 0), but positive at high AH (Δflu/ΔAH > 0), consistent with a U-shaped response of influenza survival to absolute humidity. The negative effect on incidence at low AH and positive effect at high AH appear roughly equivalent when the data are normalized to average total reported cases in a year in that country. The same analysis for temperature (Fig. 5C) does not exhibit such a clear state-dependent (nonlinear) effect. Temperature changes can have a positive (Δflu/ΔT > 0) or negative (Δflu/ΔT < 0) effect on influenza at the same temperature. Indeed, the molecular arguments for a U-shaped effect of absolute humidity on influenza also predict that temperature should be a control on the balance between the positive and negative effects of absolute humidity (7). With this in mind, we look at the effect of absolute humidity on influenza (Δflu/ΔAH) as a function of temperature (Fig. 5D). This is perhaps the most interesting picture to emerge, as there are a number of features that corroborate and elaborate existing PNAS | November 15, 2016 | vol. 113 | no. 46 | 13083 SEE COMMENTARY In Fig. 1A, however, the driver has much stronger seasonality, and is therefore more predictable. The seasonal cycle, being identical from year to year, is trivial to predict; meaning the stronger the effect of the seasonal cycle, the easier the driver is to predict. Thus, the absolute level of ρCCM is substantially higher in Fig. 1A than Fig. 1B, even though the strength of causal effect is likely unchanged. Therefore, in this case, the skill of CCM (ρCCM) should not be used as a relative measure of causal strength when comparing CCM across latitudes, as the climate time series in tropical and temperate countries do not have the same basic levels of predictability. ECOLOGY anomalies. Causal forcing is established when cross-map prediction is significantly better for the real environmental driver than it is for the null surrogates (11). Fig. 3 shows box-and-whisker plots of the null distributions for cross-map skill (ρCCM) ordered according to distance from the equator (absolute latitude). The measured CCM skill values are plotted on top as red circles that are filled if the value is significantly different from the null distribution (P ≤ 0.05), and open otherwise. Absolute humidity (AH) and temperature (T) both show significant forcing in countries across latitude. The results have very high metasignificance (Fisher’s method): P < 1.6 × 10−6 for AH and P < 2.9 × 10−3 for T. There is even higher metasignificant evidence for forcing across latitudes by relative humidity: P < 1.8 × 10−14. However, paired Wilcox tests (nonparametric generalization of Student’s t test) in the prediction skill confirm that the cross-map effect with respect to RH is weaker on average than with either AH or T (P < 0.02), but that the strength of effects of AH and T are not significantly different from one another. Finally, there is some evidence that precipitation may be a causal variable in a few countries (global metasignificance, P < 7.3 × 10−3; tropics only, P < 0.023). If all other things are equal, the magnitude of cross-map skill (ρCCM) can indicate the strength of causal effect. On this basis, one might conclude that because the absolute level of cross-map skill is lower in the tropics, the causal effect of climate (absolute humidity or temperature) is weaker. However, this would be incorrect, as “all things equal” does not apply across latitudes because of the differences in seasonality, and hence differences in the baseline predictability in these systems. This dichotomy is illustrated with the model shown in Fig. 1. Both model realizations have the same ultimate magnitude of the effect of AH on flu. −0.03 0.00 −0.03 AH+T RH AH AH+T RH Prcp T Prcp AH+T T RH −0.06 AH −0.06 0.00 0.03 Prcp −0.03 0.03 Forecast Improvement 0.00 Temperate 0.06 ( ) ( ) 0.03 Forecast Improvement Forecast Improvement C Tropical 0.06 T B Global 0.06 ( ) A Fig. 4. Forecast improvement with multivariate EDM. Causal effect is demonstrated if EDM forecast skill (ρ) improves when a driver variable is included in the EDM model. This is quantified by Δρ = ρ(with driver) − ρ(without driver), where ρ is the Pearson’s correlation between observations and EDM predictions. Including either absolute humidity (AH) or temperature (T) leads to significant (P < 0.001) improvement in forecast skill both globally (A) and the tropics specifically (B). The significance is more marginal when looking solely at temperate countries (C) (P < 0.07 for AH; P < 0.03 for T). In the tropics, including both (AH +T) is marginally better than either AH or T alone, suggesting possible compound effects of temperature and humidity. ideas. We observe the following: temperature has a relatively loose control on (Δflu/ΔAH) when it is below ∼70 °F, but the effect on (Δflu/ΔAH) is consistently negative; the positive effect of absolute humidity appears more strongly controlled by temperature and appears to be restricted to a narrow band of temperature between 75 °F and 85 °F; at the highest temperatures, the effect of absolute humidity goes to zero in exact concordance with the laboratory finding that aerosol transmission of influenza is blocked at 30 °C (86 °F) (14). These results suggest that the balance between positive and negative effects of absolute humidity appears to shift somewhere between 70 °F and 75 °F. This is especially clear if we look back at the plot in Fig. 5B that shows the effect of absolute humidity on influenza across the global range of absolute humidity. Fig. 6 shows the same data, but now the points are split between two panels, based on temperature. On the left are observations where temperature was below 75 °F; on the right are observations where the temperature was between 75 °F and 85 °F. The red lines represent the 0.1 and 0.9 quantile regressions. The 0.005 Effect of AH on Flu Flu/ AH Effect of AH on Flu Flu/ AH B 0.01 0 -0.005 -0.01 10 20 30 40 50 Discussion Prior population-level analyses have focused on explanations for influenza seasonality, using correlations. However, correlation is a limited tool for understanding causality in nonlinear systems such as those exhibiting complex host–pathogen dynamics. Building on previous work on dynamic resonance (10), we show that persistent correlations between influenza and environment should only be expected where seasonality is strongest; that is, in temperate countries (as is clear from Fig. 2). This motivates adopting an EDM framework to address the general problem of identifying external drivers of nonlinear dynamics. We are able to provide clear tests at the population level of alternative hypotheses about global climatic drivers of influenza and show that there are general rules that span temperate and tropical latitudes. We find clear evidence that temperature and 0.01 Positive relationship Negative relationship between AH and Flu between AH and Flu A quantile regressions show that the measured effect of AH on influenza is almost always negative when T < 75 °F, and almost always positive when 75 °F ≤ T ≤ 85 °F. 0.005 0 -0.005 -0.01 0 60 5 Country (ranked by |Lat|) 10 15 20 25 Absolute Humidity AH Distance From Equator C D 0.01 Effect of AH on Flu Flu/ AH Effect of T on Flu Flu/ T 0.005 0 -0.005 -0.01 20 40 60 80 100 0.01 0.005 0 -0.005 -0.01 Temperature T 13084 | www.pnas.org/cgi/doi/10.1073/pnas.1607747113 20 40 60 Temperature T 80 100 Fig. 5. Scenario exploration with multivariate EDM gives a measure of the effect of environment on influenza infection by predicting the change in influenza (Δflu) that results from a small change in absolute humidity (ΔAH) or temperature (ΔT). Analysis covers countries with at least 208 observations (4 y) and area Global environmental drivers of influenza. In temperate countries, influenza outbreaks are well correlated to seasonal changes in temperature and absolute humidity. However, tropical countries ... 1MB Sizes 0 Downloads 8 Views

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(PDF) Animal Protein Intake and Risk of Inflammatory Bowel Disease: The E3N Prospective Study PDF | Diet composition has long been suspected to contribute to inflammatory bowel disease (IBD), but has not been thoroughly assessed, and has been... | Find, read and cite all the research you need on ResearchGate Animal Protein Intake and Risk of Inflammatory Bowel Disease: The E3N Prospective Study May 2010 The American Journal of Gastroenterology 105(10):2195-201 DOI: 10.1038/ajg.2010.192 Authors: Françoise Clavel-Chapelon Abstract and Figures Diet composition has long been suspected to contribute to inflammatory bowel disease (IBD), but has not been thoroughly assessed, and has been assessed only in retrospective studies that are prone to recall bias. The aim of the present study was to evaluate the role of dietary macronutrients in the etiology of IBD in a large prospective cohort. The Etude Épidémiologique des femmes de la Mutuelle Générale de l'Education Nationale cohort consists of women living in France, aged 40-65 years, and free of major diseases at inclusion. A self-administered questionnaire was used to record dietary habits at baseline. Questionnaires on disease occurrence and lifestyle factors were completed every 24 months. IBDs were assessed in each questionnaire until June 2005, and subsequently validated using clinical and pathological criteria. We estimated the association between nutrients or foods and IBD using Cox proportional hazards models adjusted for energy intake. Among 67,581 participants (705,445 person-years, mean follow-up since completion of the baseline dietary questionnaire 10.4 years), we validated 77 incident IBD cases. High total protein intake, specifically animal protein, was associated with a significantly increased risk of IBD, (hazards ratio for the third vs. first tertile and 95% confidence interval being 3.31 and 1.41-7.77 (P trend=0.007), and 3.03 and 1.45-6.34 (P trend=0.005) for total and animal protein, respectively). Among sources of animal protein, high consumption of meat or fish but not of eggs or dairy products was associated with IBD risk. High protein intake is associated with an increased risk of incident IBD in French middle-aged women. . Baseline characteristics of IBD cases and non-cases in the E3N cohort study … . HR estimates and 95 % confi dence intervals for IBD in relation to type of protein in the E3N cohort ( n = 67,581), 1993 -2005 … Figures - uploaded by Prévost Jantchou Content may be subject to copyright. INFLAMMA TORY BOWEL DISEASE INTRODUCTION Crohn ’ s disease (CD) and ulcerative coli tis (UC) are chronic in ammatory disorders of the gastro intestinal tract.  e patho- genic mechanisms invol ve epithelial cell barrier dysfunction, defects in the innate immune system that a ect its interaction with commensal bacteria, abnormal compositio n of the bowel microbiota, and dysregula ted T-cell function promoting chronic intestinal in ammation ( 1,2 ). Genetic predisposition is well documented, particularly in CD, in which several susceptib ility genes a ecting bacterial clearance, the IL23 path way, and T -cell function have been identi ed ( 3,4 ).  ere is also some evidence that environmen t al factors may have a role in the pa thogenesis of in a mmatory b owel disease (IBD). First, the incidence of IBD has increased dramatically since W orld Wa r II, which is inconsistent with a purely genetic disease ( 5 ). Se cond, migrants who move from a low-incidence area to a high-incidence area have a high risk of IBD ( 6,7 ). Few environmental factors ha ve been linked reproducibly with susceptibili ty in IBD ( 8 ). Among them, smok- ing is the best documented; quitting smoking is a risk factor for UC, whereas continuing smoking is a risk factor f or CD ( 9 ). Diet composition has long been suspected to contribute t o IBD and has been assessed in many studies. With respect to macronutri- ents, several retrospective case – control studies have found that high intakes of sucrose and fat wer e ass ociated with CD and / or UC ( 10 – 12 ). However , these ass ociations were pro ne to recall bias, because of the retrospective design of the studies ( 13 ). In the present prospective study , we tested the hypothesis that the composition of the macr onutrient content of the diet is Animal Protein Intake and Risk of Infl ammatory Bowel Disease: The E3N Prospective Study Pr é vost Jantchou , MD 1 , Sophie Morois , MSc 1 , Fran ç oise Clavel-Chapelon , PhD 1 , Marie-Christine Boutron-Ruault , MD, PhD 1 and Franck Carbonnel , MD, PhD 1 , 2 OBJECTIVES: Diet composition has long been suspected to contribute to infl ammatory bowel disease (IBD), but has not been thoroughly assessed, and has been assessed only in retrospective studies that are prone to recall bias. The aim of the present study was to evaluate the role of dietary macronutrients in the etiology of IBD in a large prospective cohort. METHODS: The Etude É pid é miologique des femmes de la Mutuelle G é n é rale de l ’ Education Nationale cohort consists of women living in France, aged 40 – 65 years, and free of major diseases at inclusion. A self-administered questionnaire was used to record dietary habits at baseline. Questionnaires on disease occurrence and lifestyle factors were completed every 24 months. IBDs were assessed in each questionnaire until June 2005, and subsequently validated using clinical and pathological criteria. We estimated the association between nutrients or foods and IBD using Cox proportional hazards models adjusted for energy intake. RESULTS: Among 67,581 participants (705,445 person-years, mean follow-up since completion of the baseline dietary questionnaire 10.4 years), we validated 77 incident IBD cases. High total protein intake, specifi cally animal protein, was associated with a signifi cantly increased risk of IBD, (hazards ratio for the third vs. fi rst tertile and 95 % confi dence interval being 3.31 and 1.41 – 7.77 ( P trend = 0.007), and 3.03 and 1.45 – 6.34 ( P trend = 0.005) for total and animal protein, respectively). Among sources of animal protein, high consumption of meat or fi sh but not of eggs or dairy products was associated with IBD risk. CONCLUSIONS: High protein intake is associated with an increased risk of incident IBD in French middle-aged wo men. SUPPLEMENTARY MATERIAL is linked to the online version of the paper at http://www.nature.com/ajg. Am J Gastroent erol 2010; 105:2195–2201; doi: 10.1038/ajg.2010.192; published online 11 May 2010 1 INSERM, UMRS 1018, Team 9, Centre for Research in Epidemiology and Population Health, Institut Gustave Roussy , Université Paris Sud, Villejuif Cedex, France; 2 Service d’Hépatogastroentérologie, Centre Hospitalier Universitaire de Bicêtre, Assistance Publique-Hôpitaux de Paris, Universi té Paris Sud, Le Kremlin Bicêtre, France. Correspondence: Franck Carbonnel, MD, PhD, Service d’Hépatogastroentérologie, Centre Hospitalier Universitaire de Bicêtre, Assistance Publique-Hôpitaux de Paris, Université Paris Sud, 78 Rue du Général Leclerc, 94275 Le Kremlin Bicêtre, France. E-mail: [email protected] Received 4 January 2010; accepted 6 April 2010 The American J ournal of GASTROENTER OLOGY VOLUME 105 | OCTOBER 2010 www.amjgastro.com 2196 INFLAMMA TORY BOWEL DISEASE Jantchou et al. associated with IBD risk.  us, we investigated the rela tionship between pre-illness diet composition and risk o f incident IBD in a large prospective cohort of wo men w ho were free of major dis- eases at enrollment. METHODS  e Etude É pid é miologique des femmes de la Mutuelle G é n é rale de l ’ Education Nationale (E3N) is a prospective coho r t study conducted in France to study hormo nal and dietar y risk factors for the most common diseases, especially cancer , in women.  e cohort was established in 1990 and consisted of 98,995 women living in France, aged 40 – 65 years at baseline and insured with the Mutuelle G é n é rale de l ’ É ducation Nationale, which is a national health insurance scheme for teachers and co-work ers ( 14 ). E3N is also the French counterpart of the EPIC (Eur opean Prospective Investigation in to Cancer and Nutrition) study . Physical activity , reproductive factors, use of hormonal trea t- ments, tobacco consump tion, anthropometric measurements, personal history of disease, family history of cancer, and o ther factors were recorded in self-administer ed questionnaires that were completed a pproximately every 24 months. Each question- naire inquired about the occurrence of personal medical even ts, and there were speci c questions abo ut CD and UC occurrence in each questionnaire until J uly 2005. All study subjects signed an informed consent fo rm, in compliance with the rules of the French Na tional Commission for Computed Data and Indi vidual Freedom (Commission N ationale Informatique et Libert é s), from which we obtained a pproval. Dietary data collection Dietary data were collected once, between J une 1993 and July 1995, using a two-part questionnaire.  e  rst part contained questions on the usual amoun ts and frequency of consumption of foods from the various food groups, while the second included qualitative questions specifying individ ual foo ds within the food groups.  e questionnaire was used to assess the consumption of 208 food items, beverages, and the recipes used to prepar e them. It was accompanied b y a booklet of photographs illustrating por- tion sizes. Both the questionnaire and the booklet were validated ( 15 ). A high proportion of subjects (76 % for foods and 72 % for nutrients) were classi ed in the same or adjacent quintile for the dietary questionnaire and the mean of 12-monthly 24-h recalls.  e reproducib ility of the dietar y questionnaire was tested a er 1 year.  e Spearman correlation coe cient for reproducibility was 0.69 for dietary protein, 0.59 for carboh ydrates, and 0.73 for fat.  e diet histor y questionnaire was sent to 95,644 women, with two reminders to non-responders. I n all, 77,613 completed ques- tionnaires (81.1 % ) were received. A er exclusion of 978 question- naires because of absence of consent t o external he alth follow-up by the health insurer in case of dropout, 2,050 q uestionnaires because of miscoded answers, 8 blank questionnaires, a nd 46 duplicates, 74,531 questio nnaires were available for analysis. Women wi th extreme values (in the top or bottom 1 % ) for the energy intake / energy requirement ratio (calculated by taking age, weight, and height into acco unt) were excluded.  e mean daily intake of nutrients was assessed using a food com position table derived from the French na tional database. Validation of cases and non-cases Cases were  rst identi ed through the questionnaires, in which the women checked the item “ Crohn ’ s diseas e ” or “ ulcerative colitis. ” A er exclusion of miscoded answers, a letter was sent to the potential cases in order to con rm their answers and to obtain the names of their physicians.  en a brief question- naire was sent to the gastroenter ologist or general practitioner, requesting for informa t ion about diagnostic procedures,  nal IBD diagnosis a er follo w-up, location, number of  ares, and medical and surgical treatments. All cas es of IBD were reviewed by two of the a uthors (P .J. and F .C.), in order to con rm or rule out the diagnosis. De nition of IBD was based on clinical, radiological, endoscopic, and his- tological criteria. W e classi ed cases as certain or possible. Pos- sible IBD cases were those of women, which were consider ed and treated as IBD , but did not meet all inclusion criteria a er a careful review . IBD cas es for which radiological or endoscopic data did not enable us to classify them as CD or UC were consid- ered as indeterminate colitis (IC). A s IBD presentation, and thus classi cation, may vary with time, we retained the diagnosis at the latest follow-up . Non-cases were de ned as subjects who never reported a per- sonal history of IBD and were free of cancer a t bas eline. Because dent cancer were censored a t the date of cancer diagnosis.  us, subjects contributed person-time up to the da te of IBD diagnosis, date of cancer at an y site (other than basal cell skin carcinoma or in situ colorectal cancer), date of last com pleted questionnaire, or July 2005, whichever occurred  rst. Among the 74,531 women with available dietary questionnaires, we excluded 4,654 women because of p revalent cancer, 810 who were lost to follow-up a  er replying to the dietary questionnaire, 1,364 with extreme values of energy intake, and 5 with no inf orma- tion available. Statistical analysis  e association between dietary factors and IBD was estimated using Cox proportional hazards models with age as the time scale. Age at diagnosis of IBD or at cen soring date was used as the end-of-study time variable. Dietary factors that were co nsid- ered included the macronu trients (carbohydrates, proteins, ani- mal and vegetal proteins, an d lipids) and food sources of animal protein: meat (red an d w hite meat), eggs, dairy products (milk and cheese), and  sh ( sh and seafood). Diet ary intake was ana- lyzed in tertiles. T o calculate P values for trends across tertiles, participants were assigned a score ra nging from 1 to 3 according to their tertile of intake and this variable was enter e d as a continu- cated that for a bila teral test and alpha risk set at the 5 % level, we had a 78 % , 93 % , and 98 % power to detect a hazards ratio (HR) of 2.5, 3.0, or 3.5, respectively , in our main sample using tertiles for © 2010 by the American College of Gastroenterology The American J ournal of GASTROENTER OLOGY 2197 INFLAMMA TORY BOWEL DISEASE Protein Intake and IBD the exposure variable; the corresponding  gures were 38 % , 54 % , and 69 % , respectively , for the CD group, and 59 % , 78 % , and 90 % , respectively , for the UC group. When analyzing macronu t rients, adjustment for ener g y intake was made with the energy partition method, considering energy from carbohydrates, from li pids, and from proteins as three separate mutually adj usted variables ( 16 ). Else, when con- sidering subtype of protein intake (veg etable or animal) or food sources of protein in take, covariates were mutually adjust ed while adjustment on other sources o f energ y intake was car- ried out by adding energy intake from lip ids and carbohydrates (non-protein energy) as a covariate in the Cox model. In o rder to assess the potential impact of dietary changes due to undiag- nosed IBD, we carried out sensitivi ty analyses excluding cases diagnosed within 6 or 12 months a er dietary assessment. We mass index at baseline, total daily alcohol intake (all as co n- tinuous variables), ph ysical activity (tertiles of weekly energy expenditure), oral contraceptive in take, menopause hormonal treatment, smoking status at baseline (former / never / current), and the level of education as p roxy for the socio-economic status. Analyses were carried out fo r all IBD cas es, as well as separately for CD and UC. A heter ogeneity test was performed to compare the HRs associated with CD and UC risk usin g the W ald statistics ( 17 ). S ensitivity analyses were carried out a  er exclusion of 16 patien ts for whom IBD diagnosis was consid- ered only possible (5 CD, 10 UC, an d 1 IC cases). All analyses were performed using SAS so ware, version 9 (SAS Insti tute, Cary, NC); all statistical tests w ere two-sided. For all analyses, P values < 0.05 were considered statistically signi c ant. Table 1 . Baseline characteristics of IBD cases and non-cases in the E3N cohort study Baseline demographics and lifestyle characteristics Total incident IBD cases ( n =77) a Crohn’s disease ( n = 30) Ulcerative colitis ( n = 43) Non-cases ( n = 67,504) Age at dietary questionnaire (year) Mean 51.2 50.9 51.4 52.8 Range 43.5 – 67.6 43.5 – 67.6 44.1 – 63.6 41.8 – 72.0 Age at IBD diagnosis Mean 56.3 55.6 56.7 — Range 46.0 – 72.5 43.5 – 67.5 46.0 – 71.0 Time to diagnostic (years) Median 4.5 3.7 5.1 — Interquartile range 2.2 – 7.9 1.5 – 8.1 2.5 – 7.9 Anthropometric factors, mean (s.d.) Height (cm) 162 (5.4) 163.8 (5.0) 160.6 (5.2) 161 (5.7) Weight (kg) 60.3 (8.8) 61.2 (9.2) 59.1 (8.4) 60.0 (9.4) Body mass index (kg / m 2 ) b 23.0 (3.5) 22.8 (3.6) 22.9 (3.5) 23.0 (3.3) Smoking status, n ( % ) Never smoker 39 (50.7) 16 (53.3) 22 (51.2) 37,911 (56.2) Former smoker 21 (27.3) 8 (26.7) 12 (27.9) 20,325 (30.1) Current smoker 17 (22.1) 6 (20.0) 9 (20.9) 9,268 (13.7) Exogenous hormone use, n ( % ) Ever used HRT c 59 (76.6) 23 (76.7) 33 (76.7) 47,848 (70.9) Ever used oral contraceptive 49 (63.6) 19 (63.3) 27 (62.7) 41,177 (61.0) Total physical activity d , n ( % ) Weakly active 28 (36.4) 14 (46.7) 12 (27.9) 22,313 (33.3) Moderately active 22 (28.6) 9 (30.0) 12 (27.9) 22,384 (33.4) Active 27 (35.1) 7 (23.3) 19 (44.2) 22,345 (33.4) HRT, hormone replacement therapy; IBD, infl ammatory bowel disease. a Total IBD cases: 30 CD cases, 43 UC cases, and 4 indeterminate colitis. b Weight (kg) / height (m) 2 . c HRT: among postmenopausal women only . d Continuous variables are presented as mean (standard deviation). Categorical variables are presented as n ( % ). Unknown values were excluded from the calculations: physical activity, 0.6 % . The American J ournal of GASTROENTER OLOGY VOLUME 105 | OCTOBER 2010 www.amjgastro.com 2198 INFLAMMA TORY BOWEL DISEASE Jantchou et al. Table 2 . Mutually adjusted HR estimates and 95 % confi dence intervals for IBD in relation to macronutrient intake in the E3N cohort ( n = 67,581), 1993 – 2005 Nutrient Tertile 1 Tertile 2 Tertile 3 P trend IBD Protein 1 ( — ) 2.46 (1.15 – 5.28) 3.31 (1.41 – 7.77) 0.007 Carbohydrate 1 ( — ) 0.76 (0.43 – 1.37) 0.68 (0.37 – 1.27) 0.26 Fat 1 ( — ) 1.41 (0.70 – 2.84) 1.24 (0.57 – 2.72) 0.77 UC Protein 1 ( — ) 2.83 (1.07 – 7.48) 3.24 (1.07 – 9.84) 0.06 Carbohydrate 1 ( — ) 0.42 (0.19 – 0.94) 0.51 (0.24 – 1.08) 0.12 Fat 1 ( — ) 1.01 (0.41 – 2.47) 1.47 (0.56 – 3.84) 0.34 CD Protein 1 ( — ) 1.96 (0.58 – 6.68) 3.34 (0.90 – 12.4) 0.04 Carbohydrate 1 ( — ) 2.01 (0.70 – 5.72) 1.31 (0.42 – 4.14) 0.46 Fat 1 ( — ) 2.27 (0.69 – 7.45) 0.98 (0.25 – 3.88) 0.88 CD, Crohn’s disease; IBD, infl ammatory bowel disease; HR, hazard ratio; UC, ulcerative colitis. All nutrient tertiles were calculated from daily intake expressed in absolute amount of energy provided by the nutrient (Kcal / day). RESUL TS A total of 67,581 participants contributed 705,445 person-years in a mean follow-up of 10.4 years since co mpletion of the dietary questionnaire. Among the en tire cohort, 458 women checked the item “ Crohn ’ s disease ” or “ ulcerative colitis ” on the self-adminis- tered questionnaire. A er investigation, we validated the diagnosis of IBD in 194 women. Among them, 77 women wer e diagnos ed as having IBD a er inclusion in the study (incident cases) with a median of 54.5 months follo w-up (interquartile range: 26.5 – 95.5).  e 77 women with incident IBD were compar ed with the 67,504 non-cases. On the basis of available data, 43 cases were classi ed as UC, 30 cases as CD, and 4 cases as IC. In all, 16 cases wer e con- sidered as possible and 61 cases were classi ed as certain (includ- ing 33 UC cases, 25 CD cases and 3 cases with IC).  e bas eline characteristics of cases and non-cases are reported in Table 1 . HRs of IBD according to the dietary intake of macronutrien ts (expressed as the absolute amoun t of energy provided by each macronutrient) ar e presented in Table 2 . In a mutually adjusted model, total protein intake, bu t neither carbohydrates nor lipids, was statistically positively associated with IBD risk ( T able 2 ). When protein intake was exp ressed per kilogram body weight, mean intakes p er tertile were 1.08 g / kg (range: 0.30 – 1.33), 1.52 g / kg (range: 1.34 – 1.71), and 2.07 g / kg (range: 1.72 – 4.46). High intake of protein per kilogram body weight was also signi cantly associ- ated with IBD risk (HR for the third vs.  rst tertile of intake = 2.63; 95 % CI, 1.23 – 5.59; P for linear trend across tertiles = 0.008).  e positive association between high protein in t ake and IBD risk was restricted to animal protein in ta ke, as there was no signi cant association with vegetable prot ein ( T able 3 ). Although no hetero- geneity test was statistically signi cant when comparing UC a nd Table 3 . HR estimates and 95 % confi dence intervals for IBD in relation to type of protein in the E3N cohort ( n = 67,581), 1993 – 2005 Nutrient Tertile1 Tertile 2 Tertile 3 P trend IBD Animal protein 1 ( — ) 2.62 (1.29 – 5.30) 3.03 (1.45 – 6.34) 0.005 Vegetable protein 1 ( — ) 0.91 (0.47 – 1.77) 1.31 (0.59 – 2.88) 0.44 UC Animal protein 1 ( — ) 1.62 (0.65 – 4.02) 3.29 (1.34 – 8.04) 0.005 Vegetable protein 1 ( — ) 0.59 (0.23 – 1.54) 1.70 (0.59 – 4.81) 0.33 CD Animal protein 1 ( — ) 4.51 (1.28 – 15.83) 2.70 (0.69 – 10.52) 0.33 Vegetable protein 1 ( — ) 1.15 (0.39 – 3.35) 1.04 (0.28 – 3.80) 0.98 CD, Crohn’s disease; IBD, infl ammatory bowel disease; HR, hazard ratio; UC, ulcerative colitis. Model including energy from animal protein, energy from vegetable protein, and non-protein energy (i.e., energy from carbohydrates and lipids as ordered tertiles). © 2010 by the American College of Gastroenterology The American J ournal of GASTROENTER OLOGY 2199 INFLAMMA TORY BOWEL DISEASE Protein Intake and IBD egg and dairy products were not, potentially beca us e of insuf-  cient power. In this study of wo men with moderate-to-high consumption of p rotein, and especially animal protein (99 % of women consumed more than the mean n utritional requirement of 0.66 g / kg / day and 90 % more than the recommended daily amount of 0.83 g / kg / day) ( 18 ), high protein intake was associ- ated with a 3.3-fold increased risk of IBD .  is would make high- protein diet a major risk facto r of IBD.  e observed incidence of CD and UC in this cohort lies within the expected range in this age class: 4.2 per 100,000 for CD and 6.1 per 100,000 for UC ( 19,20 ). HRs were similar for UC and CD regarding total prot ein, although a dose – e ect relationship with animal pr otein was only observed for UC.  e lack of statistical signi cance fo r animal protein in CD could be due to a lack of po wer. Future studies should con rm this potential association between CD and pro- tein intake, and investiga te whether ilea l and colonic CD are equally associated with dietary protein in t ake. Our dietary survey has several strengths. Dietary question- naires were complet ed s everal years before the diagnosis of IBD .  e questionnaire has been validated and fo und to be reproducible ( 15 ). We ad justed the models for energy intake using the energy partition method, which has been largely used and advocat ed for the study of macronutrien ts ( 16 ). Using this method, we did not  nd any signi cant association between lipid o r carbohydrate intakes and the risk of CD or UC. A recent pr ospec tive study from EPIC showed a signi cant positive association between the dietary content in linoleic acid and UC incidence , and a negative one with docosohexaenoic acid intake, but not with to t al lipid intake, in agreement with our  ndings ( 21 ). Our study has some limitations. F irst, it can be argued that the potential interval between the acquisition o f t he dietary data and the onset of disease is long enough for substantial chan ges to occur in dietary habits. However , it has been shown that dietary CD, the shapes of associations between to ta l and animal protein and risk of IBD tended to di er slightly between UC and CD ( Tables 2 and 3 ). Regarding CD, a dose – e ect relationship was observed for total protein ( P = 0.04) but not for animal prot ein ( P = 0.33). Regard- ing UC, a strong dose – e ect relationship was observed with animal protein ( P = 0.005), whereas there was only a borderline statistically signi cant association with total protein ( P = 0.06). Analyses of food sources of animal protein showed tha t hig h consumptions of mea t or  sh, but not of eggs or dairy products, were positively associated with IBD risk ( Ta b l e 4 ). Further adjustment on daily alcoho l intake, physical activity, body mass index, tobacco smoking, menopause hormone ther- apy , or socio-economic status did not substantially modify the association between protein intake a nd IBD (see Supple mentary Appendix 1 and 2 online). When excluding the eight cases that occurred within 6 months (four UC, three CD, and o ne IC), the HR for the third vs.  rst ter- tile of animal protein in ta ke was 2.77 (95 % CI, 1.15 – 6.68); w hen excluding the 10 cases that occurred within 1 year (six UC, three CD, and one I C), t he corresponding HR became 2.74 (95 % CI, 1.30 – 5.78). Another sensitivity analysis was performed on the 61 certain cases of IBD and provided similar resul ts (HR for the third vs.  rst tertile of intake, 3.63; 95 % CI, 1.32 – 10.01 ( P trend = 0.01); and 3.13; 95 % CI, 1.34 – 7.31 ( P trend = 0.01) for total and animal protein, respectivel y). DISCUSSION  e main  ndin g of this prospect ive cohort study is that high protein intake was associated with an incr ease d risk of IBD.  e positive association between high protein in t ake and IBD risk was restricted to animal protein in t ake. Among animal protein sources, both  sh and meat were associated wi th r isk, whereas Table 4 . HR estimates and 95 % confi dence intervals for IBD in relation to different groups of food protein intake in the E3N cohort ( n = 67,581), 1993 – 2005 Tertile 1 Tertile 2 Tertile 3 P trend Meat (g / day) b 31.8 83.2 144.4 Number of IBD 15 25 37 Energy-adjusted HR a 1 ( — ) 1.45 (0.76 – 2.75) 1.87 (1.00 – 3.49) 0.02 Fish / sea products (g / day) b 10.7 26.4 59.1 Number of IBD 16 29 32 Energy-adjusted HR a 1 ( — ) 1.63 (0.88 – 3.01) 1.83 (1.00 – 3.36) 0.05 Eggs (g / day) b 6.9 19.5 46.1 Number of IBD 19 33 25 Energy-adjusted HR a 1 ( — ) 1.31 (0.74 – 2.31) 0.97 (0.52 – 1.78) 0.91 Dairy products (g / day) b 131.3 264.6 511.4 Number of IBD 24 27 26 Energy-adjusted HR a 1 ( — ) 1.02 (0.58 – 1.77) 0.94 (0.53 – 1.67) 0.93 HR, hazard ratio; IBD, infl ammatory bowel disease. a Mutually adjusted and adjusted on alcohol-free energy intake. b Mean value of daily consumption for each tertile. The American J ournal of GASTROENTER OLOGY VOLUME 105 | OCTOBER 2010 www.amjgastro.com 2200 INFLAMMA TORY BOWEL DISEASE Jantchou et al. habits change very little ov er time in middle-aged adults ( 22 ); in addition, changes in dietary habits would r esult in non-di erential misclassi cation of dietary exposure, and thus in diluting the e ect rather than increasing it. Some misclassi ca tion of dietary exposure is a constant featur e of dietar y surveys, and our moderately high correlation coe cients for reproducibility a er 1 year are in favor o f some degree of errors in assessing the diet. Again, this would ra ther dilute the true association. Second, this study included women aged 40 – 65 years, whereas IBD typically peaks between 20 – 35 years; further prospective studies should determine whether the results of this study could be reproduced in yo unger and male subjects. below a relative risk o f 2.5, esp ecially when considering separately CD and UC. Finally , because of the limited number of cases, we could not perform analyses based on quartiles or quin tiles, which would have better cap tured a true dose – e ect relations hip. Our results are consistent with the hypothesis fo rmulated by Shoda et al. (23), who found a temporal relationship between the rising incidence of CD in Japan a nd increasing dietary intake of animal protein ( r = 0.908). Since W orld W ar II, animal protein intake has increased not only in Ja pan but also in all developed countries ( 24 ); as an example, a 50 % increase has been reported in Belgium between the mid-1950s to 1978, with a per capita meat consumption fro m 60.8 to 98.0 kg / year ( 25 ).  e increase in animal protein consum ption may therefore explain some of the increased incidence of IBD since W o rld War II. Some (bu t not all) retrospective case – control studies have described a higher con- sumption of meat ( 26 ) and  sh ( 12 ) in CD patients as compared with controls. Mo reover, a follow-up st udy in 191 UC patients showed a signi cant association between high meat in t ake and risk of relapse of UC ( 27 ). On the contrary, a p rospec tive study that included some centers fro m t he EPIC study failed to  nd any association between any type of macr onutrient and UC risk ( 28 ). However, there w ere several di erences between that study tein considered as a percentage o f total energ y including alco- hol, heterogeneity between centers a nd dietar y questionnaires, and the inclusion of both men and women. I t needs to be furt her investigated as to what exten t these features would explain the di erent  ndings. Fro m our results and those of other studies, it may be speculated that reducin g protein intake to the  rst ter- tile ( < 67 g / day) could decrease both the incidence and activity of IBD. A prospective randomized stud y should be under taken to test this hypothesis.  ere is biological plausibility fo r a p ositive association between animal protein intake and IBD . A variable proportion of heme and amino acids, contained in animal pro teins, are not absorbed by the small bowel and reach the colonic lumen, wher e the y are metabo- lized by the micro ora ( 29 ).  is results in a number of end prod- ucts, which include hydrogen sul de, phenolic compounds, an d amines and ammonia, some of which ar e potentially toxic to the colon ( 29 ). For instance, it has been suggested that sul de, in the presence of nitric oxide pr o duced by anaerobic bacteria, ma y alter the cell membrane of the colonocyte, and lead to the loss of barrier function and the immune cascade as observed in UC ( 30 ). It has been shown that, in healthy volun teers, an increase in dietary pro- tein leads to changes in the colonic metabolism of pr oteins, which sulfur substances, and urinary p -cresol ( 31 ).  us, high a nimal protein consum ption might increase IBD risk through accrued formation of end pr o ducts by the colonic micro ora, such as hydrogen sul de, which are toxic for the colon. Addition al mecha- nisms, such as the impact of animal pro tein intake upon dysbiosis or in ammatory response, can also be suspected ( 32 ). In summary, our results may help better understand the r ole of diet in IBD risk. If con rmed, they can lead to prot ective strate- gies, especially in families at risk of IBD, a nd possibly to advice for preventing re lapse. ACKNOWLEDGMENTS  e authors are indebted to all the women o f the cohort for providing the data.  ey are grateful to the physicians and gastroenterologists for providing the clinical, endoscopy , and pathology reports, and also to Lyan Hoa ng, Marie Fangon, Maryvonne Niravong, an d C eline Kernaleguen for their technical assistance and Ms P amela Alber t for English assistance. CONFLICT OF INTEREST Guarantor of the article: Franck Carbonnel, MD, PhD. Speci c author contributions : Pr é vost Jantchou contributed to study design and implementa tion, data collection, design and implementation of the sta tistical analysis, interpretation of results, and Gastroenterol Clin Biol 2006 ; 30 : 859 – 67 . 9 . C osnes J . Tobacco and IBD: relevance in the understanding of disease mecha- nisms and clinical practice . Best Pract Res Clin Gastroenterol 2004 ; 18 : 481 – 96 . 1 0 . P e r s s o n P G , A h l b o m A , H e l l e r s G . D i e t a n d i n  ammatory bowel disease: a case-control study . Epidemiology 1992 ; 3 : 47 – 52 . 11 . Reif S , Klein I , Lubin F et al. Pre-illness dietary factors in in ammatory bowel disease . Gut 1997 ; 40 : 754 – 60 . 1 2 . S a k a m o t o N , K o n o S , W a k a i K et al. Dietary risk factors for in am matory bowel disease: a multicenter case-control study in J apan . In amm Bowel Dis 2005 ; 11 : 154 – 63 . 13 . Ekbom A . Environmental risk factors (excluding tobacco and microorgan- isms): a critical analysis of old and new h ypotheses . Best Pract Res Clin Gastroenterol 2004 ; 18 : 497 – 508 . 1 4 . C l a v e l - C h a p e l o n F , v a n L i e r e M J , G i u b o u t C et al. E3N, a French cohort study on cancer risk factors . Eur J C ancer Prev 1997 ; 6 : 473 – 8 . 1 5 . v a n L i e r e M J , L u c a s F , C l a v e l F et al. Relative validity and reproducib ility of a French dietar y history questionnaire . Int J Epidemiol 1997 ; 26 (Suppl 1) : S128 – 36 . 16 . Howe GR , Miller AB , Jain M . Re: “ Total energy intake: implications for epidemiologic analyses ” . Am J Epidemiol 1986 ; 124 : 157 – 9 . 1 7 . G r e e n l a n d S , R o t h m a n K J . I n t r o d u c t i o n t o s t r a t i  ed analysis . In: Rothman KJ, Greenland S (eds). Modern Epidemiology , 2nd edn. Lippincott-Raven: Philadelphia, P A , 1998 , pp. 53 – 79 . ... Soluble fiber from fruits and vegetables inhibits the transcription of inflammatory mediators and maintains the integrity of the epithelial barrier (Galvez et al. 2005;Rose et al. 2007). For meat and fish, which are rich in protein, total protein intake is positively associated with IBD development (Jantchou et al. 2010) . The results of this study further support the protective effect of vegetables and fruits on IBD. ... Drinking water quality and inflammatory bowel disease: a prospective cohort study ENVIRON SCI POLLUT R Shuduo Zhou Pengfei Chai Xuejie Dong Kun Chen Environmental factors, such as drinking water and diets, play an important role in the development of inflammatory bowel disease (IBD). This study aimed to investigate the associations of metal elements and disinfectants in drinking water with the risk of inflammatory bowel disease (IBD) and to assess whether diet influences these associations. We conducted a prospective cohort study including 22,824 participants free from IBD from the Yinzhou cohort study in the 2016–2022 period with an average follow-up of 5.24 years. The metal and disinfectant concentrations were measured in local pipeline terminal tap water samples. Cox regression models adjusted for multi-level covariates were used to estimate adjusted hazard ratios (aHRs) and 95% confidence intervals (95% CIs). During an average follow-up period of 5.24 years, 46 cases of IBD were identified. For every 1 standard deviation (SD) increase in the concentration of manganese, mercury, selenium, sulfur tetraoxide (SO4), chlorine, and nitrate nitrogen (NO3_N) were associated with a higher risk of IBD with the HRs of 1.45 (95% CI: 1.14 to 1.84), 1.51 (95% CI: 1.24–1.82), 1.29 (95% CI: 1.03–1.61), 1.52 (95% CI: 1.26–1.83), 1.26 (95% CI: 1.18–1.34), and 1.66 (95% CI: 1.32–2.09), whereas zinc and fluorine were inversely associated with IBD with the HRs of 0.42 (95% CI: 0.24 to 0.73) and 0.68 (95% CI: 0.54–0.84), respectively. Stronger associations were observed in females, higher income groups, low education groups, former drinkers, and participants who never drink tea. Diets have a moderating effect on the associations of metal and nonmetal elements with the risk of IBD. We found significant associations between exposure to metals and disinfectants and IBD. Diets regulated the associations to some extent. ... Red meat is one of the ingredients of a typical western diet (Thøgersen and Bertram, 2021). In human studies, red meat consumption has been associated with UC, by inducing or exacerbating this disease (Jantchou et al., 2010; Ge et al., 2015;Rashvand et al., 2015) due to increased proteolytic activity by the colonic bacteria resulting from higher substrate availability (Gilbert et al., 2018). Although the exact mechanism behind this effect is yet to be elucidated (Van Hecke et al., 2021), microbial fermentation of undigested dietary and endogenous protein sources results in various metabolites, several of which, e.g., biogenic amines, NH 4 + , indoles, phenols, and H 2 S, are considered detrimental for gut health as they are capable of stimulating the immune system and starting a cascade of inflammatory responses in the colon (Windey et al., 2012;Gilbert et al., 2018;Panah et al., 2021). ... A westernized diet changed the colonic bacterial composition and metabolite concentration in a dextran sulfate sodium pig model for ulcerative colitis Article Full-text available Apr 2023 Farhad M. Panah Katrine D Nielsen Gavin Leslie Simpson Nuria Canibe Introduction: Ulcerative colitis (UC) is characterized by chronic inflammation in the colonic epithelium and has a blurred etiology. A western diet and microbial dysbiosis in the colon were reported to play a role in UC development. In this study, we investigated the effect of a westernized diet, i.e., increasing fat and protein content by including ground beef, on the colonic bacterial composition in a dextran sulfate sodium (DexSS) challenged pig study. Methods: The experiment was carried out in three complete blocks following a 2×2 factorial design including 24 six-week old pigs, fed either a standard diet (CT) or the standard diet substituted with 15% ground beef to simulate a typical westernized diet (WD). Colitis was induced in half of the pigs on each dietary treatment by oral administration of DexSS (DSS and WD+DSS, respectively). Samples from proximal and distal colon and feces were collected. Results and discussion: Bacterial alpha diversity was unaffected by experimental block, and sample type. In proximal colon, WD group had similar alpha diversity to CT group and the WD+DSS group showed the lowest alpha diversity compared to the other treatment groups. There was a significant interaction between western diet and DexSS for beta diversity, based on Bray-Curtis dissimilarly. The westernized diet and DexSS resulted in three and seven differentially abundant phyla, 21 and 65 species, respectively, mainly associated with the Firmicutes and Bacteroidota phyla followed by Spirochaetota, Desulfobacterota, and Proteobacteria. The concentration of short-chain fatty acids (SCFA) was lowest in the distal colon. Treatment had a slight effect on the estimates for microbial metabolites that might have valuable biological relevance for future studies. The concentration of putrescine in the colon and feces and that of total biogenic amines was highest in the WD+DSS group. We conclude that a westernized diet could be a potential risk factor and an exacerbating agent for UC by reducing the abundance of SCFA-producing bacteria, increasing the abundance of pathogens such as Helicobacter trogontum, and by increasing the concentration of microbial proteolytic-derived metabolites in the colon. ... Several epidemiological, observational, prospective, and retrospective case-control studies have been conducted (Shoda et al., 1996;Sakamoto et al., 2005;Hart et al., 2008; Jantchou et al., 2010; Maconi et al., 2010;Racine et al., 2016;Limdi, 2018). They aimed to characterize the role of diet as a risk factor for the development of IBD. ... Vegetable oils and oilseeds Chapter Mar 2023 Aleksandra Tarasiuk Mikołaj Świerczyński Maciej Sałaga ... Several studies report that diet plays an important role in the development of IBDs. A relationship between animal protein consumption and IBDs has already been discussed (Jantchou et al., 2010; Windey et al., 2012;Virta et al., 2013). Recently, a Chinese study evaluated the effects of milk protein, potato protein, and their mixture on DSS-induced UC ( Table 9). ... Cereal grains and vegetables Chapter Apr 2023 Roberto de Paula do Nascimento Mariana Alves Nathan Noguera Mário Maróstica Jr Inflammatory bowel diseases (IBDs) are idiopathic chronic conditions of the gastrointestinal tract that can lead to life-threatening complications. Although their treatments can be conveniently managed nowadays, a considerable portion of patients still complains about the adverse effects, lack of efficacy, and/or high costs of drugs and surgeries. In the pursuit of finding safer, cheaper, but still effective options, researchers have been conducting experimental studies, mostly in the last 6 years, on the potential of cereal grains and vegetables against IBDs. These crops, which are probably the most consumed types of plant-associated products among populations worldwide, have been reported as relevant sources of nutrients and bioactive compounds, especially proteins, fibers, and polyphenols, with promising antiinflammatory, antioxidative, and antidysbiotic effects on colitis. All things considered, this chapter aims to present and discuss the implications of various cereal grains and vegetables on IBDs, taking as reference preclinical and clinical experiments. ... Consumption of animal-derived proteins, particularly red meat and dairy products, can lead to an increase in the abundance of bile-tolerant anaerobic bacteria, such as Bacteroidetes, Alistipes, and Bilophila, inducing an increase in the production of trimethylamine N-oxide, a compound known for its pro-atherogenic potential [9]. Furthermore, a study has shown that high consumption of proteins of animal origin could increase the risk of inflammatory bowel disease (IBD) through an accumulated production of hydrogen sulfide (H 2 S) by bacteria (for example Desulfovibrio spp.) that reduces sulphate from food inorganic sulphates and sulfur amino acids [78] . However, in terms of vegetable proteins, a study demonstrated how the consumption of pea proteins led to an increase in Bifidobacterium and Lactobacillus, and a reduction in pathogenic Bacteroides fragilis and Clostridium perfrigens [79]. ... How Do Diet Patterns, Single Foods, Prebiotics and Probiotics Impact Gut Microbiota? Mar 2023 Microbiol Res Andrea Piccioni Marcello Covino Marcello Candelli Giuseppe Merra The human gastrointestinal tract hosts a complex and dynamic population of commensal bacterial species, which have coevolved with the host, generating a symbiotic relationship. Some compounds present in foods, such as polyols, prebiotic fibers, or phenolic compounds, are poorly metabolized and absorbed by the host before the transformation guided by the colonic microbiota. By influencing gut microbiota, diet plays a fundamental role in understanding the beneficial effects of the gut microbiota on the host, including its long-term metabolism. The idea that probiotics can act not only by influencing the colonizing microbiota opens the door to a wider range of probiotic possibilities, encouraging innovation in the field. Furthermore, it has been shown both that some probiotics increase phagocytosis or the activity of natural killer cells. Current prebiotics are mainly based on carbohydrates, but other substances, such as polyphenols and polyunsaturated fatty acids, could exert prebiotic effects. A prebiotic substance has been defined as ‘a substrate that is selectively used by host microorganisms that confer a health benefit’, and so can interact with the gut microbiota through competition for nutrients, antagonism, cross-feeding, and support for microbiota stability. Influencing its composition in terms of richness and diversity, food components have a key impact on the intestinal microbiota. Eating habits can strongly influence the composition of the intestinal microbiota. A healthy intestinal microbiota is essential for maintaining general health, and diet is one of the major modulators of this fascinating world of microorganisms. This must give us one more reason to adopt a healthy lifestyle. View Show abstract

https://www.researchgate.net/publication/44590778_Animal_Protein_Intake_and_Risk_of_Inflammatory_Bowel_Disease_The_E3N_Prospective_Study

Search Results - NHS Parkinson's UK Redcar Area Support Group;Southtees SENDIASS;Carers Together Foundation;Redcar and Cleveland Council;The Grenfell Club;Zoe'S Place Baby Hospice Middlesbrough;Middlesbrough Multiple Sclerosis Therapy Centre;Carers Together Middlesbrough;Living with Secondary Breast Cancer - Middlesbrough;North East Asbestos Support & Awareness Group Results for Carer centres and services in Westfield Showing 1-10 of 97 results Please check travel times before starting your journey.Distances are given in a straight line and may not be the quickest to travel to Local National Carer-centres-and-services details Address & contact details Information supplied by Target group Parkinson's UK Redcar Area Support Group Address & contact detailsTel: 0344 225 360425K Community CentreAyton DriveRedcarTeessideTS10 4LR0.2 miles away Information supplied by Offer information, friendship and support to local people with Parkinson's, their families and carers. Also organise regular events and social activities - join and meet other people affected by Parkinson's in your area. Target group People with Parkinson's, their families and carers. Southtees SENDIASS Address & contact detailsTel: 01642 759073 / 07925 149157Redcar Children's CentreKeilder CloseRedcarTeessideTS10 4HS0.5 miles away Information supplied by The information, Advice & Support service has developed from the Parent Partnership Service. The aim of the service is to support parents, carers and young people (16-25) with Special Educational Needs and Disability. The Information, Advice & Support Service is free, impartial and confidential a... Continue reading overview Target group Parents/carers of pupils with special educational needs. Carers Together Foundation Address & contact detailsTel: 01642 488977The Innovation CentreVienna CourtKirkleatham Business ParkRedcarTeessideTS10 5SH0.8 miles away Information supplied by Carers Together Foundation is an independent registered charity which is dedicated to improving the quality of life for carers who care for someone living in South Tees (Middlesbrough, and Redcar and Cleveland). We are committed to informing, supporting and representing the needs of carers. We ... Continue reading overview Target group Carers who care for someone living in Redcar and Cleveland. Redcar and Cleveland Council Address & contact detailsTel: 01642 774774Adult & Children's ServicesSeafield HouseKirkleatham StreetRedcarTeessideTS10 1SP0.9 miles away Information supplied by Provides social services Target group Adults, children, young people, older people and those who care for them The Grenfell Club Address & contact detailsTel: 01642 471747Ridley StreetRedcarTeessideTS10 1RF0.9 miles away Information supplied by The Grenfell Club provides leisure activities for children and their families or carers. The club opens five nights a week offering a welcoming and friendly atmosphere. Members enjoy a wide variety of activities including, arts and crafts, cookery, computing, a multi-sensory room, regular disco... Continue reading overview Target group n/a Data not available Zoe'S Place Baby Hospice Middlesbrough Address & contact detailsTel: 01642 457985Crossbeck HouseHigh StreetNormanbyMiddlesbroughTeessideTS6 9DA4.4 miles away Information supplied by Zoe's Place Baby hospice Middlesbrough is a registered charity providing palliative, respite and end of life care to babies and infants aged from birth to five years suffering from life-limiting or life-threatening conditions. This care will be provided in a caring, safe, home from home environment. Target group General public Middlesbrough Multiple Sclerosis Therapy Centre Address & contact detailsTel: 01642 461673Unit B1 ,Stadium CourtWallis Road, Skippers Lane Ind.Est.South BankMiddlesbroughTeessideTS6 6JB4.7 miles away Information supplied by The Multiple Sclerosis Therapy Centre was opened in 1985 by people with Multiple Sclerosis. Its aim is to serve people affected by MS in Tees Valley, North Yorkshire, County Durham, Tyne and Wear and into Northumbria. The centre provides a range of essential therapies aimed at limiting the proble... Continue reading overview Target group People with Multiple Sclerosis their carers and their families. We also provide support and therapy to stroke victims, ME sufferers and people with other neurological disorders. Carers Together Middlesbrough Address & contact detailsTel: 01642 488977The TAD CentreOrmesby RoadMiddlesbroughTeessideTS3 7SF5.8 miles away Information supplied by Carers Together in Middlesborough is an independent registered charity which is dedicated to improving the quality of life for carers who care for someone living in Redcar and Cleveland. We are committed to informing, supporting and representing the needs of carers. We have also played a signif... Continue reading overview Target group Carers who care for someone living in Middlesborough Living with Secondary Breast Cancer - Middlesbrough Address & contact detailsTel: 0345 077 18935.9 miles away Information supplied by Our Living with Secondary Breast Cancer meet ups in Middlesbrough usually take place monthly, and you can attend as few or as many as you like. All sessions are facilitated by a counsellor and once a quarter we invite a guest speaker, for example a Clinical Nurse Specialist or Benefits Advisor. Q... Continue reading overview Target group n/a Data not available North East Asbestos Support & Awareness Group Address & contact detailsTel: 01642 240044119-121 Marton RoadMiddlesbroughTeessideTS1 2DU6.3 miles away Information supplied by The North East Asbestos Support and Awareness Group is a not-for-profit organisation that provides a comprehensive advice service for asbestos sufferers and their families. NASAG is supported by volunteers and our services are offered completely free of charge. Target group n/a Data not available

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Noise in Ultrashort Elastic Membrane Nanotube | Request PDF Request PDF | On Dec 9, 2022, K. A. Ivanova and others published Noise in Ultrashort Elastic Membrane Nanotube | Find, read and cite all the research you need on ResearchGate Article Noise in Ultrashort Elastic Membrane Nanotube December 2022 Biochemistry (Moscow) Supplement Series A Membrane and Cell Biology16(4):320-327 DOI: 10.1134/S1990747822050063 Authors: <here is a image f2f62089ad6ff61c-d19572ff1996375a> K. A. Ivanova K. A. Ivanova <here is a image 1144e1be9de06691-b3c79ed0cfa37a80> Pavel Bashkirov Russian Academy of Sciences Electrical Low-Frequency 1/ f γ Noise Due to Surface Diffusion of Scatterers on an Ultra-low-Noise Graphene Platform Article Full-text available Sep 2021 Masahiro Kamada <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Antti Laitinen Weijun Zeng <here is a image d0c1a8c735f162bc-ecce6fc2f70a1179> Pertti Hakonen Low-frequency 1/f γ noise is ubiquitous, even in high-end electronic devices. Recently, it was found that adsorbed O2 molecules provide the dominant contribution to flux noise in superconducting quantum interference devices. To clarify the basic principles of such adsorbate noise, we have investigated low-frequency noise, while the mobility of surface adsorbates is varied by temperature. We measured low-frequency current noise in suspended monolayer graphene Corbino samples under the influence of adsorbed Ne atoms. Owing to the extremely small intrinsic noise of suspended graphene, we could resolve a combination of 1/f γ and Lorentzian noise induced by the presence of Ne. We find that the 1/f γ noise is caused by surface diffusion of Ne atoms and by temporary formation of few-Ne-atom clusters. Our results support the idea that clustering dynamics of defects is relevant for understanding of 1/f noise in metallic systems. Nonlinear material and ionic transport through membrane nanotubes Article Full-text available Jun 2021 BBA-BIOMEMBRANES Dmitrii V. Ivchenkov Peter I. Kuzmin <here is a image 650b3144c88f8d37-934f18985504ef73> Timur Galimzyanov <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Vadim A. Frolov Membrane nanotubes (NTs) and their networks play an important role in intracellular membrane transport and intercellular communications. The transport characteristics of the NT lumen resemble those of conventional solid-state nanopores. However, unlike the rigid pores, the soft membrane wall of the NT can be deformed by forces driving the transport through the NT lumen. This intrinsic coupling between the NT geometry and transport properties remains poorly explored. Using synchronized fluorescence microscopy and conductance measurements, we revealed that the NT shape was changed by both electric and hydrostatic forces driving the ionic and solute fluxes through the NT lumen. Far from the shape instability, the strength of the force effect is determined by the lateral membrane tension and is scaled with membrane elasticity so that the NT can be operated as a linear elastic sensor. Near shape instabilities, the transport forces triggered large-scale shape transformations, both stochastic and periodic. The periodic oscillations were coupled to a vesicle passage along the NT axis, resembling peristaltic transport. The oscillations were parametrically controlled by the electric field, making NT a highly nonlinear nanofluidic circuitry element with biological and technological implications. Comparing Current Noise in Biological and Solid-State Nanopores Article Full-text available Feb 2020 <here is a image 4f4ba2423d437680-de5723e840c43d45> Alessio Fragasso <here is a image ea794360d30e7a3e-7f106db0be31644e> Sonja Schmid Cees Dekker Nanopores bear great potential as single-molecule tools for bioanalytical sensing and sequencing, due to their exceptional sensing capabilities, high-throughput, and low cost. The detection principle relies on detecting small differences in the ionic current as biomolecules traverse the nanopore. A major bottleneck for the further progress of this technology is the noise that is present in the ionic current recordings, because it limits the signal-to-noise ratio (SNR) and thereby the effective time resolution of the experiment. Here, we review the main types of noise at low and high frequencies and discuss the underlying physics. Moreover, we compare biological and solid-state nanopores in terms of the SNR, the important figure of merit, by measuring translocations of a short ssDNA through a selected set of nanopores under typical experimental conditions. We find that SiN x solid-state nanopores provide the highest SNR, due to the large currents at which they can be operated and the relatively low noise at high frequencies. However, the real game-changer for many applications is a controlled slowdown of the translocation speed, which for MspA was shown to increase the SNR > 160-fold. Finally, we discuss practical approaches for lowering the noise for optimal experimental performance and further development of the nanopore technology. Noise in nanopore sensors: Sources, models, reduction, and benchmarking Article Full-text available Dec 2019 Shengfa Liang <here is a image eeb154778751100e-dc01be971592152f> Feibin Xiang <here is a image c963708723a2f91d-fbc119538791a117> Zifan Tang <here is a image 80565bcd2eea114d-2f66e0690271d985> Weihua Guan Label-free nanopore sensors have emerged as a new generation technology of DNA sequencing and have been widely used for single molecule analysis. Since the first α-hemolysin biological nanopore, various types of nanopores made of different materials have been under extensive development. Noise represents a common challenge among all types of nanopore sensors. The nanopore noise can be decomposed into four components in the frequency domain (1/f noise, white noise, dielectric noise, and amplifier noise). In this work, we reviewed and summarized the physical models, origins, and reduction methods for each of these noise components. For the first time, we quantitatively benchmarked the root mean square (RMS) noise levels for different types of nanopores, demonstrating a clear material-dependent RMS noise. We anticipate this review article will enhance the understanding of nanopore sensor noises and provide an informative tutorial for developing future nanopore sensors with a high signal-to-noise ratio. Fluid surface coatings for solid-state nanopores: comparison of phospholipid bilayers and archaea-Inspired lipid monolayers Article Full-text available Apr 2019 <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Geoffray Leriche <here is a image 1f5b2c46def3b872-16b36a04f094bd04> Takaoki Koyanagi <here is a image a741c6735d96ae60-12749d4310394c92> Olivia Eggenberger <here is a image b4205e4d65cef25b-f9dbdb6ae6ceeef8> Michael Mayer In the context of sensing and characterizing single proteins with synthetic nanopores, lipid bilayer coatings provide at least four benefits: first, they minimize unwanted protein adhesion to the pore walls by exposing a zwitterionic, fluid surface. Second, they can slow down protein translocation and rotation by the opportunity to tether proteins with a lipid anchor to the fluid bilayer coating. Third, they provide the possibility to impart analyte specificity by including lipid anchors with a specific receptor or ligand in the coating. Fourth, they offer a method for tuning nanopore diameters by choice of the length of the lipid's acyl chains. The work presented here compares four properties of various lipid compositions with regard to their suitability as nanopore coatings for protein sensing experiments: (1) electrical noise during current recordings through solid-state nanopores before and after lipid coating, (2) long-term stability of the recorded current baseline and, by inference, of the coating, (3) viscosity of the coating as quantified by the lateral diffusion coefficient of lipids in the coating, and (4) the success rate of generating a suitable coating for quantitative nanopore-based resistive pulse recordings. We surveyed lipid coatings prepared from bolaamphiphilic, monolayer-forming lipids inspired by extremophile archaea and compared them to typical bilayer-forming phosphatidylcholine lipids containing various fractions of curvature-inducing lipids or cholesterol. We found that coatings from archaea-inspired lipids provide several advantages compared to conventional phospholipids; the stable, low noise baseline qualities and high viscosity make these membranes especially suitable for analysis that estimates physical protein parameters such as the net charge of proteins as they enable translocation events with sufficiently long duration to time-resolve dwell time distributions completely. The work presented here reveals that the ease or difficulty of coating a nanopore with lipid membranes did not depend significantly on the composition of the lipid mixture, but rather on the geometry and surface chemistry of the nanopore in the solid state substrate. In particular, annealing substrates containing the nanopore increased the success rate of generating stable lipid coatings. Detection of DNA molecules in a lipid nanotube channel in the low Ion strength conditions Article Full-text available Jan 2017 BIOL MEMBRANY <here is a image 380416004276da08-89c6810f91fec6ee> K. V. Chekashkina <here is a image 650b3144c88f8d37-934f18985504ef73> Timur Galimzyanov Peter I. Kuzmin <here is a image 1144e1be9de06691-b3c79ed0cfa37a80> Pavel Bashkirov investigation of the transport phenomena in the nanoscopic channels/pores with the diameter smaller than 100 nm is of utmost importance for various biological, medical, and technical applications. Presently, the main line of development of nanofiuidics is creation of biosensors capable of detecting single molecules and manipulating them. Detection of molecules is based on the measurement of electric current through a channel of appropriate size: When the molecule enters the channel, diameter of which is comparable with the molecule size, the ion current reduces. In order to improve transport properties of such channels, their walls are often coated with a lipid bilayer, which behaves as two-dimensional liquid and thus is capable of supporting transport phenomena. In the present work, we utilized this property of lipid membranes for development of a method for detecting and controlling transport of single-stranded DNA through channels formed by membrane cylinders with the luminal radii of 5-7 nm. We have demonstrated that in the conditions of small ion strength, the appearance of a DNA molecule inside such channel is accompanied by an increase of its ion conductivity and can be controlled by the polarity of applied voltage. The amplitude of the ion current increase allows evaluating the amount of DNA molecules inside the channels. It was also demonstrated that upon adsorption of DNA molecules on the lipid bilayer surface, the membrane cylinder behaves as a voltage-sensitive selective ion channel. Detection of DNA molecules in a lipid nanotube channel in the low ion strength conditions Article Full-text available Jul 2017 <here is a image 650b3144c88f8d37-934f18985504ef73> Timur Galimzyanov Peter I. Kuzmin <here is a image 380416004276da08-89c6810f91fec6ee> K. V. Chekashkina <here is a image 1144e1be9de06691-b3c79ed0cfa37a80> Pavel Bashkirov Investigation of the transport phenomena in the nanoscopic channels/pores with the diameter smaller than 100 nm is of utmost importance for various biological, medical, and technical applications. Presently, the main line of development of nanofluidics is creation of biosensors capable of detecting single molecules and manipulating them. Detection of molecules is based on the measurement of electric current through a channel of appropriate size: when the molecule enters the channel, which diameter is comparable with the molecule size, the ion current reduces. In order to improve transport properties of such channels, their walls are often coated with a lipid bilayer, which behaves as two-dimensional liquid and thus is capable of supporting transport phenomena. In the present work, we utilized this property of lipid membranes for the development of a method for detecting and controlling transport of single-stranded DNA through channels formed by membrane cylinders with the luminal radii of 5–7 nm. We have demonstrated that in the conditions of small ion strength, the appearance of a DNA molecule inside such channel is accompanied by an increase of its ion conductivity and can be controlled by the polarity of the applied voltage. The amplitude of the ion current increase allows evaluating the amount of DNA molecules inside the channels. It was also demonstrated that upon adsorption of DNA molecules on the lipid bilayer surface, the membrane cylinder behaves as a voltage-sensitive selective ion channel. Metabolomics reveals distinct neurochemical profiles associated with stress resilience Article Full-text available Dec 2017 <here is a image f6698e4bc64a397a-8f6b5d16a81d91e2> Brooke Nichole Dulka <here is a image a09c20ab41e80e58-d5a9e0642de3c3d6> Allen K Bourdon <here is a image e1925c2713d24051-fbc467fa70c71822> Catie Clinard <here is a image 582b06c5a75240c1-581ae76616565645> Matthew A Cooper Acute social defeat represents a naturalistic form of conditioned fear and is an excellent model in which to investigate the biological basis of stress resilience. While there is growing interest in identifying biomarkers of stress resilience, until recently, it has not been feasible to associate levels of large numbers of neurochemicals and metabolites to stress-related phenotypes. The objective of the present study was to use an untargeted metabolomics approach to identify known and unknown neurochemicals in select brain regions that distinguish susceptible and resistant individuals in two rodent models of acute social defeat. In the first experiment, male mice were first phenotyped as resistant or susceptible. Then, mice were subjected to acute social defeat, and tissues were immediately collected from the ventral medial prefrontal cortex (vmPFC), basolateral/central amygdala (BLA/CeA), nucleus accumbens (NAc), and dorsal hippocampus (dHPC). Ultra-high performance liquid chromatography coupled with high resolution mass spectrometry (UPLC-HRMS) was used for the detection of water-soluble neurochemicals. In the second experiment, male Syrian hamsters were paired in daily agonistic encounters for 2 weeks, during which they formed stable dominant-subordinate relationships. Then, 24 h after the last dominance encounter, animals were exposed to acute social defeat stress. Immediately after social defeat, tissue was collected from the vmPFC, BLA/CeA, NAc, and dHPC for analysis using UPLC-HRMS. Although no single biomarker characterized stress-related phenotypes in both species, commonalities were found. For instance, in both model systems, animals resistant to social defeat stress also show increased concentration of molecules to protect against oxidative stress in the NAc and vmPFC. Additionally, in both mice and hamsters, unidentified spectral features were preliminarily annotated as potential targets for future experiments. Overall, these findings suggest that a metabolomics approach can identify functional groups of neurochemicals that may serve as novel targets for the diagnosis, treatment, or prevention of stress-related mental illness. Integration of solid-state nanopores into a functional device designed for electrical and optical cross-monitoring Article Full-text available Jul 2017 BIOMED MICRODEVICES <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Raphaël Marchand <here is a image e8bdd2eb1de0e3dc-0c0fbac9732d4027> Christophe Thibault <here is a image 152134f0eb6016f6-46b2ad7af2311f0b> F. Carcenac Emmanuelle Trévisiol We present a new strategy for fabricating a silicon nanopore device allowing straightforward fluidic integration and electrical as well as optical monitoring. The device presents nanopores of diameters 10 nm to 160 nm, and could therefore be used to obtain solvent-free free-standing lipid bilayers from small unilamellar vesicles (SUV) or large unilamellar vesicles (LUV). The silicon chip fabrication process only requires front side processing of a silicon-on-insulator (SOI) substrate. A polydimethylsiloxane (PDMS) microfluidic interface is assembled on the silicon chip for fluidic handling and electrical addressing. We detail the electrical specifications of our device and some perspectives showing that the use of an SOI substrate is a convenient way to reduce the electrical noise in a silicon nanopore device without the need of a photolitographic patterned passivation layer. We then demonstrate simultaneous electrical and optical monitoring by capturing negatively charged fluorescent nanoparticles. Finally, in the perspective of solvent-free free-standing lipid bilayers, we show that incubation of SUV results in a drastic increase of the device electrical resistance, which is likely due to the formation of a free-standing lipid bilayer sealing the nanopores. Graphical abstractᅟ The Oxford Nanopore MinION: Delivery of nanopore sequencing to the genomics community Article Full-text available Nov 2016 GENOME BIOL <here is a image e629470c65e4b0dd-5a54cb3820f8ab5f> Miten Jain <here is a image 6a1fe9a144a2e281-d110ece3974eda4d> Hugh Olsen Benedict Paten <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Mark Akeson Nanopore DNA strand sequencing has emerged as a competitive, portable technology. Reads exceeding 150 kilobases have been achieved, as have in-field detection and analysis of clinical pathogens. We summarize key technical features of the Oxford Nanopore MinION, the dominant platform currently available. We then discuss pioneering applications executed by the genomics community. Fabrication of Low Noise Borosilicate Glass Nanopores for Single Molecule Sensing Article Full-text available Jun 2016 PLOS ONE <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Jayesh Arun Bafna <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Gautam Soni We show low-cost fabrication and characterization of borosilicate glass nanopores for single molecule sensing. Nanopores with diameters of ~100 nm were fabricated in borosilicate glass capillaries using laser assisted glass puller. We further achieve controlled reduction and nanometer-size control in pore diameter by sculpting them under constant electron beam exposure. We successfully fabricate pore diameters down to 6 nm. We next show electrical characterization and low-noise behavior of these borosilicate nanopores and compare their taper geometries. We show, for the first time, a comprehensive characterization of glass nanopore conductance across six-orders of magnitude (1M-1μM) of salt conditions, highlighting the role of buffer conditions. Finally, we demonstrate single molecule sensing capabilities of these devices with real-time translocation experiments of individual λ-DNA molecules. We observe distinct current blockage signatures of linear as well as folded DNA molecules as they undergo voltage-driven translocation through the glass nanopores. We find increased signal to noise for single molecule detection for higher trans-nanopore driving voltages. We propose these nanopores will expand the realm of applications for nanopore platform. Emerging Insights into the Roles of Membrane Tethers from Analysis of Whole Organisms: The Tip of an Iceberg? Article Full-text available Feb 2016 <here is a image c8c368d43ef91954-57e4989516c0864d> Wei Hong Toh Paul A. Gleeson Membrane tethers have been identified throughout different compartments of the endomembrane system. It is now well established that a number of membrane tethers mediate docking of membrane carriers in anterograde and retrograde transport and in regulating the organization of membrane compartments. Much of our information on membrane tethers have been obtained from the analysis of individual membrane tethers in cultured cells. In the future it will be important to better appreciate the network of interactions mediated by tethers and the potential co-ordination of their collective functions in vivo. There are now a number of studies which have analyzed membrane tethers in tissues and organisms which are providing new insights into the role of this class of membrane protein at the physiological level. Here we review recent advances in the understanding of the function of membrane tethers from knock outs (or knock downs) in whole organisms and from mutations in tethers associated with disease. Membrane tethering Article Full-text available Sep 2014 Pei Zhi Cheryl Chia Paul A. Gleeson Membrane trafficking depends on transport vesicles and carriers docking and fusing with the target organelle for the delivery of cargo. Membrane tethers and small guanosine triphosphatases (GTPases) mediate the docking of transport vesicles/carriers to enhance the efficiency of the subsequent SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor)-mediated fusion event with the target membrane bilayer. Different classes of membrane tethers and their specific intracellular location throughout the endomembrane system are now well defined. Recent biochemical and structural studies have led to a deeper understanding of the mechanism by which membrane tethers mediate docking of membrane carriers as well as an appreciation of the role of tethers in coordinating the correct SNARE complex and in regulating the organization of membrane compartments. This review will summarize the properties and roles of membrane tethers of both secretory and endocytic systems. A sub-nanometre view of how membrane curvature and composition modulate lipid packing and protein recruitment Article Full-text available Sep 2014 <here is a image ed5a2523881274ca-5e1ab18bd61f8de1> Stefano Vanni <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Hisaaki Hirose <here is a image 57a40d522b9dcaaf-fc9741646f6feb68> Helene Barelli <here is a image 1ce35f78ddc6efd6-ac61f1362252fd9e> Romain Gautier Two parameters of biological membranes, curvature and lipid composition, direct the recruitment of many peripheral proteins to cellular organelles. Although these traits are often studied independently, it is their combination that generates the unique interfacial properties of cellular membranes. Here, we use a combination of in vivo, in vitro and in silico approaches to provide a comprehensive map of how these parameters modulate membrane adhesive properties. The correlation between the membrane partitioning of model amphipathic helices and the distribution of lipid-packing defects in membranes of different shape and composition explains how macroscopic membrane properties modulate protein recruitment by changing the molecular topography of the membrane interfacial region. Furthermore, our results suggest that the range of conditions that can be obtained in a cellular context is remarkably large because lipid composition and curvature have, under most circumstances, cumulative effects. Sensing Membrane Stresses by Protein Insertions Apr 2014 PLOS COMPUT BIOL <here is a image 16608a4db424ef77-df47bcad67cc8f9c> Felix Campelo <here is a image 1dbc0414975da84b-f98ee37bc38f7457> Michael M Kozlov Author Summary Selective targeting of soluble proteins to cellular membranes relies on different mechanisms such as receptor-mediated recruitment or direct binding to specific lipids. A new paradigm has been recently proposed, according to which membrane binding of some proteins is driven by the geometrical and physical properties of the membranes, namely the membrane curvature and lipid packing in the external membrane monolayer. Specifically, several proteins referred to as the membrane curvature sensors have been shown to preferentially bind strongly curved membranes. This mode of protein binding is especially relevant for such fundamental cell processes as endocytosis and carrier generation from ER and Golgi Complex, which involve shaping initially flat membranes into strongly curved ones. A subset of the curvature sensors contains amphipathic or hydrophobic domains that shallowly insert into the membrane. Here we explore computationally the detailed physical mechanism underlying the membrane binding by such proteins and demonstrate that their membrane affinity is not determined by the curvature per se but rather by the membrane stress, independently of the way the stress has been generated. Hence, the significance of our work is in elucidating the relationship between the membrane binding of peripheral proteins carrying shallowly inserting domains and the membrane stresses. Lipid bilayer coated Al2O3 nanopore sensors: Towards a hybrid biological solid-state nanopore Apr 2011 BIOMED MICRODEVICES Bala Murali Venkatesan <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> James Polans <here is a image 2616130cf68264ab-95b29a086d3aa154> Jeffrey Comer Rashid Bashir Solid-state nanopore sensors are highly versatile platforms for the rapid, label-free electrical detection and analysis of single molecules, applicable to next generation DNA sequencing. The versatility of this technology allows for both large scale device integration and interfacing with biological systems. Here we report on the development of a hybrid biological solid-state nanopore platform that incorporates a highly mobile lipid bilayer on a single solid-state Al(2)O(3) nanopore sensor, for the potential reconstitution of ion channels and biological nanopores. Such a system seeks to combine the superior electrical, thermal, and mechanical stability of Al(2)O(3) solid-state nanopores with the chemical specificity of biological nanopores. Bilayers on Al(2)O(3) exhibit higher diffusivity than those formed on TiO(2) and SiO(2) substrates, attributed to the presence of a thick hydration layer on Al(2)O(3), a key requirement to preserving the biological functionality of reconstituted membrane proteins. Molecular dynamics simulations demonstrate that the electrostatic repulsion between the dipole of the DOPC headgroup and the positively charged Al(2)O(3) surface may be responsible for the enhanced thickness of this hydration layer. Lipid bilayer coated Al(2)O(3) nanopore sensors exhibit excellent electrical properties and enhanced mechanical stability (GΩ seals for over 50 h), making this technology ideal for use in ion channel electrophysiology, the screening of ion channel active drugs and future integration with biological nanopores such as α-hemolysin and MspA for rapid single molecule DNA sequencing. This technology can find broad application in bio-nanotechnology. Controlling protein translocation through nanopores with bio-inspired fluid walls Feb 2011 Nat. Nanotech. <here is a image b893ed4bdb49146b-5a5dda41ac7347f7> Erik Yusko <here is a image c3ea020a69c897de-132f81b4574d339c> Jay Johnson <here is a image 4cb53294bbba5a93-d65f7e6dea21c661> Sheereen Majd <here is a image b4205e4d65cef25b-f9dbdb6ae6ceeef8> Michael Mayer Synthetic nanopores have been used to study individual biomolecules in high throughput, but their performance as sensors does not match that of biological ion channels. Challenges include control of nanopore diameters and surface chemistry, modification of the translocation times of single-molecule analytes through nanopores, and prevention of non-specific interactions with pore walls. Here, inspired by the olfactory sensilla of insect antennae, we show that coating nanopores with a fluid lipid bilayer tailors their surface chemistry and allows fine-tuning and dynamic variation of pore diameters in subnanometre increments. Incorporation of mobile ligands in the lipid bilayer conferred specificity and slowed the translocation of targeted proteins sufficiently to time-resolve translocation events of individual proteins. Lipid coatings also prevented pores from clogging, eliminated non-specific binding and enabled the translocation of amyloid-beta (Aβ) oligomers and fibrils. Through combined analysis of their translocation time, volume, charge, shape and ligand affinity, different proteins were identified. Characterization of Individual Polynucleotide Molecules Using a Membrane Channel Dec 1996 John J. Kasianowicz <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Eric Brandin <here is a image 19e6b9ca083d83cf-3fe396e6f6f36a7e> Daniel Branton <here is a image 4ac7b1560f34ace5-4de82df975d17ddb> David Deamer We show that an electric field can drive single-stranded RNA and DNA molecules through a 2.6-nm diameter ion channel in a lipid bilayer membrane. Because the channel diameter can accommodate only a single strand of RNA or DNA, each polymer traverses the membrane as an extended chain that partially blocks the channel. The passage of each molecule is detected as a transient decrease of ionic current whose duration is proportional to polymer length. Channel blockades can therefore be used to measure polynucleotide length. With further improvements, the method could in principle provide direct, high-speed detection of the sequence of bases in single molecules of DNA or RNA. Shape bistability of a membrane neck: A toggle switch to control vesicle content release Full-text available Aug 2003 <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Vadim A. Frolov <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Vladimir Lizunov <here is a image 4b91d04e118e803f-8cf3fec7f5c27196> Antonina Dunina-Barkovskaya <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Joshua Zimmerberg Shape dynamics and permeability of a membrane neck connecting a vesicle and plasma membrane are considered. The neck is modeled by a lipid membrane tubule extended between two parallel axisymmetric rings. Within a range of lengths, defined by system geometry and mechanical properties of the membrane, the tubule has two stable shapes: catenoidal microtubule and cylindrical nanotubule. The permeabilities of these two shapes, measured as ionic conductivity of the tubule interior, differ by up to four orders of magnitude. Near the critical length the transitions between the shapes occur within less than a millisecond. Theoretical estimates show that the shape switching is controlled by a single parameter, the tubule length. Thus the tubule connection can operate as a conductivity microswitch, toggling the release of vesicle content in such cellular processes as "kiss-and-run" exocytosis. In support of this notion, bistable behavior of membrane connections between vesicles and the cell plasma membrane in macrophages is demonstrated. Noise in Solid-State Nanopores Full-text available Jan 2008 P NATL ACAD SCI USA Ralph M M Smeets Ulrich F. Keyser <here is a image 311294d4e542d7ee-191ee4eddd87fca5> Nynke Dekker Cees Dekker We study ionic current fluctuations in solid-state nanopores over a wide frequency range and present a complete description of the noise characteristics. At low frequencies (f approximately < 100 Hz) we observe 1/f-type of noise. We analyze this low-frequency noise at different salt concentrations and find that the noise power remarkably scales linearly with the inverse number of charge carriers, in agreement with Hooge's relation. We find a Hooge parameter alpha = (1.1 +/- 0.1) x 10(-4). In the high-frequency regime (f approximately > 1 kHz), we can model the increase in current power spectral density with frequency through a calculation of the Johnson noise. Finally, we use these results to compute the signal-to-noise ratio for DNA translocation for different salt concentrations and nanopore diameters, yielding the parameters for optimal detection efficiency. The application of single molecule nanopore sensing for quantitative analysis May 2022 <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Yanfang Wu J. Justin Gooding Nanopore-based sensors typically work by monitoring transient pulses in conductance via current-time traces as molecules translocate through the nanopore. The unique property of being able to monitor single molecules gives nanopore sensors the potential as quantitative sensors based on the counting of single molecules. This review provides an overview of the concepts and fabrication of nanopore sensors as well as nanopore sensing with a view toward using nanopore sensors for quantitative analysis. We first introduce the classification of nanopores and highlight their applications in molecular identification with some pioneering studies. The review then shifts focus to recent strategies to extend nanopore sensors to devices that can rapidly and accurately quantify the amount of an analyte of interest. Finally, future prospects are provided and briefly discussed. The aim of this review is to aid in understanding recent advances, challenges, and prospects for nanopore sensors for quantitative analysis. Molecular Shape Solution for Mesoscopic Remodeling of Cellular Membranes May 2022 Javier Vera Lillo <here is a image 1144e1be9de06691-b3c79ed0cfa37a80> Pavel Bashkirov Peter I. Kuzmin <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Vadim A. Frolov Cellular membranes self-assemble from and interact with various molecular species. Each molecule locally shapes the lipid bilayer, the soft elastic core of cellular membranes. The dynamic architecture of intracellular membrane systems is based on elastic transformations and lateral redistribution of these elementary shapes, driven by chemical and curvature stress gradients. The minimization of the total elastic stress by such redistribution composes the most basic, primordial mechanism of membrane curvature-composition coupling (CCC). Although CCC is generally considered in the context of dynamic compositional heterogeneity of cellular membrane systems, in this article we discuss a broader involvement of CCC in controlling membrane deformations. We focus specifically on the mesoscale membrane transformations in open, reservoir-governed systems, such as membrane budding, tubulation, and the emergence of highly curved sites of membrane fusion and fission. We reveal that the reshuffling of molecular shapes constitutes an independent deformation mode with complex rheological properties. This mode controls effective elasticity of local deformations as well as stationary elastic stress, thus emerging as a major regulator of intracellular membrane remodeling. Expected final online publication date for the Annual Review of Biophysics, Volume 51 is May 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates. Three-Dimensional Packing Defects in Lipid Membrane as a Function of Membrane Order Dec 2020 <here is a image fb9930a25b8ca568-0fb68db0b8a59752> Madhusmita Tripathy Subasini Thangamani <here is a image 0fc1b64576af4a5a-91393e7a48ae0bbc> Anand Srivastava Lipid membrane packing defects are considered to be an essential parameter that regulates specific membrane binding of several peripheral proteins. In the absence of direct experimental characterization, lipid packing defects and their role in the binding of peripheral proteins are generally investigated through computational studies, which have been immensely successful in unraveling the key steps of the membrane-binding process. However, packing defects are calculated using two-dimensional (2D) projections and the crucial information on their depths is generally overlooked. Here, we present a simple yet computationally efficient algorithm, which identifies these defects in three dimensions. We validate the algorithm on a number of model membrane systems that are previously studied using 2D defect calculations and find that the defect size and the defect depth may not always be directly correlated. We employ the algorithm to understand the nature of packing defects in flat bilayer membranes exhibiting liquid-ordered (Lo), liquid-disordered (Ld), and co-existing (Lo/Ld) phases. Our results indicate the presence of shallower, smaller, and spatially localized defects in the Lo phase membranes as compared to the defects in Ld and mixed Lo/Ld phase membranes. Such analyses can elucidate the molecular-scale mechanisms that drive the preferential localization of certain proteins to either of the liquid phases or their interface. We also analyze the membrane sensing and anchoring process of a peptide in terms of the three-dimensional defects, which provides additional insights into the process with respect to depth distributions across the bilayer leaflets. Reconstitution and real-time quantification of membrane remodeling by single proteins and protein complexes Jun 2020 <here is a image 1144e1be9de06691-b3c79ed0cfa37a80> Pavel Bashkirov Peter I. Kuzmin Ksenia Chekashkina <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Vadim A. Frolov Cellular membrane processes, from signal transduction to membrane fusion and fission, depend on acute membrane deformations produced by small and short-lived protein complexes working in conditions far from equilibrium. Real-time monitoring and quantitative assessment of such deformations are challenging; hence, mechanistic analyses of the protein action are commonly based on ensemble averaging, which masks important mechanistic details of the action. In this protocol, we describe how to reconstruct and quantify membrane remodeling by individual proteins and small protein complexes in vitro, using an ultra-short (80- to 400-nm) lipid nanotube (usNT) template. We use the luminal conductance of the usNT as the real-time reporter of the protein interaction(s) with the usNT. We explain how to make and calibrate the usNT template to achieve subnanometer precision in the geometrical assessment of the molecular footprints on the nanotube membrane. We next demonstrate how membrane deformations driven by purified proteins implicated in cellular membrane remodeling can be analyzed at a single-molecule level. The preparation of one usNT takes ~1 h, and the shortest procedure yielding the basic geometrical parameters of a small protein complex takes 10 h. Electrophysiological Methods for Detection of Membrane and Hemifission by Dynamin 1 Chapter Jun 2020 Ksenia Chekashkina <here is a image 6eaf68591ede8cea-8cfb257e34f4a104> Anna Shnyrova <here is a image 1144e1be9de06691-b3c79ed0cfa37a80> Pavel Bashkirov <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Vadim A. Frolov Membrane fusion and fission are indispensable parts of intracellular membrane recycling and transport. Electrophysiological techniques have been instrumental in discovering and studying fusion and fission pores, the key intermediates shared by both processes. In cells, electrical admittance measurements are used to assess in real time the dynamics of the pore conductance, reflecting the nanoscale transformations of the pore, simultaneously with membrane leakage. Here, we described how this technique is adapted to in vitro mechanistic analyses of membrane fission by dynamin 1 (Dyn1), the protein orchestrating membrane fission in endocytosis. We reconstitute the fission reaction using purified Dyn1 and biomimetic lipid membrane nanotubes of defined geometry. We provide a comprehensive protocol describing simultaneous measurements of the ionic conductance through the nanotube lumen and across the nanotube wall, enabling spatiotemporal correlation between the nanotube constriction by Dyn1, leading to fission and membrane leakage. We present examples of “leaky” and “tight” fission reactions, specify the resolution limits of our method, and discuss how our results support the hemi-fission conjecture. Origin of Nonequilibrium 1/f Noise in Solid-State Nanopores Mar 2020 Shihao Su <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Xun Guo Yanjun Fu Jianming Xue Nanopore devices are applied in many fields like molecule sensing and DNA sequencing, and the detection precision is primarily determined by the 1/f noise. The mechanism of 1/f noise in nanopores is still not clearly understood, especially the nonequilibrium 1/f noise in rectifying nanopores. Hereby, we propose that the 1/f noise in solid-state nanopores is originated from the electrolyte ions trapping-detrapping process on the inner surface of the nanopores, which can nonlinearly affect the ion number inside the rectifying nanopore due to the specific ion enrichment/depletion effect. Our model can not only quantitatively explain the nonlinear dependence of 1/f noise on the applied voltage, i.e., the nonequilibrium 1/f noise, for current rectifying nanopores, but also give a unified explanation on the influences of the electrolyte concentration, pH value, and the geometry of the nanopore. According to the model, we find a new flattening phenomenon of1/f noise in conical nanopores, and it is further confirmed by our experimental results. Our research can be helpful to understand and reduce 1/f noise in other nanopore devices, especially where the enrichment or depletion of ion exists. Estimation of Shape, Volume, and Dipole Moment of Individual Proteins Freely Transiting a Synthetic Nanopore Apr 2019 Jared Houghtaling <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Cuifeng Ying <here is a image a741c6735d96ae60-12749d4310394c92> Olivia Eggenberger <here is a image b4205e4d65cef25b-f9dbdb6ae6ceeef8> Michael Mayer This paper demonstrates that high-bandwidth current recordings in combination with low-noise silicon nitride nanopores make it possible to determine the molecular volume, approximate shape, and dipole moment of single native proteins in solution without the need for labeling or other chemical modifications of these proteins. The analysis is based on current modulations caused by the translation and rotation of single proteins through a uniform electric field inside of a nanopore. We applied this technique to nine proteins and show that these measured protein parameters agree well with reference values, but only if the nanopore walls were coated with a non-stick fluid lipid bilayer. One potential challenge with this approach is that an untethered protein is able to diffuse laterally while transiting a nanopore, which generates increasingly asymmetric disruptions in the electric field as it approaches the nanopore walls. These off-axis effects add an additional noise-like element to the electrical recordings, which can be exacerbated by non-specific interactions with pore walls that are not coated by a fluid lipid bilayer. We performed finite element simulations to quantify the influence of these effects on subsequent analyses. Examining the size, approximate shape, and dipole moment of unperturbed, native proteins in aqueous solution on a single-molecule level in real time while they translocate through a nanopore may enable applications such as monitoring the assembly or disassembly of transient protein complexes based on changes in their shape, volume, or dipole moment. Wafer scale fabrication of fused silica chips for low-noise recording of resistive pulses through nanopores Mar 2019 <here is a image 9fb55181bd5fa86b-f5f47520198fa9a7> Lennart de Vreede <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Cuifeng Ying Jared Houghtaling <here is a image b4205e4d65cef25b-f9dbdb6ae6ceeef8> Michael Mayer This paper presents a maskless method to manufacture fused silica chips for low-noise resistive-pulse sensing. The fabrication includes wafer-scale density modification of fused silica with a femtosecond-pulsed laser, low-pressure chemical vapor deposition (LPVCD) of silicon nitride (SiN x ) and accelerated chemical wet etching of the laser-exposed regions. This procedure leads to a freestanding SiN x window, which is permanently attached to a fused silica support chip and the resulting chips are robust towards Piranha cleaning at ∼80 °C. After parallel chip manufacturing, we created a single nanopore in each chip by focused helium-ion beam or by controlled breakdown. Compared to silicon chips, the resulting fused silica nanopore chips resulted in a four-fold improvement of both the signal-to-noise ratio and the capture rate for signals from the translocation of IgG1 proteins at a recording bandwidth of 50 kHz. At a bandwidth of ∼1 MHz, the noise from the fused silica nanopore chips was three- to six-fold reduced compared to silicon chips. In contrast to silicon chips, fused silica chips showed no laser-induced current noise - a significant benefit for experiments that strive to combine nanopore-based electrical and optical measurements. Formation of Single Nanopores with Diameters of 20-50 Nanometers in Silicon Nitride Membranes Using Laser-Assisted Controlled Breakdown Oct 2018 Cuifeng Ying Jared Houghtaling <here is a image a741c6735d96ae60-12749d4310394c92> Olivia Eggenberger <here is a image b4205e4d65cef25b-f9dbdb6ae6ceeef8> Michael Mayer Nanopores with diameters from 20-50 nm in silicon nitride (SiNx) windows are useful for single-molecule studies of globular macromolecules. While controlled breakdown (CBD) is gaining popularity as a method for fabricating nanopores with reproducible size control and broad accessibility, attempts to fabricate large nanopores with diameters exceeding ~20 nm via breakdown often result in undesirable formation of multiple nanopores in SiNx membranes. To reduce the probability of producing multiple pores, we combined two strategies: laser-assisted breakdown and controlled pore enlargement by limiting the applied voltage. We suggest that laser-induced heating increases the rate of controlled breakdown because of differences in nanopore conductance measured in electrolyte solutions of high or low ionic strengths and in the presence or absence of laser illumination. Moreover, we demonstrate that conductance values before and after coating the nanopores with a lipid bilayer may indicate fabrication of a single nanopore versus multiple nanopores. Complementary flux measurements of Ca2+ through the nanopore typically confirmed assessments of single or multiple nanopores that we obtained using the coating method. Finally, we show that thermal annealing of CBD pores significantly increased the success rate of coating and reduced the current noise before and after lipid coating. We characterize the geometry of these nanopores by analyzing the individual resistive pulses produced by translocations of spherical proteins and demonstrate the usefulness of these nanopores for estimating the approximate molecular shape of IgG proteins. Recent Progress in Solid‐State Nanopores Sep 2018 <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Kidan Lee Hyung-Jun Kim Kyeong-Beom Park Ki-Bum Kim The solid‐state nanopore has attracted much attention as a next‐generation DNA sequencing tool or a single‐molecule biosensor platform with its high sensitivity of biomolecule detection. The platform has advantages of processability, robustness of the device, and flexibility in the nanopore dimensions as compared with the protein nanopore, but with the limitation of insufficient spatial and temporal resolution to be utilized in DNA sequencing. Here, the fundamental principles of the solid‐state nanopore are summarized to illustrate the novelty of the device, and improvements in the performance of the platform in terms of device fabrication are explained. The efforts to reduce the electrical noise of solid‐state nanopore devices, and thus to enhance the sensitivity of detection, are presented along with detailed descriptions of the noise properties of the solid‐state nanopore. Applications of 2D materials including graphene, h‐BN, and MoS2 as a nanopore membrane to enhance the spatial resolution of nanopore detection, and organic coatings on the nanopore membranes for the addition of chemical functionality to the nanopore are summarized. Finally, the recently reported applications of the solid‐state nanopore are categorized and described according to the target biomolecules: DNA‐bound proteins, modified DNA structures, proteins, and protein oligomers. The solid‐state nanopore has attracted much attention as a next‐generation DNA sequencing tool or a single‐molecule biosensor platform with its high sensitivity of biomolecule detection. The history, fundamental principles, improvements in the performance, and the recently reported applications of the solid‐state nanopore are highlighted, with a focus on devices, materials, and fabrication methods. An impedance model for the low-frequency noise originating from the dynamic hydrogen ion reactivity at the solid/liquid interface Jul 2017 SENSOR ACTUAT B-CHEM <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Da Zhang <here is a image c12e60df6d03e126-c376d15f910a571d> Paul Solomon <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Zhen Zhang <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Shi-Li Zhang Understanding the dynamics of hydrogen ion reactivity at the solid/liquid interface is of paramount importance for applications involving ion sensing in electrolytes. However, the correlation of this interfacial process to noise generation is poorly characterized. Here, the relationship is unveiled by characterizing the interfacial process with impedance spectroscopy assisted by a dedicated electrochemical impedance model. The model incorporates both thermodynamic and kinetic properties of the amphoteric hydrogen ion site-binding reactions with the surface OH groups. It further takes into consideration the distributed nature of the characteristic energy of the binding sites. The simulated impedance matches the experimental data better with an energy distribution of the kinetic parameters than with that of the thermodynamic ones. Since the potentiometric low-frequency noise (LFN) originating from the solid/liquid interface correlates excellently with the real part of its electrochemical impedance spectrum, this work establishes a method for evaluating sensing surface quality aimed at mitigating LFN. Generalized Noise Study of Solid-State Nanopores at Low Frequencies <here is a image f936cef17699d142-ec9e0b1807df9062> Chenyu Wen <here is a image b88af5c6bec4068d-b4241c4411949a9c> Shuangshuang Zeng <here is a image 36f8cb8130eed485-63a7c808851aa44a> Kai Arstila <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Shi-Li Zhang The nanopore technology has been extensively investigated for analysis of biomolecules, and a success story in this field concerns DNA sequencing using a nanopore chip featuring an array of hundreds of biological nanopores (BioNs). Solid-state nanopores (SSNs) have been explored to attain longer lifetime and higher integration density than what BioNs can offer, but SSNs are generally considered to generate higher noise whose origin remains to be confirmed. Here, we systematically study low-frequency (including thermal and flicker) noise characteristics of SSNs measuring 7 to 200 nm in diameter drilled through a 20-nm thick SiNx membrane by focused ion milling. Both bulk and surface ionic currents in the nanopore are found to contribute to the flicker noise, with their respective contributions determined by salt concentration and pH value in electrolytes as well as bias conditions. Increasing salt concentration at constant pH and voltage bias leads to increase in the bulk ionic current and noise therefrom. Changing pH at constant salt concentration and current bias results in variation of surface charge density, and hence alteration of surface ionic current and noise. In addition, the noise from Ag/AgCl electrodes can become predominant when the pore size is large and/or the salt concentration is high. Analysis of our comprehensive experimental results leads to the establishment of a generalized nanopore noise model. The model not only gives an excellent account of the experimental observations, but can also be used for evaluation of various noise components in much smaller nanopores currently not experimentally available. Noise and its reduction in graphene based nanopore devices NANOTECHNOLOGY <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Ashvani Kumar Kyeong-Beom Park Hyun-Mi Kim Ki-Bum Kim Ionic current fluctuations in graphene nanopore devices are a ubiquitous phenomenon and are responsible for degraded spatial and temporal resolution. Here, we descriptively investigate the impact of different substrate materials (Si and quartz) and membrane thicknesses on noise characteristics of graphene nanopore devices. To mitigate the membrane fluctuations and pin-hole defects, a SiNx membrane is transferred onto the substrate and a pore of approximately 70 nm in diameter is perforated prior to the graphene transfer. Comprehensive noise study reveals that the few layer graphene transferred onto the quartz substrate possesses low noise level and higher signal to noise ratio as compared to single layer graphene, without deteriorating the spatial resolution. The findings here point to improvement of graphene based nanopore devices for exciting opportunities in future single-molecule genomic screening devices. Nontrivial Behavior of Water in the Vicinity and Inside Lipid Bilayers As Probed by Molecular Dynamics Simulations Sep 2013 ACS NANO <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Nikolay A Krylov Vladimir Pentkovsky <here is a image 71064a0661fafc4a-747231d9a912e996> Roman G Efremov Atomic-scale diffusion of water in the presence of several lipid bilayers mimicking biomembranes is characterized via unconstrained molecular dynamics (MD) simulations. Although overall water dynamics corresponds well to literature data - efficient braking near polar head groups of lipids - a number of interesting and biologically relevant details observed in this work have not been sufficiently discussed so far. Thus, waters "sense" the membrane unexpectedly early - before water density begins to decrease. In this "transitional zone" velocity distributions of water and their H-bonding patterns deviate from those in the bulk solution. Boundaries of this zone are well preserved even despite local (<1 nm size) perturbation of lipid bilayer, thus indicating decoupling of the surface and bulk dynamics of water. This is in excellent agreement with recent experimental data. Near the membrane surface water movement becomes anisotropic - solvent molecules preferentially move outward the bilayer. Deep in the membrane interior, the velocities can even exceed those in the bulk solvent and undergo large-scale fluctuations. Analysis of MD trajectories of individual waters in the middle part of acyl chain region of lipids reveal a number of interesting rare phenomena, like fast (c.a. 50 ps) breakthrough across the membrane or long-time (up to 750 ps) "roaming" between lipid leaflets. Analysis of these events was accomplished to delineate the mechanisms of spontaneous water permeation inside the hydrophobic membrane core. It was shown that such a nontrivial dynamics of water in the "alien" environment is driven by dynamic heterogeneities of the local bilayer structure and formation of transient atomic-scale "defects" in it. The picture observed in lipid bilayers is drastically different from that in a primitive membrane mimic - hydrated cyclohexane slab. Possible biological impact of such phenomena in equilibrated lipid bilayers is discussed. Nanopore-Spanning Lipid Bilayers on Silicon Nitride Membranes That Seal and Selectively Transport Ions Mar 2013 LANGMUIR <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Christopher E Korman <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Mischa Megens <here is a image ba943aa7b14d5c71-4ba1c17e8a549db9> Caroline M Ajo-Franklin <here is a image a656c51457c6b05f-db1108ff94ea6a75> David Horsley We report the formation of POPC lipid bilayers that span 130 nm pores in a freestanding silicon nitride film supported on a silicon substrate. These solvent-free lipid membranes self-assemble on organosilane-treated Si3N4 via the fusion of 200 nm unilamellar vesicles. Membrane fluidity is verified by fluorescence recovery after photobleaching (FRAP), and membrane resistance in excess of 1 GΩ is demonstrated using electrical impedance spectroscopy (EIS). An array of 40 000 membranes maintained high impedance over 72 h, followed by rupture of most of the membranes by 82 h. Membrane incorporation of gramicidin, a model ion channel, resulted in increased membrane conductance. This membrane conductance was diminished when the gramicidin channels were blocked with CaCl2, indicating that the change in membrane conductance results from gramicidin-mediated ion transport. These very stable, biologically functional pore-spanning membranes open many possibilities for silicon-based ion-channel devices for applications such as biosensors and high-throughput drug screening. Variation of Lipid Membrane Composition Caused by Strong Bending Jun 2011 <here is a image 380416004276da08-89c6810f91fec6ee> K. V. Chekashkina S. A. Akimov <here is a image 1144e1be9de06691-b3c79ed0cfa37a80> Pavel Bashkirov <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Vadim A. Frolov Dynamic coupling between the morphology and molecular composition of cellular membranes is crucial for formation of the intracellular organelles and transport vesicles. Most of the membrane proteins and lipids discriminate membrane curvatures. However, it remains unclear whether the curvature alone is sufficient to support heterogeneous distribution of lipids. Here we demonstrate that the curvature-driven redistribution of phospholipids, such as dioleoylphosphatidylethanolamine (DOPE), requires strong membrane bending. We used cylindrical lipid nanotubes (NTs) pulled from planar lipid membranes with lateral tension of ∼1 dyn/cm. Such high tensions forced extreme curvatures of the NT membrane, with luminal radius approaching the thickness of the lipid bilayer, 5nm. When the NT contained lipid species with high spontaneous curvature (SC), such as DOPE, we observed slow reduction of its radius. This reduction indicated the redistribution of DOPE between the inner and outer monolayers of the NT. Accordingly, the SC of DOPE was recovered from the measured changes in the radii: the SC value, calculated under the assumption that the DOPE content is coupled to the monolayer curvature, was ∼0.4 nm−1, consistent with the published data. Thus, redistribution of lipids should be taken into account in calculations of composition and material properties of strongly deformed membrane structures, such as intermediate structures arising in the processes of membrane fusion and fission. Keywordslipid membrane–splay–nanotube–lipid redistribution Fission of membrane nanotube induced by osmotic pressure <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Alexey Evseev <here is a image 1144e1be9de06691-b3c79ed0cfa37a80> Pavel Bashkirov The final stage of endocytosis is fission of a thin membrane neck, or nanotube (NT), connecting cell membrane with a forming vesicle. We studied this process using a model system consisting of NT pulled out from a flat bilayer lipid membrane. Fission of NT was induced by an increase of osmotic pressure created by local application of a concentrated salt solution in the vicinity of NT. Superfusion of NT with distilled water instead of the concentrated salt solution led to the NT expansion. This observation demonstrates the reversibility of the NT expansion-compression process under the osmotic pressure. The overall picture of fission is similar to that described earlier for the NT fission with participation of dynamin GTPase. In both cases, in order for fission to occur, it is necessary to compress the NT to a critical radius. The critical radius estimated for the osmotic pressure-induced fission exceeds the value obtained for the fission occurring in the presence of the protein. Fission under osmotic pressure, akin the dynamin-promoted fission, proceeds without leaky defects. Solid-State Nanopore Sensors for Nucleic Acid Analysis Sep 2011 Nat. Nanotech. Bala Murali Venkatesan Rashid Bashir Nanopore analysis is an emerging technique that involves using a voltage to drive molecules through a nanoscale pore in a membrane between two electrolytes, and monitoring how the ionic current through the nanopore changes as single molecules pass through it. This approach allows charged polymers (including single-stranded DNA, double-stranded DNA and RNA) to be analysed with subnanometre resolution and without the need for labels or amplification. Recent advances suggest that nanopore-based sensors could be competitive with other third-generation DNA sequencing technologies, and may be able to rapidly and reliably sequence the human genome for under $1,000. In this article we review the use of nanopore technology in DNA sequencing, genetics and medical diagnostics. Noise and Bandwidth of Current Recordings from Submicrometer Pores and Nanopores Jun 2008 ACS NANO Jeffrey D Uram Kevin Ke <here is a image b4205e4d65cef25b-f9dbdb6ae6ceeef8> Michael Mayer Nanopores and submicrometer pores have recently been explored for applications ranging from detection of single molecules, assemblies of nanoparticles, nucleic acids, occurrence of chemical reactions, and unfolding of proteins. Most of these applications rely on monitoring electrical current through these pores, hence the noise and signal bandwidth of these current recordings are critical for achieving accurate and sensitive measurements. In this report, we present a detailed theoretical and experimental study on the noise and signal bandwidth of current recordings from glass and polyethylene terephthalate (PET) membranes that contain a single submicrometer pore or nanopore. We examined the theoretical signal bandwidth of two different pore geometries, and we measured the signal bandwidth of the electronics used to record the ionic current. We also investigated the theoretical noise generated by the substrate material, the pore, and the electronics used to record the current. Employing a combination of theory and experimental results, we were able to predict the noise in current traces recorded from glass and PET pores with no applied voltage with an error of less than 12% in a range of signal bandwidths from 1 to 40 kHz. In approximately half of all experiments, application of a voltage did not significantly increase the noise. In the other half of experiments, however, application of a voltage resulted in an additional source of noise. For these pores, predictions of the noise were usually still accurate within 35% error at signal bandwidths of at least 10 kHz. The power spectra of this extra noise suggested a 1/f(alpha) origin with best fits to the power spectrum for alpha = 0.4-0.8. This work provides the theoretical background and experimental data for understanding the bandwidth requirements and the main sources of noise in current recordings; it will be useful for minimizing noise and achieving accurate recordings. Action Potential Phenomena in Experimental Bimolecular Lipid Membranes P. Mueller Donald O. Rudin A MOLECULAR mechanism which develops multivibrator kinetics such as those shown in Figs. 1 and 2 has been constructed in experimental bimolecular lipid membranes by the methods given here. These electrokinetic phenomena are indistinguishable from cellular action potentials and include thresholds, refractoriness, delayed rectification, bistable flip-flop kinetics, anodal break excitation, self-regenerative changes of membrane potential accompanied by decreases in conductance, and spontaneous rhythmic firing, the frequency of which can be precisely controlled by varying the temperature and the applied electrical polarization. They are also reversibly blocked by local anaesthetics such as cocaine at physiological concentrations (2 per cent) and are modified in various ways by acridine and phenothiazine derivatives including chlorpromazine at concentrations of about 1 gamma/c.c. Bending energy of vesicle membranes: General expressions for the first, second, and third variation of the shape energy and applications to spheres and cylinders Jun 1989 <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Zhang-Can Ou-Yang W. Helfrich

https://www.researchgate.net/publication/366160571_Noise_in_Ultrashort_Elastic_Membrane_Nanotube

Buprenorphine Disrupts Sleep and Decreases Adenosine Concentrations in Sleep-regulating Brain Regions of Sprague Dawley Rat | Anesthesiology | American Society of Anesthesiologists Buprenorphine Disrupts Sleep and Decreases Adenosine Concentrations in Sleep-regulating Brain Regions of Sprague Dawley Rat ; Elizabeth A. Gauthier, B.S. * Medical Student. Sarah E. Guzick, B.S. Sarah E. Guzick, B.S. † Graduate Student. Chad M. Brummett, M.D. Chad M. Brummett, M.D. ‡ Assistant Professor. Helen A. Baghdoyan, Ph.D. ; Helen A. Baghdoyan, Ph.D. § Professor, Department of Anesthesiology, University of Michigan, Ann Arbor, Michigan. Ralph Lydic, Ph.D. Ralph Lydic, Ph.D. § Professor, Department of Anesthesiology, University of Michigan, Ann Arbor, Michigan. Anesthesiology October 2011, Vol. 115, 743–753. https://doi.org/10.1097/ALN.0b013e31822e9f85 Background Buprenorphine, a partial μ-opioid receptor agonist and κ-opioid receptor antagonist, is an effective analgesic. The effects of buprenorphine on sleep have not been well characterized. This study tested the hypothesis that an antinociceptive dose of buprenorphine decreases sleep and decreases adenosine concentrations in regions of the basal forebrain and pontine brainstem that regulate sleep. Methods Male Sprague Dawley rats were implanted with intravenous catheters and electrodes for recording states of wakefulness and sleep. Buprenorphine (1 mg/kg) was administered systemically via an indwelling catheter and sleep-wake states were recorded for 24 h. In additional rats, buprenorphine was delivered by microdialysis to the pontine reticular formation and substantia innominata of the basal forebrain while adenosine was simultaneously measured. Results An antinociceptive dose of buprenorphine caused a significant increase in wakefulness (25.2%) and a decrease in nonrapid eye movement sleep (-22.1%) and rapid eye movement sleep (-3.1%). Buprenorphine also increased electroencephalographic delta power during nonrapid eye movement sleep. Coadministration of the sedative-hypnotic eszopiclone diminished the buprenorphine-induced decrease in sleep. Dialysis delivery of buprenorphine significantly decreased adenosine concentrations in the pontine reticular formation (-14.6%) and substantia innominata (-36.7%). Intravenous administration of buprenorphine significantly decreased (-20%) adenosine in the substantia innominata. Conclusions Buprenorphine significantly increased time spent awake, decreased nonrapid eye movement sleep, and increased latency to sleep onset. These disruptions in sleep architecture were mitigated by coadministration of the nonbenzodiazepine sedative-hypnotic eszopiclone. The buprenorphine-induced decrease in adenosine concentrations in basal forebrain and pontine reticular formation is consistent with the interpretation that decreasing adenosine in sleep-regulating brain regions is one mechanism by which opioids disrupt sleep. Topics: adenosine , buprenorphine , sleep , brain What We Already Know about This Topic Opiate-induced sleep disruption can contribute to hyperalgesia Endogenous adenosine promotes sleep and decreases nociception Antinociceptive doses of buprenorphine disrupted normal sleep architecture in rats, an effect that was attenuated by the sedative-hypnotic eszopiclone Buprenorphine also decreased adenosine levels in brain regions known to modulate sleep and nociception OPIOIDS are used effectively in the treatment of chronic and acute pain, and the extensive use of opioids encourages efforts to develop countermeasures to combat unwanted side effects.1,2Opioids disrupt sleep,3,–,7and sleep disruption can contribute to hyperalgesia,8,–,16impaired immune function,17and postoperative cognitive impairment.18,19 Adenosine is an endogenous neuromodulator that significantly enhances sleep20and diminishes nociception.21Sleep is increased by increasing adenosine in the pontine reticular formation (PnO)22,–,24and in the substantia innominata (SI) area of the basal forebrain.20,25Adenosine concentrations in the PnO and SI are decreased by the μ-opioid receptor agonists morphine and fentanyl.26 Buprenorphine, a partial μ-opioid receptor agonist and κ-opioid receptor antagonist, is an effective analgesic, but no previous studies have quantified the effects of buprenorphine on sleep architecture7,27,28or on adenosine concentrations in the PnO and SI. This study was designed to test the hypothesis that buprenorphine decreases sleep and adenosine concentrations in PnO and SI, brain regions known to modulate sleep and nociception. Materials and Methods Animals Adult, male Crl:CD(SD) (Sprague Dawley) rats (n = 26) purchased from Charles River Laboratories (Wilmington, MA) were used for all studies. Rats weighing 250–350 g were used because brains from rats in this weight range are known to fit the rat stereotaxic atlas.29Male rats were chosen to facilitate comparison of the current results to previous data obtained from males.26,30,–,32Rats were housed in a 12:12-h light–dark cycle (lights on from 8:00 to 20:00) with access to food and water ad libitum. Procedures were reviewed and approved by the University of Michigan Committee on the Use and Care of Animals. Every phase of this study adhered to the Guide for the Care and Use of Laboratory Animals(Eighth Edition, National Academy of Sciences Press, Washington DC, 2011).‖ Surgical Procedures Rats were anesthetized with 3% isoflurane (Hospira, Inc., Lake Forest, IL). The jugular vein was exposed, and a catheter (12 cm of Micro-Renathane tubing [MRE–040], Braintree Scientific, Braintree, MA) was inserted in the direction of the heart. The other end of the catheter was tunneled subcutaneously and implanted between the scapulas. A back-mounted flange guide cannula (8I 1000BM10; Plastics One, Roanoke, VA) and dummy cannula (8IC313DCCACC; Plastics One) were secured with the catheter in the midscapular position. This procedure provided subsequent venous access. Implantation of the jugular vein catheter was followed immediately by implantation of electrodes for recording sleep. Rats were moved to a Kopf Model 962 small animal stereotaxic instrument fitted with a Model 906 rat anesthesia mask (David Kopf Instruments, Tujunga, CA), and anesthesia was maintained with isoflurane (2.0%). Three electrodes (8IE36320SPCE, Plastics One) for recording cortical electroencephalogram were placed 2.0 mm posterior and 1.3 mm lateral to bregma, 2.0 mm posterior and 1.5 mm lateral to bregma, and 1.0 mm anterior and 1.5 mm lateral to bregma.29Two electrodes (4 cm of AG 7/40T Medwire, Mt. Vernon, NY) for electromyogram recordings were placed in the dorsal neck muscle, and a third electrode was placed under the skin of the neck muscle as a reference. The nonimplanted ends of the electroencephalogram and electromyogram electrodes were soldered to electrical contact pins (E363/0; Plastics One) that were plugged into a plastic pedestal (8K00022980IF; Plastics One). Three stainless steel anchor screws (MPX-0080–02P-C; Small Parts Inc., Miami Lakes, FL) were placed in the skull to secure the electrodes. Dental acrylic was used to construct a head cap covering the electrodes and to anchor the electrical connector and electrodes to the skull. Rats were then removed from the stereotaxic frame and monitored during recovery from anesthesia. Once ambulatory, animals were returned to their home cages. Behavioral Conditioning for Sleep Recordings Rats were given 1 week for surgical recovery and then conditioned for an additional week to 10 days to sleeping in a Raturn (Bioanalytical Systems, West Lafayette, IN) recording chamber. During conditioning, the implanted electrodes were attached by a cable (363–441/six 80CM 6TCM; Plastics One) to amplifiers and a computer for digital recording of electroencephalogram and electromyogram signals. Rats had free access to food and water while in the recording chambers. Nociceptive Testing An initial series of experiments was conducted to confirm that the 1 mg/kg dose of buprenorphine produced antinociception as reported previously.33Procedures for thermal nociceptive testing have been described in detail.34,35Briefly, rats were conditioned to being placed in the plexiglass chamber of a Hargreaves Paw Withdrawal unit (Model 336T; IITC Life Science, Woodland Hills, CA) 1 h each day for the week before data collection. The Model 400 (IITC Life Science) heated glass stand and base was set to 30°C for the last 10 min of each conditioning session, and both hind paws of the rat were exposed to the heat stimulus.36Five baseline measurements were taken after the habituation time. As soon as baseline measurements were recorded, saline or buprenorphine hydrochloride (Sigma–Aldrich, Saint Louis, MO; 1 mg/kg) was administered viathe jugular vein catheter. The injection volume was 1 ml. Measures of paw withdrawal latency (PWL) were taken at 10, 20, 30, 60, 90, and 120 min after saline or buprenorphine administration. A cutoff time of 15 s was set to prevent tissue damage of the hind paw. Drug Administration and Recordings of Sleep–Wake States A second series of experiments was designed to quantify the effect of intravenously administered buprenorphine on states of sleep and wakefulness. Buprenorphine was dissolved in sterile saline (pH 5.8 ± 0.2) and administered intravenously in a 1-ml volume at a dose of 1 mg/kg. Saline injection provided a negative control condition. Recordings of sleep and wakefulness began at 08:00 at the initiation of the light phase of the light–dark cycle. Rats are nocturnal and light onset corresponds to the rat subjective night. To determine whether buprenorphine caused sleep disruption, as do other opioids,26,37this study was designed to deliver buprenorphine at light onset. Rats were placed in the recording chamber, and the electromyogram and electroencephalogram electrodes were attached viaswivel cable to the amplifiers and computer. All injections were administered during a 4-min interval. The data acquisition software was started when drug or vehicle administration began. The electroencephalogram signals were filtered between 0.3 and 30 Hz and amplified. Each rat (n = 7) received one injection of buprenorphine and one injection of saline separated by at least 1 week. The rats were then allowed to sleep and wake spontaneously for the remainder of the 24-h recording. At the end of the recording interval, rats were returned to the vivarium. Every 10 s of the 24-h recording was scored as wakefulness, nonrapid eye movement (NREM) sleep, or rapid eye movement (REM) sleep. All sleep recordings were also scored by a second individual who was blinded to the injection condition. There was 93% agreement between the two sleep scorers. A third series of experiments was designed to coadminister the nonbenzodiazepine sedative-hypnotic eszopiclone (Toronto Research Chemicals, Toronto, Canada) with buprenorphine to quantify the effect on sleep and wakefulness. Eszopiclone is a benzodiazepine receptor agonist with a nonbenzodiazepine structure, marketed as Lunesta® (Sunovion Pharmaceuticals, Marlborough, MA). Eszopiclone is the (S)-isomer of the cyclopyrrolone zopiclone and is indicated for the treatment of insomnia.38As discussed in detail elsewhere,26patients who experience pain often report poor sleep. Clinically used doses of opioids significantly disrupt sleep,37and disordered sleep exacerbates pain.26,39,40These data raise the question of whether enhancement of sleep by a sedative-hypnotic would have a beneficial effect of diminishing opioid-induced sleep disruption. If so, this would encourage future studies aiming to determine whether combining opioid and sedative-hypnotic therapy could diminish pain. Eszopiclone was dissolved in sterile saline and dimethyl sulfoxide (1%; pH 6.0 ± 0.2) and administered intravenously (3 mg/kg). Buprenorphine (1 mg/kg) was then delivered viathe same IV cannula. For these studies, rats (n = 4) received an injection of eszopiclone followed immediately by an injection of saline or buprenorphine. Measurement of Brain Adenosine Concentrations during Microdialysis Delivery of Buprenorphine A fourth set of experiments sought to identify brain regions through which buprenorphine decreased sleep. Normal cholinergic neurotransmission is essential for maintaining wakefulness, and opioids disrupt cholinergic neurotransmission in the SI region of the basal forebrain.30Adenosine is known to promote sleep, and previous studies have shown that adenosine concentrations in the PnO are decreased by fentanyl and morphine.26Both fentanyl and morphine cause sleep disruption. Therefore, the current experiments also used in vivomicrodialysis and high-performance liquid chromatography to measure adenosine concentrations in the PnO and SI during dialysis delivery of buprenorphine. Buprenorphine (100 μM) was prepared the morning of each experiment. The drug was dissolved in Ringer's solution (pH 5.8–6.2) comprised of 146 nM NaCl, 4.0 mM KCl, 2.4 mM CaCl 2, and 10 μM erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA; Sigma–Aldrich), which is an adenosine deaminase inhibitor. Each rat was placed in an induction chamber and anesthetized with 4% isoflurane (Hospira, Inc.) in 100% oxygen. After 5 min, the rat was moved from the chamber into a stereotaxic frame and fitted with a rat anesthesia mask, as described. The isoflurane concentration was reduced to 2.5%. A midline scalp incision was then made to expose lambda and bregma. A rotary tool (Dremel, Racine, WI) was used to make a small craniotomy, through which a dialysis probe was placed in the brain. A rat brain atlas29was used to position a CMA-11 microdialysis probe (Cuprophane membrane: 1 mm long, 0.24 mm in diameter, 6-kDa cutoff; CMA Microdialysis, North Chelmsford, MA) in the PnO or in the SI. Stereotaxic coordinates for the PnO were 8.4 mm posterior to bregma, 1.0 mm lateral to the midline, and 9.2 mm below bregma. The coordinates for the SI were 1.6 mm posterior to bregma, 2.5 mm lateral to the midline, and 8.7 mm below bregma. The delivered isoflurane concentration was held at 1.5% and measured continuously throughout the duration of the experiment. A water blanket and recirculating heat pump (Gaymar Industries, Orchard Park, NY) were used to maintain body temperature at 37°C throughout data collection and recovery. Histologic Localization of Dialysis Sites Four to 5 days after the microdialysis experiment, each rat was deeply anesthetized and decapitated. Brains were removed, cut into 40-μm thick coronal sections with a cryostat (Leica Microsystems, Nussloch, Germany), and slide-mounted serially. Slides with brain sections containing the SI and PnO were fixed in paraformaldehyde vapor (80°C) and stained with cresyl violet. All sections were then digitized using a Nikon Super Coolscan 4000 scanner (Tokyo, Japan). Coronal sections were compared with plates in a rat brain atlas,29and the dialysis sites were localized to either the PnO or the SI. Statistical Analysis Statistical programs used for data analysis included Prism 5 (Graph Pad Software, Inc., La Jolla, CA) and SAS v9.2 (SAS Institute Inc., Cary, NC). All data were tested to ensure they met the assumptions of the underlying statistical model. PWL was converted to percent maximum possible effect (% MPE) using the following equation: % MPE = (PWL experimental − PWL baseline)/(15 s − PWL baseline) × 100. Repeated measures, two-way analysis of variance (ANOVA) was used to analyze results for changes over time and changes caused by the drug, and Bonferroni post hoctests were used to detect differences at specific time points. Every 10 s of the 24-h sleep and wakefulness recording was scored as wakefulness, NREM sleep, or REM sleep. Dependent measures included percent of time spent in each state, latency to onset of the first episode of NREM sleep and REM sleep, number of episodes, average episode duration, and number of transitions between states. To avoid the problem of inflated degrees of freedom resulting from the large number of 10-s epochs analyzed, the sleep–wake data were averaged for each rat. Dependent measures of sleep and wakefulness were analyzed by repeated measures two-way ANOVA and paired ttests using Bonferroni correction. As described in detail elsewhere,24,43,44fast Fourier transform of the electroencephalogram was performed to determine whether the electroencephalogram was altered by buprenorphine. Electroencephalographic power was analyzed by repeated measures two-way ANOVA and post hoctests for comparison at every 0.5-Hz frequency band (wakefulness and REM sleep 5.0–10.0 Hz; NREM sleep 0.5–5.0 Hz). For each experiment, adenosine measures during dialysis with Ringer's solution (control) were compared with adenosine concentrations during dialysis delivery of buprenorphine. This design allowed each experiment to contribute one mean adenosine value derived from four control (Ringer's solution) samples and one mean adenosine value derived from four measures obtained during administration of buprenorphine. These values were then averaged across multiple experiments and analyzed individually for PnO and SI brain regions using paired ttests. A probability value of P< 0.05 was considered statistically significant. Results Buprenorphine was Antinociceptive Figure 1depicts % MPE for PWL as a function of time after IV administration of saline and buprenorphine. ANOVA revealed that buprenorphine caused significant ( P= 0.0072) antinociception. Bonferroni post hoccomparisons indicated that buprenorphine significantly ( P< 0.05) increased % MPE at 20, 30, 60, and 120 min after injection. This antinociceptive dose of buprenorphine was used for subsequent studies of sleep and wakefulness. Fig. 1. Buprenorphine increased paw withdrawal latency (PWL) after thermal stimulus. The ordinate shows PWL expressed as percent maximum possible effect (% MPE). The abscissa shows time course of % MPE relative to injection at time zero. Asterisks (*) indicate a significant increase in % MPE after intravenous administration of buprenorphine (1 mg/kg) compared with saline (vehicle control). Fig. 1. Buprenorphine increased paw withdrawal latency (PWL) after thermal stimulus. The ordinate shows PWL expressed as percent maximum possible effect (% MPE). The abscissa shows time course of % MPE relative to injection at time zero. Asterisks (*) indicate a significant increase in % MPE after intravenous administration of buprenorphine (1 mg/kg) compared with saline (vehicle control). Buprenorphine Altered the Temporal Organization of Sleep and Wakefulness Figure 2illustrates the temporal distribution of wakefulness, NREM sleep, and REM sleep for 24 h after IV administration of saline (control) and buprenorphine.Figure 3summarizes group data for the light phase (first 12 h after injection) showing buprenorphine-induced changes in the temporal organization of sleep and wakefulness. ANOVA indicated a significant ( P< 0.01) effect of buprenorphine on percent of time spent in states of wakefulness, NREM sleep, and REM sleep, as well as a significant ( P< 0.0001) drug-by-state interaction (fig. 3A). Paired ttests with Bonferroni correction showed that buprenorphine significantly ( P< 0.001) increased the percentage of time spent in waking (25.2%) and significantly decreased the amount of time spent in NREM sleep (−22.1%) and REM sleep (−3.1%). Buprenorphine significantly delayed the onset of NREM sleep and REM sleep (fig. 3C). Fig. 2. Representative 24-h plots from one rat illustrating the buprenorphine-induced disruption of sleep architecture. The ordinate plots the temporal distribution of wakefulness (Wake), nonrapid eye movement sleep (NREM), and rapid eye movement sleep (REM) for 24 h after intravenous administration of saline ( A) and 1 mg/kg buprenorphine ( B). The injections and recordings of sleep and wakefulness began at 08:00 (time 0 on ordinate) at the initiation of the light phase (white horizontal rectangle) of a 12:12 h light–dark cycle. During the 12-h light phase (abscissa 0–12), Aillustrates extended blocks of NREM sleep typically displayed by these nocturnal animals. In contrast, Bshows that during this same interval buprenorphine decreased NREM sleep and REM sleep. Fig. 3. Buprenorphine altered the temporal organization of sleep and wakefulness, and eszopiclone countered buprenorphine-induced sleep disruption. Panels A, C, E, G, and Isummarize the effect of buprenorphine relative to saline (control) on five dependent measures averaged for seven rats recorded during the light portion (rat sleep phase) of the light–dark cycle. Panels B, D, F, H, and Jsummarize the temporal changes in sleep and wakefulness after coadministration of buprenorphine and eszopiclone (n = 4). Eszopiclone counteracted most of the sleep disruption caused by buprenorphine. Asterisks (*) indicate significant differences compared with saline across states of wakefulness (Wake), nonrapid eye movement sleep (NREM), and rapid eye movement sleep (REM). Fig. 3. Buprenorphine altered the temporal organization of sleep and wakefulness, and eszopiclone countered buprenorphine-induced sleep disruption. Panels A, C, E, G, and Isummarize the effect of buprenorphine relative to saline (control) on five dependent measures averaged for seven rats recorded during the light portion (rat sleep phase) of the light–dark cycle. Panels B, D, F, H, and Jsummarize the temporal changes in sleep and wakefulness after coadministration of buprenorphine and eszopiclone (n = 4). Eszopiclone counteracted most of the sleep disruption caused by buprenorphine. Asterisks (*) indicate significant differences compared with saline across states of wakefulness (Wake), nonrapid eye movement sleep (NREM), and rapid eye movement sleep (REM). There was a significant ( P< 0.0001) drug main-effect and state-by-drug interaction ( P< 0.0001) for the number of sleep–wake episodes (fig. 3E). Buprenorphine decreased the number of episodes of wakefulness (−88.2%), NREM sleep (−89.5%), and REM sleep (−90.8%).Figure 3G shows that buprenorphine significantly ( P< 0.0001) altered the duration of sleep–wake episodes. The average duration of wakefulness was significantly increased (529.6%), and the duration of sleep epochs was decreased for both NREM sleep (−30.8%) and REM sleep (−87.5%).Figure 3I shows that buprenorphine also significantly ( P< 0.0001) decreased the number of transitions (−89.8%) between states. Figure 4plots the percentage state for each drug condition during the 12-h dark phase (rat subjective day) of the light–dark cycle that followed the 12-h light phase depicted byfigure 3. Within the dark phase, when rats are normally awake and active, the time spent awake was significantly ( P= 0.012) decreased by buprenorphine. The buprenorphine condition within the dark phase revealed significantly ( P= 0.0019) more NREM sleep and a nonsignificant decrease in REM sleep compared with the saline condition. Fig. 4. The percentage of time during the 12-h dark phase that rats (n = 7) spent in states of wakefulness (Wake), nonrapid eye movement sleep (NREM), and rapid eye movement sleep (REM). These recordings were obtained during hours 12 through 24 after administration of either saline (control) or buprenorphine. Asterisks (*) indicate significant differences compared with saline. These results illustrate that after the buprenorphine-induced inhibition of sleep during the light phase (Fig. 3A) there was a significant rebound increase in NREM sleep and REM sleep during the dark phase, when these rats normally would be awake. Fig. 4. The percentage of time during the 12-h dark phase that rats (n = 7) spent in states of wakefulness (Wake), nonrapid eye movement sleep (NREM), and rapid eye movement sleep (REM). These recordings were obtained during hours 12 through 24 after administration of either saline (control) or buprenorphine. Asterisks (*) indicate significant differences compared with saline. These results illustrate that after the buprenorphine-induced inhibition of sleep during the light phase (Fig. 3A) there was a significant rebound increase in NREM sleep and REM sleep during the dark phase, when these rats normally would be awake. The effect of buprenorphine on states of sleep and wakefulness can also be visualized by comparing the light phase (fig. 3A) and dark phase (fig. 4) results. NREM sleep after buprenorphine increased significantly ( P= 0.0003) from an average of 5.5% in the light phase (fig. 3A) to 27.4% in the dark phase (fig. 4). There was also a significant ( P= 0.003) rebound increase in REM sleep, from an average of 0.33% after buprenorphine during the light phase (fig. 3A) to approximately 4% after buprenorphine during the dark phase (fig. 4). Eszopiclone Decreased the Sleep Disruption Caused by Buprenorphine The five illustrations in the right column offigure 3summarize the results of experiments designed to determine whether the sedative-hypnotic eszopiclone countered the buprenorphine-induced inhibition of sleep. Eszopiclone when coadministered with buprenorphine prevented the significant increase in wakefulness (fig. 3B) caused by buprenorphine alone (fig. 3A). Similarly, the significant buprenorphine-induced decrease in NREM sleep and REM sleep (fig. 3A) was prevented by coadministration of eszopiclone (fig. 3B). Eszopiclone blocked the significant increase in latency to sleep onset (comparefig. 3C and D). Eszopiclone partially reversed the buprenorphine-induced decrease in the number of wakefulness and NREM sleep episodes (comparefig. 3E and F). The 530% increase in average duration of waking episodes caused by buprenorphine (fig. 3G) was reduced to a 171% increase by coadministration of eszopiclone (fig. 3H). Eszopiclone blocked the significant decrease in number of state transitions caused by buprenorphine (comparefig. 3I and J). Buprenorphine Increased Electroencephalogram Delta Power during NREM Sleep Figure 5A–C illustrates electroencephalogram power recorded across states of sleep and wakefulness after IV administration of buprenorphine to awake, freely moving rats. Buprenorphine did not alter electroencephalogram power during wakefulness or REM sleep (fig. 5, A and C) but did increase electroencephalogram power in the delta frequency range during NREM sleep (fig. 5B). ANOVA revealed a significant ( P= 0.007) buprenorphine main effect on electroencephalogram frequency bands ranging from 0.5 to 5.0 Hz in 0.5-Hz increments (fig. 5B). The fast Fourier transform analyses were conducted for electroencephalogram measures obtained during the 12-h light period ( i.e., rat's subjective night) that immediately followed buprenorphine administration. Asfigures 2and3show, buprenorphine depressed NREM sleep for 6–8 h. Measurement of the increase in electroencephalogram delta power was conducted for as long as 12-h after buprenorphine administration. A future study will be needed to determine whether, and for how long beyond 12-h, electroencephalogram delta power is increased by buprenorphine. Fig. 5. Electroencephalographic power (ordinate) plotted as a function of frequency (abscissa) averaged for five rats across states of sleep and wakefulness. During wakefulness ( A) and rapid eye movement (REM) sleep ( C), buprenorphine did not change electroencephalographic power. In contrast, during NREM sleep that occurred after buprenorphine administration ( B) there was a significant increase in electroencephalogram delta power ( black horizontal barabove 2.0–3.0 Hz). EEG = electroencephalogram; NREM = nonrapid eye movement. Fig. 5. Electroencephalographic power (ordinate) plotted as a function of frequency (abscissa) averaged for five rats across states of sleep and wakefulness. During wakefulness ( A) and rapid eye movement (REM) sleep ( C), buprenorphine did not change electroencephalographic power. In contrast, during NREM sleep that occurred after buprenorphine administration ( B) there was a significant increase in electroencephalogram delta power ( black horizontal barabove 2.0–3.0 Hz). EEG = electroencephalogram; NREM = nonrapid eye movement. Buprenorphine Decreased Adenosine Concentrations in PnO and SI Histologic analyses confirmed that all microdialysis sites were localized to the PnO or SI (fig. 6A).Figure 6B shows the results of one representative experiment. Adenosine concentrations in the SI are plotted as a function of time during dialysis with Ringer's solution (121–180 min after probe placement) followed by dialysis delivery of buprenorphine (181–240 min after probe placement).Figure 6, C and D, confirms chromatographic identification of adenosine.Figure 6C illustrates chromatograms produced by five known concentrations of adenosine.Figure 6D shows chromatograms reflecting brain adenosine (dialyzed Ringer's solution), a negative control (nondialyzed Ringer's solution), a positive control (brain adenosine sampled during dialysis delivery of the adenosine deaminase inhibitor EHNA), and an adenosine standard. Fig. 6. Microdialysis delivery of buprenorphine decreased adenosine concentrations in the basal forebrain and pontine reticular formation oral part (PnO). ( A) Cresyl-violet stained coronal sections show representative dialysis sites in the PnO and in the substantia innominata (SI) region of the basal forebrain. Arrows indicate the location of each microdialysis membrane. ( B) Time course of a representative experiment showing adenosine concentrations in the SI before (Ringer's solution) and during dialysis delivery of buprenorphine. Each bar represents one sample (30 μl) acquired during 15 min of dialysis. ( C) Chromatograms produced by ultraviolet measurement during the detection of adenosine concentrations ranging from 0 to 200 nM. Each microdialysis experiment began by creating a 5-point standard curve. The standard curve was used to convert ultraviolet absorbance values to concentration of brain adenosine for each microdialysis sample. ( D) Chromatograms resulting from four different measurement conditions. The black traceillustrates the chromatogram produced when the instrument was injected with Ringer's solution that did not pass through the microdialysis probe (negative control). The green traceshows a chromatogram reflecting brain adenosine measured during brain microdialysis with Ringer's solution. The blue traceis a chromatogram produced by measurement of a 100-nM adenosine standard. The red tracerepresents a brain sample obtained during dialysis with Ringer's solution + 1 mM erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), an adenosine deaminase inhibitor that increased adenosine (positive control). The key point is that all chromatograms showed the same elution time for adenosine. Thus, the magnitude of the standards ( C) and the elution times ( D) confirm the presence of adenosine in the brain samples. Fig. 6. Microdialysis delivery of buprenorphine decreased adenosine concentrations in the basal forebrain and pontine reticular formation oral part (PnO). ( A) Cresyl-violet stained coronal sections show representative dialysis sites in the PnO and in the substantia innominata (SI) region of the basal forebrain. Arrows indicate the location of each microdialysis membrane. ( B) Time course of a representative experiment showing adenosine concentrations in the SI before (Ringer's solution) and during dialysis delivery of buprenorphine. Each bar represents one sample (30 μl) acquired during 15 min of dialysis. ( C) Chromatograms produced by ultraviolet measurement during the detection of adenosine concentrations ranging from 0 to 200 nM. Each microdialysis experiment began by creating a 5-point standard curve. The standard curve was used to convert ultraviolet absorbance values to concentration of brain adenosine for each microdialysis sample. ( D) Chromatograms resulting from four different measurement conditions. The black traceillustrates the chromatogram produced when the instrument was injected with Ringer's solution that did not pass through the microdialysis probe (negative control). The green traceshows a chromatogram reflecting brain adenosine measured during brain microdialysis with Ringer's solution. The blue traceis a chromatogram produced by measurement of a 100-nM adenosine standard. The red tracerepresents a brain sample obtained during dialysis with Ringer's solution + 1 mM erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), an adenosine deaminase inhibitor that increased adenosine (positive control). The key point is that all chromatograms showed the same elution time for adenosine. Thus, the magnitude of the standards ( C) and the elution times ( D) confirm the presence of adenosine in the brain samples. Figure 7A summarizes a final set of experiments that quantified adenosine concentrations in SI and PnO as a function of route of buprenorphine administration. Microdialysis delivery of buprenorphine significantly ( P= 0.03) decreased adenosine concentrations in PnO (−14.8%) and significantly ( P= 0.0004) decreased adenosine concentrations in the SI region of the basal forebrain (−36.7%).Figure 7B plots adenosine concentrations in the SI before and after IV administration of buprenorphine to isoflurane-anesthetized rat. Buprenorphine significantly ( P< 0.0001) decreased (−20.3%) adenosine concentrations in the SI. Fig. 7. Central and systemic delivery of buprenorphine decreased brain adenosine concentrations. Microdialysis delivery of buprenorphine to the pontine reticular formation (PnO) oral part (n = 5 rats) or to the substantia innominata (SI) (n = 5 rats) decreased adenosine concentrations in the PnO or SI, respectively ( A). When administered systemically (n = 5), buprenorphine also decreased adenosine concentrations in the SI ( B). Asterisks (*) in Aindicate a significant decrease in adenosine concentrations during microdialysis delivery of buprenorphine compared with Ringer's solution (control). Asterisk (*) in Bindicates a significant decrease in adenosine concentrations in the SI caused by intravenous (IV) delivery of buprenorphine. Thus, SI adenosine concentrations were decreased by microdialysis delivery of buprenorphine to the SI and by IV buprenorphine ( B). Fig. 7. Central and systemic delivery of buprenorphine decreased brain adenosine concentrations. Microdialysis delivery of buprenorphine to the pontine reticular formation (PnO) oral part (n = 5 rats) or to the substantia innominata (SI) (n = 5 rats) decreased adenosine concentrations in the PnO or SI, respectively ( A). When administered systemically (n = 5), buprenorphine also decreased adenosine concentrations in the SI ( B). Asterisks (*) in Aindicate a significant decrease in adenosine concentrations during microdialysis delivery of buprenorphine compared with Ringer's solution (control). Asterisk (*) in Bindicates a significant decrease in adenosine concentrations in the SI caused by intravenous (IV) delivery of buprenorphine. Thus, SI adenosine concentrations were decreased by microdialysis delivery of buprenorphine to the SI and by IV buprenorphine ( B). Discussion Buprenorphine can be efficacious in the treatment of opioid and heroin addiction,45,–,47and there is increasing interest in the use of buprenorphine for pain management.27,28The analgesic effects of buprenorphine are mediated, in part, viaagonist actions at the μ-opioid receptor.48This is the first study presenting electrographic data that demonstrate significant sleep disturbance (fig. 3) caused by an antinociceptive dose of buprenorphine (fig. 1). The current finding that an antinociceptive dose of buprenorphine disrupts sleep is discussed relative to the relationship between sleep and nociception, the potential for developing countermeasures for opioid-induced sleep disruption, and the underlying mechanisms. Buprenorphine Disrupted Sleep Some data suggest that buprenorphine is superior to traditional opioids for the treatment of pain because of its reported analgesic and antihyperalgesic effects with fewer side effects (low incidence of respiratory depression and less constipation).28Buprenorphine shares some similar pharmacodynamic properties with traditional opioids, and patient-report data indicate benefits from buprenorphine therapy. Freye et al.27found that self-report sleep quality rated as “good” or “very good” increased from 14% to 74% when patient regimens were transitioned from high-dose oral morphine to transdermal buprenorphine. Transdermal buprenorphine has been compared with placebo for ability to decrease pain and promote sleep, and patients randomized to receive buprenorphine report less pain and improved sleep.49Specifically, subjects who received buprenorphine reported less trouble falling asleep, decreased requirement for sleeping medication, and decreased awakening at night because of pain. Another study found a nonsignificant trend of improved sleep favoring transdermal buprenorphine over extended-release tramadol tablets for the treatment of osteoarthritis.50A known limitation of such studies is that self-assessment of sleep quality may not show faithful concordance with objective, electrographic measures of sleep.51 There is a growing appreciation for the interrelationship between sleep and pain.39Sleep deprivation in healthy normal individuals decreases pain perception thresholds.52The chronic effects of μ-opioid receptor agonists on sleep in patients with pain are not understood completely.39Opioids cause sleep disturbance4,37,53and the current results demonstrate that buprenorphine increases wakefulness and disrupts the temporal organization of sleep (figs. 2–4). Sleep, like breathing, is an endogenously generated biologic rhythm. Just as rhythmic switching from inspiration to expiration is essential for gas exchange, the ability of sleep to produce reports of rest and recovery requires a normal temporal organization. Buprenorphine caused a decrease in the number (fig. 3E) and an increase in the duration (fig. 3G) of wakefulness episodes. The decreased number of state transitions (fig. 3I) reflects the buprenorphine-induced disruption of sleep continuity.Figure 4summarizes the percentage of time spent in states of sleep and wakefulness during the 12-h dark phase when rats are normally active. These dark-phase recordings were continuous with thefigure 3data during the 12-h light phase. Thus, thefigure 4data show that for 12–24 h after administration of buprenorphine, there was a rebound increase in sleep at a time when nocturnal rodents are normally most active. The potential clinical relevance of buprenorphine-induced disruption of sleep continuity derives from repeated sleep disruption negatively affecting neurocognitive function as severely as does total sleep deprivation.54 Opioid-induced sleep disruption has the potential to negatively affect patient care because sleep deprivation is known to decrease pain threshold.15,16,52This study did not address the impact of pain or the treatment of pain on sleep disturbance. Some believe that medications from the agonist-antagonist class, such as buprenorphine, may be less associated with the adverse effects of traditional μ-agonists; however, the current data indicate that the sleep-disrupting effects of buprenorphine are similar to those of other opioids.26,30,37,53The current results encourage studies designed to objectively quantify the effects of buprenorphine on sleep in humans. The U.S. Food and Drug Administration approved buprenorphine for the treatment of opioid addiction. Suboxone® (buprenorphine-naloxone sublingual tablet, Reckitt Benckiser Pharmaceuticals Inc., Richmond, VA), Subutex® (buprenorphine sublingual tablet, Reckitt Benckiser Pharmaceuticals Inc.), and transdermal buprenorphine (not available in the United States) are increasingly used to treat pain. The current finding of buprenorphine-induced sleep disruption also is relevant to evidence indicating that sleep disturbance can lead to a higher potential for addiction relapse.55,–,57 Electroencephalogram delta waves (0.5–4 Hz) are one of three rhythmic waveforms characteristic of NREM sleep.58Delta waves provide an index of sleep intensity,59,60and electroencephalogram power in the delta frequency increases during recovery sleep that follows sleep deprivation.61,62The finding that buprenorphine caused an increase in electroencephalogram delta power during NREM sleep (fig. 5B) is consistent with subjective reports that buprenorphine improves sleep7,27and with evidence that opioids increase delta power in humans.63A search of the Medline database from 2001 to 2011 revealed no polysomnographic data characterizing the effect of buprenorphine on human sleep. Eszopiclone Was an Effective Countermeasure that Prevented Sleep Disruption by Buprenorphine Buprenorphine significantly disrupted the temporal organization of sleep (fig. 3, panels A, C, E, G, and I). Most aspects of sleep disturbance were prevented by concomitant treatment with the nonbenzodiazepine hypnotic eszopiclone (fig. 3, panels B, D, F, H, and J). Sedative-hypnotics are a standard treatment for insomnia, but their effects in the treatment of pain- and opioid-induced sleep disturbance remain poorly understood. The use of sedative-hypnotics may not be a mainstay of addiction therapy, but the current results (fig. 3, panels B, D, F, H, and J) indicate their potential to prevent buprenorphine-induced sleep disturbance. Eszopiclone as an adjunctive agent coadministered with the antidepressant fluoxetine resulted in a faster onset and greater magnitude of the desired antidepressant effect.64An exciting area open to future study is to determine whether hypnotics can be used as an effective countermeasure for opioid-induced sleep disruption. Limitations, Potential Clinical Relevance, and Conclusions The current study was designed to quantify the effects of buprenorphine on sleep and adenosine concentrations. The results are limited to documenting that buprenorphine, similar to morphine and fentanyl, disrupted sleep and decreased adenosine concentrations in sleep-related brain regions. The results do not imply that the effects of buprenorphine were mediated only by μ-opioid receptors. Buprenorphine may have disrupted sleep and decreased adenosine, in part, by acting as a κ antagonist. The analgesic21and sleep-promoting65effects of adenosine are well known and suggest adenosine as a molecule of potential clinical relevance for anesthesiology. There is good agreement between preclinical and clinical data that opioids disrupt sleep,37a finding confirmed by administering opioids to pain-free humans.53The restorative effects of sleep require normal temporal organization of sleep. Unfortunately, morphine and fentanyl slow the electroencephalogram during wakefulness, increase lighter stage 2 NREM sleep, decrease stage 3 and 4 NREM sleep, and decrease or eliminate REM sleep. Disruption of normal sleep impairs immune function, exacerbates pain,39and, particularly in older patients, can be a precipitating factor for postoperative delirium.66 In conclusion, the results show for the first time that buprenorphine disrupted normal sleep architecture and decreased adenosine concentrations in sleep-regulating regions of the basal forebrain and PnO (figs. 6and7). The buprenorphine results are consistent with the discovery that fentanyl and morphine decrease adenosine concentrations in basal forebrain and PnO.26The current study extends the previous findings by providing mechanistic insights into a brain site and a molecule by which buprenorphine disrupts sleep. Novel insights were obtained by holding the site of adenosine measurement constant within the SI region of the basal forebrain while varying the route of buprenorphine delivery. The results show that both microdialysis delivery to the SI and systemic administration of buprenorphine caused a significant decrease in adenosine in the SI. As demonstrated elsewhere,30,44when effects caused by drug delivery to a specific brain region replicate the effects caused by systemic delivery, it is logical to infer that the actions of systemically administered drugs are mediated, in part, by that brain region and by the neurotransmitter molecule being measured. Thus, the neurochemical results, combined with the sleep-disrupting effect of buprenorphine, support the interpretation that one mechanism by which buprenorphine disrupts sleep is decreasing adenosine concentrations in the SI region of the basal forebrain. For expert assistance, the authors thank Sha Jiang, B.S., Research Associate, Mary A. Norat, B.S., Senior Research Associate, and Sarah L. Watson, B.S., Senior Research Associate, Department of Anesthesiology; and Kathy Welch, M.A., M.P.H., Statistician Staff Specialist, Center for Statistical Consultation and Research, University of Michigan, Ann Arbor, Michigan. References 1. Furlan AD, Sandoval JA, Mailis-Gagnon A, Tunks E: Opioids for chronic noncancer pain: A meta-analysis of effectiveness and side effects. CMAJ 2006; 174:1589–94 2. Noble M, Tregear SJ, Treadwell JR, Schoelles K: Long-term opioid therapy for chronic noncancer pain: A systematic review and meta-analysis of efficacy and safety. J Pain Symptom Manage 2008; 35:214–28 3. Cronin A, Keifer JC, Baghdoyan HA, Lydic R: Opioid inhibition of rapid eye movement sleep by a specific mu receptor agonist. Br J Anaesth 1995; 74:188–92 4. Dimsdale JE, Norman D, DeJardin D, Wallace MS: The effect of opioids on sleep architecture. J Clin Sleep Med 2007; 3:33–6 5. Keifer JC, Baghdoyan HA, Lydic R: Sleep disruption and increased apneas after pontine microinjection of morphine. ANESTHESIOLOGY 1992; 77:973–82 6. Shaw IR, Lavigne G, Mayer P, Choinière M: Acute intravenous administration of morphine perturbs sleep architecture in healthy pain-free young adults: A preliminary study. Sleep 2005; 28:677–82 51. Baker FC, Maloney S, Driver HS: A comparison of subjective estimates of sleep with objective polysomnographic data in healthy men and women. J Psychosom Res 1999; 47:335–41 52. Chhangani BS, Roehrs TA, Harris EJ, Hyde M, Drake C, Hudgel DW, Roth T: Pain sensitivity in sleepy pain-free normals. Sleep 2009; 32:1011–7 53. Bonafide CP, Aucutt-Walter N, Divittore N, King T, Bixler EO, Cronin AJ: Remifentanil inhibits rapid eye movement sleep but not the nocturnal melatonin surge in humans. ANESTHESIOLOGY 2008; 108:627–33 54. Van Dongen HP, Maislin G, Mullington JM, Dinges DF: The cumulative cost of additional wakefulness: Dose-response effects on neurobehavioral functions and sleep physiology from chronic sleep restriction and total sleep deprivation. Sleep 2003; 26:117–26 55. Arnedt JT, Conroy DA, Brower KJ: Treatment options for sleep disturbances during alcohol recovery. J Addict Dis 2007; 26:41–54 56. Brower KJ, Perron BE: Sleep disturbance as a universal risk factor for relapse in addictions to psychoactive substances. Med Hypotheses 2010; 74:928–33 57. 58. 59. Friedman L, Bergmann BM, Rechtschaffen A: Effects of sleep deprivation on sleepiness, sleep intensity, and subsequent sleep in the rat. Sleep 1979; 1:369–91 60. 61. Feinberg I, Floyd TC, March JD: Effects of sleep loss on delta (0.3–3 Hz) EEG and eye movement density: New observations and hypotheses. Electroencephalogr Clin Neurophysiol 1987; 67:217–21 63. Greenwald MK, Roehrs TA: Mu-opioid self-administration vs passive administration in heroin abusers produces differential EEG activation. Neuropsychopharmacology 2005; 30:212–21 64. Fava M, McCall WV, Krystal A, Wessel T, Rubens R, Caron J, Amato D, Roth T: Eszopiclone co-administered with fluoxetine in patients with insomnia coexisting with major depressive disorder. Biol Psychiatry 2006; 59:1052–60 65. Porkka-Heiskanen T, Alanko L, Kalinchuk A, Stenberg D: Adenosine and sleep. Sleep Med Rev 2002; 6:321–32 66. Rudolph JL, Marcantonio ER: Postoperative delirium: Acute change with long-term implications. Anesth Analg 2011; 112:1201–11

https://pubs.asahq.org/anesthesiology/article/115/4/743/12866/Buprenorphine-Disrupts-Sleep-and-Decreases?searchresult=1

Linux Graphics, a Tale of Three Drivers [LWN.net] Linux Graphics, a Tale of Three Drivers Linux Graphics, a Tale of Three Drivers I read the article and I suspect many would agree... that an open-source driver is indeed better. I recently moved *BACK* from running an Intel GMA950 (X3100) onboard graphics card to an nVidia based card (GeForce 8800 GT). As such I've moved from an open source driver back to the binary blog known as nVidia drivers. Why? Whilst I would prefer running a card that has opensource drivers, I found that many graphics intensive 3D apps (not games) crawled under the X3100, making them either unable to run or so slow they were unusable. I'm personally haven't run an ATI card, so I cannot state how well their Radeon-HD cards compare, particularly with open source drivers, yet the state of their open source drivers/performance. If Intel want to become dominant on the Linux desktop in graphics drivers I would recommend: 1. They start producing graphics chipsets that can handle something more than low-end 3D. 2. Dedicated add-on cards instead of onboard graphics cards would also be advisable, or at least the option. Onboard graphics cards for laptops seems reasonable, but just doesn't cut it for desktops. They are woefully under powered. Whilst I'm stuck on a binary-only driver and it isn't ideal, I'm at least seeing the 3D performance some of these desktop apps require. I'll stay on the nVidia train for the time being until Intel can produce a card that matches the performance of even the 7000 series cards. In a perfect world nVidia would open-source up their graphics drivers, but I'm not holding my breath. Linux Graphics, a Tale of Three Drivers Whilst I would prefer running a card that has opensource drivers, I found that many graphics intensive 3D apps (not games) crawled under the X3100, making them either unable to run or so slow they were unusable. Which apps are you speaking of? Linux Graphics, a Tale of Three Drivers > Which apps are you speaking of? I don't speak for the original poster, but I can attest to similar problems arising from the molecular visualization programs used routinely in my lab. Most are built on OpenGL. Performance using Intel graphics chips is adequate but not great. Aside from uninspiring performance per se, it is also annoying that even the latest intel drivers still cause the machines to lock up hard occasionally. So, reluctantly, I'm back to fitting out new machines with nVidia cards even if they were originally configured with Intel graphics. If you want example apps to download and try, I suggest PyMol http://pymol.sourceforge.net/ ccp4mg http://www.ysbl.york.ac.uk/~ccp4mg/ Coot http://www.ysbl.york.ac.uk/~emsley/coot/ VMD http://www.ks.uiuc.edu/Research/vmd/ Linux Graphics, a Tale of Three Drivers I had the same experience running the *ahem* productivity application known as World of Warcraft under wine. The performance was not impressive, although I didn't care that much, and then the system would hard lock for around 30 seconds and reset. I blame thermal issues, but it was using the heatsink etc as supplied from manufacturer, and applying much more prodigious heatsinks did not resolve it. Linux Graphics, a Tale of Three Drivers IIRC Google Earth does not run with the free nvidia driver. You need the binary driver with 3D acceleration to run the application. But I guess for most uses Google Maps is good enough. Linux Graphics, a Tale of Three Drivers I very much support / only accept using the open source drivers, but I do acknowledge the lack of performance in Intel graphics. I think the article was a bit (not too much, but somewhat) biased. Some points I think are worth mentioning: - Intel drivers are good, especially the new 2.3 series, but not perfect - they still require manually forcing overlay support (instead of textured video) and greedy fallback EXA functionality to perform at useful speed in all situations I encounter. Implemented like that in Ubuntu. - Intel is, while fixing drivers, fixing X and architecture at the same time. I believe that by the end of this year full EXA acceleration can be enabled without problems. - Intel's 3D performance is only at most 50% from what the hw would allow, because of the delays in implementing TTM/GEM stuff in kernel and batchbuffer functionality (without regressions) in the DDX driver. It will get better from the current situation. But, all in all, Intel is the best we have in terms of open source drivers, and they're coming better all the time. ATI's open drivers are quite good, but before AMD opened specifications there were bad problems with specific chipsets, together with some lack of functionality (like sw rendering if anti-aliased lines were enabled in any OpenGL program), which hindered the quality. The next round of driver releases (DDX, Mesa) will have a lot of those fixed, and AMD's R300-R500 (9500 - X1950) might become the preferred choice for many since they outperform Intel solutions so greatly. The ATI's open drivers do not reach the full potential of the cards yet either, though. The X2000 - X4000 series 3D support is probably not ready this year yet, or maybe towards the end of this year compiz might be running... dunno. ATI's binary drivers have sucked, but have finally been improving to a state they actually might eg. not crash on a 64-bit computer or when using both (!) VGA and DVI outputs like in my case, but I wouldn't install those anyway. The open drivers are better in every other aspect except for the raw 3D speed. Lastly, the reality is that for many casual users NVIDIA's binary drivers seem to deliver the best experience. 3D performance is reaching the full potential of the hardware, and they are relatively stable at least for some. The same casual users blame the non-functionality like lack of Randr1.2 support (ie. ordinary tools for changing resolution do not work) on the "Linux", not the drivers. And likewise if the system crashes, which it does on many configurations. And likewise if the NVIDIA's drivers do not support latest X, it's X's fault. But generally for those that the drivers work, they're getting speedy 3D. So, it's easy to understand the cheer for NVIDIA from time to time. But Intel is starting to kick serious ass in any non-heavy 3D use, ie. normal computer use, while ATI has reached binary driver parity with NVIDIA (from what I've heard) and they're starting to have some good open source drivers finally. When the X2000 - X4000 get 3D support, they can be easily recommended over NVIDIA as they have better support for eg. standard X features like Randr1.2, and they work out-of-the-box on any distribution without the need to install drivers. But that'll be next year. Linux Graphics, a Tale of Three Drivers Hope you are right and I am just impatient, but the current state of things regarding intel graphics drivers in Linux is quite SAD, as one can discover in this launchpad bug: https://bugs.launchpad.net/ubuntu/+source/xserver-xorg-vi... And you can even see in this video: http://www.youtube.com/watch?v=7cWW-12gxdY Just as a summary:Ubuntu 7.04&7.10: glxgears was around 1500FPSUbuntu 8.10: glxgears drops to around 500FPSBut wait! there is still the latest versions to come! the ones with GEM that were going to save us:Ubuntu 9.04 Alpha1: glxgears does around 300FPS!!! Great! Linux Graphics, a Tale of Three Drivers http://wiki.cchtml.com/index.php/Glxgears_is_not_a_Benchmark

https://lwn.net/Articles/287238/

Complete rupture of the triceps brachii muscle | British Journal of Sports Medicine Case Report Complete rupture of the triceps brachii muscle Free R K Singh , J Pooley Upper Limb Unit, Department of Orthopaedics, Queen Elizabeth Hospital, Gateshead NE9 6SX, UK Correspondence to:
 Dr Singh, Upper Limb Unit, Department of Orthopaedics, Queen Elizabeth Hospital, Gateshead NE9 6SX, UK; [email protected] Abstract The case is reported of a professional ice hockey player who sustained an intramuscular rupture of all three heads of the triceps brachii muscle. After surgical repair, he made a complete recovery and was able to return to professional ice hockey. triceps brachii muscle rupture http://dx.doi.org/10.1136/bjsm.36.6.467 triceps brachii muscle rupture CASE REPORT A 31 year old Norwegian professional ice hockey player, playing in the United Kingdom at the time, presented for evaluation of non-dominant left arm weakness of three weeks duration. He had sustained an injury to his left arm during a game of ice hockey when he was hit directly across the middle and posterior part of the arm by an opponent’s stick, after which he lost balance and went into a wall with his stick held in his hand at waist height. He was unable to continue with the game because of what he described as a tearing sensation and severe pain in his arm. He was assessed by the team physiotherapist and doctor, and was given a broad arm sling with advice to start gentle supervised physiotherapy as the pain settled. Gradually the pain settled but he still had considerable residual weakness of arm extension. On physical examination, a palpable gap in the substance of the triceps muscle was found and he was unable to hold the arm in an extended position. He denied the use of anabolic steroids. A diagnosis of intramuscular rupture of the triceps was made and a magnetic resonance imaging scan requested. This was reported as normal and did not show any conclusive evidence of rupture of the triceps. The patient underwent surgical exploration through a midline posterior incision. An incision through the triceps aponeurosis revealed the site of the rupture involving mainly the long and medial heads of the triceps but also extending into the lateral head. The muscle ends were contracted and oedematous, with considerable distortion of the anatomy (fig 1). Figure 1 Rupture of the triceps brachii muscle involving mainly the long and medial heads but also extending into the lateral head. In an attempt to define the anatomy, the intermuscular aponeurosis, separating the lateral head of triceps from the long and medial heads, was mobilised (fig 2). The muscle bellies were further mobilised as best as possible (fig 3). Repair was carried out by suturing the torn ends of the muscle together, with difficulty because of the oedema and retraction of the cut ends, but mainly because of the intermuscular aponeurosis. Finally, the triceps aponeurosis was replaced and sutured back into place (fig 4). Figure 2 The intermuscular aponeurosis (held by the forceps) separating the lateral head of the triceps from the long and medial heads was mobilised. Figure 3 Mobilisation of the torn muscle bellies. Figure 4 The final repair. The torn ends of the muscle have been sutured together but mainly to the intermuscular aponeurosis. The triceps aponeurosis (held by the forceps) is being replaced and sutured back into place. The patient returned to Norway where further follow up and supervised physiotherapy was undertaken. Six months after the operation, he had gone back to playing ice hockey at a professional level. DISCUSSION Disruption of the triceps can occur in one of three anatomic locations: the tendon attachment to bone, the musculotendinous junction, or in the muscle substance, in decreasing order of frequency. Tarsney 2stated that cases involving disruption of the triceps at its insertion into the olecranon are more accurately termed “avulsion of the triceps” with “rupture” being used for intramuscular or musculotendinous disruption of the triceps. Numerous authors have noted the rarity of these injuries. 1, 3– 9The most common mechanism of injury resulting in avulsion of the triceps tendon has been reported to be a fall on the outstretched hand with the elbow in mid flexion, with or without a concomitant direct blow to the posterior aspect of the elbow. 1Farrar and Lippert 10stated that the most common mechanism of injury was a deceleration stress superimposed on a contracted triceps muscle, with or without a concomitant blow to the posterior aspect of the elbow. There are increasing reports of major tendon avulsion and rupture in patients with chronic renal failure and secondary hyperparathyroidism. 11– 16Wagner and Cooney 8reported a case of rupture of the triceps at the musculotendinous junction in a patient with diabetes mellitus, nephrolithiasis, essential hypertension, and obesity. Another group of patients presenting with rupture and avulsion of the triceps tendon are those with a history of using anabolic steroids. Bach et al 4reported the case of a 33 year old Olympic weightlifter who ruptured the long and lateral heads of the triceps at the musculotendinous junction. His past medical history was significant for anabolic steroid usage for four months preceding the injury. Sollender et al 17reported four cases of triceps tendon rupture in weightlifters, two of whom had received local steroid injections for pain in the triceps. All four had taken oral anabolic steroids before the injury. The first reported case of intramuscular rupture of the triceps brachii muscle was by Penhallow 18in a 35 year old man who was struck by a large box above the elbow. Surgical exploration showed a complete rupture of the long head and partial rupture of the medial head of the triceps brachii muscle, which was repaired. Since then, a further five cases have been reported 19– 22(summarised in table 1). None of these patients were reported to be suffering from any previous medical problems or gave a history of anabolic steroid use. This is in contrast with cases of avulsion of the triceps tendon or rupture at the musculotendinous junction which are increasingly being seen in association with various disease conditions and use of anabolic steroids (see above). Table 1 Intramuscular rupture of the triceps brachii muscle Diagnosis in all cases could be made on clinical grounds. The pathognomonic features are inability or weakness of extension of the elbow and a palpable gap in the substance of the muscle. Depending on the duration of the injury, bruising, ecchymosis, and localised tenderness may be present. Further imaging studies may be performed to help to define the exact extent of the injury, but the results should be viewed with caution as is well illustrated in our case where the magnetic resonance imaging scan was reported as normal with no evidence of rupture of the triceps muscle. Only one of the six cases reported can be presumed to be of a complete rupture of all three heads of the triceps. 19A further two cases involved the long head, 20, 21one of the medial head 22and one of uncertain extent but definitely partial as the patient had good power on triceps testing. 22Penhallow 18had found a rupture of the long and partial medial heads during surgery. In our case the rupture involved all three heads, which was repaired. Two of the cases were treated surgically and a further three non-operatively. One patient underwent surgical exploration but no repair was carried out as the muscle was already found to be scarred and adhesed 22(table 1). Too few cases have been reported to allow a conclusion to be drawn about whether operative or non-operative treatment is the best option for intramuscular rupture of the triceps brachii muscle. We would agree with O’Driscoll 20that non-operative treatment is effective and can be recommended for patients who do not require great endurance strength in elbow extension, but would hasten to add that this may be true only in cases of partial rupture, and that in all cases of confirmed or suspected complete intramuscular rupture of the triceps surgical repair would seem to be essential to restore functional extension strength to the elbow. In conclusion, intramuscular rupture of the triceps brachii muscle is a rare injury usually seen in young, otherwise healthy people, involved in sporting activities. Including this case, only seven cases have been reported in the English literature, with only two being of a complete rupture of all three heads. Take home message Intramuscular rupture of the triceps brachii muscle is seen in young healthy people involved in sporting activities Diagnosis can be made on clinical grounds but further imaging studies should be performed to establish the full extent of the injury The decision to treat conservatively or operatively should be made on the basis of the extent of the injury and the functional requirements of the patient REFERENCES ↵ Anzel SH , Covey KW, Weiner AD, et al . Disruption of muscles and tendons: an analysis of 1014 cases. Surgery 1959 ; 45 : 406 –14. ↵ Tarsney FT . Rupture and avulsion of the triceps. Clin Orthop 1972 ; 83 : 177 –83. ↵ Anderson KJ , LeCocq JF. Rupture of the triceps tendon. J Bone Joint Surg [Am] 1957 ; 39 : 444 –6. ↵ Bach BR Jr , Warren RF, Wickiewicz TL. Triceps rupture: a case report and literature review. Am J Sports Med 1987 ; 15 : 285 –9. Bennet BS . Triceps tendon rupture: case report and a method of repair. J Bone Joint Surg [Am] 1962 ; 44 : 741 –4. Inhofe PD , Moneim MS. Late presentation of triceps rupture: a case report and review of literature. Am J Orthop 1996 ; 25 : 790 –2. Pantazopoulos T , Exarchou E, Stavrou Z, et al . Avulsion of the triceps tendon. J Trauma 1975 ; 15 : 827 –9. ↵ Wagner JR , Cooney WP. Rupture of the triceps muscle at the musculotendinous junction: a case report. J Hand Surg 1997 ; 22 : 342 –3. ↵ Waugh RL , Hatchcock TA, Elliot JL.Ruptures of muscles and tendons. Surgery 1949 ; 25 : 370 –91. ↵ Farrar EL , Lippert FG. Avulsion of the triceps tendon. Clin Orthop 1981 ; 161 : 242 –6. ↵ Cirincione RJ , Baker BE. Tendon ruptures with secondary hyper-parathyroidism. J Bone Joint Surg [Am] 1975 ; 57 : 852 –3. de Waal Malefijt MC , Beeker TW. Avulsion of the triceps tendon in secondary hyperparathyroidism: a case report. Acta Orthop Scand 1987 ; 58 : 434 –5. Mankin HJ . Rickets, osteomalacia and renal osteodystrophy. J Bone Joint Surg [Am] 1974 ; 56 : 352 –80. Murphy KJ , McPhee I. Tears of major tendons in chronic acidosis with elastosis. J Bone Joint Surg [Am] 1965 ; 47 : 1253 –8. Preston FS , Adicoff A. Hyperparathyroidism with avulsion of three major tendons: report of a case. N Engl J Med 1962 ; 266 : 968 –71. Google Scholar ↵ Searfoss R , Tripi J, Bowers W. Triceps brachii rupture: a case report. J Trauma 1976 ; 16 : 244 –6. ↵ Sollender JL , Rayan GM, Barden GA. Triceps tendon rupture in weight lifters. J Shoulder Elbow Surg 1998 ; 7 : 151 –3. ↵ Penhallow DP . Report of a case of ruptured triceps due to direct violence. N Y Med J 1910 ; 91 : 76 –7. ↵ Montgomery AH . Two cases of muscle injury. Surgical Clinics of Chicago 1920 ; 4 : 871 –7. ↵ O’Driscoll SW . Intramuscular triceps rupture. Can J Surg 1992 ; 35 : 203 –6. ↵ Sheps D , Black B, Reed M, et al . Rupture of the long head of the triceps muscle in a child: case report and review of the literature. J Trauma 1997 ; 42 : 318 –20. ↵ Aso K ,Torisu T. Muscle belly tear of the triceps. Am J Sports Med 1984 ; 12 : 485 –7.

https://bjsm.bmj.com/content/36/6/467.full

Fukushima Technical Discussion Open Thread | Brave New Climate It was suggested in a comment -- and I agree -- that the previous open threads on the Fukushima Daiichi Nuclear Accident were becoming difficult to read, because they are such a mixture of technical details and philosophical discourse. That is, it's generally a bad idea to cater to two different audiences in one comment… Fukushima Technical Discussion Open Thread It wassuggested in a comment— and I agree — that the previous open threads on the Fukushima Daiichi Nuclear Accident were becoming difficult to read, because they are such a mixture of technical details and philosophical discourse. That is, it’s generally a bad idea to cater to two different audiences in one comment thread. So, I will split them up. Pleaserestrictall discussion here to technical information, analysis, criticisms and questions on FD — no philosophising or excursions into whether nuclear power is ‘good’ or ‘bad’ or the implications of FD for the future of nuclear power (except for new technical developments, e.g. safety standards), etc. You may impart your deep wisdom on how the world should work on the other open thread I’m about to open. Besides the above guidelines, the other rules of the Open Threadson BNC apply.Read herefor details. To kick off discussion, below is the latest FEPCstatus report (I’ll update this as new reports come in). You will also be interested in: — JAIF Updates#35and#36 —NISA Major Parameters 0600 March 29 —NISA Summary Conditions 0600 March 29 —————- Radiation Levels At 11:45PM (JST) on March 28, TEPCO announced that plutonium 238, 239 and 240 were detected in the soil sampled on March 21st and 22nd at five spots in Fukushima Daiichi Nuclear Power Station. Concentration of detected plutonium 238, 239 and 240 are the same level of the fallout observed in Japan at the atmospheric nuclear tests in the past and poses no major impact on human health. At 6:30PM on March 29, radiation level at main gate (approximately 3,281 feet from Unit 2 reactor building) of Fukushima Daiichi Nuclear Power Station: 177 micro Sv/hour. At 6:30PM on March 29, radiation level at west gate (approximately 3,609 feet from Unit 2 reactor building) of Fukushima Daiichi Nuclear Power Station: 120.2 micro Sv/hour. Measurement results of environmental radioactivity level around Fukushima Nuclear Power Station announced at 7:00PM on March 29 are shown in the attached PDF file. English version is available at: http://www.mext.go.jp/english/radioactivity_level/detail/1304082.htm For comparison, a human receives 2,400 micro Sv per year from natural radiation in the form of sunlight, radon, and other sources. One chest CT scan generates 6,900 micro Sv per scan. Fukushima Daiichi Unit 1 reactor At 1:00PM on March 29, pressure inside the reactor core: 0.371MPa. At 1:00PM on March 29, water level inside the reactor core: 1.65 meters below the top of the fuel rods. At 1:00PM on March 29, pressure inside the primary containment vessel: 0.265MPaabs. At 1:00PM on March 29, the temperature of the reactor vessel measured at the water supply nozzle: 570.9 degrees Fahrenheit As of 3:00PM on March 29, transferring the water found at the turbine building to the condenser continues. As of 4:00PM on March 29, the injection of freshwater into the reactor core continues. Fukushima Daiichi Unit 2 reactor At 1:00PM on March 29, the temperature of the spent fuel pool: 114.8 degrees Fahrenheit. At 1:00PM on March 29, pressure inside the reactor core: -0.025MPa. At 1:00PM on March 29, water level inside the reactor core: 1.5 meters below the top of the fuel rods. At 1:00PM on March 29, pressure inside the primary containment vessel: 0.1MPaabs. As of 4:00PM on March 29, the injection of freshwater into the reactor core continues. As of 7:00PM on March 29, approximately 96 tons of water in total has been injected into the spent fuel storage pool. Fukushima Daiichi Unit 3 reactor At 12:00PM on March 29, pressure inside the reactor core: 0.029MPa. At 12:00PM on March 29, water level inside the reactor core: 1.85 meters below the top of the fuel rods. At 12:00PM on March 29, pressure inside the primary containment vessel: 0.1075MPaabs. At 2:17PM on March 29, TEPCO began to shoot freshwater aimed at the spent fuel pool, with a specialized vehicle normally used for pumping concrete, until 6:18PM (approximately 100 tons in total). As of 4:00PM on March 29, the injection of freshwater into the reactor core continues. As of 7:00PM on March 29, approximately 4,697 tons of water in total has been shot to the spent fuel storage pool. Fukushima Daiichi Unit 4 reactor At 11:50AM on March 29, lighting was restored in the Central Control Room. As of 7:00PM on March 29, approximately 960 tons of water in total has been shot to the spent fuel storage pool. Fukushima Daiichi Unit 5 reactor At 2:00PM on March 29, the temperature of the spent fuel pool: 101.5 degrees Fahrenheit. Fukushima Daiichi Unit 6 reactor At 2:00PM on March 29, the temperature of the spent fuel pool: 70.7 degrees Fahrenheit. Fukushima Daiichi Common Spent Fuel Pool At 3:10PM on March 28, the temperature of the spent fuel pool: 95 degrees Fahrenheit. As of 7:00PM on March 29, approximately 130 tons of water in total has been injected to the spent fuel storage pool. (1) A complete loss of station power occurs,accompanied by complete loss of emergency corecooling systems. The control rods areautomatically inserted at the beginning of theaccident, terminating the fission reaction. (2) No active efforts are made by the operators tomitigate the accident. (3) A short time after the loss of power and corecooling the upper core becomes uncovered. (3) The control rods melt in a part of the uppercore, probably near to the center and top. Thiscan happen without a simultaneous melting of thefuel rods, if the control rods are made of boroncarbide, since it has a lower melting point thanZircaloy fuel cladding. Control rods are relocatedin this way, out of a part of the upper core,leaving that area with close to normal reactorgeometry, but no control rods. But there is nowater present, only steam; and therefore there isstill no fission reaction. (4) Just at this point, and before fuel rods inthe upper core begin to melt and relocatedownwards together with the control rods, theelectrical power is suddenly restored to theplant. (5) The partially damaged core is then flooded bycold fresh water, due to the restarting of theemergency core cooling pumps. Fresh water is themoderator in a BWR. So re-critcality can now occurin that part of the core which has been cleared ofcontrol rods, but still has close to normalreactor geometry. (6) The fission reaction restarts in part of thedamaged core (re-criticality). Since the control rods are now gone, the factorsthat will limit the re-initiated fission reactionare the natural ones that always operate in a LWR:namely void formation and Doppler broadening. That is: as the fission reaction speeds up, itgenerates more heat. The additional heat boilswater, creating steam bubbles in the liquidphase. Steam is less dense than liquid water, soit doesn’t slow neutrons nearly asefficiently. This void formation in turn slows thefission reaction down. Doppler broadening of the neutron spectrum alsooccurs due to heat generated by the fissionreaction. Neutrons which hit nuclei in the fuel,or in the cladding, or that hit oxygen nuclei inthe water scatter elastically if they aren’tabsorbed and don’t cause fission reactions. Butdue to heat, the nuclei are also movingaround. Such elastically scattered neutrons gainor lose a little energy as they scatter, and thiswill change the overall energy spectrum of theneutrons. The neutrons will gain energy onaverage, as the scattering material heats up. Butthermal neutron induced fission cross-sectionsalso drop significantly with increasing neutronenergies, so the rate of the fission reaction isin turn reduced as temperature increases. Depending on the rate at which fresh cold water ispumped into the damaged core region, faster beingworse, and also depending on other details, therecan occur a big pulse of reactor power in thesmall fraction of the core that is partiallydamaged. Depending on how big a pulse and how thatenergy is dissipated, the reactor power can eitheroscillate and finally die away, continueoscillating, or possibly die down to some constantlow fraction of the total reactor power. This is potentially a big problem, if theoperators don’t do anything at all to try tomitigate the situation. But the whole sequence of events can be avoided,though, if the operators inject borated instead offresh water into the damaged core once power comesback, or as soon as they have the ability to doso. Borated water acts as a neutron poison (likethe control rods) and will prevent re-criticality. That was actually done in this accident, as Iunderstand it. The operators pumped boratedseawater into the RPVs, over the damaged cores. I imagine that uncertaintly about the extent ofdamage to the reactor cores and worry about theabove scenario may have been the main reason forinjecting the borated seawater into the cores. More complete melting of the reactor cores makesrecriticality very much less likely, because thefuel then relocates downward in the RPV togetherwith the melted fuel rods. Moreover, the geometryof completely melted fuel is extremelyunfavourable for forming a critical configurationwith reespect to thermal neutrons: there is noroom to fit the moderator inside a blob of moltenfuel cladding, control rods, and fuel pellets, andany water that gets trapped or sits on top wouldsimply be boiled away by the heat. A critical configuration with respect to fastneutrons isn’t a realistic possibility, due to thelow enriched fuel that is used in light waterreactors. Critical masses with fast neutrons aresimply too large (even for the optimal sphericalgeometry). Notice that the above scenario for re-criticalityhas to occur pretty early on in the accident. It’svery hard for me to imagine a scenario in whichre-criticality occurs at late times, or once thefuel has become significantly damaged. I suppose conceivably, such a thing might occur,for a vey small fraction of the core, if enoughshattered or disintegrated fuel pellets somehowsurvived melting, and found their way into a poolof fresh water, say in the suppression pool. Butsuch a reaction would be limited by the samemechanisms mentioned above, as well as probablyothers, like the ability of such fuel to flow awayfrom the critical region. If you go to the latest JAIF report (http://www.jaif.or.jp/english/news_images/pdf/ENGNEWS01_1302343542P.pdf), water injection to containment vessel is a reporting item. Status reads : to be confirmed (R1), to be decided (seawater) (R2), and, to be confirmed (R3). George: I do seem to remember seeing that item about injecting water to containment in some of the earlier JAIF updates, but I wasn’t sure whether injection to containment meant just a slow spray of water into the containment, possibly for cooling purposes, or an actual attempt to flood the containment. I certainly agree with you that if containment flooding were to be accomplished, then all of the major drainage paths out of the PCV, below the level of the bottom of the RPV would need to be closed off as tightly as possible. etudiant, on 11 April 2011 at 12:54 AM said: Given the estimate of a decade long D&D program, these emissions look to remain a factor for some time There are lot’s of unanswered questions at the moment, but for radiation to get far it needs a transport mechanism. The first job will be to deny a transport mechanism. [Comment deleted. Violation of citation policy] MODERATORDo not post just a throwaway remark and a link. Citation policy is this: BARRY BROOKThe commenting rules are not meant to be confusing, they’re meant to be logical. This is not a forum for cut-and-pasting slabs of text, with no other comment other than a link. Tell people why you think they should be interesting in reading this, and what it means for this discussion. Otherwise, you’re not thinking and not contributing. Simple as that. Citing literature and other sources: appropriate and interesting citations and links within comments are welcomed, but please DO NOT cite material that you have not yourself read, digested and understood. As a general rule, please introduce any and every link or reference with a short description of the material, your judgement on its quality, and the specific reason you are including it (i.e. how it is relevant to the discussion). Question: when does TEPCO run out of qualified workers? I mean workers who have both required skills and useful remaining radiation allowance. I would like to know how fast the existing TEPCO manpower pool is being depleted (I mean their remaining radiological allowances). A week ago WNN reported that 21 out of 370 workers had reached 100 milliseverts [thanks to Hank Roberts 4/11/11 2:01]: http://www.world-nuclear-news.org/RS_Deaths_confirmed_at_Fukushima_Daiichi_0304111.html It would help to have an idea what part of the remaining work requires high-skill and high-site-experience vs. just cleanup labour. Is extensive site-experience even a big issue? The labour supply is defined by the radiological allowance. Is there a consensus on what the worker allowance should be? Steve Darden, on 12 April 2011 at 8:07 PM said: Question: when does TEPCO run out of qualified workers? I mean workers who have both required skills and useful remaining radiation allowance> A fair portion of the maintenance and upgrades that are done at NPP’s is performed during refueling outages. Since refueling only occurs every 18 months or so there is an industry pool of ‘temporary help’. 2011 US Columbia Operating Station budget – includes $10 million for temporary help and overtime during refueling outage. Click to access Final%202011%20Columbia%20Generating%20Station.pdf “(3) The control rods melt in a part of the uppercore, probably near to the center and top. Thiscan happen without a simultaneous melting of thefuel rods, if the control rods are made of boroncarbide, since it has a lower melting point thanZircaloy fuel cladding.” false The boron carbide has the melting point 2763°C –http://en.wikipedia.org/wiki/Boron_carbide, Zircaloy has melting point much lower – The pure Zirconium itself has 1855°C melting point –http://en.wikipedia.org/wiki/Zirconium, (and is alloyed by Sn – melting point 231°C – and or Niobium – melting point 2477°C – to form Zircaloys) and the Zirconium even at much lower temperatures already violently reacts with any water present. So the described mechanism of the recriticality is a patent nonsense and I don’t wonder that it is again not sourced by any link whatsoever – as was several times called for by the moderater here… Personally I think the recriticality claims about the Fukushima are red herrings (I was looking into it after there were the weird Te-129 readings which subsequently showed being errors) because even if the whole cores would melt through the cladding and end on the bottom of the RPV’s there’s not the unpoisoned (borated etc.) moderator (crucial, without it the chain reaction is impossible in the low enriched fuel) for it to regain criticality) The same for the spent fuel pools, moreover there are the fuel rods packed in the boron carbide racks. You’re right: it’s false, at least the way I summarized thescenario. But, not being an expert, I’ve summarized it wrongly. The meltingof the B4C is said to occur at 1500K, due to a reaction of thecarbide with stainless steel. Possibly stainless steel may be inproximity with control rods in these reactors. So the described mechanism of the recriticality is a patentnonsense and I don’t wonder that it is again not sourced by anylink whatsoever – as was several times called for by themoderater here… There are many sources describing the scenario forrecriticality. Here’s a link to the SARA study: ftp://ftp.cordis.europa.eu/pub/fp5-euratom/docs/09-sara.pdf Recriticality has been studied for a total loss ofelectric power accident scenario. In a BWR, the B4C control rodswould melt at about 1500K (due to a eutectic reaction between B4Cand stainlesss steel) and relocate from the core before the fuelwould during core uncovery and heat-up. If electrical powerreturns during this time-window, the duration of which ispredicted to be in the range of a few minutes to about 40nminutes, it is likely that the core will be reflooded usingwater from Emergency Core Cooling Systems (ECCS) or feed watersupplies. Since this water is normally unborated, recriticalityis possible … Here’s a link to a report produced at ORNL: IDENTIFICATIONOF AND ASSESSMENT OF BWR IN-VESSEL SEVERE ACCIDENT MITIGATION STRATEGIES which describes the injection of borated water as a strategy toprevent recriticality in a BWR accident with severe core damagewhen ” … control blades have melted and relocated fromthe core.” (from section 5. of this report) NUREG/CR5653 documents the conclusion that there is apotentiality for recriticality in BWRs, if core reflood occurs. See: http://www.nrc.gov/reading-rm/doc-collections/nuregs/staff/sr0933/sec3/155r2.html Personally I think the recriticality claims about theFukushima are red herrings (I was looking into it after therewere the weird Te-129 readings which subsequently showed beingerrors) because even if the whole cores would melt through thecladding and end on the bottom of the RPV’s there’s not theunpoisoned (borated etc.) moderator (crucial, without it thechain reaction is impossible in the low enriched fuel) for it toregain criticality) The same for the spent fuel pools, moreoverthere are the fuel rods packed in the boron carbideracks. I agree with you about all of the late recriticality claims, andI would add that the early recriticality possibility seems torequire a perfect storm. But it has, at least, been considered apossibility. Thanks David and Jan. g Here’s the relevant NRC snippet on recrit. Thanks to David. ISSUE 155.4: IMPROVE CRITICALITY CALCULATIONS DESCRIPTION The Board believed that doubts still remained as to whether the TMI-2 core became critical, or was very close to critical, during the TMI-2 accident and recommended that the NRC establish guidelines that deal with criticality following a severe reactor accident.1362 These guidelines should take into account abnormal geometries and possible core conditions that could result from the accident. The Board believed that the accident scenario developed by the TMI-2 licensee was sufficiently detailed that a series of geometric configurations could be simulated for criticality calculations. Variables that could be estimated reasonably well included the presence of water, oxidation of cladding, melting and movement of fuel, melting of poison rods, and movement of poison. CONCLUSION The safety concern was addressed by DSR/RES in SARP Task 4.3: Investigate the Possibility and Consequences of Recriticality in Degraded BWR Cores.1382 The staff’s study was documented in NUREG/CR-56531379 in which it was concluded that there was the potential for recriticality in BWRs, if core reflood occurs after control blade melting has begun but prior to significant fuel rod melting. However, a recriticality event would most likely not generate a pressure pulse significant enough to fail the vessel. Two strategies were identified that would aid in regaining control of the reactor and terminate the recriticality event before containment failure pressures are reached: (1) initiation of boron injection at or before the time of core reflood, if the potential for control blade melting exists; and (2) initiation of RHR suppression pool cooling to remove the heat load generated by the recriticality event and extend the time available for boration. The issue was not considered to be a major concern for PWRs because of their design that includes a safety injection system for supplying borated water to the core. Furthermore, it was concluded in NUREG/CR-58561417 that, during a severe accident, an unmoderated recriticality of the molten, consolidated portion of a degrading core cannot occur at U235 enrichments characteristic of a PWR. Based on the staff’s efforts in addressing the safety concerns in the SARP, this issue was DROPPED from further pursuit as a new and separate issue. In an RES evaluation,1564 it was concluded that consideration of a 20-year license renewal period did not change the priority of the issue. Cheers, Gregory :) harrywr2, on 12 April 2011 at 11:19 PM said “…industry pool of ‘temporary help’.” Thanks Harry – that’s comforting. “Sorry I have to spell it out — I thought it would be interesting that the Union of Concerned Scientists stated that nitrogen injection has never been attempted before (to their knowledge) after a reactor has been running.” Nitrogen in the containment is quite standard feature of the newer BWR designs by default to prevent hydrogen combustion in case of a LOCA accident and yes it must be sometimes injected in the CPV during operation to maintain the pressure. se e.g. here: I[Comment deleted. Violation of the citation rule] @ David Kahana I’m not much convinced about the mechanism of the recriticality you describe anyway:The B4C-steel eutectic melting would be impossible in cases the RPV would be completely dry, because the Zircaloy rapid oxidation would be impossible in such a case. If it would still not be completely dry as is most likely and Zircaloy starts to burn with steam then even if triggering the control rod eutectic it would have much higher temperature – partly because the exothermic violent reaction with the steam, partly because of the decay heat of the fuel inside, so it would most probably be the fuel rods not the control rods which will loose integrity first.I think the theorizing about this scenatio is based on tests like this:http://cat.inist.fr/?aModele=afficheN&cpsidt=15755028But they have major flaw – no fuel with very high after scram decay heat is involved inside the Zircaloy tubes. Btw. Only a crazy would flood the RPV with even partially melted core with a fresh unborated water. It would be better to leave the RPV dry and flood the CPV instead – as the document you link suggests. @ David B. BensonIn the case of the No4 it is quite sure – there was no fuel in the RPV whatsoever. This is only tangentially related to Fukushima, but it does illustrate the difficulty of doing any kind of sensible risk analysis based on likely tsunami height. http://en.wikipedia.org/wiki/Megatsunami I would be interested in a technical discussion on environmental aspects of Fukushima. Has anyone compiled available data into coherent picture? Particularly interested in total Cs deposition as this will be the important one going forward. Sea water more difficult but should be less of an issue due to high salts which should inhibit significant uptake in marine organisms. Fukushima prefecture is starting to look ugly. Depends whether highs are coherent or spot samples. Anyone? Does anyone know how much spent fuel is stored at Fukushima in casks? I assumed that since there is a separate spent fuel pool that there was no cask storage but this article suggests otherwisehttp://www.technologyreview.com/energy/37388/?nlid=4364&a=f An 18 March update on radiation levels around Fukushima from the DoE at their blog,http://blog.energy.gov/content/situation-japan/ Although it’s helpful in some ways, I find it a little irritating that they have only given a map of the estimated one-year dose, they have saturated their colour scale rather early at 20mSv, and they have included doses in the month just gone, likely more than half the dose. Is anyone aware of an accessible (less processed) version of current dose rates? Athttp://www.jaif.or.jp/english/there are occasional, most recently April 19th, PDFs with the titleTrend of Radiation in the Environment around Fukushima Daiichi NPS PDF. Also similar updates for radioactivity in the nearby sea. ( How fire can be domesticated) The following is a quote from a WSJ article in this mornings WSJ “Under its strategic plan unveiled Sunday, Tepco is planning to fill the containment vessels of the plant’s No. 1 and No. 3 reactor units with water and employ heat-exchanging systems that could include an air-cooling solution. Similar plans are in place for the heavily damaged No. 2 reactor unit but must await the repair of leaks that are allowing the escape of highly radioactive water contaminated by the damaged fuel rods.” The entire article can be found here.http://online.wsj.com/article/SB10001424052748703916004576270643486719236.html As read from the quote above, Tepco is planning to fill the “containment” vessels. This must be an error. I think they are just going to cool the water in the PRV by creating a new closed loop cooling system for the RPV and cooling this loop via an air cooled heat exchanger similer to french design. I would assume also that the air cooling would consist of an evap cooling tower. Can anyone clear up what is going on here?? Thx,GSB A better description can be found at IEEE: TEPCO Announces a “Roadmap to Restoration” at Fukushima Dai-1 A link is given there to theTEPCO description of the roadmap. I don’t detect a mention of filling the containment vessels with water. Mention is made of increasing the level of water to cover the “active fuel”. 1 Maintain Stable Cooling– Nitrogen gas injection– Flooding up to top of active fuel– Examination and implementation ofheat exchange function. 2 (Unit 2) Cool the reactor while controlling the increase of accumulated water until the PCV is sealed. Sounds more like they want to increase the water levels in the RPVs and for unit 2 to control the additional release of water until they can fix the hole in the suppression pool … Oops, sorry, spoke too soon. In the second pdf at the TEPCO site: Current Status [1] (Units 1 to 3) Cooling Achieved by water injection while there is partial damage to fuel pellets. -Continued injection of fresh water and further cooling measures are required. Countermeasure [1]: Injecting fresh water into theRPV by pumps.Risk[1]: Possibility of hydrogen explosion due to condensation of steam in the PCV when cooled, leading to increased hydrogen concentration. Countermeasure [2]: Injecting Nitrogen gas into the PCV (start from unit 1.) Countermeasure [3]: Consideration of flooding the PCV up to the top of active fuel. Thx for the response David, Perhaps they are anticipating a situation where they can not stop the leakage from the PRV in R2. I don’t know how they would repair leaking control rod seals. Pump seals could more easily be fixed. At any rate it is something “under consideration”. I would think that R1 and R2 would be cooled without flooding the CV’s. Set up a new closed loop on the PRV cooling water using an “air cooled” heat exchanger and that this air cooled heat exchanger would use air cooled water towers. Can anyone agree or disagree?? Oops. Revise last statement to R1 and R3. I would think that R1 and R3 would be cooled without flooding the CV’s. Set up a new closed loop on the PRV cooling water using an “air cooled” heat exchanger and that this air cooled heat exchanger would use air cooled water towers. Chhers, George. It would seem crazy to flood the PCV of R2 if there is a leak in the torus which is what I though they suspected … that would only make the leak worse. So maybe they’re thinking about that only for R1 and R3? For R2, it would seem they’ld have to fix the hole first. And that fix will have to be able to take the hydrostatic pressure, before they could flood it.I also don’t see how they could accomplish that fix with very high radiation levels that may exist near the leak. It would seem to me to be better to cut the flow rate to the RPV in R2 to the minimum necessary to continue cooling so as to limit the leakage, and like you say, try to set up some kind of air cooling with a closed loop, putting the heat exchanger in a lower radiation area. I think the other problem is that the CV may not be able to stand another earthquake when said CV is filled w/ water. So in summary I just don’t see flooding ANY of the CV’s w/ water as a likely route they would take. Perhaps we will get some video from the Robot. Apparently it gets it’s camera all fogged up w/ steam when they try to go inside the CV of R2. David,Cyril R posted this in the other thread. We are keeping the CV flooding issue here in this blog. CyrilR quote: Cyril R., on 20 April 2011 at 3:34 AM said: George, the latest JAIF update says ‘to be confirmed’ in the category water injection to containment vessel, for units 1 and 3. For unit 2, it says ‘to be decided (seawater)’. There is also the AREVA presentation which says they flooded at least 1 unit (unit 1 probably). George, yes, that’s a very good point aboutthe additional stress on the PCV and thechance of another earthquake. I did see the Areva presentation, and itcertainly suggested that flooding of R1 PCV hadbeen done. There were also hints that R1 hadhad a LOCA caused by the earthquake before thetsunami hit — it was in a video presentation by a Hitachi engineer. I’ll try to find a link. I understand why they would want to flood the containment in such a case, but I imagined itwould only be up to the level of the PRV bottomto try to prevent melting through. About the JAIF updates: I’ve never been quite clearwhat they mean by “water injection to containment.” Is there a separate water spray system inside the drywell, to be used for cooling to condense steam? Or do they really mean filling the whole thing up? I’ld really love to see the video from the robot. I hopeit works. Does anyone know how much spent fuel is stored at Fukushima in casks? Don’t know how much fuel is stored in casks but Tepco has a picture of a cask and cask mover on their website. Link with presentation from Dr. Matthias Braun (AREVA) for reference to the statement that PCVs flooding: It says at least one contaiment (unit 1) flooded. The JAIF references say ‘to be confirmed’ at containment flooding. Perhaps mistranslation but my definition of ‘to be confirmed’ is ‘we’ve done it but haven’t verified the exact effect’. Pump seals could more easily be fixed. At any rate it is something “under consideration”. I would think that R1 and R2 would be cooled without flooding the CV’s. In my simple mind, I think the problem is that the seals that are leaking are located inside the containment vessel which would make the inside of the containment vessel inaccessible. As far as the cooling system, it appears they are talking about an air cooling heat exchanger…I.E. A large radiator rather then getting the sea water pumps and condensers functioning again. Thx Cyril for the link, an excellent presentation. I found the reactor core isolation pump/cooling slide especially interesting. The reactor was able to run it’s cooling loop pump off the turbine that used reactor steam as an energy source. No outside source required for the pump, just battery power….pretty ingenious. Also R1 went the longest w/o cooling. Perhaps 1 or more of the wet wells ARE flooded as shown in the slide. Thanks GRLC. Only Iitate is above 2microsieverts/hr of those monitoring stations, and a rough calculation shows that the dose from 15-19 Mar at that location is equivalent to a year at the current dose rate (which will drop further). So for evacuation-lifting purposes, almost all areas should be re-opened unless there is a realistic threat of further releases from Fukushima Daiichi on the same scale as mid March. Certainly people should be allowed back to check on their houses and possessions, where the roads etc. permit. @George Bower – your PCV/containment flooding questionI confirmed the PCV flooding strategy in my commentson the 18th: TEPCO’s briefing slides (4) are availablehere. My speculation that they would flood the PCV appears to be correct: Countermeasure [9]: Flood the PCV up to the top of active fuel. I agree with David – if they achieve their objectives in 9 months I will be impressed and relieved. My understanding is that TEPCO knows little about the actual state of the machinery inside the buildings. And they have no experience of doing such construction in a hot environment – with or without robots. The TEPCO plan describes the strategy for R1 and R3 fairly clearly: first fix the leaks, seal the containment, then flood to at least the top of the fuel and recirculate and cool as you outlined. For R2 the torus leaks must first be repaired (the “sticky cement” plan?). The annotated TEPCO diagram was excellent. I’ve found two well-informed sources of analysis on the Daiichi stabilization plan. KBMAN worked 25 years ago on the same generation BWR/Type I containment. Hislatest on the TEPCO planincludes this One of the primary steps mentioned in their plan is to flood the containment vessels of units 1 and 3 up to the level of the fuel to guarantee the fuel remains covered and to assist in cooling. They are also looking at building external heat exchangers to be used in conjunction with filtration systems to be able to recycle the water being pumped through the reactors and also cool the water in the containments. This step also helps to provide more effective cooling for the fuel sediment on the bottom of the reactors by conducting heat away from the bottom of the reactor vessel. They also wish to do this for unit 2 but must first repair the damage to the containment vessel. There is mention of using “sticky cement” to make this repair, I’m not sure exactly what they mean by that. This is presumably the leak caused by the explosion in or near the torus during the first week of the emergency. Meanwhile, the leaks in the containment at unit 2 appear to be flowing directly to the service trench in the turbine building. They pumped thousands of gallons only to have the level rise back up again. They may end up needing to seal up the secondary leaks and treat the entire system – trench and all – as the “reactor vessel”. Stop adding new water and instead process the water from the trench and inject it back into the reactor. Will Davis offers very timely analysis.His firston the TEPCO announced plan discussed the PCV flooding strategy. The detailed releases by TEPCO do also indicate the desire to flood the PCV’s or Primary Containment Vessels (dry wells) up to the top of the active fuel region. The few illustrations released so far are not of what you might call a highly technical nature but they seem to indicate at least feed to the plant through the normal feed line and then removal of the water from the suppression pool. There is also one that shows a totally separate closed circuit cooling system that circulates only through the pressure vessel and a heat exchanger. Clearly the plans are in a state of flux and we’ll have to see how this shakes out. To help you understand why some of these considerations are of interest to us, look at the following illustration of an earlier style BWR reactor pressure vessel and internals. You’ve seen this before on this site, but it’s been marked up for this article… Thanks for your rough calculations and observation that “almost all areas should be re-opened unless there is a realistic threat of further releases”. The latest DOE dose-map slide I’ve seen was headed First-Year…commencing March 16. So that’s good, it does not sum up the earlier exposures. But as I read the map everything hotter than the blue will continue to be an exclusion zone. Am I missing something? Steve, I haven’t seen the future plans of the definition of the evacuation zone anywhere. I doubt they exist in any formal sense. For reference, the actual current evacuation zone is the inner circle on that presentation. But I don’t understand your “that’s good” remark – the significant impact on these areas was from 15 March onward, so practically all the initial higher-level dose rates are included in the calculation for that map. I’d like to see a map that starts its year from now, or at least from the start of April. It would look very different. The continuation of an exclusion zone when access would not be harmful is a repeat of the mass psychology mistakes identified after Chernobyl. This land is not blighted or uninhabitable and should not be treated as such. But I don’t understand your “that’s good” remark – the significant impact on these areas was from 15 March onward, so practically all the initial higher-level dose rates are included in the calculation for that map. I’d like to see a map that starts its year from now, or at least from the start of April. It would look very different. My bad – I wanted DOE to be doing what you said. That “must be” why I saw March 16 and thought April 16. There are alternative age-related hypotheses for synapse meltdowns like that… The continuation of an exclusion zone when access would not be harmful is a repeat of the mass psychology mistakes identified after Chernobyl. This land is not blighted or uninhabitable and should not be treated as such. I agree. I’ve not done my homework to verify what the projected dose would be. My guess is the authorities will continue the currently defined zone until they are confident there will not be any more “excitement” with new releases from NPP1. I.e., they are excluding in fear of new releases rather than based on existing contamination. Steve Darden, Article on long term exposure standardhttp://www.asahi.com/english/TKY201104110139.html 2.5 uSv=0.25mRem.The Japanese set the ‘long term’ exposure standard to 20 mSv per year or 2.5 uSv/hr.(using 8,000 hour year) The DOE Map from April 7th is more informative of current dose rate.http://www.slideshare.net/energy/radiation-monitoring-data-from-fukushima-area-04072011

https://bravenewclimate.com/2011/03/30/fukushima-technical-discussion-open-thread/?like_comment=125132&_wpnonce=84ec74e5ef

Galaxies | Free Full-Text | Debate on the Physics of Galactic Rotation and the Existence of Dark Matter This Special Issue was motivated by the disparate explanations of galactic dynamics promulgated by different philosophical camps [...] Debate on the Physics of Galactic Rotation and the Existence of Dark Matter by Anne M. Hofmeister and Robert E. Criss * Department of Earth and Planetary Science, Washington University, St. Louis, MO 63130, USA Author to whom correspondence should be addressed. Galaxies 2020 , 8 (3), 54; https://doi.org/10.3390/galaxies8030054 Received: 8 June 2020 / Revised: 14 June 2020 / Accepted: 1 July 2020 / Published: 15 July 2020 Keywords: spiral galaxies ; rotation curves ; gravitation ; dark matter This Special Issue was motivated by the disparate explanations of galactic dynamics promulgated by different philosophical camps. The most populous camp advocates Newtonian orbital models where stars revolve in a galactic disk; these models require huge amounts of dark matter to reconcile the observed rotation curves (RCs) of stars that are smoothly distributed in spiral galaxies with the behavior exhibited by the few planets independently orbiting in the Solar system. A competing view called MOND offers “modified Newtonian dynamics” to explain galactic rotation without dark matter; these models require modification of the classical law of gravitation, but not in a way that is consistent with general relativity. These ideas are irreconcilable and consensus does not exist. Consequently, a few individuals and small research groups have pursued alternate means to explain galactic rotation without requiring either dark matter or non-Newtonian physics. The overarching purpose of this Special Issue is to stimulate discussion and provide alternative ideas as a step towards an improved understanding of galactic dynamics. As originally conceived, a main goal was to assemble brief reviews by advocates of diverse viewpoints that summarize their preferred models while providing quantitative comparisons and cogent criticisms of other models. Our hope was to foster constructive criticism and communication between the various proponents. The second goal was to solicit novel explanations for galactic rotation, comprehensive analyses of RC, or new evidence for the existence of dark matter, which would shed light on various possibilities. A broad solicitation for contributions was issued, and numerous advocates of prevailing and novel models were specifically invited. Most of the manuscripts we received did not adhere to the requested format. However, all papers met the second goal of offering comprehensive analyses, new evidence, or novel approaches. One paper reviews the literature on MOND and applies this type of analysis to a new database [ 1 ]. Surface brightness vs. radius ( r ) extracted from a new database is flat and distance, which shows ordinary matter, is more abundant than assumed in many models [ 2 ]. Three quite different Newtonian inverse models are presented, which extract density (ρ) from RC. Results for ρ( r ), obtained for a thick disk numerically [ 3 ] and for the equatorial plane via matrix inversion [ 4 ], bracket those obtained analytically for a spinning oblate shape [ 5 ]. None of these three inverse models require dark matter, and neither does a forward Newtonian model which considers a log normal distribution of mass [ 6 ]. Despite our broad solicitation and many invitations, only one paper argues for non-baryonic halos [ 7 ]. This author also models dark matter near the Earth [ 8 ]. Thus, the desired debate was limited. Two explanations for this come to mind. One possibility is a recent development in particle physics, where multiple heavy hadron collider experiments in 2019 failed to yield evidence for non-baryonic matter. This non-result, which followed the discounting of neutrinos as halo constituents from 2015 observational collaborations, eliminated the other contender for non-baryonic matter, as no other means for its direct detection currently exist. The lack of debate may also reflect some troubling trends of our times. Research in the physical sciences has become highly compartmentalized, impeding effective communication outside one’s group. Over-specialization is driven by ongoing realities. One is that obtaining funding now requires researchers to strongly advocate their own work, rather than fairly assess their work in the context of current knowledge. In addition, the long-standing master–apprentice relationship in graduate school preferentially populates the research factions that are most successful in obtaining funding, favoring those with consensus views over those that innovate. Statistics of the reviewer pool and its anticipated feedback are among the obvious reasons for this outcome. Because the desired debate did not materialize, we reviewed the mathematical underpinnings of all RC models and also closely examined the methods by which Doppler measurements are converted to RCs [ 9 ]. We present evidence that “warps” represent a perpendicular axis of rotation, in accordance with Jacobi’s formulation for triaxial ellipsoids and SB morphology. We hope that the reader will examine the various alternatives presented in this issue with a look to the future. What needs to be done next to better understand the dynamics of spiral galaxies? Our opinion is that the Doppler data need reprocessing to address the facts that spiral galaxies are not optically thin in the z-direction and that density grades outwards in z, while also accounting for the possibility of a second spin axis. Improvements in data processing, and refinements in existing inverse and forward models of RC, are needed before we can effectively address the evolutionary behavior of the amazing giants in our universe. This research received no external funding. Acknowledgments We thank the editors and staff of Galaxies for their considerable efforts in bringing the special issue to fruition. Conflicts of Interest The authors declare no conflict of interest. References McGaugh, S.S. Predictions and Outcomes for the Dynamics of Rotating Galaxies. Galaxies 2020 , 8 , 35. [ Google Scholar ] [ CrossRef ] Sipols, A.; Pavlovich, A. Surface Brightness Plateau in S4G Galaxies. Galaxies 2020 , 8 , 48. [ Google Scholar ] [ CrossRef ] Feng, J.Q. Rotating Disk Galaxies without Dark Matter Based on Scientific Reasoning. Galaxies 2020 , 8 , 9. [ Google Scholar ] [ CrossRef ][ Green Version ] Sipols, A.; Pavlovich, A. Dark matter dogma: A study of 214 galaxies. Galaxies 2020 , 8 , 36. [ Google Scholar ] [ CrossRef ] Criss, R.E.; Hofmeister, A.M. Density Profiles of 51 Galaxies from Parameter-Free Inverse Models of Their Measured Rotation Curves. Galaxies 2020 , 8 , 19. [ Google Scholar ] [ CrossRef ][ Green Version ] Marr, J.H. Entropy and Mass Distribution in Disc Galaxies. Galaxies 2020 , 8 , 12. [ Google Scholar ] [ CrossRef ][ Green Version ] Sofue, Y. Rotation curve of Milky Way. Galaxies 2020 , 8 , 37. [ Google Scholar ] [ CrossRef ] Sofue, Y. Gravitational Focusing of Low-Velocity Dark Matter on the Earth’s Surface. Galaxies 2020 , 8 , 42. [ Google Scholar ] [ CrossRef ] Hofmeister, A.M.; Criss, R.E. Debated Models for Galactic Rotation Curves: A Review and Mathematical Assessment. Galaxies 2020 , 8 , 47. [ Google Scholar ] [ CrossRef ] MDPI and ACS Style Hofmeister, A.M.; Criss, R.E. Debate on the Physics of Galactic Rotation and the Existence of Dark Matter. Galaxies 2020, 8, 54. https://doi.org/10.3390/galaxies8030054 AMA Style Hofmeister AM, Criss RE. Debate on the Physics of Galactic Rotation and the Existence of Dark Matter. Galaxies. 2020; 8(3):54. https://doi.org/10.3390/galaxies8030054 Chicago/Turabian Style Hofmeister, Anne M., and Robert E. Criss. 2020. "Debate on the Physics of Galactic Rotation and the Existence of Dark Matter" Galaxies8, no. 3: 54. https://doi.org/10.3390/galaxies8030054 MDPI and ACS Style AMA Style Hofmeister AM, Criss RE. Debate on the Physics of Galactic Rotation and the Existence of Dark Matter. Galaxies. 2020; 8(3):54. https://doi.org/10.3390/galaxies8030054 Chicago/Turabian Style Hofmeister, Anne M., and Robert E. Criss. 2020. "Debate on the Physics of Galactic Rotation and the Existence of Dark Matter" Galaxies8, no. 3: 54. https://doi.org/10.3390/galaxies8030054

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Jiang Lin | Electricity Markets and Policy Group Jiang Lin Dr. Jiang Lin is the Nat Simons Presidential Chair in China Energy Policy at the Lawrence Berkeley National Lab, a Staff Scientist at its Department of Electricity Market and Policy, and an Adjunct Professor at the Department of Agricultural and Resource Economics at the University of California at Berkeley. Dr. Lin's research is focused on energy and climate policy, energy and emissions pathways with a focus on non-CO2 GHGs (methane, F-gas, etc.), electricity market and planning, low-carbon economic transition, and appliance efficiency issues in China. From 2016-2020, he was a co-Director of the Berkeley-Tsinghua Joint Research Center on Energy and Climate Change, a collaborative initiative between Berkeley Lab, the University of California-Berkeley, and Tsinghua University in China. From 2007-2016, Dr. Lin was the Director of the Energy Foundation's China Sustainable Energy Program (2007-2013) and Senior Vice President for Strategy and Analysis (2014-2016). Dr. Lin managed the growth of Energy Foundation China into one of the largest international NGOs devoted to promoting clean energy and climate solutions in China. Before joining the Energy Foundation, Dr. Lin was previously at LBNL from 1994-2007, researching the Appliance Standards and China Energy Groups. Dr. Lin has a Ph.D. in Demography from the University of California-Berkeley, an MS in Population Studies, and a BS from the Department of Cybernetics Engineering from Xi'an Jiaotong University, China. [email protected] (link sends e-mail) (510) 495-8886 1 Cyclotron Road, Berkeley, CA 94720 USA Jiang Lin (active tab) Publications Dr. Jiang Lin is the Nat Simons Presidential Chair in China Energy Policy at the Lawrence Berkeley National Lab, a Staff Scientist at its Department of Electricity Market and Policy, and an Adjunct Professor at the Department of Agricultural and Resource Economics at the University of California at Berkeley. Dr. Lin's research is focused on energy and climate policy, energy and emissions pathways with a focus on non-CO2 GHGs (methane, F-gas, etc.), electricity market and planning, low-carbon economic transition, and appliance efficiency issues in China. From 2016-2020, he was a co-Director of the Berkeley-Tsinghua Joint Research Center on Energy and Climate Change, a collaborative initiative between Berkeley Lab, the University of California-Berkeley, and Tsinghua University in China. From 2007-2016, Dr. Lin was the Director of the Energy Foundation's China Sustainable Energy Program (2007-2013) and Senior Vice President for Strategy and Analysis (2014-2016). Dr. Lin managed the growth of Energy Foundation China into one of the largest international NGOs devoted to promoting clean energy and climate solutions in China. Before joining the Energy Foundation, Dr. Lin was previously at LBNL from 1994-2007, researching the Appliance Standards and China Energy Groups. Dr. Lin has a Ph.D. in Demography from the University of California-Berkeley, an MS in Population Studies, and a BS from the Department of Cybernetics Engineering from Xi'an Jiaotong University, China. ETA Staff Page (link is external) [email protected] (link sends e-mail) (510) 495-8886 1 Cyclotron Road, Berkeley, CA 94720 USA 2023 Chen, Yidan , Jiang Lin , David Roland-Holst , Xu Liu , and Can Wang . " Declining Renewable Costs, Emissions Trading, and Economic Growth: China’s Power System at the Crossroads ." Energies (2023). Shiraishi, Kenji , Won Young Park , Nikit Abhyankar , Umed Paliwal , Nina Khanna , Toru Morotomi , Jiang Lin , and Amol A Phadke . The 2035 Japan Report: Plummeting Costs of Solar, Wind, and Batteries Can Accelerate Japan’s Clean and Independent Electricity Future . 2023. LBNL-2001526. Related Files Report (English) PDF (6.92 MB) Summary (Japanese) PDF (3.17 MB) Luo, Qian , Fernando Garcia-Menendez , Haozhe Yang , Ranjit Deshmukh , Gang He , Jiang Lin , and Jeremiah X Johnson . " The Health and Climate Benefits of Economic Dispatch in China’s Power System ." Environmental Science & Technology 57.7 (2023) 2898–2906. Wang, Wenjun , Nina Khanna , Jiang Lin , and Xu Liu . " Black carbon emissions and reduction potential in China: 2015–2050 ." Journal of Environmental Management 329 (2023). 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Luke 4:6 (KJV) - Barnes' Book Notes And the devil said unto him, All this power will I give thee, and the glory of them: for that is delivered unto me; and to whomsoever I will I give it. Luke 4:6  ( King James Version ) Book Notes: Preface to Luke Little is "certainly" known concerning the time and place of writing this Gospel, or concerning the author. The first time we have any mention of the author is in his own history,Acts 16:10-11. He was then the companion of Paul in his travels, and it is evident that he often attended Paul in his journeys, compareActs 16:11-17;Acts 21:1-6. In each of these places the author of "the Acts " speaks of "his" being in company with Paul. That the same person was the writer of this Gospel is also clear fromActs 1:1. From this circumstance the ancients regarded this Gospel as in factthe Gospelwhich Paul had preached. They affirm that Luke recorded what the apostle preached. Thus, Irenaeus says, "Luke, the companion of Paul, put down in a book the gospel preached by him." He also says, "Luke was not only a companion, but also a fellow-labourer of the apostles, especially of Paul." Origen, speaking of the Gospels, says, "The third is that according to Luke, the gospel commended by Paul, published for the sake of the Gentile converts." The testimony of the fathers is uniform that it was written by Luke, the companion of Paul, and was, therefore, regarded by them as really the gospel which Paul preached. It is not known Where it was written. Jerome says it was composed in Achaia. There seems to be some probability that it was written to persons that were well acquainted with Jewish manners, as the author does not stop to explain the peculiar customs of the Jews, as some of the other evangelists have done. Respecting the time when it was written nothing very definite is known. All that can with certainty be ascertained is that it was written before the death of Paul (65 a.d.), for it was written before the ActsActs 1:1, and that book only brings down the life of Paul to his imprisonment at Rome, and previous to his going into Spain. It has been made a matter of inquiry whether Luke was a Gentile or a Jew. On this subject there is no positive testimony. Jerome and others of the fathers say that he was a Syrian, and born at Antioch. The most probable opinion seems to be that he was a proselyte to the Jewish religion, though descended from Gentile parents. For this opinion two reasons may be assigned of some weight. First, he was intimately acquainted, as appears by the Gospel and the Acts , with the Jewish rites, customs, opinions, and prejudices; and he wrote in their "dialect," that is, with much of the Hebrew phraseology, in a style similar to the other evangelists, from which it appears that he was accustomed to the Jewish religion, and was, therefore, probably a proselyte. Yet the "preface" to his Gospel, as critics have remarked, is pure classic Greek, unlike the Greek that was used by native Jews; from which it seems not improbable that he was by birth and education a Gentile. second. InActs 21:27, it is said that the Asiatic Jews excited the multitude against Paul because he had introduced "Gentiles" into the temple, thus defiling it. InActs 21:28it is said that the Gentile to whom they had reference was "Trophimus," an Ephesian. Yet "Luke" was also at that time with Paul. If he had been regarded as "a Gentile" it is probable that they would have made complaint respecting "him" as well as "Trophimus;" from which it is supposed that he was a Jewish proselyte. But again, in the Epistle to the Colossians,Colossians 4:9-11, we find Paul saying that Aristarchus, and Marcus, andBarnabas, and Justus saluted them, "who are," he adds, "of the circumcision," that is, Jews by birth. InColossians 4:14he says that "Luke," the beloved physician, and Demas also saluted them; from which it is inferred that they were "not of the circumcision," but were by birth Gentiles. Most writers suppose that Luke, the writer of this Gospel, was intended in the above place in Colossians. If so, his profession was that of "a physician;" and it has been remarked that his descriptions of diseases are more accurate and circumstantial, and have more of "technical" correctness than those of the other evangelists. Luke does not profess to have been an eye-witness of what he recorded. SeeLuke 1:2-3. It is clear, therefore, that he was not one of the seventy disciples, nor one of the two who went to Emmaus, as has been sometimes supposed. Nor was he an apostle. By the fathers he is uniformly called the "companion" of the apostles, and especially of Paul. If he was not one of the apostles, and if he was not one of those expressly commissioned by our Lord to whom the promise of the infallible teaching of the Holy Ghost was given, the question arises by what authority his Gospel and the Acts have a place in the sacred canon, or what evidence is there that he was divinely inspired? In regard to this question the following considerations may give satisfaction: 1. They were received by all the churches on the same footing as the first three Gospels. There is not a dissenting voice in regard to their authenticity and authority. The value of this argument is this - that if they had been spurious, or without authority, the fathers were the proper persons to know it. 2. They were published during the lives of the apostles Peter, Paul, and John, and were received during their lives as books of sacred authority. If the writings of Luke were not inspired, and had no authority, those apostles could easily have destroyed their credit, and we have reason to think it would have been done. 3. It is the united testimony of the fathers that this Gospel was submitted to Paul, and received his express approbation. It was regarded as the substance of his preaching, and if it received his approbation it comes to "us" on the authority of his name. Indeed, if this be the case, it rests on the same authority as the epistles of Paul himself. 4. It bears the same marks of inspiration as the other books. It is simple, pure, yet sublime; there is nothing unworthy ofGod; and it is elevated far above the writings of any uninspired man. 6. If it be objected that, not being an apostle, he did not come within the "promise" of inspirationJohn 14:26;John 16:13-14made to the apostles, it may be replied that this was also the case with Paul; yet no small part of the New Testament is composed of his writings. The evidence of the inspiration of the writings of Luke and Paul is to be judged, not only by that "promise," but by the early reception of the churches; by the testimony of the fathers as to the judgment of "inspired men" when living, and by the internal character of the works. Luke has all these equally with the other evangelists.

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Bandha Branch Post Office, Sihora 22, Katni, Madhya Pradesh Bandha Post Office, Sihora Bandha post office is located at Bandha, Sihora, Katni of Madhya Pradesh state. It is a branch office (B.O.). A Post Office (PO) / Dak Ghar is a facility in charge of sorting, processing, and delivering mail to recipients. POs are usually regulated and funded by the Government of India (GOI). Pin code of Bandha PO is483222. This Postoffice falls under Jabalpur postal division of the Madhya Pradesh postal circle. The related head P.O. for this branch office is Katni head post office and the related sub-post office (S.O.) for this branch office is Gosalpur post office. Bandha dak ghar offers all the postal services like delivery of mails & parcels, money transfer, banking, insurance and retail services. It also provides other services including passport applications, P.O. Box distribution, and other delivery services in Bandha. Types of Post Offices Post offices are basically classified into 3 types, namely – Head Post Office, Sub-Post Office including E.D. Sub-Office and Branch Postoffice. Bandha P.O. is a Branch Post Office. So far as the public is concerned, there is basically no difference in the character of the service rendered by Sub-Post Offices and Head-Post Offices except in regard to a few Post Office Savings Bank (SB) transactions. Certain Sub Post Offices do not undertake all types of postal business. Facilities are generally provided at Branch Post Offices for the main items of postal work like delivery and dispatch of mails, booking of registered articles and parcels accepting SB deposits and effecting SB withdrawals, and issue and payment of money orders, though in a restricted manner. Post Office Type Head Post Office Sub-Post Offices including E.D. Sub-Offices Branch Post Office Bandha Post Office & Its Pin Code Branch Office Information Bandha Post Office Services Mail Services Parcels Retail Services Premium Services Speed Post India Post Speed Post Tracking Tracking System India Post Tracking Number Formats Express Parcel Post Media Post Greetings Post Logistics Post ePost Office Financial Services Savings Bank (SB) Account Recurring Deposit (RD) Account Monthly Income Scheme (MIS) Monthly Public Provident Fund (PPF) Time Deposit (TD) Senior Citizen Saving Scheme (SCSS) National Savings Certificate (NSC) Kisan Vikas Patra (KVP) Sukanya Samriddhi Accounts (SSA) Post Office Timings India Post Tracking Bandha Post Office Recruitment Location Map Contact Details About India Post Bandha Post Office & Its Pin Code Often Post Offices are named after the town / village / location they serve. The Bandha Post Office has the Postal Index Number or Pin Code 483222. A Pincode is a 6 digit post code of postal numbering system used by India Post. The first digit indicates one of the regions. The first 2 digits together indicate the sub region or one of the postal circles. The first 3 digits together indicate a sorting / revenue district. The last 3 digits refer to the delivery post office type. P.O. Name Bandha PO Pincode 483 222 The first digit of 483222 Pin Code '4' represents the region, to which this Post Office of Bandha belongs to. The first two digits of the Pin Code '48' represent the sub region, i.e, Madhya Pradesh. The first 3 digits '483' represent the post-office revenue district, i.e, Jabalpur. The last 3 digits, i.e, '222' represent the Bandha Delivery Branch Office. Branch Office Information The Bandha Post Office is a branch office. The Delivery Status for this PO is that it has delivery facility. Postal division name for this Dak Ghar is Jabalpur, which falls under Indore region. The circle name for this PO is Madhya Pradesh and it falls under Sihora Taluka and Katni District. The state in which this Dakghar is situated or located is Madhya Pradesh. The related head postoffice is Katni post office and the related sub post office is Gosalpur post-office. The phone number of Bandha post office is unavailable at present. PO Type Branch Office Delivery Status Delivery Postal Division Jabalpur Postal Region Indore Postal Circle Madhya Pradesh Town / City / Tehsil / Taluka / Mandal Sihora District Katni State Madhya Pradesh Related Sub PO Gosalpur Sub Office Related Head PO Katni Head Post Office Bandha Post Office Services Traditionally the primary function of Bandha post office was collection, processing, transmission and delivery of mails but as of today, a Post Office offers many other vital services in addition to its traditional services. The additional services provided by a Dak Ghar include – Mail Services, Financial Services, Retail Services and Premium Services. Mail Services Mail Services are the basic services provided by Bandha P.O. Mails and mail services include all or any postal articles whose contents are in the form of message which may include Letters, Postcards, Inland letter cards, packets or parcels, Ordinary mails etc. Parcels Mail Service also includes transmission and delivery of Parcels. A parcel can be anything ranging from a single written letter or anything addressed to an addressee. No parcel shall be by any chance be in a shape, way of packing or any other feature, such that it cannot be carried or transmitted by post or cause serious inconvenience or risk. Every parcel (including service parcels) that needs to be transmitted by post must be handed over at the window of the post office. Any parcel found in a letter box will be treated and charged as a registered parcel. Delivery services are provided by some selected delivery and branch post offices. This dakghar have the facility of delivery, thus the people of Bandha and nearby localities can avail all the types of mail services. Retail Services Post offices in India serve in various ways and Bandha Post Office offer most of the retail services. They offer the facility to accept or collect constomer bills like telephone or mobile bills, electricity bills for Government and private organizations through Retail Post. Some of the aditional agency services that Post offices offers through retail services are as follows - Telephone revenue collection, e-Ticketing for Road Transport Corporations and Airlines, Sale of UPSC forms, university applications, Sale of Passport application forms, Sale of Gold Coins, Forex Services, Sale of SIM and recharge coupons, Sale of India Telephone cards, e-Ticketing of Railway tickets etc. The postal customers of Bandha can pay their bills and avail other retail services from this Dak Ghar. Premium Services Most of the premium services can be availed by the Bandha peoples and nearby living people. The premium services provided by Bandha Post Office are - Speed Post, Business Post, Express Parcel Post, Media Post, Greeting Post, and Logistics Post. Speed Post Speed Post is a time bound service in express delivery of letters and parcels. The max weight up to which an article or parcel be sent is 35 kgs between any two specified stations in India. Speed Post delivers 'Value for money' to everyone and everywhere, delivering Speed Post upto 50 grams @ INR 35 across the country and local Speed Post upto 50 grams @ INR 15, excluding applicable Service Tax. Kindly check official website for updated Speed Post service charges. India Post Speed Post Tracking Speed Post offers a facility of on-line tracking and tracing that guarantees reliability, speed and customer friendly service. Using a 13 digit barcode that makes a Speed Post consignment unique and identifiable. A web-based technology (www.indiapost.gov.in/speednettracking.aspx) helps the Bandha customers track Speed Post consignments from booking to delivery. Tracking System Except Speed Post, India Post also allows people to track their order information for certain products like Parcels, Insured letters, Speed Post, Registered Post, Electronic Money Orders (EMO) and Electronic value payable parcel (EVPPs) etc. The tracking number is available on the receipt given at Bandha Post Office. Using the tracking number postal customers can find out the date and time of dispatch of an article at various locations. The time of booking and the time of delivery of article. India Post Tracking Number Formats Different types of postal service have different kinds of tracking number formats. The tracking number for Express Parcel is a 13 digit alphanumeric format. The format for Express Parcel is XX000000000XX. The tracking number for a Registered Mail is a 13 digit alphanumeric number and its format is RX123456789IN. But a Electronic Money Order (EMO) has a 18 digit tracking number and its format is 000000000000000000. For domestic Speed Post (EMS) there is a 13 digit alphanumeric tracking number with the format EE123456789IN. Bharatiya Dak Ghar Seva Tracking Number Format Number of Digits Electronic Money Order (eMO) 000000000000000000 18 Express Parcel XX000000000XX 13 International EMS Artilces to be delivered in India EE123456789XX 13 Registered Mail RX123456789IN 13 Speed Post (EMS) Domestic EE123456789IN 13 Express Parcel Post In Express Parcel Post, the Bandha postal customer gets time bound delivery of parcels. These parcels will be transmitted through air or any other fastest mean available at that time. Minimum chargeable weight for which Express Parcel consignments will be booked is 0.5 Kg. Maximum weight of Express Parcel consignments which shall be booked across the Post Office counter by a retail customer shall be 20 Kg and maximum weight that can be booked by corporate customer is 35 kgs. Media Post India Post offers a unique way or concept to help the Indian corporate organisations and the Government organizations reach potential customers through media post. Through media post people can advertise on postcards, letters, aerogramme, postal stationary etc. Customers get to see the logo or message of the respective corporate or government organizations. The Aerogramme even gives the organizations the opportunity to make their product have a global impact. Greetings Post Greeting Post is yet another innovative or unique step by India Post. It consists of a card with an envelope with pre-printed and pre attached postage stamp on the envelope. The stamp on the envelope is a replica of the design that appears upon the card but in miniature form. Thus there is no need affix postage stamps on the envelope implicitly saving your time of going to post offices and standing in the queue. All the rules and that are applicable for the postage dues will also be applicable to the Greeting Post. Logistics Post Logistics Post manages the entire transmission and distribution side of the parcels. It deals with collection of goods, storage of goods, carriage and distribution of the various parcels or goods, from order preparation to order fulfilment. And that too at the minimum possible price. Logistics Post services provides the Bandha postal customer with cost-effective and efficient distribution across the entire country. ePost Office The advent of internet made communication very rapid through emails. But, the internet has not yet reached most of the rural parts of India. To change this division between rural & urban life, and to get the benefit of internet technology to Bandha people's lives, Indian Postal Department has introduced e-post. e-post is a service in which personalized handwritten messages of customers are scanned and sent as email through internet. And at the destination address office, these messages are again printed, enveloped and delivered through postmen at the postal addresses. E-post centres are established in the Post Offices, covering a large geographical area including major cities and districts. These e-post centres are well equipped with internet connection, scanners, printers and other necessary hardware equipment. However, this e-post service doesn’t particularly need a e-post centre, but can this facility can be availed at any normal Post Office or you can visit www.epostoffice.gov.in to access postal services on your desktop, laptop or even on mobile. If a message is booked at Bandha post office, the post is scanned and sent to an e-post centre by e-mail and a mail received at e-post centre is printed and sent to nearby Post Office for dispatch. A Bandha customer can also avail these services of an e-post, at his/ her home. All he/ she has do is to register as a user at www.epostoffice.gov.in website. After registration, a user can use e-post by scanning and sending messages, printing and receive messages. The message to be scanned must not be written in a paper not more A4. There is no limit for sending number of sheets of messages in e-post. E-Post Office offers certain services like – Philately, Postal Life Insurance, Electronic Indian Postal Order, Information Services, Track & Trace and Complaints & Guidelines services. Philately Philately service deals with collection, sale and study of postage stamps. Philately includes lot of services Philately Information, Stamp issue Program, Stamps List and Buy Stamps service. Postal Life Insurance (PLI) A service offered by the Government to pay a given amount of money on the death of an individual to his prescribed nominee. The amount may also be paid to the person himself, in case he survives that maturity period. The two services offered under Postal Life Insurance are – Pay Premium service and PLI information. Electronic Indian Postal Order eIPO or Electronic Indian Postal Order is a facility to purchase an Indian Postal Order electronically by paying a fee on-line through e-Post Office. This service is launched by the Department of Posts, Ministry of Communications & IT, Government of India. eIPO can now be used by Indian Citizens living in India for paying online fee, whoever seeks information under the RTI Act, 2005. eIPO offers 2 types of services – eIPO information and payment of online fees. Information Services This helps Bandha customers to get information regarding certain products like – Pin Code search, Speed post, Banking, Insurance, Business Post, Logistics Post, IMTS and many more other services. Track & Trace The track & trace service is very helpful as it aids in getting information of our valuables. Track & Trace service offers 5 different services – Pin Code search, EMO tracking, Speed Post tracking, WNX tracking and International mail service. Complaints & Guidelines Using e-post office service Bandha postal costumer can access services based on – complaint registration, complaint status and guidelines on complaints. ePost Office Website www.epostoffice.gov.in Financial Services The customers of Bandha can enjoy the various savings schemes available in this post office that prove to be highly beneficial for the people living in Bandha area. The Financial service offered by PO includes Savings and Postal Life Insurance (PLI). There are various options available to save and invest with post-offices. The commonly used ones include - Savings account, Recurring Deposit, Monthly Income Scheme, Monthly Public Provident Fund, Time Deposit, Senior Citizen Saving Scheme, National Savings Certificate, Kisan Vikas Patra and Sukanya Samriddhi Yojana. Post Office also offers Insurance product through Postal Life Insurance (PLI) and Rural Postal Life Insurance (RPLI) schemes that offer low premium and high bonus. Post Office Financial Services Kisan Vikas Patra (KVP) Monthly Income Scheme (MIS) Monthly Public Provident Fund (PPF) National Savings Certificate (NSC) Recurring Deposit (RD) Account Savings Bank (SB) Account Senior Citizen Saving Scheme (SCSS) Sukanya Samriddhi Accounts (SSA) Time Deposit (TD) Savings Bank (SB) Account A Savings bank account serves the need of regular deposits for its customers as well as withdrawals. Cheque facility is also avail by Bandha postal consumers. Recurring Deposit (RD) Account A post office offers a monthly investment option with handsome return at the time period with an option to extend the investment period. Insurance facility is also available with certain conditions. Monthly Income Scheme (MIS) MIS offers a fixed investment technique for five or more years with monthly interest payment to the account holder. There is also a facility of automatic crediting of interest to SB account of the Bandha postal customer. Monthly Public Provident Fund (PPF) This service offers intermittent deposits subject to a particular limit for a time period of 15 years with income tax exemptions, on the investment. It also offers loan and withdrawal facilities for the postal customers. Time Deposit (TD) Fixed deposit option for periods ranging from one, two, three to five years with facility to draw yearly interest offered at compounded rates. Automatic credit facility of interest to SB account. Senior Citizen Saving Scheme (SCSS) Offers fixed investment option for senior citizens for a period of five years, which can be extended, at a higher rate of interest that are paid in quarterly instalments. National Savings Certificate (NSC) NSC is offered with a fixed investment for 5 or 10 years on certificates of various denominations. Pledging facility available for availing loan from Banks. Kisan Vikas Patra (KVP) Kisan Vikas Patra is a saving certificate scheme in which the amount Invested doubles in 110 months (i.e. 9 years & 2 months). It is available in denominations of Rs 1,000, 5000, 10,000 and Rs 50,000. Minimum deposit is Rs 1000/- and there is no maximum limit. The KVP certificate can be purchased by any adult for himself or on the behalf of a minor. This certificate can also be transferred from one account holder to another and from one post office to another. This certificate can be en-cashed only after 2 and 1/2 years from the date of issue. Sukanya Samriddhi Accounts (SSA) Sukanya Samriddhi Account Yojana offers a small deposit investment for the girl children as an initiative under 'Beti Bachao Beti Padhao' campaign. This yojana is to facilitate girl children proper education and carefree marriage expenses. One of the main benefits of this scheme is that it is very affordable and offers one of the highest interest rates. Currently its interest rate is set as 8.6% per annum that is again compounded yearly. The minimum deposit allowed in a financial year is INR. 1000/-and Maximum is INR. 1,50,000/-. Subsequent deposits can be made in multiples of INR 100/-. Deposits can be made all at a time. No limit is set on number of deposits either for a month or a financial year. A legal Guardian can open an account in the name of a Girl Child. Account can be closed only after completion of 21 years of the respective child. The normal Premature closure allowed is after completion of 18 years only if that girl is getting married. Post Office Timings The official working hours of Post Offices vary from one another, but the general Post Office opening time starts from 09:00 AM or 10:00 AM and the closing time is 06:00 PM or 07:00 PM respectively. The working days are from Monday to Saturday, Sunday being a holiday. This doesn't include the public holidays or the extended working hours. You can verify the working hours of Bandha Branch Post Office from the official resources. India Post Tracking Online tracking of India Post allowed Bandha people to access their postal article tracking information and confirm the delivery of their postal article by using the tracking number assigned to them at the time of Booking. They can find the tracking number on the Postal acknowledgement handed over to them at the Bandha Branch Post Office counter at the time of postal article booking. Following items can track through the www.indiapost.gov.in/articleTracking.aspx official website. Business Parcel Business Parcel COD Electronic Money Order (e-MO) Electronic Value Payable Parcel (eVPP) Express Parcel Express Parcel COD Insured Letter Insured Parcel Insured Value Payable Letter Insured Value Payable Parcel International EMS Registered Letter Registered Packets Registered Parcel Registered Periodicals Speed Post Value Payable Letter Value Payable Parcel The India Post tracking system is updated at regular intervals to give the Bandha postal customers with the most up to date information available about the location and status of their postal article. They'll be able to find out the following: When their postal article was booked When their postal article was dispatched at various locations during its Journey When their postal article was received at various locations during its Journey When their postal article was delivered, or When a delivery intimation notice was issued to notify the recipient that the postal article is available for delivery Bandha Post Office Recruitment For latest Bandha post office recruitment kindly visit www.indiapost.gov.in/recruitment.aspx. Location Map Bandha Branch Post Office is located in Bandha, Sihora, Katni. Contact Details All the queries or complaints regarding Bill Mail Service, Booking Packets, Business Post, Direct Post, Flat Rate Box, Indian Postal Orders, Inland Letters, Instant Money Orders, Insurance of Postal Articles, Insurance of Postal Parcels, Letters, Logistics Posts, MO Videsh, Money Orders, Parcels, Post Office Savings Bank, Postal Life Insurance, Postcards, Registration of Postal Articles, Registration of Postal Parcels, Rural Postal Life Insurance, Saving Certificates, Small Saving Schemes, Speed Post, Value Payable Post etc. services in Bandha Post Office, can be resolved at Bandha Branch Post Office. You can send letters to "Postmaster, Bandha Branch Post Office, Bandha, Sihora, Katni, Madhya Pradesh, India, Pincode: 483 222". The official website of the Berhampur University Sub Office is www.indiapost.gov.in. Bandha Branch Office Address: Bandha Branch Post Office, Bandha, Sihora, Katni, Madhya Pradesh, India Pin Code: 483222 Website: www.indiapost.gov.in About India Post India Post is a government-operated postal system, which is part of the Ministry of Communications and Information Technology of the Government of India. It has the largest Postal Network in the India with over 154882 Post Offices. There are around 139182 Post Offices in the rural India and 15700 Post Offices in urban India. The individual post office serves an area of 21.22 Sq. Km. and a population of 8221 people. The slogan of India Post is Dak Seva Jan Seva. There are 25464 departmental post offices and 129418 extra-departmental branch post offices in India. Bandha Post Office Summary Dak Ghar Name Bandha Branch Post Office Pincode 483222 Dakghar Type Branch Office Post Office Delivery Status Delivery Branch Office Postal Division Jabalpur Postal Region Indore Postal Circle Madhya Pradesh Location Bandha Town / City / Tehsil / Taluka / Mandal Sihora District Katni State Madhya Pradesh Country India Related Sub Office Gosalpur Sub Office Related Head Office Katni Head Post Office Website www.indiapost.gov.in ePost-office Web Site Address www.epostoffice.gov.in Speed Post Tracking Website www.indiapost.gov.in/speednettracking.aspx Recruitment Web Site Address www.indiapost.gov.in/recruitment.aspx

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PEOPLE v. ROSA | 138 A.D.2d 753 | N.Y. App. Div. | Judgment | Law | CaseMine Get free access to the complete judgment in PEOPLE v. ROSA on CaseMine. PEOPLE v. ROSA 138 A.D.2d 753 Case Information CITATION CODES 138 A.D.2d 753 ATTORNEY(S) Smart Summary Important Paras As the Court of Appeals has observed, "[a] defendant has a constitutional right to a trial by a `particular jury chosen according to law, in whose selection [the defendant] has had a voice'" (People v. Buford , 69 N.Y.2d 290, 297-298 , quoting from People v. Ivery , 96 A.D.2d 712 ; see , N.Y. Const, art I, § 2; People v. Anderson , 70 N.Y.2d 729, 730 ). Moreover, in order "[t]o protect this constitutional right in criminal cases, the Legislature has enacted several procedural safeguards" (People v Buford, supra , at 298) among which is CPL 270.35, pursuant to which the jury forewoman at bar was discharged. The Court of Appeals in People v. Anderson (supra , at 730-731) further noted that the denial to a defendant of a chosen jury on an improper basis is a deprivation of the constitutional right to a jury trial, to which harmless error analysis is inapplicable. Go to CPL 270.35 provides, in pertinent part, that the trial court must discharge a juror "[i]f at any time after the trial jury has been sworn and before the rendition of its verdict, a juror is unable to continue serving by reason of illness or other incapacity, or for any other reason is unavailable for continued service". The statute, it has been observed, "does not define `illness or other incapacity' and, as a result, the language should be construed in accordance with its common, everyday meaning, thereby permitting the court, in the exercise of its discretion, to make a determination as to incapacity to serve on a case-by-case basis" (People v. Washington , 131 A.D.2d 118, 120 , lv granted 70 N.Y.2d 938 ; People v. Pierce , 97 A.D.2d 904 ; cf., People v. Burns , 118 A.D.2d 864 ; McShall v. Henderson , 526 F. Supp. 158 ). Go to The record reveals that on Friday, February 8, 1985, shortly before the close of testimony, the jury forewoman — a Seventh Day Adventist — informed the court that by virtue of her religious beliefs she would be unable to remain at the court-house after dark, although she would be able to resume deliberations the following Monday. The court expressed its intention to complete testimony, finish summations, and charge the jury that afternoon. After discussing the matter with counsel, the court stated that it preferred to discharge the forewoman if summations and the jury charge could be completed on Friday. In support of its decision, the court expressed the concern that if deliberations were delayed over the weekend, the jurors' recollections could be dimmed. The court further stated that, "given the fact I have another case that is slated to start on Monday, I am going to excuse this juror and replace her with an alternate". Go to When construed in terms of their common, everyday meaning (People v. Washington, supra , at 120), and tempered by consideration of the fundamental nature of the right involved, we are of the view that the statutory phrases "incapacity" and "unavailab[ility] for continued service" do not embrace the circumstances with which the court was confronted in this case. At bar, the forewoman was available for continued service immediately after her religious observances were completed, which would have required only a brief, weekend hiatus in submitting the case to the jury. Go to PEOPLE v. ROSA Ordered, that the judgment is reversed, on the law, and a new trial is ordered; no questions of fact have been raised or considered. The record reveals that on Friday, February 8, 1985, shortly before the close of testimony, the jury forewoman — a Seventh Day Adventist — informed the court that by virtue of her religious beliefs she would be unable to remain at the court-house after dark, although she would be able to resume deliberations the following Monday. The court expressed its intention to complete testimony, finish summations, and charge the jury that afternoon. After discussing the matter with counsel, the court stated that it preferred to discharge the forewoman if summations and the jury charge could be completed on Friday. In support of its decision, the court expressed the concern that if deliberations were delayed overthe weekend, the jurors' recollections could be dimmed. The court further stated that, "given the fact I have another case that is slated to start on Monday, I am going to excuse this juror and replace her with an alternate". In response, the defense counsel argued, inter alia, that "this has been such a short trial that there aren't so many facts that the jury is going to forget just because we have a hiatus of a weekend". The court subsequently noted that in order to accommodate the juror, it would be necessary to recess Friday afternoon and that to do so would result in "closing our courtroom for a whole afternoon for no reason, and I do not think that, in view of the fact that this Forewoman did not tell us ahead of time of the problem, that we should disrupt the trial in this way and have a long recess between the summations and the charge". Notably, there is nothing in the record which indicates that the jury forewoman was questioned in respect to the existence of potential conflicts resulting from religious beliefs. Nevertheless, on Friday afternoon, after summations had been completed, and prior to the court's charging of the jury, the forewoman was discharged over the defense counsel's objection and an alternate juror substituted in her place. The jury was charged that day, and, after deliberating for approximately 90 minutes, rendered its verdict of guilty. On appeal, the defendant argues, inter alia, that the court erred in discharging the jury forewoman. We agree. As the Court of Appeals has observed, "[a] defendant has a constitutional right to a trial by a `particular jury chosen according to law, in whose selection [the defendant] has had a voice'" (People v. Buford, 69 N.Y.2d 290, 297-298, quoting from People v. Ivery, 96 A.D.2d 712; see, N.Y. Const, art I, § 2; People v. Anderson, 70 N.Y.2d 729, 730). Moreover, in order "[t]o protect this constitutional right in criminal cases, the Legislature has enacted several procedural safeguards" (People v Buford, supra, at 298) among which is CPL 270.35, pursuant to which the jury forewoman at bar was discharged. The Court of Appeals in People v. Anderson (supra, at 730-731) further noted that the denial to a defendant of a chosen jury on an improper basis is a deprivation of the constitutional right to a jury trial, to which harmless error analysis is inapplicable. CPL 270.35 provides, in pertinent part, that the trial court must discharge a juror "[i]f at any time after the trial jury has been sworn and before the rendition of its verdict, a juror is unable to continue serving by reason of illness or otherincapacity, or for any other reason is unavailable for continued service". The statute, it has been observed, "does not define `illness or other incapacity' and, as a result, the language should be construed in accordance with its common, everyday meaning, thereby permitting the court, in the exercise of its discretion, to make a determination as to incapacity to serve on a case-by-case basis" (People v. Washington, 131 A.D.2d 118, 120, lv granted70 N.Y.2d 938; People v. Pierce, 97 A.D.2d 904; cf., People v. Burns, 118 A.D.2d 864; McShall v. Henderson, 526 F. Supp. 158). When construed in terms of their common, everyday meaning (People v. Washington, supra, at 120), and tempered by consideration of the fundamental nature of the right involved, we are of the view that the statutory phrases "incapacity" and "unavailab[ility] for continued service" do not embrace the circumstances with which the court was confronted in this case. At bar, the forewoman was available for continued service immediately after her religious observances were completed, which would have required only a brief, weekend hiatus in submitting the case to the jury. Moreover, while the objectives of maintaining calendar control and otherwise ensuring the efficient resolution of the court's business are certainly laudable goals, minor scheduling difficulties involving the utilization of courtroom space or the brief delay which may have ensued had the forewoman been accommodated are not meaningful counterweights to the defendant's constitutional right to trial by a jury in whose selection he has participated. Further, this case is distinguishable from those in which the nature and extent of the juror's incapacity or unavailability for continued service is not readily ascertainable at the time the issue of discharge arises (see, e.g., People v Washington, supra; cf., People v. Hewlett, 133 A.D.2d 417) or where the period of unavailability is such that a reasonable accommodation cannot be fashioned so as to facilitate retention of the juror without unduly delaying the trial proceedings (see, e.g., People v. Burns, supra).In light of the foregoing, the brief period of unavailability resulting from the forewoman's religious observance in this particular case cannot be characterized as a disability resulting in "incapacity" or "unavailab[ility] for continued service" within the meaning of CPL 270.35. Finally, the issues to be resolved in this case — in which only one count of the indictment was submitted to the jury — were neither factually nor legally complex, as the court itself remarked prior to administering its charge. Indeed, the recordreveals that the taking of testimony commenced only one day before the case was ultimately submitted to the jury and that the jury rendered its verdict after deliberating for only 90 minutes. Accordingly, there was no danger that the jurors' recollections would be dimmed by delaying the commencement of their deliberations for the weekend. We have reviewed the defendant's remaining contentions and find them to be without merit. Rubin, J.P., Kooper, Sullivan and Harwood, JJ., concur.

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BCM-HGSC | Eric Boerwinkle, Ph.D. | page 7 Associate Director, Baylor College of Medicine Human Genome Sequencing Center Contact information [email protected] Other positions Dean, UTHealth School of Public Health M. David Low Chair in Public Health Kozmetsky Family Chair in Human Genetics Professor, Human Genetics Center and Dept. of Epidemiology Research interests The research interests of Dr. Boerwinkle encompass the Eric Boerwinkle, Ph.D. Associate Director, Baylor College of Medicine Human Genome Sequencing Center Contact information [email protected] (link sends e-mail) Other positions Dean, UTHealth School of Public HealthM. David Low Chair in Public HealthKozmetsky Family Chair in Human GeneticsProfessor, Human Genetics Center and Dept. of Epidemiology Research interests The research interests of Dr. Boerwinkle encompass the genetic analysis of the common chronic diseases in humans, including coronary artery disease, hypertension, and non-insulin dependent (type II) diabetes. Dr. Boerwinkle received his B.S. in Biology from the University of Cincinnati in 1980, an M.A. in Statistics (1984), and M.S. and Ph.D. in Human Genetics (1985) from the University of Michigan, Ann Arbor where he served as Senior Research Associate in the Department of Human Genetics from 1985-1986. He joined the University of Texas-Houston Center for Demographic/ Population Genetics in 1986 as a Research Assistant and became Assistant Professor in the same year. In 1991 he joined the Department of Human Genetics at the School of Public Health, University of Texas-Houston Health Science Center as Associate Professor, in 1996 was promoted to Professor, and in 1997, Director of the Human Genetics Center. He became a faculty member of the Institute of Molecular Medicine in 1996 and became Professor and Director of the Research Center for Human Genetics. Dr. Boerwinkle is a member of the American Diabetes Association and the American Society of Human Genetics. The research interests of Dr. Boerwinkle encompass the genetic analysis of common chronic diseases in humans, including coronary artery disease, hypertension, and non-insulin dependent (type II) diabetes. This work includes localizing genes which contribute to disease risk, identification of potentially functional mutations within these genes, testing these candidate functional mutations in experimental systems, defining the impact of gene variation on the epidemiology of disease, and determining the extent to which these genes interact with environmental factors to contribute to disease risk. Activities include both statistical analysis and laboratory work. A large part of Dr. Boerwinkle's current research effort consist of localizing genes contributing to disease risk using modern genome-wide mapping methods. Success depends on keeping up with the latest genomic technical advances. The laboratory is set-up and operating as a high through-put sequencing and genotyping facility in which speed, accuracy and efficiency are monitored continuously. However, we are constantly seeking out more efficient methods to collect and manage genetic information. Dr. Boerwinkle and colleagues have completed the world's first genome-wide analyses for a variety of CAD risk factors, including diabetes and hypertension. These investigations have lead to the identification of novel susceptibility genes in both cases. Dr. Boerwinkle is particularly interested in methods for identifying potentially functional mutations within a gene region. This seemingly simple objective is made difficult because the functional mutations are expected to have small effects and are imbedded in a sea of silent genetic variation. Once nearly all of the variation is catalogued directly by DNA sequencing, individuals are genotyped for each variable site. Both novel and traditional statistical methods are applied to relate the array of genetic information to a wealth of phenotypic data. This algorithm generates "candidate functional mutations" that are then tested in an in vitro or mouse model system. Once a functional mutation has been identified, Dr. Boerwinkle's group evaluates the ability of the variable site to predict the onset of disease (e.g. myocardial infarction or stroke) above and beyond traditional risk factors. This work is carried out as part of multiple prospective studies of cardiovascular disease and its risk factors in tens of thousands of individuals representing the major American ethnic groups. Finally, he is working on experimental designs for studying genotype by environment interaction in humans. In particular, we are working on the extent to which interindividual variation in lipid lowering and anti-hypertensive medications are influenced by genetic factors. The practical objective of the research is to use genetic information to identify individuals at increase risk of disease and to design more efficacious interventions. Genetic studies are defining, at the molecular level, novel mechanisms of disease risk, onset and progression. Dr. Boerwinkle and collaborators address the localization of genes which contribute to disease risk in cardiovascular diseases, hypertension and diabetes. The methodology used involves screening of families having the disease and linking the presence of disease with known markers of the human genome. In this manner, the genomic region in which relevant mutations are located can be mapped and the relevant DNA sequenced. By assessing the structural change the mutation may have caused in the gene product (protein), it is possible to infer how it may affect biological function. In order to determine experimentally whether a mutation is functional, it is necessary to introduce the mutated gene into an animal, usually a mouse, and assess its biological effects on the animal's phenotype. Dr. Boerwinkle has participated in multiple notable discoveries since joining the Institute. Only two will be highlighted here. First, Dr. Boerwinkle's group has completed the first ever genome-wide search for genes contributing to inter-individual blood pressure levels. This initial effort has lead to the identification of an important gene (an adrenergic receptor) which influences blood pressure levels and the risk to hypertension. This is the first time that such a genome-wide approach has led to the identification of a susceptibility gene to a major cardiovascular disease risk factor. Second, Dr. Boerwinkle has participated in similar efforts to identify genes contributing to the risk of developing non-insulin dependent (type II) diabetes. In this case, however, there were no genes in the region that were suspects for the disease. A team of collaborating investigators have painstakingly characterized the genetic region and identified the mutated gene (in this case a protease). This is the first time that anyone has ever positionally cloned a gene contributing to any common chronic disease. This work is of obvious potential clinical importance. It may lead to improved prediction of those at increased risk of disease and the design of more efficacious intervention strategies. The technologies and information from the human genome project provide new tools for lessening the burden of ill-health. Dr. Boerwinkle's accomplishments in developing an internationally recognized team of investigators targeting the genetics of cardiovascular disease and its risk factors ensures a productive future and further discoveries. Publications 2019 Tin A , Yu B , Ma J , Masushita K , Daya N , Hoogeveen RC , et al. . Reproducibility and Variability of Protein Analytes Measured Using a Multiplexed Modified Aptamer Assay. J Appl Lab Med. 2019;4(1):30-39. PubMed DOI Google Scholar Tagged de Vries PS , Brown MR , Bentley AR , Sung YJ , Winkler TW , Ntalla I , et al. . Multiancestry Genome-Wide Association Study of Lipid Levels Incorporating Gene-Alcohol Interactions. Am J Epidemiol. 2019;188(6):1033-1054. PubMed DOI Google Scholar Tagged Yu B , Zanetti KA , Temprosa M , Albanes D , Appel N , Barrera CBarrios , et al. . The Consortium of Metabolomics Studies (COMETS): Metabolomics in 47 Prospective Cohort Studies. Am J Epidemiol. 2019;188(6):991-1012. PubMed DOI Google Scholar Tagged Flannick J , Mercader JM , Fuchsberger C , Udler MS , Mahajan A , Wessel J , et al. . Exome sequencing of 20,791 cases of type 2 diabetes and 24,440 controls. Nature. 2019;570(7759):71-76. PubMed DOI Google Scholar Tagged Li Z , Li X , Liu Y , Shen J , Chen H , Zhou H , et al. . Dynamic Scan Procedure for Detecting Rare-Variant Association Regions in Whole-Genome Sequencing Studies. Am J Hum Genet. 2019;104(5):802-814. PubMed DOI Google Scholar Tagged Loomis SJ , Köttgen A , Li M , Tin A , Coresh J , Boerwinkle E , et al. . Rare variants in SLC5A10 are associated with serum 1,5-anhydroglucitol (1,5-AG) in the Atherosclerosis Risk in Communities (ARIC) Study. Sci Rep. 2019;9(1):5941. PubMed DOI Google Scholar Tagged Petty LE , Highland HM , Gamazon ER , Hu H , Karhade M , Chen H-H , et al. . Functionally oriented analysis of cardiometabolic traits in a trans-ethnic sample. Hum Mol Genet. 2019;28(7):1212-1224. PubMed DOI Google Scholar Tagged Bentley AR , Sung YJ , Brown MR , Winkler TW , Kraja AT , Ntalla I , et al. . Multi-ancestry genome-wide gene-smoking interaction study of 387,272 individuals identifies new loci associated with serum lipids. Nat Genet. 2019;51(4):636-648. PubMed DOI Google Scholar Tagged Posey JE , O'Donnell-Luria AH , Chong JX , Harel T , Jhangiani SN , Akdemir ZHCoban , et al. . Insights into genetics, human biology and disease gleaned from family based genomic studies. Genet Med. 2019;21(4):798-812. PubMed DOI Google Scholar Tagged Li AH , Hanchard NA , Azamian M , D'Alessandro LCA , Coban-Akdemir Z , Lopez KN , et al. . Genetic architecture of laterality defects revealed by whole exome sequencing. Eur J Hum Genet. 2019;27(4):563-573. PubMed DOI Google Scholar Tagged Liu Y , Chen S , Li Z , Morrison AC , Boerwinkle E , Lin X . ACAT: A Fast and Powerful p Value Combination Method for Rare-Variant Analysis in Sequencing Studies. Am J Hum Genet. 2019;104(3):410-421. PubMed DOI Google Scholar Tagged Justice AE , Karaderi T , Highland HM , Young KL , Graff M , Lu Y , et al. . Protein-coding variants implicate novel genes related to lipid homeostasis contributing to body-fat distribution. Nat Genet. 2019;51(3):452-469. PubMed DOI Google Scholar Tagged McGuire AL , Majumder MA , Villanueva AG , Bardill J , Bollinger JM , Boerwinkle E , et al. . Importance of Participant-Centricity and Trust for a Sustainable Medical Information Commons. J Law Med Ethics. 2019;47(1):12-20. PubMed DOI Google Scholar Tagged Kunkle BW , Grenier-Boley B , Sims R , Bis JC , Damotte V , Naj AC , et al. . Genetic meta-analysis of diagnosed Alzheimer's disease identifies new risk loci and implicates Aβ, tau, immunity and lipid processing. Nat Genet. 2019;51(3):414-430. PubMed DOI Google Scholar Tagged de Vries PS , Sabater-Lleal M , Huffman JE , Marten J , Song C , Pankratz N , et al. . A genome-wide association study identifies new loci for factor VII and implicates factor VII in ischemic stroke etiology. Blood. 2019;133(9):967-977. PubMed DOI Google Scholar Tagged Chen H , Huffman JE , Brody JA , Wang C , Lee S , Li Z , et al. . Efficient Variant Set Mixed Model Association Tests for Continuous and Binary Traits in Large-Scale Whole-Genome Sequencing Studies. Am J Hum Genet. 2019;104(2):260-274. PubMed DOI Google Scholar Tagged Sabater-Lleal M , Huffman JE , de Vries PS , Marten J , Mastrangelo MA , Song C , et al. . Genome-Wide Association Transethnic Meta-Analyses Identifies Novel Associations Regulating Coagulation Factor VIII and von Willebrand Factor Plasma Levels. Circulation. 2019;139(5):620-635. PubMed DOI Google Scholar Tagged Kilpeläinen TO , Bentley AR , Noordam R , Sung YJu , Schwander K , Winkler TW , et al. . Multi-ancestry study of blood lipid levels identifies four loci interacting with physical activity. Nat Commun. 2019;10(1):376. PubMed DOI Google Scholar Tagged Selvaraj S , Seidelmann S , Silvestre OM , Claggett B , Ndumele CE , Cheng S , et al. . HFE H63D Polymorphism and the Risk for Systemic Hypertension, Myocardial Remodeling, and Adverse Cardiovascular Events in the ARIC Study. Hypertension. 2019;73(1):68-74. PubMed DOI Google Scholar Tagged Floyd JS , Bloch KM , Brody JA , Maroteau C , Siddiqui MK , Gregory R , et al. . Pharmacogenomics of statin-related myopathy: Meta-analysis of rare variants from whole-exome sequencing. PLoS One. 2019;14(6):e0218115. PubMed DOI Google Scholar Tagged

https://www.hgsc.bcm.edu/people/boerwinkle-e?f%5Bauthor%5D=12793&page=6

Electronic structure of NaCo_ {2} O_ {4} Download Citation | Electronic structure of NaCo_ {2} O_ {4} | First principles density functional calculations of the electronic and magnetic properties of the layered oxide NaCo2O4 have been performed using... | Find, read and cite all the research you need on ResearchGate Electronic structure of NaCo_ {2} O_ {4} May 2000 Physical Review B 61(20) DOI: 10.1103/PhysRevB.61.13397 To read the full-text of this research, you can request a copy directly from the author. Abstract First principles density functional calculations of the electronic and magnetic properties of the layered oxide NaCo2O4 have been performed using the general potential linearized augmented plane wave method. The electronic transport properties are discussed in terms of the calculated paramagnetic band structure. It is found that the thermopower and specific heat data above 10 K are well accounted for with only moderate renormalization in spite of the fact that this is a strongly correlated system characterized by W≪U. Weak instabilities of itinerant magnetic character are found. The low temperature properties (T below 10 K) are discussed in terms of these. ... Key features of this process are an extensive TM-O covalent interaction ("hybridization") in combination with the splitting of the t 2g states stabilizing the material. Such a mechanism requires sufficient structural flexibility such as a height reduction of the cobalt-oxygen slab [17] and/or a distortion of the CoO 6 octahedron [55] in order to allow for increased covalent interaction with the O2p states (σ-and π-bonding) [12,58] and/or changes in the crystal field [55,56]. ... ... Key features of this process are an extensive TM-O covalent interaction ("hybridization") in combination with the splitting of the t 2g states stabilizing the material. Such a mechanism requires sufficient structural flexibility such as a height reduction of the cobalt-oxygen slab [17] and/or a distortion of the CoO 6 octahedron [55] in order to allow for increased covalent interaction with the O2p states (σ-and π-bonding) [12,58] and/or changes in the crystal field [55, 56]. ... ... Upon charging, both Co3d-derived states and O2p band broaden and overlap; additionally, new states form at around 8 eV. The broadening of the states at the valence band edge has been observed before[52][53][54] and is here attributed to the splitting of the t 2g states, with the states at lower binding energy having dominant a 1g character [55] . The additional states at higher binding energies are attributed to the formation of the peroxo-dimers.The evolution of the electronic states in the valence region upon charging is shown inFigure 4a. ... The role of covalent bonding and anionic redox for the performance of sodium cobaltate electrode materials Article Full-text available Feb 2021 Conrad Guhl Jochen Rohrer Philipp Kehne René Hausbrand It is common belief that conventional layered oxide battery cathode materials operate on a cationic redox mechanism, and that oxygen oxidation triggered by charge compensation results in oxygen loss and limited practical capacity. In this work, thin films of sodium cobaltate demonstrate a mixed redox behavior with strong oxygen participation well within the reversible intercalation range. This unexpected behavior is attributed to an increasing degree of covalency of the transition metal-oxygen (TM-O) bonds during deintercalation in combination with a high structural flexibility of the material. These insights are based on electronic structure data obtained using an operando solid state cell, allowing analysis with unpreceded control, and are supported by density functional theory (DFT) calculations. They demonstrate that the spectrum of possible charge compensation mechanisms is broader than commonly believed, and indicate that tuning TM-O covalency is a key factor for increasing the capacity also of conventional layered materials. ... As observed from the panels of Fig. 6(d), EDMs taken along the Γ-K direction, the bands dispersing from the Fermi level down to 1 eV are only accessible with p-polarized light, while the bands below 1 eV are accessible with s-polarized light. We can conclude from this observation that the a 1g band that is dominating in the vicinity of E F has even parity [30] , while the e g bands dominating below 1 eV have the odd parity with the respect to our measuring geometry (see Experimental Methods section). By subtracting s-polarized data with the p-polarized as shown in p−s panel of Fig. 6(d), we can clearly disentangle the evenparity states (blue colored) from the odd parity states (red colored). ... ... As observed in this study and reported in the literature, near the Fermi level only one band disperses from E F down to a binding energy of 0.4 eV [29,46]. The same has been confirmed from the DFT calculations as well, especially, in AHL plane [30] . ... ... With the help of average Fermi vector (k F =0.51±0.02Å −1 ) and Fermi velocity, we estimated the Seebeck coefficient using the Boltzman theory [7, 30] , ... Anomalous band renormalization due to high energy kink in the colossal thermoelectric material K0.65RhO 2 Article Full-text available May 2021 Susmita Changdar Grigory Shipunov Nesta Benno Joseph Thirupathaiah Setti We report on low-energy electronic structure and electronic correlations of K$_{0.65}$RhO$_2$, studied using high-resolution angle-resolved photoemission spectroscopy (ARPES) technique and density functional theory (DFT) calculations. We observe a highly correlated hole pocket on the Fermi surface. We further notice that the correlations are momentum dependent. Most importantly, two $kinks$ at binding energies of 75 meV and 195 meV have been observed from the band dispersion in the vicinity of the Fermi level. While the low energy $kink$ at 75 meV can be understood as a result of the electron-phonon interaction, the presence of high energy $kink$ at 195 meV is totally a new discovery of this system leading to an anomalous band renormalization. Based on systematic analysis of our experimental data, we propose high frequency bosonic excitations as a plausible origin of the high energy anomaly. Further, we notice that the high energy anomaly has important implications in obtaining the colossal thermoelectric power of K$_{0.65}$RhO$_2$. ... To model the electronic structure, we use a three-band tight-binding model for the t 2g bands as formulated by Mochizuki et al. [14] which describes the low-energy characteristics of LDA band structure calculations [57] quite well. This model includes a crystal field term accounting for the trigonal deformation of the CoO 6 octahedra because of the plane height reduction. ... ... where t 3 is the unit of energy. Setting ∆ CF = 0.4 reproduces LDA band structure calculations [57] well, particularly around the Fermi level. The value of ∆ CF significantly influences the Fermi surface topology. ... An efficient fluctuation exchange approach to low-temperature spin fluctuations and superconductivity: from the Hubbard model to Na$_x$CoO$_2\cdot y$H$_2$O Preprint Dec 2020 Niklas Witt E. G. C. P. van Loon Takuya Nomoto Tim O. Wehling Superconductivity arises mostly at energy and temperature scales that are much smaller than the typical bare electronic energies. Since the computational effort of diagrammatic many-body techniques increases with the number of required Matsubara frequencies and thus with the inverse temperature, phase transitions that occur at low temperatures are typically hard to address numerically. In this work, we implement a fluctuation exchange (FLEX) approach to spin fluctuations and superconductivity using the "intermediate representation basis" (IR) [Shinaoka et al., PRB 96, 2017] for Matsubara Green functions. This FLEX+IR approach is numerically very efficient and enables us to reach temperatures on the order of $10^{-4}$ in units of the electronic band width in multi-orbital systems. After benchmarking the method in the doped repulsive Hubbard model on the square lattice, we study the possibility of spin-fluctuation-mediated superconductivity in the hydrated sodium cobalt material Na$_x$CoO$_2\cdot y$H$_2$O reaching the scale of the experimental transition temperature $T_{\mathrm{c}}=4.5$ K and below. ... Finally, figure 14 shows a comparison of the valence band region for the four investigated thin films. The line shape of the valence band for both CoCr 2 O 4 thin films consists of a sharp feature located between the Fermi level (E F , BE = 0 eV) and 2 eV BE, which can mainly be ascribed to a superposition of Cr 3d and Co 3d partial density of states [50] [51] [52]. This peak is separated from a wider structured region between~2 eV and~8 eV, where O 2p states are expected to be located [51]. ... ... The line shape of the valence band for both CoCr 2 O 4 thin films consists of a sharp feature located between the Fermi level (E F , BE = 0 eV) and 2 eV BE, which can mainly be ascribed to a superposition of Cr 3d and Co 3d partial density of states [50][51][52]. This peak is separated from a wider structured region between~2 eV and~8 eV, where O 2p states are expected to be located [51] . The broad region between~8 eV and~12 eV is likely to derive from Co/Cr 4s states. ... Role of Ni substitution on structural, magnetic and electronic properties of epitaxial CoCr2O4 spinel thin films Cubic spinel CoCr2O4has attained recent attention due to its multiferroic properties. However, the Co site substitution effect on the structural and magnetic properties has rarely been studied in thin film form. In this work, the structural and magnetic properties of Co1-xNixCr2O4(x = 0, 0.5) epitaxial thin films deposited on MgAl2O4(100) and MgO (100) substrates to manipulate the nature of strain in the films using pulsed laser deposition (PLD) technique are presented. The epitaxial nature of the films was manifested through X-ray diffraction (XRD), reciprocal space mapping (RSM) and Rutherford backscattering spectrometry (RBS) measurements. Raman measurements revealed a disappearance of characteristic A1gand F2gmodes of the CoCr2O4with increase in the Ni content. Atomic force microscopy (AFM) and field emission scanning electron microscopy (FE-SEM) studies show a modification of the surface morphology upon Ni substitution. Magnetic measurements disclose that the ferrimagnetic Curie temperature (TC) of the CoCr2O4in thin film grown on MgAl2O4(100) and MgO (100) substrates were found to be 100.6 ± 0.5 K and 93.8 ± 0.2 K, respectively. With Ni substitution the TC values were found to be enhanced to 104.5 ± 0.4 K for MgAl2O4(100) and 108.5 ± 0.6 K for MgO (100) substrates. X-ray photoelectron spectroscopy (XPS) suggests Cr3+oxidation states in the films, while Co ions are present in a mixed Co2+/Co3+oxidation state. The substitution of Ni at Co site significantly modifies the line shape of the core level as well as the valence band. Ni ions are also found to be in a mixed 2+/3+ oxidation state. O 1s core level display asymmetry related to possible defects like oxygen vacancies in the films. ... However, such method resulted in energy bands that are less dispersive. [34] Nevertheless, the reliability of Takeuchi's theory has been proved by other groups, theoretically [34] and experimentally. [35] For degenerated semiconductors (parabolic band, energy-independent scattering approximation [36] ), Equation (2) is further modified by carrier concentration and effective mass of the carrier [37] ... ... However, such method resulted in energy bands that are less dispersive. [34] Nevertheless, the reliability of Takeuchi's theory has been proved by other groups, theoretically [34] and experimentally. [35] For degenerated semiconductors (parabolic band, energy-independent scattering approximation [36] ), Equation (2) is further modified by carrier concentration and effective mass of the carrier [37] ... Recent Advances of Layered Thermoelectric Materials ... The formal valence of Co ions deduced from the chemical composition Bi 2 A 2 Co 2 O 8 is 3+ , leaving 6 electrons in Co-3d orbitals. In the edge sharing hexagonal CoO 2 layer, the trigonal distortion of CoO 2 octahedra reduces the O h local point group to a d 3 subgroup, further splitting the t 2g orbitals into an a 1g singlet and an ′ e g doublet 23 . The previous local density approximation (LDA) study concentrated on the nonmagnetic state of Bi 2 A 2 Co 2 O 8 16 . ... ... The Hubbard U and Hund's exchange coupling J H are usually extracted from fitting the core-electron satellite spectra of transition-metal oxides to a cluster model, and the values vary depending on the crystal structures and the valence of Co-ions. The values, 5 eV < U < 8 eV, J H = 1 eV, recommended by Singh 23 were widely cited for cobalt based perovskite. Korotin et al. 30 obtained similar values (U = 7.8 eV, J H = 0.92 eV) in discussing the IS state associated with the high temperature magnetic structure of LaCoO 3 . ... How does the spin-state of Co ions affect the insulator-metal transition in Bi2A2Co2O8 (A = Ca, Sr, Ba)? The misfit layered Bi2A2Co2O8 (A = Ca, Sr, Ba) compounds experience an insulator to metal transition as A’s ionic radius increases. This feature is contradictory to the conventional wisdom that larger lattice constant favors insulating rather than metallic state, and is also difficult to be reconciled using the Anderson weak localization theory. In this paper, we show from the first-principles calculation that an insulator-metal transition takes place from a nonmagnetic low-spin state of Co3+ ions to a hexagonally arranged intermediate-spin low-spin mixed-state in CoO2 plane when ionic radius increases from Ca to Ba. The predicted low-spin state of Bi2Ca2Co2O8 and Bi2Sr2Co2O8 and intermediate-spin low-spin mixed-state of Bi2Ba2Co2O8 are consistent not only with their measured transport properties, but also with the magnetic-field suppressed specific-heat peak observed at the transition temperature. In agreement with experiments, strong electronic correlation is required to stabilize the low-spin insulator and intermediate-spin low-spin metal. View ... d n electronic configuration spin state ground multiplet term ( 2S+1 Γ) degeneracy of 2S+1 Γ (Γ)) When a material can be well described as band picture (mean-field approximation), the Boltzmann equation regime works well. Based on the band calculation and the Boltzmann equation, Singh reproduced a large thermopower (∼ 110 µV/K) of NaCo 2 O 4 at 300 K [28] . Even if band picture does not work well (ex. ... Thermopower in transition-metal perovskites High-temperature thermopower is interpreted as entropy that a carrier carries. Owing to spin and orbital degrees of freedom, a transition metal perovskite exhibits large thermopower at high temperatures. In this paper, we revisit the high-temperature thermopower in the perovskites to shed light on the degrees of freedom. Thus, we theoretically derive an expression of thermopower in one-dimensional octahedral-MX6-clusters chain using linear-response theory and electronic structure calculation of the chain based on the tight-binding approximation. The derived expression of the thermopower is consistent with the extended Heikes formula and well reproduced experimental data of several perovskite oxides at high temperatures. In this expression, a degeneracy of many electron states in octahedral ligand field (which is characterized by multiplet term) appears instead of the spin and orbital degeneracies. Complementarity in between our expression and the extended Heikes formula is discussed. ... [18] The Rashba spin splitting leads to a distinctive feature in the electronic density of states (DOS) due to the unique Fermi surface topology [19]. Consequently, the power factor is found to be large because ( 2 σ) of the sharp peak in DOS [20] . Thus, the Rashba spin splitting can potentially be a very effective mechanism for thermoelectricity. ... Realizing high Near-Room-Temperature Thermoelectric Performance in n-type Ag2Se through Rashba Effect and Entropy Engineering Although there are enormous numbers of high-temperature thermoelectric materials present, designing a near-room-temperature especially n-type thermoelectric material with high zT is extremely challenging. Generally, pristine Ag2Se exhibits unusual low thermal conductivity along with high electrical conductivity and Seebeck coefficient, which leads to high thermoelectric performance (n-type) at room temperature. Herein, we report a pseudoternary phase, Ag2Se0.5Te0.25S0.25, which shows improved thermoelectric performance (zT ~ 2.1 at 400 K). Density functional theory reveals that the Rashba type of spin-dependent band spitting originated because of Te-doping, enhancing carrier mobility. Using density functional perturbation theory, we hereby realize that the intrinsic carrier mobility is not only controlled by carrier effective mass, neither deformation potential theory, instead it is substantially limited by longitudinal optical phonon scattering. In fact, locally off-centered S atoms and rising configurational entropy via substitution of Te and S atoms in Ag2Se significantly reduce the lattice thermal conductivity (klat ~ 0.34 at 400 K). In order to accurately obtain electrical as well as thermal transport coefficient, we adopt deformation potential theory based on Boltzmann transport formalism. The combined consequence of the Rashba effect coupled with configurational entropy synergistically results in such high thermoelectric performance with the development of new n-type thermoelectric material working at the near-room-temperature regime. View ... 29 Electronic-structure calculation performed later on found that this compound locates at an itinerant magnetism instability with large electronic density of states derived from 3d orbitals. 30 Within the itinerant picture, the a(H) profile of NaCo 2 O 4 is also approachable by considering magnetic-field tuning to spin polarization at the Fermi level. 31 Here, spin entropy is involved in shaping the electronic structure, as concluded from recent angle-resolved photoemission spectroscopy experiments. ... Generic Seebeck effect from spin entropy How magnetism affects the Seebeck effect is an important issue widely concerned in the thermoelectric community yet remaining elusive. Based on a thermodynamic analysis of spin degrees of freedom on varied d-electron based ferro- and anti-ferromagnets, we demonstrate that in itinerant or partially itinerant magnetic compounds there exists a generic spin contribution to the Seebeck effect over an extended temperature range from slightly below to well above the magnetic transition temperature. This contribution is interpreted as resulting from transport spin entropy of (partially) delocalized conducting d electrons with strong thermal spin fluctuations, even semiquantitatively in a single-band case, in addition to the conventional diffusion part arising from their kinetic degrees of freedom. As a highly generic effect, the spin-dependent Seebeck effect might pave a feasible way to efficient “magnetic thermoelectrics”. ... Based on the energy-independent carrier relaxation time and parabolic band approximations, the Seebeck coefficient of a TE material is approximately proportional to the density-of-states effective mass (m * D ). 52, 53 Higher m * D originate from sharp peaks in the density of state (DOS) near band edges which indicate flat and dispersionless bands. 54,55 Therefore, in this section, we examine the DOS of the 3d TM and 4d TM containing TMFe 2 O 4 compounds. ... High-Performance Thermoelectric Oxides Based on Spinel Structure Julio Gutierrez Moreno Marco Fronzi M. Hussein N. Assadi High-performance thermoelectric oxides could offer a great energy solution for integrated and embedded applications in sensing and electronics industries. Oxides, however, often suffer from low Seebeck coefficient when compared with other classes of thermoelectric materials. In search of high-performance thermoelectric oxides, we present a comprehensive density functional investigation, based on GGA+U formalism, surveying the 3d and 4d transition-metal-containing ferrites of the spinel structure. Consequently, we predict MnFe2O4 and RhFe2O4 have Seebeck coefficients of ~±600 μV K−1 at near room temperature, achieved by light hole and electron doping. Furthermore, CrFe2O4 and MoFe2O4 have even higher ambient Seebeck coefficients at ~±700 μV K−1. In the latter compounds, the Seebeck coefficient is approximately a flat function of temperature up to ~700 K, offering a tremendous operational convenience. Additionally, MoFe2O4 doped with 1019 holes/cm3 has a calculated thermoelectric power factor of 689.81 μW K−2 m−1 at 300 K, and 455.67 μW K−2 m−1 at 600 K. The thermoelectric properties predicted here can bring these thermoelectric oxides to applications at lower temperatures traditionally fulfilled by more toxic and otherwise burdensome materials. ... The band structures of stoichiometric ARhO 2 are similar to that of the isostructural NaCoO 2 . 39 The d orbital of Rh splits into fully empty eg and fully occupied t 2g orbitals in the octahedron ... First-principles calculations of structural and electronic properties of layered A x RhO 2 (A = Li, Na, K, Rb, Cs) Article Full-text available Mar 2020 Rui Guo Zhi-An Zhu Y. B. Chen Jian Zhou Layered transition metal oxides, such as NaxCoO2, are known for their various interesting physical phenomena, which are mainly due to the strong correlation of the transition elements and tunable concentration of alkali metals. Here, we have systematically investigated the structural and electronic properties of 4d layered transition metal oxides AxRhO2 (A = Li, Na, K, Rb, Cs) by first-principles calculations. It is found that when the concentration (x) of alkali ions in AxRhO2 increases, the in-plane lattice constant (a) increases while the out-of-plane one (c) decreases. In the case of stoichiometric ARhO2 (i.e., x = 1), both lattice constants (a and c) increase when the alkali ions changes from Li to Cs. The calculated electronic band structures and density of states indicate that all the stoichiometric ARhO2 compounds are indirect band-gap semiconductors with band gaps ranging from 3 eV to 3.6 eV. Finally, we calculate the Fermi surfaces of KxRhO2 and demonstrate the Lifshitz transition, which could be triggered by potassiation/depotassiation in experiments. Despite the structural similarity between these materials, we have observed the difference in their band structures at the valence band maximum, which will possibly result in a different behavior of the Lifshitz transition. Our calculations point out the similarities and the subtle differences between different alkali rhodates, which give some useful information for future experimental works on these materials. ... Before proceeding to the next section, we briefly review the Boltzmann transport theory, which was used in calculations presented later in this review. Calculation of transport properties using the DFT band structure 42) is nowadays a very popular approach. In the Boltzmann transport theory, the transport coefficients K are represented as follows: ... Theoretical Aspects of the Study on the Thermoelectric Properties of Pnictogen-Dichalcogenide Layered Compounds Article Jan 2019 J PHYS SOC JPN Masayuki Ochi Hidetomo Usui Kazuhiko Kuroki Recent experimental studies revealed that BiCh2 (Ch = S, Se) layered compounds, which have been well known as superconductors, also exhibit high thermoelectric performance. We review theoretical investigations on the thermoelectric properties of PnCh2 (Pn = Bi, Sb, As) layered compounds. Their electronic structure shows quasi-one-dimensionality, which is favorable for realizing high power factor. This quasi-one-dimensionality originates from the anisotropy of the px,y orbitals on the PnCh2 square lattice. First-principles calculation suggests that atomic substitution can enhance the one-dimensionality, and thus the power factor. Another important aspect is the lattice instability, which can result in the strong anharmonicity of phonons and the low lattice thermal conductivity. ... ARPES studies indicate that the e ′ g levels are completely filled and therefore should be electronically inactive [612,613], although this is somewhat controversial [614]. Band calculations find a larger Fermi surface due to a 1g levels, and a smaller Fermi surface due to the e ′ g levels [615] . The physical behavior of anhydrous Li x CoO 2 and Na x CoO 2 are strongly x-dependent (see Fig. B.60(b)) [608]. ... From charge- and spin-ordering to superconductivity in the organic charge-transfer solids Article Feb 2018 PHYS REP R. Torsten Clay Sumit Mazumdar ... [68] As an alternative interpretation, Singh used an itinerant picture and the 2D of the system to explain the good TE properties. [69] There have been excellent reviews of these oxide systems with particular spin states. [16,66,70] The spin Seebeck effect is an interesting phenomenon derived from spin current, which can generate voltage through the inverse spin Hall effect. ... Novel Principles and Nanostructuring Methods for Enhanced Thermoelectrics Article Sep 2017 Takao Mori Thermoelectrics (TE), the direct solid-state conversion of waste heat to electricity, is a promising field with potential wide-scale application for power generation. Intrinsic conflicts in the requirements for high electrical conductivity but (a) low thermal conductivity and (b) a large Seebeck coefficient have made enhancing TE performance difficult. Several recent striking advances in the field are reviewed. In regard to the former conflict, notable bottom-up nanostructuring methods for phonon-selective scattering are discovered, namely using nanosheets, dislocations, and most strikingly a process to fabricate nano-micropores leading to a 100% enhancement in the figure of merit (ZT ≈ 1.6) for rare-earth-free skutterudites. Porous materials are hitherto considered as having poor TE performance, so this is a new paradigm. In regard to the latter conflict, nanocomposite materials with hybrid effects and use of magnetism are emerging as novel bottom-up methods to enhance TE. Material informatics efforts to identify high-ZT materials are also reviewed. ... The formal valence of Co ions deduced from the chemical composition Bi 2 A 2 Co 2 O 8 is 3+, leaving six electrons in Co 3d orbitals. In the edge-sharing hexagonal CoO 2 layer, the trigonal distortion of CoO 2 octahedra reduces the O h local point group to a d 3 subgroup, in which the t 2g orbitals are further split into an a 1g singlet and an e g doublet [16] . ... Theoretical study of the thickness dependence of the metal-insulator transition in Bi2Sr2Co2O8nanosheets Article Full-text available Jul 2017 Xiaokun Huang Weiyi Zhang The bulk Bi2Sr2Co2O8 crystal is an insulator at low temperature and experiences an insulator-metal transition at TC≈60K. The new experiment showed that TC increases to 140 K for a mechanically exfoliated nanosheet of four blocks and is beyond room temperature for nanosheets thinner than four blocks. We show that the thickness-dependent insulator-metal transition observed in Bi2Sr2Co2O8 nanosheets can naturally be explained by the strongly correlated low-spin-state insulator (LS state) and intermediate-spin-low-spin-state metal (IS-LS state). In particular, the energy difference between the LS state and IS-LS state qualitatively reproduces the trend of the transition temperature with the nanosheet thickness. The predicted transition temperature of a nanosheet with three blocks is only slightly above room temperature, a result that can be used to check our proposed mechanism. Further experiments on the distinct magnetotransport properties and spin-fluctuation behaviors of the LS state and IS-LS state are also very helpful to resolve the issue. The weak interblock binding is also consistent with the layer-resolved partial densities of states. ... Then, we constructed the tight-binding model with the obtained hopping parameters among the Wannier functions, and analyzed the transport properties using this model. For this purpose, we employed the Boltzmann transport theory [55] , where the transport coefficients K ν are represented as follows: ... Prediction of the High Thermoelectric Performance of Pnictogen Dichalcogenide Layered Compounds with Quasi-One-Dimensional Gapped Dirac-like Band Dispersion Article Dec 2017 Masayuki Ochi Hidetomo Usui Kazuhiko Kuroki Thermoelectric power generation has been recognized as one of the most important technologies, and high-performance thermoelectric materials have long been pursued. However, because of the large number of candidate materials, this quest is extremely challenging, and it has become clear that a firm theoretical concept from the viewpoint of band-structure engineering is needed. We theoretically demonstrate that pnictogen dichalcogenide layered compounds, which originally attracted attention as a family of superconductors and have recently been investigated as thermoelectric materials, can exhibit very high thermoelectric performance with elemental substitution. Specifically, we clarify a promising guiding principle for material design and find that LaOAsSe2, a material that has yet to be synthesized, has a power factor that is 6 times as large as that of the known compound LaOBiS2 and can exhibit a very large ZT under some plausible assumptions. This large enhancement of the thermoelectric performance originates from the quasi-one-dimensional gapped Dirac-like band dispersion, which is realized by the square-lattice network. We offer one ideal limit of the band structure for thermoelectric materials. Because our target materials have high controllability of constituent elements and feasibility of carrier doping, experimental studies along this line are eagerly awaited. ... 63 Three-dimensional layered complex metal nitrides AMN 2 (A = Sr or Na; M = Zr, Hf, Nb, Ta) with α-NaFeO 2 crystal structure are particularly interesting because they are used for energy conversion and storage applications, For example, Na x CoO 2 exhibits excellent thermoelectric properties 65 and is a base material for superconductors, 66 and LiCoO 2 is used as a cathode material in lithium ion batteries. 59 α-NaFeO 2 materials exhibit isotropic transport, 65, 67, 68 which results in a large thermoelectric effect. In terms of the thermoelectricity for three-dimensional AMN 2 compounds, Isao Ohkubo and Takao Mori made a great contribution for better understanding of thermoelectric transport of the layered complex nitrides. ... WASTE ENERGY HARVESTING USING III-NITRIDE MATERIALS Chapter Full-text available Jun 2017 Ehsan Ghafari Evan Witkoske Y. Liu Na Lu ... 9 The CRTA methodology has been successfully applied to other oxide thermoelectric materials. 10, 11 Within this methodology, τ is cancelled in the expression of the Seebeck coefficient, and thus can be directly evaluated from the first-principles band structure. Spin degrees of freedom have been neglected in these calculations. ... Baran BSCO SI Chem Mater Jakub D. Baran Demie Kepaptsoglou Marco Molinari ... A unique feature is that the carrier concentration is large enough to be of the order of 10 22 cm −3 , which could reduce the resistivity, but it is unlikely that the large thermopower can be understood within a simple one-electron picture. 4 Theoretical explanations beyond the conventional design rules have been proposed, [5] [6][7][8][9] and good thermoelectric oxides are still being actively sought world-wide. 10 The dimensionless figure of merit zT determining the conversion efficiency at temperature T is plotted as a function of T for various oxides in Fig. 1. [11][12][13][14][15][16][17][18][19][20][21][22] Some oxides indeed show reasonably large values of zT at high temperatures, and at least we can say that oxides are not out of the question. ... Research Update: Oxide thermoelectrics: Beyond the conventional design rules Materials’ design for high-performance thermoelectric oxides is discussed. Since chemical stability at high temperature in air is a considerable advantage in oxides, we evaluate thermoelectric power factor in the high temperature limit. We show that highly disordered materials can be good thermoelectric materials at high temperatures, and the effects of strong correlation can further enhance the figure of merit by adding thermopower arising from the spin and orbital degrees of freedom. We also discuss the Kelvin formula as a promising expression for strongly correlated materials and show that the calculation based on the Kelvin formula can be directly compared with the cross-layer thermopower of layered materials. ... 2, 13 The anisotropic transport in Na x CoO 2 is consistent with the electronic structure calculations, which show a cylindrical hole Fermi surface, indicating a quasi-two-dimensional electronic structure. 30 In contrast, the constant energy surfaces at the electron carrier concentrations to achieve maximum power factors for NaTaN 2 and NaNbN 2 are not cylindrical (not shown), and this indicates that the electronic structures are three-dimensional. This is consistent with the weak anisotropic thermoelectric transport properties. ... Anisotropic thermoelectric properties in layered complex nitrides with α-NaFeO2-type structure Isao Ohkubo Takao Mori Electronic structures and thermoelectric transport properties of α-NaFeO2-type d 0-electron layered complex nitrides AMN2 (A = Sr or Na; M = Zr, Hf, Nb, Ta) were evaluated using density-functional theory and Boltzmann theory calculations. Despite the layered crystal structure, all materials had three-dimensional electronic structures. Sr(Zr, Hf)N2 exhibited isotropic electronic transport properties because of the contribution of the Sr 4d orbitals to the conduction band minimums (CBMs) in addition to that of the Zr 4d (Hf 5d) orbitals. Na(Nb,Ta)N2 showed weak anisotropic electronic transport properties due to the main contribution of the Nb 4d (Ta 5d) and N 2p orbitals to the CBMs and no contribution of the Na orbitals. Understanding the Highly Electrocatalytic Active Mixed Triple Conducting Na x Ca 3– x Co 4 O 9–δ Oxygen Electrode Materials Kwangho Park Hyun-Kyu Kim Jun Young Park Hohan Bae Despite their high electrocatalytic activity for oxygen electrode reactions, the low phase stability and high thermal expansion of perovskite structured materials have created difficulties in cell fabrication scale‐up and long‐term operational stability of reversible ceramic cells. Herein, an exceptionally high‐performance electrocatalyst is presented based on a misfit‐layered structure, Na0.15Ca2.85Co4O9–δ (NCCO). NCCO cells enable exceptional fuel cell performance down to 400 °C, with peak power densities of 0.18–5.15 W cm−2 at 400–800 °C, as well as electrolysis performance of minus current density 5.96–15.07 A·cm−2 (at 1.4 V) at 600–750 °C, exceeding the values of all previously described reversible (oxygen and proton) ceramic cells. Furthermore, the durability of NCCO cells is demonstrated for over 900 h at high current densities of 1 and 2 A cm−2 in fuel cells and –0.5 and –4 A cm−2 electrolysis cell modes under load cycle and constant current reversible operation, respectively. Doping with basic monovalent Na+ ions in the Ca‐site in Ca3Co4O9+δ generates a high density of extra charge carrier species with the increased Co oxidation state and facilitates the proton uptake and diffusion properties of misfit‐layered materials. This finding can deliver a new opportunity to develop innovative bifunctional oxygen electrode catalysts, while providing more favorable reaction pathways for the diffusion of charged species. Misfit‐layered Na0.15Ca2.85Co4O9–δ (NCCO) is introduced as a highly active and durable bifunctional air/steam electrode material, moving away from the conventional perovskite materials. The misfit‐layered NCCO cell achieves high performance for reversible protonic ceramic cells with excellent long‐term electrochemical stability, while opening up new potential for finding technical breakthroughs for a variety of next‐generation electrochemical devices. Structure and Electronic Structure Evolution of P2-Na x CoO 2 Phases from X-ray Diffraction and 23 Na Magic Angle Spinning Nuclear Magnetic Resonance Yohan Biecher Anthony Baux François Fauth Dany Carlier The Drude model in correlated systems * L. Degiorgi The Drude model provides a useful phenomenological approach for the discussion of the metallic properties in conducting materials, particularly from the perspective of their electrodynamic response. We first introduce the model and its generalization, which accounts for the frequency dependence of the scattering rate and effective mass of the quasiparticles. Three selected examples, from the optical research on the topic of strongly correlated metals, will illustrate the variety of issues and broad spectrum of informations, which can be addressed and achieved within this approach. Electronic Properties and Lattice Dynamics of LixCoO2 and NaxCoO2 (x = 0, 0.5, 1) Studied by Hybrid Density Functional Theory Nina Mattila Antti J. Karttunen A systematic hybrid density functional theory study on the electronic and vibrational properties of Mx CoO2 compounds with M = Li, Na and x=0,0.5,1 is reported. The used DFT‐PBE0 method describes the structural parameters of the studied compounds well in comparison to experimental data. All studied magnetic species are treated as ferromagnets and the Co(IV) atoms possess a magnetic moment of 1.2 μB. At 0 K, CoO2 favors a monoclinic structure very close to trigonal symmetry and behaves as a Mott insulator. The electronic band gap increases as x increases from 0 to 1. The simulated IR and Raman spectra together with full phonon dispersion relations show that the intercalation of Li and Na ions affects the lattice dynamics of CoO2 in a different way. This article is protected by copyright. All rights reserved. Thermopower in transition-metal perovskites Wataru Kobayashi High-temperature thermopower is interpreted as entropy that a carrier carries. Owing to spin and orbital degrees of freedom, a transition-metal perovskite exhibits large thermopower at high temperatures. In this paper, we revisit the high-temperature thermopower in the perovskites to shed light on the degrees of freedom. Thus, we theoretically derive an expression of thermopower in one-dimensional octahedral-MX6-cluster chain using linear-response theory and electronic structure calculation of the chain based on the tight-binding approximation. The derived expression of the thermopower is consistent with the extended Heikes formula and well reproduces experimental data of several perovskite oxides at high temperatures. In this expression, a degeneracy of many-electron states in an octahedral ligand field (which is characterized by the multiplet term) appears instead of the spin and orbital degeneracies. Complementarity in between our expression and the extended Heikes formula is discussed. Efficient fluctuation-exchange approach to low-temperature spin fluctuations and superconductivity: From the Hubbard model to Na x CoO 2 · y H 2 O Niklas Witt E. G. C. P. van Loon Takuya Nomoto Tim O. Wehling Superconductivity arises mostly at energy and temperature scales that are much smaller than the typical bare electronic energies. Since the computational effort of diagrammatic many-body techniques increases with the number of required Matsubara frequencies and thus with the inverse temperature, phase transitions that occur at low temperatures are typically hard to address numerically. In this work, we implement a fluctuation exchange (FLEX) approach to spin fluctuations and superconductivity using the “intermediate representation basis” (IR) [Shinaoka et al., Phys. Rev. B 96, 035147 (2017)] for Matsubara Green functions. This FLEX+IR approach is numerically very efficient and enables us to reach temperatures on the order of 10−4 in units of the electronic bandwidth in multiorbital systems. After benchmarking the method in the doped repulsive Hubbard model on the square lattice, we study the possibility of spin-fluctuation-mediated superconductivity in the hydrated sodium cobalt material NaxCoO2·yH2O reaching the scale of the experimental transition temperature Tc=4.5K and below. Electronic Properties of Single-Layer CoO 2 /Au(111) Ann Julie Utne Holt Sahar Pakdel J. Rodríguez-Fernández Charlotte E. Sanders Thermoelectric Properties of Cobalt Oxides and Other Doped Mott Insulators Chapter May 2021 Wataru Koshibae One of the most important topics of cobalt oxides is the large thermopower (absolute Seebeck coefficient), which has been attracted attention due to their potential application as thermoelectric conversion material. In this chapter, the basics of thermoelectrics and thermoelectric response in cobalt oxides and related transition metal oxides are introduced. Thermopower is corresponding to the carried entropy by the electric current. In the strongly correlated electron systems, the spin and orbital degrees of freedom contribute to the entropy flow. The role of spin and orbital degrees of freedom on the thermoelectric effect is theoretically discussed. Transition from perovskite to misfit-layered structure materials: A highly oxygen deficient and stable oxygen electrode catalysts Article Full-text available Mar 2021 Muhammad Saqib In-Gyu Choi Hohan Bae Jun-Young Park Despite recent substantial progress in reversible protonic ceramic cells (RPCCs) it remains essential to further develop oxygen electrode materials that show superior activity and stability for oxygen reduction and evolution... Thermoelectric properties beyond the standard Boltzmann model in oxides: A focus on the ruthenates Chapter Jan 2021 Florent Pawula Ramzy Daou Sylvie Hébert Antoine Maignan Investigation into the thermoelectric properties of oxides in the last 20 years has shown that optimization of thermoelectric power depends on original parameters that cannot be simply described by Boltzmann transport equations, as the entropy part of the Seebeck coefficient (S) plays a major role. The optimization of S strongly depends on carrier doping and also on the specific orbital filling of these carriers, on the presence of electronic correlations, and on carrier spins and magnetic exchanges between them. The peculiarities observed mostly in p-type oxides are presented here, and the Seebeck coefficient in different ruthenium oxides will be discussed. In these ruthenates with different crystallographic structures and electronic and magnetic ground states, and for which spin-orbit coupling plays a major role, the Seebeck coefficient is shown to converge toward a common value at very high temperatures, which depends on the spin entropy only. This highlights the dominant role of entropy in these p-type oxides, and demonstrates how spin and magnetism can strongly affect the thermopower of oxides. Recent results have shown that apart from oxides, magnetism can also be a tuning parameter in other chalcogenides such as sulfides. Large thermoelectric power variations in epitaxial thin films of layered perovskite GdBaCo2O5.5±δ with different preferred orientation and strain Arindom Chatterjee Belen Ballesteros Jose Santiso The growth of thin epitaxial films of the layered perovskite material GdBaCo2O5.5±δ (GBCO) on different single crystal substrates SrTiO3 (STO), (LaAlO3)0.3(Sr2TaAlO6)0.7 (LSAT) and LaAlO3 (LAO) is described as a way to study changes in the thermoelectric properties by means of the induced epitaxial strain. In addition to the changes in strain the films grow with considerably different preferred orientations and domain microstructure: GBCO films on STO are purely c-axis oriented (c-perpendicular) with an average 0.18% in-plane tensile strain; GBCO on LSAT are composed of domains with mixed orientation (c-parallel and c-perpendicular) with an average 0.71% in-plane compressive strain; while on LAO it is b-axis oriented (c-parallel) with an average 0.89% in-plane compressive strain. These differences result in important changes in the sign and temperature dependence of the thermopower while the electrical conductivity remains almost isotropic. In general, compressively strained films show negative thermopower (n-type) while tensile strained film show positive thermopower (p-type) at low temperatures, probing the adaptive nature of GdBaCo2O5.5±δ compound. These results point to the spontaneous generation of oxygen vacancies to accommodate epitaxial stress as responsible for this effect. High figure-of-merit in the heavy-fermion UN 2 system for radioisotope thermoelectric applications Z. Z. Zhou Dengdong Fan Huijun Liu Jing Liu The design of uranium-based thermoelectric (TE) materials presents a novel and intriguing strategy for directly converting nuclear heat into electrical power. Using high-level first-principles approach combined with accurate solution of Boltzmann transport equation, we demonstrate that a giant n-type power factor of 13.8 mW m-1 K-2 and a peak ZT value of 2.2 can be realized in the heavy-fermion UN2 compound at 700 K. Such promising TE performance arises from the large degeneracy (N v = 14) of heavy conduction band coupled with weak electron-phonon interactions, which is in principle governed by the strong Coulomb correlation among the partially filled U-5f electrons in the face-centered cubic structure. Collectively, our theoretical work suggests that the energetic UN2 could serve as both excellent heat source and efficient power convertor, which also uncovers an underexplored area for TE research. Exploring the possibility of enhancing the figure-of-merit (> 2) of Na0.74CoO2: A combined experimental and theoretical study Article Aug 2020 Shamim Sk Jayashree Pati R. S. Dhaka Sudhir Kumar Pandey Search of new thermoelectric (TE) materials with high figure-of-merit (ZT) is always inspired the researcher in TE field. Here, we present a combined experimental and theoretical study of TE properties on Na0.74CoO2 compound in high temperature region. The experimental Seebeck coefficient (S) is found to vary from 64 to 118 μV/K in the temperature range 300−620 K. The observed value of thermal conductivity (κ) is ~2.2 W/m-K at 300 K. In the temperature region 300−430 K, the value of κ increases up to ~2.6 W/m-K and then decreases slowly till 620 K with the corresponding value of ~2.4 W/m-K. We have also carried out the theoretical calculations and the best matching between experimental and calculated values of transport properties are observed in spin-polarized calculation within DFT + U by chosen U = 4 eV. The maximum calculated value of ZT is found to be ~0.67 at 1200 K for p-type conduction. Our computational study suggests that the possibility of n-type behaviour of the compound which can lead to a large value of ZT at higher temperature region. Electron doping of ~5.1 × 1020 cm−3 is expected to give rise the high ZT value of ~2.7 at 1200 K. Using these temperature dependent ZT values, we have calculated the maximum possible values of efficiency (η) of thermoelectric generator (TEG) made by p and n-type Na0.74CoO2. The present study suggests that one can get the efficiency of a TE cell as high as ~11% when the cold and hot end temperature are fixed at 300 and 1200 K, respectively. Such high values of ZT and efficiency suggest that Na0.74CoO2 can be used as a potential candidate for high temperature TE applications. Charge Transfer in Thermoelectric Nanocomposites: Power Factor Enhancements and Model Systems Chapter Mar 2019 Matt Beekman Sage R. Bauers Danielle Marie Hamann David C. Johnson This chapter contains sections titled: Introduction Composite Thermoelectric Materials Concept of Modulation Doping Charge Transfer and Modulation Doping in Bulk Nanocomposites Modulation Doping in Heterostructures Ferecrystals Concluding Remarks Acknowledgements Self-organized formation of chemical bonding in misfit-layered bismuth-based cobaltites Article J ELECTRON SPECTROSC Makoto Maki Sho-ichi Takakura Isamu Yamamoto Junpei Azuma Angle-resolved photoemission measurements have been performed on a commensurate system [Bi 2 Ba 2 O 4 ] 0.50 CoO 2 with various photon energies. The results show that the Bloch waves propagate through the interface between CoO 2 and Bi 2 Ba 2 O 4 structures, contrasting with a previous study in an incommensurate system [Bi 2 Sr 2 O 4 ] 0.51 CoO 2 . On the other hand, transmission electron microscopy (TEM) measurements indicate that there is a non-negligible amount of scattering between the adjacent substructures. We present a detailed comparison of TEM images for three different misfit compounds. The results demonstrate that the self-organization at the atomic level enables to stabilize the misfit structure and modifies the electronic states in those systems. Enhanced Seebeck coefficient by a filling-induced Lifshitz transition in K x RhO 2 Article Jan 2019 Naoko Ito Mayu Ishii Ryuji Okazaki We have systematically measured the transport properties in the layered rhodium oxide KxRhO2 single crystals (0.5≲x≲0.67), which is isostructural to the thermoelectric oxide NaxCoO2. We find that below x=0.64 the Seebeck coefficient is anomalously enhanced at low temperatures with increasing x, while it is proportional to the temperature as a conventional metal above x=0.65, suggesting the existence of a critical content x*≃0.65. For the origin of this anomalous behavior, we discuss a filling-induced Lifshitz transition, which is characterized by a sudden topological change in the cylindrical hole Fermi surfaces at the critical content x*. Valence transition model of the pseudogap, charge order, and superconductivity in electron-doped and hole-doped copper oxides Article Sumit Mazumdar We present a valence transition model for electron- and hole-doped cuprates, within which there occurs a discrete jump in ionicity Cu2+→Cu1+ in both families upon doping, at or near optimal doping in the conventionally prepared electron-doped compounds and at the pseudogap phase transition in the hole-doped materials. In thin films of the T′ compounds, the valence transition has occurred already in the undoped state. The phenomenology of the valence transition is closely related to that of the neutral-to-ionic transition in mixed-stack organic charge-transfer solids. Doped cuprates have negative charge-transfer gaps, just as rare-earth nickelates and BaBiO3. The unusually high ionization energy of the closed shell Cu1+ ion, taken together with the doping-driven reduction in three-dimensional Madelung energy and gain in two-dimensional delocalization energy in the negative charge transfer gap state drives the transition in the cuprates. The combined effects of strong correlations and small d−p electron hoppings ensure that the systems behave as effective 1/2-filled Cu band with the closed shell electronically inactive O2− ions in the undoped state, and as correlated two-dimensional geometrically frustrated 1/4-filled oxygen hole band, now with electronically inactive closed-shell Cu1+ ions, in the doped state. The model thus gives microscopic justification for the two-fluid models suggested by many authors. The theory gives the simplest yet most comprehensive understanding of experiments in the normal states. The robust commensurate antiferromagnetism in the conventional T′ crystals, the strong role of oxygen deficiency in driving superconductivity and charge carrier sign corresponding to holes at optimal doping are all manifestations of the same quantum state. In the hole-doped pseudogapped state, there occurs a biaxial commensurate period 4 charge density wave state consisting of O1−−Cu1+−O1− spin singlets that coexists with broken rotational C4 symmetry due to intraunit cell oxygen inequivalence. Finite domains of this broken symmetry state will exhibit two-dimensional chirality and the polar Kerr effect. Superconductivity within the model results from a destabilization of the 1/4-filled band paired Wigner crystal [Phys. Rev. B 93, 165110 (2016) and Phys. Rev. B 93, 205111 (2016)]. We posit that a similar valence transition, Ir4+→Ir3+, occurs upon electron doping Sr2IrO4. We make testable experimental predictions in cuprates including superoxygenated La2CuO4+δ and iridates. Finally, as indirect evidence for the valence bond theory of superconductivity proposed here, we note that there exist an unusually large number of unconventional superconductors that exhibit superconductivity proximate to exotic charge ordered states, whose band fillings are universally 1/4 or 3/4, exactly where the paired Wigner crystal is most stable. View Show abstract Quantifying Anharmonic Vibrations in Thermoelectric Layered Cobaltites and Their Role in Suppressing Thermal Conductivity Optimizing multiple materials properties which are simultaneously in competition with each other is one of the chief challenges in thermoelectric materials research. Introducing greater anharmonicity to vibrational modes is one strategy for suppressing phonon thermal transport in crystalline oxides without detrimentally affecting electronic conductivity, so that the overall thermoelectric efficiency can be improved. Based on perturbed molecular dynamics and associated numerical analyses, we show that CoO2 layers in layered cobaltite thermoelectrics NaxCoO2 and Ca3Co4O9 are responsible for most of the in-plane heat transport in these materials, and that the non-conducting intermediate layers in the two materials exhibit different kinds of anharmonicity. More importantly, thermal conduction is shown to be altered by modifying the structure of the intermediate layers. The simulation methods developed to quantify the effect of anharmonic atomic vibrations on thermal conductivity provide a new tool for the rational design of thermoelectric materials, and the insights gained should hasten the attainment of higher conversion efficiencies so that thermoelectrics can be put to widespread practical use. Enhanced thermopower via spin-state modification We investigated the effect of pressure on the magnetic and thermoelectric properties of Sr3.1Y0.9Co4O10+δ. The magnetization is reduced with the application of pressure, reflecting the spin-state modification of the Co3+ ions into the nonmagnetic low-spin state. Accordingly, with increasing pressure, the Seebeck coefficient is enhanced, especially at low temperatures, at which the effect of pressure on the spin state becomes significant. These results indicate that the spin-orbital entropy is a key valuable for the thermoelectric properties of the strongly correlated cobalt oxides. Transparent conducting properties of c-axis-oriented NaxCoO2 epitaxial thin films Article Oct 2018 Thermoelectric properties of a magnetic semiconductor CuFeS 2 We present theoretical calculations of the thermoelectric properties of a magnetic semi-conductor CuFeS2. The electrical contributions, namely the Seebeck coefficients, electrical conductances, and electronic thermal conductances, are investigated using first-principles calculation techniques based on density functional theory and the nonequilibrium Green's function formalism. The lattice thermal transport properties are analyzed using the nonequilbirium molecular dynamics simulation method. Using a combination of the calculated electronic contributions and the lattice thermal contribution, the thermoelectric figures of merit are calculated and analyzed in doped configurations for the theoretical prediction of the highest thermoelectic energy conversion efficiency. It is shown that dilute doping enhances the figure of merit by 2.5 times compared to 3% doping which was previously experimentally obtained to give the best thermoelectric performance among samples studied. Electronic structure and polar catastrophe at the surface of Li x CoO 2 studied by angle-resolved photoemission spectroscopy We report an angle-resolved photoemission spectroscopy (ARPES) study of LixCoO2 single crystals which have a hole-doped CoO2 triangular lattice. Similar to NaxCoO2, the Co 3da1g band crosses the Fermi level with strongly renormalized band dispersion while the Co 3deg′ bands are fully occupied in LixCoO2 (x=0.46 and 0.71). At x=0.46, the Fermi surface area is consistent with the bulk hole concentration indicating that the ARPES result represents the bulk electronic structure. On the other hand, at x=0.71, the Fermi surface area is larger than the expectation which can be associated with the inhomogeneous distribution of Li reported in the previous scanning tunneling microscopy study by Iwaya et al. [Phys. Rev. Lett. 111, 126104 (2013)]. However, the Co 3d peak is systematically shifted towards the Fermi level with hole doping excluding phase separation between hole rich and hole poor regions in the bulk. Therefore, the deviation of the Fermi surface area at x=0.71 can be attributed to hole redistribution at the surface avoiding polar catastrophe. The bulk Fermi surface of Co 3da1g is very robust around x=0.5 even in the topmost CoO2 layer due to the absence of the polar catastrophe. Magnetism and spin entropy in Ru doped Na 0.5 CoO 2 The effect of Ru doping on the magnetic coupling among Co ions and on the Seebeck effect in Na0.5CoO2 was systematically studied using density functional theory. It was found that the Ru dopant takes the 4+ oxidation state and replaces a Co(4+) ion. In addition, the remaining Co(4+) ions in Na0.5CoO2:Ru were stabilized in a low spin state. Magnetically, the Ru dopants couple in a ferrimagnetic manner with Co ions in the host lattice. Due to the higher electronic degeneracy of Ru(4+) dopants, the Seebeck coefficient in Na0.5CoO2 is predicted to be higher than that of the pristine compound. Large thermopower from dressed quasiparticles in the layered cobaltates and rhodates The origin of the large thermopower in NaxCoO2 is complicated by correlation phenomena. To disentangle the effects from multiple interactions, we use angle-resolved photoemission to study KxRhO2, an isostructural analogy of NaxCoO2 with large thermopower and weak electron correlation. Using the experimentally measured electronic structure, we demonstrate that the thermopower in KxRhO2 can be quantitatively explained within the quasiparticle framework after including an electron-phonon mass enhancement effect. Extending the analysis to the cobaltate, we find the doubling in thermopower is well accounted for by additional band renormalization from electron correlation. As such, the large thermopower emerges from the itinerant quasiparticles dressed by hierarchical electron-phonon and electron-electron interactions. Novel natural super-lattice materials with low thermal conductivity for thermoelectric applications: A first principles study A systematic study which reveals the low thermal conductivity and high thermopower on a series of natural superlattice structures in the form BaXFCh (X: Cu, Ag, Ch: S, Se, Te), LaXSO (X: Cu, Ag) and SrCuTeF are presented. Low thermal conductivity is predicted by combining elastic constants and few well established models. The electronic properties reveal the highly two dimensional nature of band structure in the valence band, and this is confirmed through effective mass calculations. The huge difference in effective mass along different crystallographic directions in valence band introduces anisotropy in the transport properties for hole doping, and ’a’ axis is found to be more favourable. In addition to these, the parameter A (S²σ/τT/ /τ), which can decouple the relaxation time is also calculated, and it reveals the possibility of good thermoelectric properties in these compounds. Our results are comparable with prototype thermoelectric materials, and show better values than traditional TE materials. Giant Enhancement in High-Temperature Thermoelectric Figure-of-Merit of Layered Cobalt Oxide, LiCoO2, Due to a Dual Strategy-Co-Substitution and Lithiation The chemical composition of LiCoO2, a layered oxide commonly used as electrode in batteries, was changed to Li1+yCo1-xNixO2 by a combination of substitution and lithiation to enhance the thermoelectric figure-of-merit at high temperatures. Substitution of Ni as well as lithiation does not change the crystal structure, R3̅m. The lattice parameters c and a are found to increase slightly but maintain a nearly constant ratio, 4.99, indicating no lattice distortion. The trivalent Co was substituted with divalent Ni to synthesize LiCo1-xNixO2 series of p-type compounds with x varying up to 0.15. The high-temperature thermopower decreases drastically from ∼600 to 300 μV K(-1), while the electrical resistivity drops by an order of magnitude from 1 × 10(-2) to 1 × 10(-3) Ω m due to substitution of 15 atom % Ni. The total thermal conductivity also decreases from ∼3 to 1.5 W m(-1) K(-1). Increasing the amount of Li in LiCo0.85Ni0.15O2 changes the thermophysical properties further and leads to enhancement of figure-of-merit. The power factor is found to change from 37.6 μW m(-1) K(-2) for the base compound to 120 μWm(-1) K(-2), a significant improvement for a p-type compound. The overall figure-of-merit as a result increases to 0.12 at ∼1100 K due to substitution and lithiation, a giant increase of ∼760% compared to 1 × 10(-2) for the pure compound LiCoO2. These substituted and lithiated compounds are found to be extremely stable even after six months and exhibit totally reproducible thermophysical properties. High-Pressure Synthesis and Electronic Properties of Cr(IV) Oxides Synthesis and electronic properties of Cr(IV) oxides with CrO6 octrahedra are overviewed focusing on the way of the connection of the octahedra. High-pressure synthesis under 6–8 GPa is effective for the preparation of them. The reaction to make some Cr(IV) oxides is found to be promoted by higher pressures even in case that it is less promoted by elevation of reaction temperatures. Magnetic and electric properties are closely related with the deep energy level of the 3d orbitals of the Cr⁴⁺ ions. For more specific description, α-Sr2CrO4 and NaCr2O4 are taken as examples of the compounds with corner- and edge-sharing octahedra, respectively. α-Sr2CrO4 has interesting correlation between spin and orbital degrees of freedom, which occurs due to the reverse split of the t2g orbitals to the expectation of the conventional crystal-field theory. NaCr2O4 shows a new type of double exchange interaction between Cr spins owing to ligand holes. Electrically, these two compounds are insulating with small energy gaps whereas some related compounds are metallic. Thus, the Cr(IV) oxides appear to be located near the border of Mott transition. View Thermoelectric Properties of Pb and Sr Doped Ca3Co4O9 Chapter Dec 2009 Hiroshi Nakatsugawa Hyeon Mook Jeong N. Gomi Rak Hee Kim IntroductionExperimentResults and DiscussionConclusion Acknowledgments Thermoelectric Properties of Bi2Sr2Co2Oy Thin Films and Single Crystals Article Feb 2017 PHYSICA B Zhenyu Diao H.N. Lee M.F. Chisholm Rongying Jin Bi2Sr2Co2O9 exhibits a misfit-layered structure with good thermoelectric properties. We have investigated the thermoelectric properties of Bi2Sr2Co2Oy in both thin-film and single-crystal forms. Among thin films grown at different temperatures, we find that both the in-plane thermoelectric power (Sab) and electrical resistivity (ρab) vary in an opposite trend, i.e., Sab is high when ρab is small. This results in large power factor (Sab²/ρab~5.5 μW/K² cm for the film grown at 700 °C), comparable to that for whiskers. For single crystals, the electrical resistivity shows metallic behavior in a large temperature range, but has higher magnitude than that of films grown at 675 °C and 700 °C. The annealing of single crystals under Ar atmosphere leads to even higher resistivity while Sab is improved. We discuss the thermoelectric performance of this material considering both oxygen concentration and phase purity. The Role of Structure and Defect Chemistry in High-Performance Thermoelectric Bismuth Strontium Cobalt Oxides Article Sep 2016 Jakub D. Baran Demie Kepaptsoglou Marco Molinari Stephen Charles Parker [Bi0.87SrO2]2[CoO2]1.82 (BSCO) is one of the best p-type thermoelectric oxides but its structural and electronic properties are still little understood. BSCO is a misfit-layered compound consisting of an incommensurate stacking of hexagonal CoO2 and double rock-salt BiSrO2 layers. Here we combine experimental and computational approaches to investigate its crystallographic and electronic structure as well as thermoelectric transport properties. Considering different approximations for the subsystems stacking we present a structural model that agrees well with both bulk and atomic-scale experimental data. This model, which suggests a level of Bi deficiency in the rock-salt layers, is then used to discuss the material?s electronic, magnetic and transport properties. We show that Bi-deficiency leads to a band-gap opening and increases p-type electronic conductivity due to the formation of Co4+ species that serve as itinerant holes within the predominantly Co3+ framework of the CoO2 layer. We validate these predictions using electron energy loss spectroscopy in the scanning transmission electron microscope. The relationship between the hole-doping mechanism and the changes of the local structure (in particular the level of Bi deficiency) is evaluated. The reliability of the simulations is supported by the calculated temperature dependence of the Seebeck coefficient, in good agreement with experimental measurements. Specific-heat evidence for strong electron correlations in the thermoelectric material (Na, Ca) Co2O4 Article Oct 1999 Phys Rev B Yoichi Ando N. Miyamoto Kouji Segawa I. Terasaki The specific heat of (Na,Ca)Co2O4 is measured at low-temperatures to determine the magnitude of the electronic specific-heat coefficient gamma, in an attempt to gain an insight into the origin of the unusually large thermoelectric power of this compound. It is found that gamma is as large as ~48 mJ/molK2, which is an order of magnitude larger than gamma of simple metals. This indicates that (Na,Ca)Co2O4 is a strongly-correlated electron system, where the strong correlation probably comes from the low-dimensionality and the frustrated spin structure. We discuss how the large thermopower and its dependence on Ca doping can be understood with the strong electron correlations. Jul 1999 Viadimir Anisimov M. A. Korotin M. Zölfl T. M. Rice The electronic structure of the first reported heavy fermion compound without f electrons, LiV2O4, was studied by an ab initio calculation method. As a result of the trigonal splitting and d-d Coulomb interaction, one electron of the d1.5 configuration of the V ion is localized and the rest partially fills a relatively broad conduction band. The effective Anderson impurity model was solved by the non-crossing-approximation method, leading to an estimation for the single-site Kondo energy scale TK. Then, we show how the so-called exhaustion phenomenon of Nozières for the Kondo lattice leads to a remarkable decrease of the heavy-fermion (or coherence) energy scale Tcoh≡T2K/D ( D is the typical bandwidth), comparable to the experimental result. Electronic structure and heavy-fermion behavior in LiV2O4 Dec 1999 Dr Jaspreet Singh P. Blaha Karlheinz Schwarz I. I. Mazin First principles density functional calculations of the electronic and magnetic properties of spinel-structure LiV2O4 have been performed using the full potential linearized augmented plane-wave method. The calculations show that the electronic structure near the Fermi energy consists of a manifold of 12 bands derived from V t2g states, weakly hybridized with O p states. While the total width of this active manifold is approximately 2 eV, it may be roughly decomposed into two groups: high velocity bands and flatter bands, although these mix in density functional calculations. The flat bands, which are the more atomiclike, lead to a high density of states and magnetic instability of local moment character. The value of the on-site exchange energy is sensitive to the exact exchange correlation parametrization used in the calculations, but is much larger than the interaction between neighboring spins, reflecting the weak coupling of the magnetic system with the high velocity bands. A scenario for the observed heavy-fermion behavior is discussed, in which conduction electrons in the dispersive bands are weakly scattered by local moments associated with strongly correlated electrons in the heavy bands. This is analogous to that in conventional Kondo-type heavy fermions, but is unusual in that both the local moments and conduction electrons come from the same d manifold. Dramatic Change of the Magnetic Response in LiV 2 O 4 : Possible Heavy Fermion to Itinerant d-Metal Transition Apr 1999 April A. Krimmel Alois Loidl H. Schober The magnetic relaxation of the d-metal oxide LiV 2 O 4 has been measured by means of quasielastic neutron scattering. At low temperatures, the observed residual linewidth of 0.5 meV and a square-root temperature dependence of the magnetic relaxation rate are canonical features of heavy-fermion systems. For T . 40 K the magnetic response changes dramatically and can be characterized as a metal close to ferromagnetic order. [S0031-9007(99)08823-7] PACS numbers: 71.27. + a, 61.12.Ex, 71.20.Be, 71.30. + h LiV 2 O 4 is a transition-metal oxide that crystallizes in the cubic spinel-type structure [1]. In this compound the vanadium ions have the formal charge V 13.5 and it was shown two decades ago that LiV 2 O 4 is metal-lic [2]. No sign of magnetic order or superconductiv-ity could be observed for T $ 0.02 K [3] as opposed to the isostructural compounds LiTi 2 O 4 [4] and ZnV 2 O 4 [5] which become superconducting T c 13.7 K and mag-netically ordered T N 40 K, respectively. In the fcc spinel structure the transition-metal ions occupy corner-sharing tetrahedral sites and, due to this inherent geomet-rical frustration, magnetic order can easily be suppressed. Indeed, the antiferromagnetic (AFM) order in ZnV 2 O 4 be-comes possible only due to a cubic-to-tetragonal structural phase transition at 50 K [5]. Based on the results of spe-cific heat, susceptibility and NMR measurements, Kondo et al. [3] proposed that LiV 2 O 4 be the first example of a d-metal compound exhibiting heavy-fermion (HF) behavior [6] with a characteristic temperature of T 30 K [3]. Subsequently, Chmaissem et al. [7] reported an anomalous temperature dependence of the lattice con-stants below T 20 K and interpreted it in terms of a strongly enhanced Grüneisen parameter, which again is a characteristic fingerprint of HF systems. 7 Li NMR re-sults, similar to those presented by Kondo et al. [3], were published by Fujiwara et al. [8] but were interpreted in terms of a spin-fluctuation theory. Quasielastic neutron scattering experiments provide a unique characterization of the dynamic properties of a magnetic system. The most fundamental property is the dynamical susceptibility x 00 Q, v, T that can be measured via the dynamic structure Electronic structure and magnetic interactions in LiV2O4 Article Full-text available Jun 1999 V. Eyert Karl-Heinz Höck Siegfried Horn Peter Riseborough We present results of all-electron electronic structure calculations for the recently discovered d electron heavy fermion compound LiV_2O_4. The augmented spherical wave calculations are based on density functional theory within the local density approximation. The electronic properties near the Fermi energy originate almost exclusively from V 3d t_{2g} states, which fall into two equally occupied subbands: While sigma-type metal-metal bonding leads to rather broad bands, small pi-type p-d overlap causes a narrow peak at E_F. Without the geometric frustration inherent in the crystal structure, spin-polarized calculations reveal an antiferromagnetic ground state and ferromagnetic order at slightly higher energy. Since direct d-d exchange interaction plays only a minor role, ordering of the localized vanadium moments can be attributed exclusively to a rather weak superexchange interaction. With the magnetic order suppressed by the geometric frustration, the remaining spin fluctuations suggest an explanation of the low temperature behaviour of the specific heat. Comment: Europhys. Lett. 46, 762-767 (1999). Additional material avalable at http://www.physik.uni-augsburg.de/~eyert/ Planewaves, Pseudopotentials and the LAPW Method Book

https://www.researchgate.net/publication/235575202_Electronic_structure_of_NaCo_2_O_4

'I couldn't bring myself to set a batsman up by giving him runs' Glenn McGrath talks about the method behind his metronomic consistency, visualisation, and why aggression isn't about sledging 'I couldn't bring myself to set a batsman up by giving him runs' Glenn McGrath talks about the method behind his metronomic consistency, visualisation, and why aggression isn't about sledging There is this line in your autobiography: "I can't ever remember having a bad dream about bowling. When I dreamt about cricket, I just bowled the ball I wanted to."That is positive reinforcement. I used to call it visualisation. The night before a game, I'd think about who I was playing, and then how I'd bowled against those guys, if I had got them out previously. While I was playing, I could recall nearly all my wickets and how I got the batsman out. If you continually watch yourself do something well, it has a positive effect. If you sat down and watched yourself bowling, batting or fielding badly, it will probably have the equal effect. I just found that worked for me. Even when I played I'd visualise at the top of my mark the ball carrying through and what I wanted to deliver. Was that how you had always been, or did you have to work on it?I don't know, I think it is something that came pretty naturally. I am quite a positive person. I always try to see the good in every situation, the good in everybody. My wife has a go at me every now and then that I can still see too much good in people sometimes, when they probably do not deserve it. I have always been the glass-half-full person. Even if we lost a game, I'd work on the positives and then think about where we could improve. Early into your career, you had a major injury and you had to work hard on your fitness. Everyone talks about physical fitness for a fast bowler. You were probably one of the strongest bowlers mentally. How important is mental strength for a fast bowler?I came back from the West Indies in 1995. I'd torn my intercostal, one of my side muscles, where you get your power from for a fast bowler. I weighed 77 kilos, which is about 25 kilos less than what I am now. I was injured, and I thought that if I want to stay playing at this level, which I was absolutely loving, I was going to have to do something differently. So I found a trainer, who was one of the toughest, and worked with him. He made me nearly unbreakable. That was my attitude with what I wanted to do. I think being physically fit and strong is hugely important for a fast bowler. On the other side of things, you have to be mentally strong as well. My strength was probably more the mental side of the game rather than the skill side. I always had that self-belief that I was good enough. You have got to believe you are good enough, otherwise there is no point to it. I was prepared to work as hard as I could. The old saying: "The harder you work, the luckier you get" is very, very true. I would never give up. I was never satisfied. I'd always want to improve and do better next game. I felt we could win from any situation no matter how bad it was. I'd say I never gave up. I loved what I did, and if you have a real love and passion for what you do, you can't help but be successful. "I probably sledged myself a lot more than I sledged the batsman, because I had such high expectations of myself" I always had a game plan, what I was looking to achieve. When I was bowling well, I had already worked out the next two overs - what I was going to bowl and where I was going to bowl. It is just that mindset - knowing your game and yourself, how you work at your best and what you are looking to achieve. I never had any doubts when I was playing. I never worried about another bowler coming in and taking my position. All the focus was on what I wanted to achieve, and how I was going to go about doing it, and I just went out and did it. Where would you say these values came from? Is it an Australian way or from the farm, from your early years?I'd like to say it is a little bit of an Australian attitude. There were some batsmen in the team who did not like it when I made predictions and targeted batsmen of the other team. Maybe it was my upbringing - the country attitude. When you grow up on a farm you are instilled with a certain work ethic from a young age. We were driving tractors, working on the land, from a young age. Everyone has a conscious decision to make from any situation. They can either look at it from a negative perspective and let it affect them or look at it from a positive one and use that. I love life and I want to make the most of it. I try to live in the now as well. Don't think too much about the past - just experiences that have taught me things. Growing up on a farm and having that freedom as a young fellow and what my parents instilled in me - it all led me in that direction. Shane Warne called your bowling method the torture technique. Drips on the forehead till the batsman gives up. Did you think of it that way?The old Chinese water torture. Just drying them up, not letting them get any easy runs, slowly building the pressure until they got out or were shot mentally. I'd like to think I did a little bit more than slowly torture them, but it is an interesting comment from Warnie. How did you zero in on that method, coming at the taile-end of an era where fast bowlers looked to intimidate batsmen? Yours wasn't physical intimidation, it was more mental.If I could have bowled 160kph or 100mph, I would have definitely been bowling that fast. Physically, I could not. But what I did do well is, I could land the ball. I had pretty good accuracy and I could get good bounce. I was not that quick, I did not swing the ball a great deal, but what I could do, I did very well. That was my strength. I only looked to get a batsman out one of three ways: bowled, lbw or caught behind. I thought it is pointless bowling middle stump because it would take all my slips out and it makes it easy for batsmen to score runs on the leg side. So off stump, or just outside, was where I wanted to bowl. I had that mental strength and I loved the challenge of bowling to guys who were classed the best. I loved bowling in pressure situations. If I miss anything in cricket, it is being in those pressure situations, where it comes down to you having to perform for the team to win. That is what I loved. "I only looked to get a batsman out one of three ways: bowled, lbw or caught behind" © Getty Images People saw some of your qualities in Mohammad Asif. He said that line was mandatory and that he hated giving runs off the pads. How important was line to you?I hated giving the batsman even a single. If the batsman hit me for four, it wasn't because he hit a good shot. It was because I had bowled the ball where he could hit me for four. So I was a bit annoyed with myself. It is all about control. Bowling the ball in the right area, hitting the deck, top of off stump, where the batsman is not sure whether to come forward or go back. A lot of people call it the corridor of uncertainty. That is what I try to stipulate when I speak to young bowlers at the MRF Pace Foundation - that it is about control. You look at Mitchell Johnson. He is still bowling 150kph, but he has got control now, and that makes him a lethal bowler. If the bowlers have control, they can bowl it where they want to. They are going to be a lot more effective, be able to build pressure, and are going to get a lot more wickets. I do not like to see guys substitute pace for control. You just need to work harder on getting that control without giving up something else. If you have got control and pace, you are a pretty dangerous bowler. That length seemed irritating even on television. What do you do with that length? Do you come forward or go back? Was that length natural?It was pretty natural. I never looked at the spot on the wicket where I wanted to bowl. It was always sort of locked in at the top of my mark that this is the type of delivery I want to bowl and it is all about feel. The last thing I wanted to do was bowl it where the batsman wanted it to come. It is that in-between length where they cannot really come forward or go back. If they go forward it is not quite there, if they go back it is not there and they nick. That length is a different length on every wicket. You have to assess the conditions, the bounce, the seam, and then you have to adjust accordingly. I think that was one thing I did. I could adjust to the wicket very quickly, find out that length in that corridor of uncertainty and try to capitalise on that. They used to call you the Metronome. How hard is it mentally to stick to control? You had decent pace. Didn't you ever feel like indulging yourself?I look at those things as a compliment. Precision is something I look upon fondly. My goal was to bowl what I classed as the perfect game, where every ball I bowled went exactly where I wanted to bowl it. That is what I was striving for. That does not mean I have to bowl every ball on the same spot. You can still intimidate them with short-pitched bowling, with aggressive fields, set a person up for an inswinging yorker, but it is just about being able to land the ball where you want to land it. Being a fast bowler at the end of the day is an aggressive thing. It is just not being aggressive with sledging. It is about body language, attitude, field placements. It is about the way you bowl. You have got to be the whole package. How important was bounce to you? Ricky Ponting has said that it is more bounce than pace that gets batsmen out.Speaking to the guys who were classed the best batsmen in the world - you mentioned Ricky there, [Brian] Lara, [Sachin] Tendulkar, [Rahul] Dravid, guys like that, they said they would rather face someone bowling at 150kph who skidded the ball on rather than someone who bowled mid-130s and got that bounce. That was one of my weapons. If I tried to bowl too fast, I'd probably go a bit low and I lost that bounce, which I felt was a more dangerous weapon than an extra 4-5kph in pace. That was my strength. I could bowl good areas and I got bounce and a bit of seam movement and that brought all my catchers into play. What was your attitude to sledging? Was it an additional weapon?Yes and no. Some batsmen, if you have a bit of chat to them, they went to water. Someone like Lara, if you had a chat to him one day, you'd get him out because he'd go to water. Next day, if you have a chat to him, that's it, you are never going to get him out. So it worked against some batsmen, for some it didn't. It is not something where you went out and said, "We're going to target this guy. We are going to sledge him." For me, personally, I probably sledged myself a lot more than I sledged the batsman, because I had such high expectations of myself. And half the time if I said something to the batsmen it was probably more out of frustration that I didn't achieve what I wanted to with that particular delivery. It is part of the game. Test cricket is a test physically, skill-wise and mentally. And probably the mental side of the game is bigger than the other two. How did you succeed in the subcontinent? Did you modify your approach, because not many overseas fast bowlers have done well here?I tried to adjust to the conditions as quickly as I could. What are the positives of being a fast bowler in India? To me, the new ball is hard. It will carry through okay, so you have to use the new ball. Then the ball gets a bit soft, it stops swinging. Just got to keep it tight, work on the ball, then you are going to get reverse swing and all of a sudden it comes back into the bowler's favour. That is all I concentrated on. Use the new ball when it is hard and when it is old, look after it, get reverse swing, and set fairly straight fields and bowl a lot straighter than what you would in Australia. That is all I tried to do, and again, I was trying and looking at the positives, or what the game plan was on these wickets, in these conditions, and how to best succeed. I still did not want to go for runs. I never set a batsman up by giving him runs. I could not bring myself to do that. Did you have to be more patient in Asia?It still comes back to execution and control. Mystats in Indiawere not too bad compared to the rest of the world. I did not worry that I was bowling in India compared to Australia and the UK. It was just a challenge that I enjoyed. To be classed a good bowler or a great bowler, you have got to be able to perform in every condition, every country, on every type of wicket. Patience, working to a game plan, bowling in partnerships - they were all part of the game. But ultimately my motivation was taking wickets. The end result was that I was looking to get that batsman out. There were reverse-swing exponents like Wasim and Waqar. And guys like you and Curtly Ambrose made seam bowling famous at that time. Was it always seam for you?Pretty much so. When I first got selected to play for Australia, a lot of people were saying you have to bowl a consistent outswinger to be successful at Test cricket. And I wanted to be successful at Test cricket so I started swinging the ball. I remember a Test I played against England at theGabbain the 1994-95 series and I was swinging the ball quite a lot. I ended up with match figures of none for 101 at the end of that match and did not play the next three games. I went back and thought, "Well, I got picked for a reason. I got picked because of the way I bowl. So I am just going to stick to that. That bounce. That seam movement. And building pressure." That was a good learning experience. Listening to other people did not work for me. I tried it and I learned from it. My strength was hitting the deck, coming from fairly high, using that bounce and hitting the seam. Sometimes that would carry straight through, sometimes it would come back in off the seam, predominantly more so than away, but that natural variation there was enough to unsettle a lot of batsmen. That fast incutter. Was that an effort ball?It was more a natural delivery. Looking at my action - because I jumped in a bit at the end - I had a strong core, which allowed me to stay tall without falling away. But it meant I had to go across myself, which lent itself to hitting the wicket and going in to the right-hander or going away from the left-hander. Did you always have a repeatable action?That is the way I bowled. I didn't have any coaching. The first coaching I'd ever had, I was 22. And I did not model myself on anyone else. That held me in good stead, because back when I was growing up, it was all "get side-on". Dennis Lillee had the classical side-on action and he was my hero growing up. So if I had had coaching when I was younger, they would have probably tried to get me side-on. Who knows where I could have been? I may not have ever played. My body just found the most natural way to bowl and it worked for me. Nowadays we know a lot more about coaching. There is front-on and side-on, even somewhere in between. As long as your hips and shoulders are in line, it does not matter where you are within that range. Your back will be fine. It is when your shoulders and hips get out of line that you have problems. "With Warnie, the partnership we had was quite amazing. Two totally different styles of bowling but two very similar bowlers in the way we went about it" © Getty Images You lost a bit of speed towards the end. How did you make up for it?It wasn't a conscious thing to lose speed. That is the way it happened. But then it is all about control and bowling where you wanted to. Look at Jason Gillespie, who was a similar style of bowler to me but a little bit quicker. A lot of batsmen would play and miss because though they picked the line, it bounced and seamed and was past the bat before they could adjust. Whereas when I hit the deck and it did something off the wicket, the batsman would see it and had time to adjust and maybe just follow it. I got a lot more edges because of that. The fact that I was not express sometimes worked in my favour. Those legendary partnerships - McGrath-Warne, McGrath-Gillespie - which one was dearer to you?They were both equally important. I loved bowling with Jason at the other end. We still have a great friendship and I always enjoyed that. With Warnie, the partnership we had was quite amazing. Two totally different styles of bowling but two very similar bowlers in the way we went about it. He and I had very good control. We could build pressure from both ends and I can definitely thank Shane for a lot of my wickets, and he has come out and said he thanks me for some of his wickets as well. To bowl with Shane at the other end was something pretty special. We won themajority of the Test matcheswe played in and took over 1000 Test wickets between us. They are not bad stats. Were you and Curtly Ambrose similar bowlers?I think similar in what we delivered. Curtly just did it so easy. He was so loose: just come up and hit the deck and he could really get bounce and seam. He was one of the bowlers I admired from among those I played against. Curtly had two or three gears where he could crank it up. He was always pretty relaxed but if you got under his skin or fired him up, all of a sudden he could bowl another 5 to 10kph quicker and then he was a real handful. Probably a good example is our results of bowling to Michael Atherton. I got Athers out19 timesin Tests and Amby got him 17 times. So the fact that we were similar bowlers was unfortunate for Athers. Probably a style of bowling he did not enjoy the most. Did it get too easy against Athers as it went on?You'd never say it was too easy. There was only one time where I got him out where I felt it was probably a bit too easy. And that was hislast Test. I wasn't bowling that quickly, it was gun-barrel straight as the batsmen say. And I was outside off stump, and he just kept playing and missing. So I thought I'd get one a bit straighter, he just nicked it to Warnie at first slip. And I thought, "it should not really be that easy." But I think probably Athers was a bit shocked. Definitely had a mental edge over him at that stage. One of my childhood memories is Lara trying to defend against you off the back foot and a bail going up in the air. What were those battles like?I enjoyed bowling against guys who were classed the best. That really tells you how good you are. I loved bowling to Sachin, Brian. That wicket you are mentioning there was in the 1999 World Cup,at Old Trafford. We were under the pump, we had to win every game to stay in the World Cup. Lot of people say it wasan amazing delivery. I think it did just enough to beat Brian's bat and just clipped the top of off. Mark Waugh said it was gun-barrel straight, that Brian just played the wrong line, and it wasn't anything special. How quickly you remembered that dismissal. Do you still remember most of your dismissals?Some dismissals I remember. Back when I had 360-370 Test wickets, I could sit here and write them all down in order and picture how I got them out. I guess that was my motivation, and my goal was taking wickets. Bit long in the tooth now and the brain does not work as well as it used to, but I can still remember certain series, certain Tests and what have you. Did it ever change for you under different captains? Or were you always the same?No, the game plan was pretty similar as it went on. It was only the West Indies series in 1995 where our game plan was to bowl aggressively to their bowlers, to bowl lot of short-pitched stuff to intimidate them, to show them that we were here to win. Abhishek Purohit is a senior sub-editor at ESPNcricinfo

https://www.thecricketmonthly.com/story/773201/-i-couldn-t-bring-myself-to-set-a-batsman-up-by-giving-him-runs

Leviticus 16 ISV KJV Parallel ISV Parallel KJV [BSB CSB ESV HCS KJV ISV NAS NET NIV NLT HEB] <table><tbody><tr><td> International Standard Version</td><td> King James Bible</td></tr></tbody></table> <table><tbody><tr><td> 1 The LORD spoke to Moses after the death of Aaron's two sons when they had approached the LORD and died.</td><td> 1 And the LORD spake unto Moses after the death of the two sons of Aaron, when they offered before the LORD, and died;</td></tr></tbody></table> <table><tbody><tr><td> 2 The LORD told Moses, "Remind your brother Aaron that at no time is he to enter the sacred place from the room that contains the curtain into the presence of the Mercy Seat on top of the ark. Otherwise, he'll die, because I will appear in a cloud at the Mercy Seat.</td><td> 2 And the LORD said unto Moses, Speak unto Aaron thy brother, that he come not at all times into the holy placewithin the vail before the mercy seat, which isupon the ark; that he die not: for I will appear in the cloud upon the mercy seat.</td></tr></tbody></table> <table><tbody><tr><td> 3 Aaron is to enter the sacred place with a young bull for a sin offering and a ram for a whole burnt offering.</td><td> 3 Thus shall Aaron come into the holy place: with a young bullock for a sin offering, and a ram for a burnt offering.</td></tr></tbody></table> <table><tbody><tr><td> 4 He is to wear a sacred linen tunic and linen undergarments that will cover his genitals. He is to clothe himself with a sash and wrap his head with a linen turban. Because they are sacred garments, he is to wash himself with water before putting them on."</td><td> 4 He shall put on the holy linen coat, and he shall have the linen breeches upon his flesh, and shall be girded with a linen girdle, and with the linen mitre shall he be attired: these areholy garments; therefore shall he wash his flesh in water, and soput them on.</td></tr></tbody></table> <table><tbody><tr><td> 5 "He is to take two male goats for a sin offering and one ram for a whole burnt offering from the assembly of the Israelis.</td><td> 5 And he shall take of the congregation of the children of Israel two kids of the goats for a sin offering, and one ram for a burnt offering.</td></tr></tbody></table> <table><tbody><tr><td> 6 Then Aaron is to bring the bull as a sin offering for himself and make atonement for himself and his household.</td><td> 6 And Aaron shall offer his bullock of the sin offering, which isfor himself, and make an atonement for himself, and for his house.</td></tr></tbody></table> <table><tbody><tr><td> 7 Then he is to take the two male goats and present them in the LORD's presence at the entrance to the Tent of Meeting.</td><td> 7 And he shall take the two goats, and present them before the LORD atthe door of the tabernacle of the congregation.</td></tr></tbody></table> <table><tbody><tr><td> 8 Aaron is to cast lots over the two male goats—one lot for the LORD and the other one for the scapegoat.</td><td> 8 And Aaron shall cast lots upon the two goats; one lot for the LORD, and the other lot for the scapegoat.</td></tr></tbody></table> <table><tbody><tr><td> 9 Aaron is then to bring the male goat on which the lot fell for the LORD and offer it as a sin offering.</td><td> 9 And Aaron shall bring the goat upon which the LORD'S lot fell, and offer him fora sin offering.</td></tr></tbody></table> <table><tbody><tr><td> 10 The male goat on which the lot fell for the scapegoat is to be brought alive into the LORD's presence to make atonement for himself. Then he is to send it into the wilderness."</td><td> 10 But the goat, on which the lot fell to be the scapegoat, shall be presented alive before the LORD, to make an atonement with him, andto let him go for a scapegoat into the wilderness.</td></tr></tbody></table> <table><tbody><tr><td> 11 "Aaron is then to bring the bull for a sin offering for himself, thus making atonement for himself and his household. He is to slaughter the ox for himself.</td><td> 11 And Aaron shall bring the bullock of the sin offering, which isfor himself, and shall make an atonement for himself, and for his house, and shall kill the bullock of the sin offering which isfor himself:</td></tr></tbody></table> <table><tbody><tr><td> 12 Then he is to take a censer and fill it with coals from the fire on the altar in the LORD's presence. With his hands full of spiced and refined incense, he is to bring it beyond the curtain.</td><td> 12 And he shall take a censer full of burning coals of fire from off the altar before the LORD, and his hands full of sweet incense beaten small, and bring itwithin the vail:</td></tr></tbody></table> <table><tbody><tr><td> 13 "Then he is to place the incense over the fire in the LORD's presence, ensuring that the smoke from the incense covers the Mercy Seat, according to regulation, so he won't die.</td><td> 13 And he shall put the incense upon the fire before the LORD, that the cloud of the incense may cover the mercy seat that isupon the testimony, that he die not:</td></tr></tbody></table> <table><tbody><tr><td> 14 He is to take blood from the ox and sprinkle it with his forefinger toward the surface of the Mercy Seat. Then he is to sprinkle the blood on the surface of the Mercy Seat with his forefinger seven times.</td><td> 14 And he shall take of the blood of the bullock, and sprinkle itwith his finger upon the mercy seat eastward; and before the mercy seat shall he sprinkle of the blood with his finger seven times.</td></tr></tbody></table> <table><tbody><tr><td> 15 "He is to slaughter the male goat as a sin offering for the people and bring its blood beyond the curtain and do with its blood as he did with the blood of the bull: He is to sprinkle it on the Mercy Seat—that is, over the surface of the Mercy Seat.</td><td> 15 Then shall he kill the goat of the sin offering, that isfor the people, and bring his blood within the vail, and do with that blood as he did with the blood of the bullock, and sprinkle it upon the mercy seat, and before the mercy seat:</td></tr></tbody></table> <table><tbody><tr><td> 16 Then he is to make atonement on the sacred place on account of the uncleanness of the Israelis, their transgressions, and all their sins. This is how he is to act in the Tent of Meeting, which will remain with them in the middle of their uncleanness.</td><td> 16 And he shall make an atonement for the holy place, because of the uncleanness of the children of Israel, and because of their transgressions in all their sins: and so shall he do for the tabernacle of the congregation, that remaineth among them in the midst of their uncleanness.</td></tr></tbody></table> <table><tbody><tr><td> 17 "No person is to be there when he enters the Tent of Meeting to make atonement in the sacred place, until he comes out and has made atonement on account of himself, his household, and the entire assembly of Israel.</td><td> 17 And there shall be no man in the tabernacle of the congregation when he goeth in to make an atonement in the holy place, until he come out, and have made an atonement for himself, and for his household, and for all the congregation of Israel.</td></tr></tbody></table> <table><tbody><tr><td> 18 When he goes to the altar in the LORD's presence to make atonement for himself, he is to take some of the blood from the bull and the male goat, place it around the horns of the altar,</td><td> 18 And he shall go out unto the altar that isbefore the LORD, and make an atonement for it; and shall take of the blood of the bullock, and of the blood of the goat, and put itupon the horns of the altar round about.</td></tr></tbody></table> <table><tbody><tr><td> 19 and sprinkle it with the blood on his forefinger seven times, cleansing and sanctifying it from Israel's sins."</td><td> 19 And he shall sprinkle of the blood upon it with his finger seven times, and cleanse it, and hallow it from the uncleanness of the children of Israel.</td></tr></tbody></table> <table><tbody><tr><td> 20 "When he has completed making atonement at the sacred place, the Tent of Meeting, and the altar, then he is to present the live male goat.</td><td> 20 And when he hath made an end of reconciling the holy place, and the tabernacle of the congregation, and the altar, he shall bring the live goat:</td></tr></tbody></table> <table><tbody><tr><td> 21 Aaron is to lay his two hands upon the head of the male goat and confess over it the sins of Israel, all their transgressions, and all their sins, thus placing them on the head of the male goat that he'll then send out to the wilderness by the hand of a man capable of carrying out this task.</td><td> 21 And Aaron shall lay both his hands upon the head of the live goat, and confess over him all the iniquities of the children of Israel, and all their transgressions in all their sins, putting them upon the head of the goat, and shall send himaway by the hand of a fit man into the wilderness:</td></tr></tbody></table> <table><tbody><tr><td> 22 The male goat will bear on itself all their sins to a solitary land as Aaron sends the goat out to the wilderness.</td><td> 22 And the goat shall bear upon him all their iniquities unto a land not inhabited: and he shall let go the goat in the wilderness.</td></tr></tbody></table> <table><tbody><tr><td> 23 "Then Aaron is to enter the Tent of Meeting, take off his white linen clothes that he had put on when he entered the sacred place, and leave them there.</td><td> 23 And Aaron shall come into the tabernacle of the congregation, and shall put off the linen garments, which he put on when he went into the holy place, and shall leave them there:</td></tr></tbody></table> <table><tbody><tr><td> 24 He is to wash his body with water at the sacred place and put on his clothes. Then he is to go out and offer a whole burnt offering for himself and a whole burnt offering for the people, thereby making atonement on account of himself and on account of the people.</td><td> 24 And he shall wash his flesh with water in the holy place, and put on his garments, and come forth, and offer his burnt offering, and the burnt offering of the people, and make an atonement for himself, and for the people.</td></tr></tbody></table> <table><tbody><tr><td> 25 "As to the fat from the burnt offering, he is to incinerate it on the altar.</td><td> 25 And the fat of the sin offering shall he burn upon the altar.</td></tr></tbody></table> <table><tbody><tr><td> 26 The one who sent away the male goat as a scapegoat is to wash his clothes and bathe his body with water. After doing so, he may enter the camp.</td><td> 26 And he that let go the goat for the scapegoat shall wash his clothes, and bathe his flesh in water, and afterward come into the camp.</td></tr></tbody></table> <table><tbody><tr><td> 27 "The bull for the sin offering and the male goat for the sin offering, whose blood was brought into the sacred place, are to be taken outside the camp. Their skin, meat, and offal are to be incinerated.</td><td> 27 And the bullock forthe sin offering, and the goat forthe sin offering, whose blood was brought in to make atonement in the holy place, shall onecarry forth without the camp; and they shall burn in the fire their skins, and their flesh, and their dung.</td></tr></tbody></table> <table><tbody><tr><td> 28 The one who burns them is to wash his clothes and bathe his body with water. After doing so, he may enter the camp."</td><td> 28 And he that burneth them shall wash his clothes, and bathe his flesh in water, and afterward he shall come into the camp.</td></tr></tbody></table> <table><tbody><tr><td> 29 "This is to be a perpetual statute for you: On the tenth day of the seventh month, you (including both the native born and the resident alien) are to humble yourselves by not doing any work,</td><td> 29 And thisshall be a statute for ever unto you: thatin the seventh month, on the tenth dayof the month, ye shall afflict your souls, and do no work at all, whether it beone of your own country, or a stranger that sojourneth among you:</td></tr></tbody></table> <table><tbody><tr><td> 30 because on that day, atonement will be made for you to cleanse you from all your sins. You are to be clean in the LORD's presence.</td><td> 30 For on that day shall the priestmake an atonement for you, to cleanse you, thatye may be clean from all your sins before the LORD.</td></tr></tbody></table> <table><tbody><tr><td> 31 It's the Sabbath of all Sabbaths for you, so humble yourselves. This is to be a perpetual statute.</td><td> 31 It shall bea sabbath of rest unto you, and ye shall afflict your souls, by a statute for ever.</td></tr></tbody></table> <table><tbody><tr><td> 32 The priest who has been anointed and consecrated to be priest after his father is to make the atonement. He is to put on the sacred linen clothing</td><td> 32 And the priest, whom he shall anoint, and whom he shall consecrate to minister in the priest's office in his father's stead, shall make the atonement, and shall put on the linen clothes, eventhe holy garments:</td></tr></tbody></table> <table><tbody><tr><td> 33 and make atonement for the sacred sanctuary, the Tent of Meeting, and the altar where atonement is carried out. He is also to make atonement for the priests and the people of the entire assembly.</td><td> 33 And he shall make an atonement for the holy sanctuary, and he shall make an atonement for the tabernacle of the congregation, and for the altar, and he shall make an atonement for the priests, and for all the people of the congregation.</td></tr></tbody></table> <table><tbody><tr><td> 34 This will be a perpetual statute for you as you make atonement once a year for the Israelis on account of all their sins." So Moses did just as the LORD had commanded him.</td><td> 34 And this shall be an everlasting statute unto you, to make an atonement for the children of Israel for all their sins once a year. And he did as the LORD commanded Moses.</td></tr></tbody></table> <table><tbody><tr><td> The Holy Bible: International Standard Version® Release 2.1 Copyright © 1996-2012 The ISV FoundationALL RIGHTS RESERVED INTERNATIONALLY.</td><td> King James Bible, text courtesy ofBibleProtector.com.</td></tr></tbody></table>

https://www.biblehub.com/p/isv/kjv/leviticus/16.shtml

Water Resources Act 1991 Mae'r cwcis ar deddfwriaeth.gov.uk yn gwneud dau beth: maent yn cofio unrhyw osodiadau rydych chi wedi'u dewis felly does dim rhaid i chi eu dewis ar bob tudalen, ac maent yn ein helpu i ddeall sut mae pobl yn pori ein gwefan, er mwyn i ni allu gwneud gwelliannau a thrwsio problemau. Mae angen eich caniatâd arnom i ddefnyddio rhai o'r cwcis hyn. Ydyn, mae'r cwcis hyn yn IAWN Dysgu mwy neu osod dewisiadau unigol ar gyfer cwcis Nac ydyn, hoffwn wrthod yr holl gwcis Yn ôl i’r edrychiad llawn Water Resources Act 1991 Section 137. SCHEDULE 16 E+W SCHEMES IMPOSING SPECIAL DRAINAGE CHARGES Submission of scheme E+W 1 (1) Before submitting a special charges scheme to either of the Ministers, the [ F1 Agency ] shall consult organisations appearing to it to represent the interests of persons engaged in agriculture in the area designated in the scheme. E+W (2) As soon as any special charges scheme has been submitted to either of the Ministers, the [ F1 Agency ] shall— (a) send copies of the scheme to— (i) the council of any county, [ F2 county borough ] district or London borough wholly or partly within the relevant area; (ii) the drainage board for any internal drainage district within the relevant area; and (iii) every organisation appearing to the [ F1 Agency ] to represent the interests of persons engaged in agriculture in the relevant area; and (b) publish, in one or more newspapers circulating in the area affected by the scheme, a notice stating— (i) that the scheme has been submitted to that Minister; (ii) that a copy of it is open to inspection at a specified place; and (iii) that representations with respect to the scheme may be made to that Minister at any time within one month after the publication of the notice. (3) Where the [ F1 Agency ] submit a special charges scheme which designates any watercourse wholly or partly within an internal drainage district, then (unless the [ F1 Agency ] is the drainage board for that district) the scheme must be accompanied either— (a) by a statement of the drainage board for that district that they have consented to the designation; or (b) by a statement that they have not consented thereto and a further statement setting out the reasons why the watercourse should nevertheless be designated for the purposes of section 137 of this Act. Textual Amendments F1 Words in Sch. 16 para. 1 substituted (subject to other provisions of the amending Act) (1.4.1996) by 1995 c. 25 , s. 120 , Sch. 22 para. 128 (with ss. 7(6) , 115 , 117 ); S.I. 1996/186 , art. 3 F2 Words in Sch. 16 para. 1(2)(a)(i) inserted (1.4.1996) by 1994 c. 19 , s. 22(5) , Sch. 11 Pt. I para. 3(10)(a) (with ss. 54(4)(7) , 55(5) , Sch. 17 paras. 22(1) , 23(2) ); S.I. 1996/396 , art. 3 , Sch. 1 Confirmation of scheme E+W 2 (1) Subject to the following provisions of this Schedule the Minister to whom a special charges scheme has been submitted may by order made by statutory instrument confirm the scheme either with or without modifications. E+W (2) Neither of the Ministers shall confirm a special charges scheme unless he is satisfied that the scheme is reasonable and financially sound, having regard to all the circumstances, and in particular to any contributions from local authorities and internal drainage boards which, if the scheme is confirmed, are likely to be available to the [ F3 Agency ] in addition to the special drainage charge authorised by the scheme. (3) An order confirming a special charges scheme may contain provisions with respect to the persons by whom all or any of the expenses incurred by either of the Ministers or by other persons in connection with the making or confirmation of the order, or the making of the scheme, are to be borne. Textual Amendments F3 Word in Sch. 16 para. 2 substituted (subject to other provisions of the amending Act) (1.4.1996) by 1995 c. 25 , s. 120 , Sch. 22 para. 128 (with ss. 7(6) , 115 , 117 ); S.I. 1996/186 , art. 3 Notice of proposed order E+W 3 (1) Before either of the Ministers makes an order confirming a special charges scheme he shall cause notice of— E+W (a) the intention to make it; (b) the place where copies of the draft order may be inspected and obtained; and (c) the period within which, and the manner in which, objections to the draft order may be made, to be published in the London Gazette and in such other manner as he thinks best adapted for informing persons affected and to be sent to the persons specified in sub-paragraph (2) below. (2) The persons referred to in sub-paragraph (1) above are— (a) every county council [ F4 , county borough council ] or London borough council in whose area any part of the area proposed to be affected by the order is situated and, if any part of that area is situated in the City of London, the Common Council of the City of London; and (b) the [ F5 Agency ] and every drainage body, navigation authority, harbour authority or conservancy authority that is known to the Minister in question to be exercising jurisdiction within the area proposed to be affected by the order. (3) In sub-paragraph (2) above “ drainage body ” has the same meaning as in section 108 of this Act. Textual Amendments F4 Words in Sch. 16 para. 3(2)(a) inserted (1.4.1996) by 1994 c. 19 , s. 22(5) , Sch. 11 Pt. I para. 3(10)(b) (with ss. 54(4)(7) , 55(5) , Sch. 17 paras. 22(1) , 23(2) ); S.I. 1996/396 , art. 3 , Sch. 1 F5 Word in Sch. 16 para. 3 substituted (subject to other provisions of the amending Act) (1.4.1996) by 1995 c. 25 , s. 120 , Sch. 22 para. 128 (with ss. 7(6) , 115 , 117 ); S.I. 1996/186 , art. 3 Determination of whether to make order E+W 4 (1) Before either of the Ministers makes an order confirming a special charges scheme, he— E+W (a) shall consider any objections duly made to the draft order; and (b) may, in any case, cause a public local inquiry to be held with respect to any objections to the draft order. (2) Each of the Ministers shall have power, in making an order confirming a special charges scheme, to make such modifications in the terms of the draft as appear to him to be desirable. Procedure and other matters after the making of an order E+W 5 (1) After either of the Ministers has made an order confirming a special charges scheme, the order (together with a notice under sub-paragraph (2) below) shall be published in such manner as he thinks best adapted for informing the persons affected. E+W (2) A notice under this sub-paragraph is a notice— (a) that the Minister in question has made the order; and (b) that the order will become final and have effect unless, within such period of not less than thirty days as may be specified in the notice, a memorial praying that the order shall be subject to special parliamentary procedure is presented to that Minister, by a person who is affected by the order and has such interest as may be prescribed by regulations made by one of the Ministers as being sufficient for the purpose. Orders subject to special parliamentary procedure E+W 6 (1) If— E+W (a) no such memorial as is mentioned in paragraph 5(2) above has been presented within the period so mentioned in respect of any order confirming a special charges scheme; or (b) every such memorial has been withdrawn, the Minister who made the order shall confirm the order and it shall thereupon have effect. (2) If such a memorial has been presented in respect of such an order and has not been withdrawn, the order shall be subject to special parliamentary procedure. (3) An order confirming a special charges scheme shall in any event be subject to special parliamentary procedure if the Minister who makes the order so directs. (4) The Minister who makes an order confirming a special charges scheme may, at any time before it has been laid before Parliament, revoke, either wholly or partially, any order that is subject to special parliamentary procedure. Notice of unconfirmed orders E+W 7 E+W As soon as may be after an unconfirmed order has effect, the Minister who made the order shall publish in the London Gazette, and in such other manner as he thinks best adapted for informing persons affected, a notice— (a) stating that the order has come into force; and (b) naming a place where a copy of it may be seen at all reasonable hours. Challenge to unconfirmed orders E+W 8 (1) If any person aggrieved by an unconfirmed order desires to question its validity on the ground— E+W (a) that it is not within the powers of this Act; or (b) that any requirement of this Act has not been complied with, he may, within six weeks of the relevant date, make an application for the purpose to the High Court. (2) Where an application is duly made to the High Court under this paragraph, the High Court, if satisfied— (a) that the order is not within the powers of this Act; or (b) that the interests of the applicant have been substantially prejudiced by any requirements of this Act not having been complied with, may quash the order either generally or in so far as it affects the applicant. (3) Except by leave of the Court of Appeal, no appeal shall lie to the House of Lords from a decision of the Court of Appeal in proceedings under this paragraph. (4) Subject to the preceding provisions of this paragraph an unconfirmed order shall not at any time be questioned in any legal proceedings whatsoever. (a) where the order is subject to special parliamentary procedure, the date on which the order becomes operative under the M1 Statutory Orders (Special Procedure) Act 1945; (b) where the order is not subject to special parliamentary procedure, the date of the publication of the notice mentioned in paragraph 7 above. Marginal Citations M1 1945 c. 18 (9 & 10 Geo 6) . Power to make regulations for purposes of Schedule E+W 9 E+W The Ministers may make regulations in relation to— (a) the publication of notices under this Schedule; (b) the holding of public local inquiries under this Schedule and procedure at those inquiries; and (c) any other matters of procedure respecting the making of orders confirming a special charges scheme. Interpretation E+W 10 (1) In this Schedule— E+W “ special charges scheme” means a scheme under section 137 of this Act; and “ unconfirmed order” means an order confirming a special charges scheme, other than one which is itself confirmed under section 6 of the Statutory Orders (Special Procedure) Act 1945. (2) Section 113 of this Act shall apply for the interpretation of this Schedule as it applies for the interpretation of Part IV of this Act.

https://www.legislation.gov.uk/cy/ukpga/1991/57/schedule/16/2004-07-05?view=plain

Histopathological Comparison Between Superficial Pressure Ulcers and Incontinence-associated Dermatitis - Full Text View - ClinicalTrials.gov Histopathological Comparison Between Superficial Pressure Ulcers and Incontinence-associated Dermatitis The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been evaluated by the U.S. Federal Government. Read our disclaimer for details. ClinicalTrials.gov Identifier: NCT03685929 Recruitment Status : Completed First Posted : September 26, 2018 Last Update Posted : November 29, 2021 Sponsor: University Hospital, Ghent Information provided by (Responsible Party): University Ghent Study Details Study Description Brief Summary: This study aims to identify differences and similarities between superficial pressure ulcers (category II and III) and incontinence-associated dermatitis (category IIA) at tissue level. Skin biopsies will be obtained from 30 patients with pressure ulcers (n = 10), incontinence-associated dermatitis (n = 10), and combined lesions (n = 10). The results from the histopathologic examination will be compared with the clinical diagnosis by wound care experts, which will be based on photographs and relevant patients' characteristics. Condition or disease Intervention/treatment Pressure Ulcer Incontinence-associated Dermatitis Irritant Contact Dermatitis Diaper Rash Procedure: Punch skin biopsy Detailed Description: Despite current preventive strategies, prevalence of pressure ulcers and incontinence-associated dermatitis (IAD) is still estimated at 5.9-6.9% in hospitalised patients, and 5.0-5.9 in home care patients. Both skin conditions may occur in the same body region but are considered as separate entities, based on differences in etiology. At tissue level, it is assumed that pressure ulcers are characterised by ischemia and tissue deformation. In contrast, IAD is assumed to be characterised by an irritative pattern. These assumptions are reflected in current preventive strategies. However, a small scale study, performed by Houwing et al. (2007), described the additional presence of an ischemic pattern in patients with IAD. More insights into the histopathologic pattern of pressure ulcers and IAD is needed to optimise current preventive strategies. Tha aim of this study is to compare superficial pressure ulcers (category II and III) and IAD (category IIA) at tissue level. Layout table for study information Study Type : Observational Actual Enrollment : 30 participants Observational Model: Case-Only Time Perspective: Cross-Sectional Official Title: Histopathological Comparison Between Superficial Pressure Ulcers and Incontinence-associated Dermatitis Actual Study Start Date : October 1, 2018 Actual Primary Completion Date : December 31, 2019 Actual Study Completion Date : December 31, 2019 Resource links provided by the National Library of Medicine MedlinePlus related topics: Biopsy Pressure Sores U.S. FDA Resources Groups and Cohorts Group/Cohort Intervention/treatment Pressure ulcers Patients with pressure ulcers category II/III at sacrum or trochanter Procedure: Punch skin biopsy Two punch biopsies will be performed in each patient, one at the border of the skin lesion, and one at the adjacent healthy skin. A disposable biopsy punch of 4 mm will be used. A biopsy punch is a small circular blade (like an apple corer) which is rotated into the skin to obtain a cylindrical specimen. Local anaesthesia will be applied and the circular wound will be closed by a single suture. Sutures will be removed after one week. Incontinence-associated dermatitis Patients with incontinence-associated dermatitis category IIA at sacrum (or trochanter) Procedure: Punch skin biopsy Two punch biopsies will be performed in each patient, one at the border of the skin lesion, and one at the adjacent healthy skin. A disposable biopsy punch of 4 mm will be used. A biopsy punch is a small circular blade (like an apple corer) which is rotated into the skin to obtain a cylindrical specimen. Local anaesthesia will be applied and the circular wound will be closed by a single suture. Sutures will be removed after one week. Combined lesion Patients with pressure ulcer category II/III and incontinence-associated dermatitis category IIA at sacrum or trochanter Procedure: Punch skin biopsy Two punch biopsies will be performed in each patient, one at the border of the skin lesion, and one at the adjacent healthy skin. A disposable biopsy punch of 4 mm will be used. A biopsy punch is a small circular blade (like an apple corer) which is rotated into the skin to obtain a cylindrical specimen. Local anaesthesia will be applied and the circular wound will be closed by a single suture. Sutures will be removed after one week. Outcome Measures Primary Outcome Measures : Prevalence of pressure ulcers and incontinence-associated dermatitis [ Time Frame: 1 day ] Presence of pressure ulcers and/or incontinence-associated dermatitis based on photographs and patient characteristics Histopathologic pattern [ Time Frame: 1 day ] Secondary Outcome Measures : Rate of agreement between visual and histopathologic diagnosis [ Time Frame: 1 day ] Agreement between diagnosis of wound care experts (based on photographs), and diagnosis by skin pathologists (based on histopathologic examination) Biospecimen Retention:   Samples Without DNA Punch skin biopsy Eligibility Criteria Information from the National Library of Medicine Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the contacts provided below. For general information, Learn About Clinical Studies. Layout table for eligibility information Ages Eligible for Study: 18 Years and older   (Adult, Older Adult) Sexes Eligible for Study: All Accepts Healthy Volunteers: No Sampling Method: Non-Probability Sample Study Population Patients admitted on following wards of a university hospital Pneumology Gastroenterology Endocrinology Nephrology Neurology Geriatrics Criteria Inclusion Criteria: Incontinent for urine, stool, or both Superficial pressure ulcer (category II or III) and/or incontinence-associated dermatitis (category IIA) Expected length of stay of 7 days following punch skin biopsy Exclusion Criteria: Skin infection at biopsy site Medical contraindication to obtain a biopsy End of life care Not able to provide informed consent Locations University Ghent Investigators Layout table for investigator information Principal Investigator: Sofie De Schepper University Hospital, Ghent Publications: Beeckman D, Van Damme N, Schoonhoven L, Van Lancker A, Kottner J, Beele H, Gray M, Woodward S, Fader M, Van den Bussche K, Van Hecke A, De Meyer D, Verhaeghe S. Interventions for preventing and treating incontinence-associated dermatitis in adults. Cochrane Database Syst Rev. 2016 Nov 10;11(11):CD011627. doi: 10.1002/14651858.CD011627.pub2. Layout table for additonal information Responsible Party: University Ghent ClinicalTrials.gov Identifier: NCT03685929 History of Changes Other Study ID Numbers: 2018/0946 First Posted: September 26, 2018 Key Record Dates Last Update Posted: November 29, 2021 Last Verified: November 2021 Individual Participant Data (IPD) Sharing Statement: Plan to Share IPD: No Layout table for additional information Studies a U.S. FDA-regulated Drug Product: No Studies a U.S. FDA-regulated Device Product: No Keywords provided by University Ghent: Pressure ulcer Incontinence-associated dermatitis Histopathology Ischemia Irritation Prevention Tissue Skin

https://clinicaltrials.gov/ct2/show/NCT03685929

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(PDF) A new modeling approach for mixture fraction statistics based on dissipation elements PDF | The probabilty density function (PDF) of the mixture fraction is of integral importance to a large number of combustion models. Here, a novel... | Find, read and cite all the research you need on ResearchGate PDF Available A new modeling approach for mixture fraction statistics based on dissipation elements September 2020 Proceedings of the Combustion Institute38(2) DOI: 10.1016/j.proci.2020.06.208 License CC BY-NC-ND 4.0 Authors: <here is a image f2f62089ad6ff61c-d19572ff1996375a> Dominik Denker Dominik Denker <here is a image ae54ee794086d28b-d9b6e73755d79a4e> Antonio Attili The University of Edinburgh <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Michael Gauding RWTH Aachen University <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Kai Niemietz RWTH Aachen University Abstract and Figures The probabilty density function (PDF) of the mixture fraction is of integral importance to a large number of combustion models. Here, a novel modelling approach for the PDF of the mixture fraction is proposed which employs dissipation elements. While being restricted to the commonly used mean and variance of the mixture fraction, this model approach individually considers contributions of the laminar regions as well as the turbulent core and the turbulent/non-turbulent interface region. The later region poses a highly intermittent part of the flow which is of high relevance to the non-premixed combustion of pure hydrocarbon fuels. The model assumptions are justified by means of the gradient trajectory based analysis of high fidelity direct numerical simulation (DNS) datasets of two turbulent inert configurations and a turbulent non-premixed jet flame. The new dissipation element based model is validated against the DNS datasets and a comparison with the beta PDF is presented. <here is a image 683751a0a2e07471-b74d3b26ed748fc0> PDFs of the arithmetic mean P ( Z m ) for (solid lines): DNS and (dash-dotted lines): modeling results. ( ): Mixing Layer, ( ): Jet Flame and ( ): Inert Jet. … <here is a image 7e7f7c0d643a56f6-b1b23d5dae5561ab> PDF of the mixture fraction P ( Z ) for fiv e cross-stream positions y / h 1/2 , with the jet half width h 1/2 , for the Jet Flame (left) and Inert Jet (right). Top to bottom: PDFs obtained from DNS, DE-based model and β-PDF. Please cite this article as: D. Denker, A. Attili and M. Gauding et al., A new modeling approach for mixture fraction statistics based on dissipation elements, Proceedings of the Combustion Institute, https://doi.org/10.1016/j.proci.2020. 06.208 … <here is a image 662f533c156d027b-a00059b85bb1dcf6> Mean density ρ obtained from the convolution of P ( Z ) with a steady state flamelet solution for the Jet Flame. (Solid line): P DNS ( Z ), (dashed line): P DE ( Z ), and (dash-dotted line): P β ( Z ). … Figures - available via license: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International ARTICLEINPRESS JID: PROCI [mNS; September 5, 2020;11:11 ] Available online at www.sciencedirect.com Proceedings ofthe Combustion Institute 000 (2020) 1–9 www.elsevier.com/locate/proci A new modeling approach for mixturefractionstatistics basedondissipation elements Dominik Denker a , ∗ , Antonio Attili a , Michael Gauding b , Kai Niemietz a , Mathis Bode a , Heinz Pitsch a a Institute for Combustion Technology, RWTH Aachen University, Aachen 52062, Germany b CORIA - CNRSUMR 6614, Saint Etienne duRouvray, France Received 5 November 2019; accepted 8 June 2020 Abstract The probabilty densityfunction (PDF) of the mixturefraction is ofintegral importance to a large number ofcombustion models. Here, a novel modelling approach for the PDF ofthe mixturefraction is proposed which employs dissipation elements. While being restricted to the commonly used mean and variance of the mixture fraction, this modelapproachindividually considerscontributions of the laminar regions as well as the turbulent core and the turbulent/non-turbulent interface region. The later region poses a highly intermit- tent part of the ow which is ofhigh relevance to the non-premixed combustion ofpure hydrocarbon fuels. The model assumptions are justied by means of the gradient trajectory based analysis of high delity direct numerical simulation(DNS)datasets of two turbulent inert congurations anda turbulent non-premixed jet ame. The new dissipation element based model is validated against the DNS datasets anda comparison with the beta PDF ispresented. © 2020 Published by Elsevier Inc. on behalf of The Combustion Institute. Keywords: Turbulent non-premixed ames; Direct numerical simulations; High Reynolds numbers; Combustion modeling 1. Introduction Many modeling approaches forpracticalengi- neering combustionapplications, suchas internal combustion enginesor combustors inaircraft engines,are based on the mixture fraction Z . An example is the amelet model [1] . In this model, the ∗ Corresponding author. E-mail address: [email protected] (D. Denker). assumption of fast, but not innitely fast, chemical time scalesleads tothin laminaramestruc- tures attached tothe larger amesurface which is corrugated byturbulence. In thecontext of non-premixed combustionin the amelet regime, all reacting scalarsare a functionof themixture fraction Z and thescalar dissipation rate χ st at the position ofthe stoichiometric mixture fraction or progressvariable. As themixturefraction is a randomlyuctuating quantityina turbulent ow, the accuratepredictionof theprobability densityfunction (PDF)of themixturefraction https://doi.org/10.1016/j.proci.2020.06.208 1540-7489 © 2020 Published by Elsevier Inc. on behalf of The Combustion Institute. Please cite this articleas: D. Denker, A. Attili and M. Gauding et al., A new modeling approach for mixture fraction statistics based on dissipation elements, Proceedings ofthe Combustion Institute, https://doi.org/10.1016/j.proci.2020. 06.208 2 D. Denker, A. Attili and M. Gauding et al. / Proceedings of theCombustion Institute xxx (xxxx) xxx ARTICLEINPRESS JID: PROCI [mNS; September 5, 2020;11:11 ] P ( Z ) is of fundamentalimportance to the quality of any simulation employing amelets or any other mixture fraction based combustion models. Therefore, extensive effortshave beenmade to understand andmodel theproperties of themix- ture fraction PDF [2–4] . In a binary mixing process ina fully turbulent ow, P ( Z ) is mostcommonly modeled employing the betafunction P β ( Z ) [5] . Conveniently, P β ( Z ) can be computed exclusively from the mean Z and variance Z  2 , which are read- ilyavailablein most Reynolds Averaged Navier Stokes (RANS) simulations. Theusage of P β ( Z ) stems in large part from the ability to matchexperimentallymeasured mixturefraction PDFs. However, inhighlyinter- mittent ows, P ( Z ) displays shapes which cannot be adequately reproduced by P β ( Z ), such as more complex multimodalshapes ora singularity at either Z = 0 or Z = 1and anintermediate local maximum. Due to limitationsof the experimental setups and measurement techniques, which are fur- ther exacerbated by the higher values the viscosity in reacting ows, fundamental turbulentcom- bustion experimentsoften use dilutedfuelsand consequentlyahigh stoichiometricmixturefrac- tion. Ye t , real worldcombustion applicationsof hydrocarbons rarely employ fuel streams with such high leveles ofdilution and therefore typically have very low values of Z st . This places the combustion in the highly intermittent region at the outer edge of theturbulent regionsin free shearows in the vicinity or even inside the turbulent/non-turbulent interface (TNTI) [6] . Effelsberg & Peters [7] arguedthatthe TNTI represents a third regime of nitethickness infree shear ows, which separates the laminarouter ow and fully turbulent inner ow. To account for the ef- fects of the TNTI layer on the statisticsof Z , they derived a composite PDF model taking the previ- ously mentionedthree regimesinto account.Ex- cellent agreement with experimental data ofround jets with a wide range ofReynolds numbers was re- ported [8] . However,this model relieson the rst four momentsof the mixturefraction Z -Z 4 . On top of the challenge ofsolving transport equations for up to Z 4 with a multitudeof further modeling assumptions required to close these transport equa- tions, the inherent nature ofthe diffusively domi- nated TNTI layer and the ill posed eddy viscosity ν t in the vicinity of laminar regions poses the question whether this is notonly impractical but potentially impossible ina RANS sense. A potential substitute for the lack of the higher momentsof Z inRANS proposed here is the use of gradienttrajectory (GT)statistics. Peters & Troulliet [9] demonstrated thereconstructionof P ( Z ) and P ( χ ) using datafrom a direct numerical simulation (DNS) of a temporally evolving mixing layer by means of GTs of the mixture fraction eld. Furthermore, GTs were used in the precise regime classicationof free shearows intoturbulent core regions,interfaceregions,and the outer ow by Mellado et al. [10] . Aphysicallymotivated method forgrouping GTs andcompartmentalizing scalarelds φ is the Dissipation Element (DE)analysisdeveloped by Wang & Peters [11,12] . DEs are space lling regions in which the scalarbehavesmonotonically,i.e. the space between two turbulenceinduced extremal points. The entiresub-regionof the scalar eld which constitutes a DE can then be parametrized by the two linked extremal points, namely the linear separation distance between the minimum and the maximum  and the scalar difference in these points φ . DE analysis was extensivelyapplied toinert turbulent ows [13–17] . A remarkable invariance of the normalized statistics of  ∗ = /  and φ ∗ = φ/ φ, with the volume averaged separation dis- tance  andscalar difference φ, towards changes in the chosen scalar eld and Reynolds number was observed.This invariance of  ∗ carries over to re- acting ows, andif the scalar eld does not possess asource term, of φ ∗ as well [18] . Indeed, the re- construction of P ( Z ) obtained from the experimen- tal measurements of aninertround jet using only DE parameterstatistics wasdemonstrated [19] . However, these earlier reconstruction studies relied on theDE parameterstatistics directly obtained from the specic DNS orexperiments; the univer- sality of the parameter statistics was not leveraged and modelingpotentials were restricted. In the rst part, aftera briefdescriptionof the numerical methods, aGT basedinvestigationof the the Z -elds obtained fromthree DNS data sets is shown and the reconstruction of P ( Z ) by means ofDE parameterstatistics willbedemonstrated. The rst congurationisamoderately turbulent inert planartemporallyevolving jet. To show the feasibility of the approach in reacting ows, a non- premixed planar temporally evolving jet ame with a comparable Reynolds number is also considered. To demonstrate thatthe inuence ofthevarious ow regimeson the mixturefractionstatistics are not exclusivelycaused by low Reynoldsnumber effects and to rule out any particularitiesof a tem- porallyevolvingconguration, the reconstruction procedureisapplied toahighly turbulent inert spatially evolving mixinglayer [20] . In the secondpart, aDE-parameter statis- tic based compositePDF approach P DE will be explained exploiting the invariances of the normal- ized DEparameters andscalingswithstatistical quantities readily availablein RANS. Additionally, the modelassumptions forthe DE parameter statistics aremotivated andjustied bymeans of the Z -elds of the three DNS cases. Finally, the modelling results for P DE ( Z ) are pre- sented andcompared to the DNS results.Unlike the beta PDF, P DE ( Z ) captures all important char- acteristics of the mixturefraction in the turbulent Please cite this articleas: D. Denker, A. Attili and M. Gauding et al., A new modeling approach for mixture fraction statistics based on dissipation elements, Proceedings ofthe Combustion Institute, https://doi.org/10.1016/j.proci.2020. 06.208 D. Denker, A. Attili and M. Gauding et al. / Proceedings ofthe Combustion Institute xxx(xxxx) xxx 3 ARTICLEINPRESS JID: PROCI [mNS; September 5, 2020;11:11 ] Table 1 Numerical and physical initial parameters ofthe DNS. Jet Flame Inert Jet Mixing Layer Re jet,0 10,0009850 – Re λ 86.4 101.1 305.1 L x 8.5 H 0 6 π H 0 473 δ ω ,0 L y 8.2 H 0 12.5 H 0 290 δ ω ,0 L z 6.4 H 0 6 π H 0 158 δ ω ,0 n gridpoints [10 9 ] 1.2 8.6 18.7  / η 0.86 1.4 1.1 core, as wellas the TNTI layer ofthe respective congurations. 2. Direct numerical simulations 2.1.Congurations The initial analysisand the nalvalidation are based on three high delity DNS data sets. The rst congurationis an inertnon-dimensionalplanar temporally evolving jet henceforth called theInert Jet. This congurationpossesses periodicbound- ary conditions inboththestream-wise x and the span-wise z directionsandfree-slipconditions in the cross-stream direction y . The stream-wise ve- locity componentand the passivescalar eldare prescribed via a hyperbolic tangentprole. In the core region, the initial velocity is perturbedwith a broadband random Gaussian velocity eld derived from a one-dimensionalturbulent energy spec- trum to speed up the laminar-turbulent transition. The initial jetReynolds numberis set to Re jet = U 0 H 0 /ν = 9850 , with the initial mean center line velocity U 0 and initial jetwidth H 0 . The resolu- tion in the core region is  / η ≈ 1.4, with the Kol- mogorov micro scale η = ν 3 / 4 ε − 1 / 4 and ε the mean turbulent kinetic energydissipation rate. Further detailsregarding the DNS congurationsand nu- merical parameters can be found in Table 1 . The secondcongurationisanon-premixed planar temporally evolving jet called the Jet Flame in the following investigation. The domain is peri- odic in thestream-wiseandspan-wise directions. Boundary conditions inthe cross-wise direction were chosen as outlets [21] . The initial Reynolds numberis set to Re jet = U 0 H 0 /ν fuel = 10,000, with kinematicviscosityof the fuel ν fuel . The turbulent Damköhlernumber Da τ = χ q H / U 0 is set to Da = 0 . 15 . The oxidizer stream in all reactingcases isair, consistingof oxygen Y O 2 , 1 = 0 . 232 andnitrogen Y N 2 , 1 = 0 . 768 . The fuel streamconsists of highlydilutedmethane with Y CH 4 , 2 = 0 . 07 toachieve a high stoichiomet- ric mixturefractionof Z st = 0 . 45 . The velocity eld in the jetcore wasinitialized withan in- stantaneous realizationof a turbulent channel ow. The Z -eld wasinitialized using an error functionprole. The resolution was chosen to re- solve the OH with at least 10 grid points at all times. The minimum Kolmogorov scale is  / η ≈ 0.86. The nalcongurationisa non-dimensional inert planar spatially evolving mixinglayer hence- forth called the Mixing Layer. Along the stream- wise direction,theboundary conditions are imposed inow at x = 0and free outow [21] at x = L x . The boundary conditions are periodic in the span-wise directionandfree-slip in the cross- wise direction. The inow isahyperbolic tangent prole for thestream-wise velocity uwith pre- scribedinitial vorticitythickness δ ω ,0 . Low ampli- tude white noise is superimposed on the hyperbolic tangent prole, resulting inthe fasterfacilitation into a fullyturbulent owdownstream fromthe inlet. The Taylor micro scalebasedReynolds number reaches up to Re λ = u  λ/ν ≈ 305 . 1 , with the turbulence intensity u  and λbeing the Taylor length scale. The resolutionin theinvestigated regionsis  / η ≤ 1.1. Further details regarding the conguration can be found in [22] . 2.2.Numerical methods The Inert Jet DNS is performed by solving the non-dimensionalunsteadyincompressible Navier- Stokes equationsemploying thein-housesolver psDNS [23] . Additionally, an advection-diffusion equation issolved fora passivescalar.Spatial derivatives are calculated bythe implicitsixth- order nite difference compact schemeintroduced by Lele [24] . The temporal integration is performed using alow storage fourth-orderRunge-Kutta method. ThePoisson equation issolved inspectral space by adapting a Helmholtzequation [25] . The Jet FlameDNS wasperformedwith the reactive, unsteady Navier-Stokes equationsin the low-Mach numberlimit using thein-house solver CIAO [26] . The transport of species mass frac- tions is described using the Hirschfelderand Cur- tiss approximation to thediffusive uxes together with a velocity-correction approach for mass con- servation [27] . Momentum equations arespatially discretizedwith afourth-order nite difference scheme. Species and temperature equations are dis- cretized with a fth-order WENO scheme [28] . The temperature and species equations are advanced by introducing the symmetric operator split of Strang [29] . The two independent operators account for transport andreaction.Time advancement is achieved using asemi-implicitCrank-Nicholson time integration. Thechemistryoperator uses a time-implicit backward difference method [30] . For further details aboutthe appliednumericalalgo- rithms andcode verication, the reader is referred to Desjardins et al. [26] . Combustion is modeled using a reducedmechanism forthe oxidationof methane comprising 28 speciesand 102 reactions Please cite this articleas: D. Denker, A. Attili and M. Gauding et al., A new modeling approach for mixture fraction statistics based on dissipation elements, Proceedings ofthe Combustion Institute, https://doi.org/10.1016/j.proci.2020. 06.208 4 D. Denker, A. Attili and M. Gauding et al. / Proceedings ofthe Combustion Institute xxx(xxxx) xxx ARTICLEINPRESS JID: PROCI [mNS; September 5, 2020;11:11 ] [31] . Additionally, the formation ofNO is included by means of the Zeldovich mechanism [32] . The Mixing Layer DNS was performedsolving the unsteady Navier-Stokes equationsin thelow Mach number limit employing the solver NGA de- veloped at Stanford University [26] . Identical to the methods employed for the Jet Flame DNS, the solverimplements a nite difference method on a spatially andtemporallystaggered gridwith the semi-implicit fractional-stepmethod [33] . 3.Gradient trajectory analysis In the following section, the profoundly differ- ent characteristics of the zonal PDFs in the turbu- lent core P TC ( Z ) and in the TNTI layer P TNT ( Z ) will be investigated using GTanalysis. The cru- cial abilityofconsidering non-localeffects in an analysis employing GTs will be exploited. 3.1.Regime decomposition A challenge faced in theanalysis of freeshear ows is the presence of both laminar and turbulent regions. Physicalpropertiesof theow change drastically from one region to the other. Therefore, itis apparent to see that precisely separating tur- bulent andlaminar partsis of primeimportance to obtain meaningfulstatistics. While different and easy-to-use methodsoflocating the TNTI exist [6,34] , the ow in the turbulent region just behind the TNTI would not be considered fully tur- bulent [35] . Simply using all material points behind the TNTI wouldpotentially lead to theinclusion of lowReynolds number effectsin the statistics. The region between the TNTI andtheturbulent core needs to be considered a third region [7] . A methodbasedonGTs foraprecise regime classication was proposed by Mellado et al. [10] . The turbulent core region of a free shear ow is de- ned by regions of the ow which possessDEs, i.e. regionsin which all materialpoints can be linked to a pair of extremal points via GTs. The TNTI re- gionis then dened by the materialpoints whose GTs cross the TNTIas dened by the methodof Bisset et al. [34] , which uses thresholds ofthe en- strophy for the TNTI detection. Statistics of the arithmetic mean Z m between the extremal points connected by GTsof a DE Z m = Z max + Z min 2 , (1) with Z max and Z min being the mixturefractionin the maximumand the minimum, respectively, and the scalar difference  Z in these points  Z = Z max − Z min , (2) show distinctly different characteristics in the two ow regimes. In Fig. 1 , this is demonstrated via the jointprobabilitydensity functions (JPDFs) of the two DEparameters in theturbulent core P TC ( Z m ,  Z ) and in the TNTI region P TNT ( Z m ,  Z ). The JPDFs display fundamentally different shapes in the two ow regimes, with P TNT ( Z m ,  Z ) display- ing a thin line at Z m =  Z/ 2 . This signies that the GTs indeed continue indenitelyonce the TNTI is crossed. Furthermore, the fundamental differences serve to highlight the importance of the considera- tion of a distinct TNTI region in a model approach for P ( Z ) in free shear ows. 3.2.Modeling the mixture fraction structure For the desired gradient-trajectory-based recon- struction ofmixturefractionstatistics, theGTs need to be properly parametrized. For this purpose, Peters and Truoillet [9] proposed the previously in- vestigated Z m and  Z on these points. Amodel for the scalar structure between these extremal points was then proposed as Z ( s ) =  Z 2 sin ( π s − π/ 2) + Z m , (3) were s is the arc-length distance of the GT from the minimum. The results ofthe model for Z ( s ) are compared to results from the three DNS cases. To this end, the normalized mixture fraction Z ∗ = ( Z − Z min ) /  Z is conditioned on the normalizedarc-length s ∗ = s / s max , with the total arc length s max from the minimum tothe maximum. In Fig. 2 a), the conditional mean  Z ∗ | s ∗  is shown for the turbulent core regions of thethreeDNScases, anda pas- sive scalar eldobtained from DNSofisotropic turbulence [36] . The normalized mixture fraction shows a perfectcollapse for all cases and displays a symmetric shape. Furthermore,the presence of heat release, variable density, andviscosity does not affect the small scale structure ofthe mixture fraction and the proposedanalytic model is a very good t of the DNS data. This nding is signicant as itdemonstrates thatthebalancebetween the turbulent generationand diffusive annihilation of extremal points and thestructureof the passive scalardoes notfundamentally differin reacting and non-reacting ows. In Fig. 2 b),  Z ∗ | s ∗  is shown for the TNTI region. Here, the endpoint for GTs beyond the TNTI is determined by the gradi- ent threshold based methodof Mellado et al. [10] . While the conditional means share characteristics among the cases, differences can beobserved; foremost between the mixinglayerand jet cong- urations. Unsurprisingly,the agreement withthe model, which was derived for the turbulent core, is not perfect anymore in the TNTI regions. However, in thefollowing investigation andreconstruction procedure, this inaccuracy is accepted in favour of retaining a fully analytical approach and the model Eq. (3) is also applied to the TNTI regions. Please cite this articleas: D. Denker, A. Attili and M. Gauding et al., A new modeling approach for mixture fraction statistics based on dissipation elements, Proceedings ofthe Combustion Institute, https://doi.org/10.1016/j.proci.2020. 06.208 D. Denker, A. Attili and M. Gauding et al. / Proceedings ofthe Combustion Institute xxx(xxxx) xxx 5 ARTICLEINPRESS JID: PROCI [mNS; September 5, 2020;11:11 ] Fig. 1. JPDFofthe DE parameters conditioned on turbulent core region (left) and TNTI region (right) in the Inert Jet case. Fig. 2.Mean normalized mixture fraction conditioned on the normalizedarc-length  Z ∗ | s ∗  a) in the turbulent core and b) the TNTI region. ( ): Isotropic Turbulence, ( ): Mixing Layer, ( ): Jet Flame, ( ): Inert Jet, ( ): model. Assuming P ( s ) to be uniform, the conditional model PDF P ( Z | Z m ,  Z ) can be derived [9] as P ( Z ) = P ( s ) | ∂ Z /∂ s | . (4) The derivative ∂ Z / ∂ s can be obtainedanalytically from Eq. (3) . Therefore, P ( Z ) can be expressed with the DEparameters P ( Z | Z m ,  Z ) = π − 1 | ( Z − Z m +  Z/ 2) 1 / 2 ( Z m +  Z/ 2 − Z ) 1 / 2 | . (5) Finally, the zonal PDF P i ( Z ) can be reconstructed by integrating the conditional model PDF with the JPDF of the DE parameters P i ( Z ) =  1 0   Z max 0 P ( Z | Z m ,  Z ) P i ( Z m ,  Z )d  Z d Z m (6) The results ofthe reconstruction procedure are shown in Fig. 3 . For all cases, excellent results in theturbulent core region and the overall ow are achieved. The reconstructed PDF in the TNTI re- gion reproduces the DNS well for large to interme- diate values of Z . For smaller values of Z, P TNT ( Z ) qualitatively matches the DNS results. The differ- ence in the reconstructed PDFsstemsfromthe deviation of theGTscrossing the TNTIfrom the universal form observed in Fig. 3 b) for the turbu- lent core. Keeping in mind that the model for Z ( s ) was obtained tomatch the turbulent core region, the deviations from the DNS casesare acceptable. 4.DE basedmodeling approach In this section, the original proposed composite PDF [7] is modied using the previously introduced PDF reconstruction andexploiting other charac- teristicsofDE parameter statistics with the result of reducing the required input. The composite PDF P DE ( Z ) is dened as P DE ( Z ) = (1 − γ ) δ ( Z ) + γ [ γ t P TC ( Z ) + (1 − γ t ) P TNT ( Z ) ] , (7) with the intermittencyfactor γ asdened by [37] and the diracfunction δ ( Z ) represent- ing the irrotational part of the ow. The turbulent intermittency factor γ tisanalogous to γand signies the probability of encountering turbulent core in theturbulent region. P TC ( Z ) and P TNT ( Z ) are thepreviouslyanalysedzonal PDFs. The intermittency factor γ requires thedenition of a threshold ofthe enstrophy ω 2 0 to distinguish between turbulent andnon-turbulent uid andis therefore linked to the TNTI. However, the results are very insensitive to the specic choice for ω 2 0 as at the TNTI achange in magnitudeof the threshold will only result in a changed position of the TNTI inthe order ofthe Kolmogorov micro scale [38] . Please cite this articleas: D. Denker, A. Attili and M. Gauding et al., A new modeling approach for mixture fraction statistics based on dissipation elements, Proceedings ofthe Combustion Institute, https://doi.org/10.1016/j.proci.2020.

https://www.researchgate.net/publication/345453790_A_new_modeling_approach_for_mixture_fraction_statistics_based_on_dissipation_elements

JCM | Free Full-Text | Radial Extracorporeal Shockwave Therapy versus Ultrasound Therapy in Adult Patients with Idiopathic Scoliosis Background: This study aimed to compare the effectiveness of radial extracorporeal shockwave and ultrasound therapies in adult patients with idiopathic scoliosis in terms of pain, disability, and quality of life. Methods: Forty-eight patients with idiopathic scoliosis were randomly divided into three groups of 16: shockwave, ultrasound, and control. The patients were evaluated at admission (day one) and at discharge (day 14) for pain, by using the visual analogue scale; for disability, by using the Oswestry disability index; and for the quality of life, with short form-36. Results: Radial extracorporeal shockwave therapy was more effective than ultrasound in reducing pain (p = 0.004) and increasing quality of life, bringing extra vitality (p = 0.003) and emotional comfort (p = 0.007) to the patient. Both shockwave therapy (p = 0.001) and ultrasound therapy (p = 0.003) were effective in reducing pain. In terms of disability, both treatments had similar effects (p = 0.439). Conclusion: Radial shockwave was significantly more effective than ultrasound in reducing pain and increasing the quality of life, bringing additional vitality and emotional comfort to the patient with idiopathic scoliosis. In terms of disability, both treatments had similar effects when associated with kinesitherapy. Radial Extracorporeal Shockwave Therapy versus Ultrasound Therapy in Adult Patients with Idiopathic Scoliosis Cristina Daia 1,2,† , Cristian Scheau 3,* , Corneliu Toader 1,4,† , Ana Maria Bumbea 5,6,† , Visarion Danut Caimac 5,6,† , Ioana Andone 1,2 , Cristina Popescu 1,2 , Aura Spanu 1,2 and Gelu Onose 1,2 Neuromuscular Department, Clinical Emergency Hospital “Bagdasar Arseni”, 041914 Bucharest, Romania 3 Department of Physiology, “Carol Davila” University of Medicine and Pharmacy, 050474 Bucharest, Romania 4 Department of Neurosurgery, National Institute of Cerebro-Vascular Diseases, 041914 Bucharest, Romania 5 Department of Medical Rehabilitation, University of Medicine and Pharmacy, 200349 Craiova, Romania 6 Neurorehabiltation Department, Clinical Neuropsychiatry Hospital, 200473 Craiova, Romania * Author to whom correspondence should be addressed. † These authors have contributed equally to this work and share first authorship. J. Clin. Med. 2021 , 10 (8), 1701; https://doi.org/10.3390/jcm10081701 Received: 24 February 2021 / Revised: 9 April 2021 / Accepted: 12 April 2021 / Published: 15 April 2021 (This article belongs to the Special Issue Improved Rehabilitation for Patients with Chronic Pain ) Abstract : Background: This study aimed to compare the effectiveness of radial extracorporeal shockwave and ultrasound therapies in adult patients with idiopathic scoliosis in terms of pain, disability, and quality of life. Methods: Forty-eight patients with idiopathic scoliosis were randomly divided into three groups of 16: shockwave, ultrasound, and control. The patients were evaluated at admission (day one) and at discharge (day 14) for pain, by using the visual analogue scale; for disability, by using the Oswestry disability index; and for the quality of life, with short form-36. Results: Radial extracorporeal shockwave therapy was more effective than ultrasound in reducing pain ( p = 0.004) and increasing quality of life, bringing extra vitality ( p = 0.003) and emotional comfort ( p = 0.007) to the patient. Both shockwave therapy ( p = 0.001) and ultrasound therapy ( p = 0.003) were effective in reducing pain. In terms of disability, both treatments had similar effects ( p = 0.439). Conclusion: Radial shockwave was significantly more effective than ultrasound in reducing pain and increasing the quality of life, bringing additional vitality and emotional comfort to the patient with idiopathic scoliosis. In terms of disability, both treatments had similar effects when associated with kinesitherapy. Keywords: radial extracorporeal shockwave therapy ; ultrasound therapy ; adult idiopathic scoliosis ; pain ; quality of life ; disability 1. Introduction Idiopathic scoliosis is a three-dimensional spinal deviation of the spinal axis with a curvature exceeding 10° on a plain anteroposterior X-ray image, with no other underlying disease identified to provoke it [ 1 ]. It is the most common spinal deformity that develops in otherwise healthy children [ 2 ]. A proportion of 23% to 51% of adolescents with idiopathic scoliosis had significant back pain [ 2 , 3 ]. Back pain is approximately twice as prevalent in adolescents with idiopathic scoliosis compared with non-scoliosis patients [ 4 ], and it decreases the patients’ quality of life [ 5 ]. Literature data regarding pain in adults with idiopathic scoliosis are scarce; however, a main cause of pain in idiopathic scoliosis is muscle contracture, mainly located in the convexity of the spine [ 6 ]. Adults with idiopathic scoliosis have back pain generated by self-sustaining muscle contractures [ 7 ] induced by movements; hence, it is important to have an effective and secure therapeutic approach with muscle-relaxing effects such as physiotherapy procedures, as in as shockwave (SW) [ 7 ] or ultrasound (US) therapies [ 8 ], which are two related procedures in terms of mechanical and physical properties. Oscillatory electro-mechanic therapy is the application of oscillation waves with high frequency and pressure, which relies on a mechanical means of propagation. In theory, this category includes ultrasonic waves and ballistic shock waves. Ultrasound is a mechanical oscillation produced by physiotherapy devices based on the inverted piezoelectric effect, which spreads within tissues with high frequencies in the range of 8 × 10 5 –1 × 10 6 Hz [ 9 , 10 , 11 ]. Therapeutic dosing of US is carried out according to power density (W/cm 2 ), application time (minutes), and pulse application form (continuous/burst) [ 9 , 10 , 11 ]. Shockwaves are a complex of “high-pressure” oscillation waves that yield around 150 mega Pascals (MPa), causing real mechanical shocks in the targeted tissues. Alternate names such as “high-pressure waves”, “ballistic waves”, or “shockwaves” were assigned for this very reason, as they cause micro-trauma [ 12 , 13 ]. There are multiple means of producing SWs. One method is based on the electrohydraulic effect, resulting in low-pressure SWs, also known as radial shockwaves (RSW). Another manner of generating SWs relies on the electromagnetic effect, the inverted piezoelectric effect, and a spark discharge, producing high-pressure SWs, labeled focused shockwaves. Shockwave therapy dosing factors in the number of shocks, application time, frequency, energy transfer, and form of application (continuous or burst) [ 12 ]. Both US and SWs produce a mechanical impact on the target tissue, causing similar physical and chemical effects, such as heat generation, cavitation, and diffusion. The two types of waves also have common clinical effects, mainly, pain relief and muscle relaxation [ 14 , 15 ]. However, the pressure level at which these effects occur is different, as SWs have a pressure peak of at least 1000 times stronger than that of US [ 16 , 17 ]. Therefore, patient pathology plays an important role in choosing the appropriate method [ 18 , 19 ]. The effectiveness of US and SWs has been compared in pathologies such as fibromyalgia/myofascial pain syndrome [ 20 ], plantar fasciitis [ 21 , 22 ], epicondylitis [ 23 , 24 ], and tendonitis of the shoulder [ 25 ], and no significant differences were noted in terms of alleviating pain, increasing the quality of life (using the short form-36 (SF-36)), or improving disability. In our paper, we aim to measure the effectiveness of these two treatment modalities in terms of improving the pain, disability, and quality of life in adult patients with idiopathic scoliosis. 2. Materials and Methods The study comprises 48 patients diagnosed with idiopathic dorsal-lumbar scoliosis, either admitted to the hospital or benefiting from ambulatory care, between February 2019 and February 2020. 2.1. Inclusion Criteria Patients over 30 years old; Low back dorsal-lumbar nonspecific musculoskeletal pain; Diagnosed with dorsal-lumbar idiopathic scoliosis of any type (mild, moderate, or severe) and any degree of curvature from 10 to 60 degrees. 2.2. Exclusion Criteria Our study employed the classical contraindications for any electrotherapy procedure [ 7 ]: Idiopathic scoliosis that required surgery intervention; Any vertebral or spinal bone marrow pathology other than idiopathic scoliosis: disc herniation, spinal cord injury, benign or malignant tumors, and so on; Fever, regardless of the cause; Acute or decompensated diseases, regardless of their nature: infectious, inflammatory, cardiovascular, respiratory, neurological, rheumatological, metabolic, and so on; Uncontrolled hypertension with systolic blood pressure higher than 150 mmHg; Any mental disorder; Any neoplasm; Osteosynthesis materials or other metals in the area of application; Patients carrying a cardiac pacemaker; Pregnant women, women during menstruation, women with intrauterine devices (IUD); Patients with altered general condition or cachexia; Patients with any pathology that could generate a disability other than idiopathic scoliosis; Patients with any type of idiopathic scoliosis other than dorsal-lumbar; Patients with any pain other than low-back dorsal-lumbar pain. 2.3. Study Design In our study, we used block randomization to obtain three balanced groups. We set block randomization for gender (male and female), scoliosis type (dorsal and lumbar; then, left and right curvatures), curves in degree categories (0–10, 11–20, 21–30, 31–40, 41–50, 51–60), and age groups (30–40, 41–50, 51–60, 61–70, 71–80) ( Table 1 ). We balanced the number of subjects in each block in order to increase the comparability between the groups. Group A ( n = 16) received US treatment (UST), applied bilaterally in the paravertebral dorsal-lumbar region. The regimen was 0.5 W/cm 2 , for 5 min, using continuous emission, dynamic application in a slow rhythm, and daily application rhythm over a total of 10 sessions. Group B ( n = 16) received RSW treatment (RSWT) to the paravertebral dorsal-lumbar area using a pressure of 0.2 MPa (2 Barr) in addition to a series of 2000 shocks in pulse emission (BURST mode) with a frequency of 10 Hz and an average energy transfer of 0.4 m J/mm 2 for a duration of 4.48 min. The application rate was every two days with a total of five sessions. Radial extracorporeal shockwave therapy was delivered every two days in order to avoid microtrauma cumulating effects. Group C ( n = 16) was the control group and did not receive physiotherapy. The total treatment time was 14 days. 2.4. Patient Rehabilitation Management Each patient signed a detailed informed consent. The study respects the ethical principles for research presented in the Declaration of Helsinki. All patients included in this study underwent adequate treatment in compliance with ethical guidelines, standing good practices, and associated comorbidities. All patients benefited from the same kinesitherapy procedures as part of the rehabilitation program. The kinesitherapy program was provided by a kinetotherapist and consisted of a sum of specific exercises named the Klapp method [ 27 ]. All patients benefited from the same exercise program, applied daily for the same period of 10 days, 60 min per day. 2.5. Patient Evaluation All patients were evaluated according to the national protocol of evaluation in non-traumatic spine pathologies [ 28 ]. The clinical and paraclinical evaluation consists of recording and describing six specific syndromes through various diagnostic methods: (1) spinal, active, and passive (posture and radiography); (2) muscle (contracture, ultrasonography [ 29 ]); (3) radicular (neurologic examination and MRI); (4) spinal cord (neurologic examination); (5) psychological (psychological examination); and (6) functional (disability evaluation) [ 28 ]. The pain experienced by the patients in our study was caused by muscle contraction, and this was a criterion for enrollment in this study. The kinesitherapy program was administrated concomitant with the interventions, for all groups. 2.6. Evaluated Parameters For all groups, the total time for evaluation was 14 days. The following parameters were assessed: Pain: using the visual analogue scale (VAS), which highlights 10 degrees of pain from 0 (no pain) to 10 points (the most severe pain) [ 30 ]. The VAS scale was performed on days 1 and 14 for all patients, before and after the application of US and RSW for the study groups. Disability: using the Oswestry disability index (ODI). The ODI scale was performed on days 1 and 14 for all patients [ 31 ]. Quality of life (QoL): using short form-36 (SF-36), which consists of 8 subscales with 36 items that evaluate the physical, social, and usual activity limitations due to physical or emotional problems, bodily pain, mental health, vitality, and general health perceptions [ 32 ]. The QoL survey was performed only on day 14 for all patients. 2.7. Statistical Analysis SPSS Statistics for Windows version 15.0 (SPSS Inc., Chicago, IL, USA) was used for statistical analysis. The distribution of patient demographic features was analyzed using descriptive statistical methods. Population normality was assessed using the Kolmogorov–Smirnov test. Levene’s test for equality of variances, t -test for equality of means, the Mann–Whitney test, and the Wilcoxon test were used for comparison of parameters. The Tamhane post hoc evaluation was employed after multiple comparisons were performed by ANOVA when equal variances were not assumed. Pearson’s chi-squared test was used for comparing categorical data. The statistical significance of p -value was considered p < 0.05. For the Kolmogorov–Smirnov test, p -value was considered p > 0.2. 3. Results The demographic analysis of the study groups showed that the patients were similar in terms of age (F = 0.07, sig. 0.926, ANOVA) and gender (0.182 Pearson chi-square, sig. 0.913) distribution ( Table 2 ). The analysis of the initial VAS in all lots shows similar values (0.485, ANOVA; 0.963, Tamhane post hoc). The evaluation of pain on the VAS scale shows that the average differences in the final VAS (compared with the initial score) were significantly lower in the RSWT lot (−6.38 ± 1.02) compared with the UST lot (−5.38 ± 0.71) and the control group (−4.50 ± 0.62), with a statistical significance of p < 0.001 and p = 0.004, respectively (Mann–Whitney). The difference in the VAS score was also significantly lower in the UST patients compared with the control patients ( Table 3 ). The analysis of the initial ODI in all lots shows a comparable value (0.465 ANOVA, and 0.981 Tamhane post hoc). Regarding the evaluation of disability using ODI (highlights), the average of the differences between the final and the initial ODI in the RSWT group (−28.06 ± 4.32) did not differ significantly from that of the UST group (−29.56 ± 6.29) or the control group (−27.31 ± 4.743) ( p = 0.361 Mann–Whitney, p = 0.439, ANOVA, post hoc Tamhane 0.955). Regarding the QoL, the SF-36 subscales showed significantly better values for the following parameters: vitality (means 21.22 versus 11.78; p = 0.003) and emotional component (means 20.84 versus 12.16; p = 0.007) in the RSWT group versus UST; no other statistically significant differences were noted ( Table 4 ). No significant immediate or delayed adverse reactions were observed following the physiotherapy procedures. In this study, we used five large block randomizations to increase the comparability between groups by keeping the ratio of the number of subjects between them almost the same. Consequently, the risk of selection bias was increased as the treatment of the subject in the block was known. Because we analyzed a new physiotherapy treatment, we chose this type of randomization in order to recognize its therapeutic benefits, starting from similar lots, to the best of our ability [ 33 ]. Although randomization minimizes the selection bias in this case–control study, it was not a blinded intervention; therefore, another limitation of this study was allowing the subsequent differential cointerventions or biased assessment of outcomes [ 34 ]. The number of cases in the analyzed groups is low; therefore, the results we share through this study should be considered as indicative while encouraging subsequent research to verify them on larger populations. For the symptomatic treatment of idiopathic scoliosis, we chose to compare two methods of physiotherapy with similar features, as they both rely on mechanical waves. Participants had low-back muscle pain caused by prolonged muscle contracture, especially from the convexity of scoliosis. Patients had no headache, sore areas, spinal pain, generalized pain, or of any other type. The selection of a clear type of pain (muscle contraction pain), due to a specific type of idiopathic scoliosis (dorsal-lumbar scoliosis), could be another limitation of the antalgic potential of the proposed physiotherapy: RSWT and UST. On the other hand, we demonstrated that RSWT and UST have significant statistical antalgic effects acting on muscle contractures; therefore, we can recommend this kind of physiotherapy for this specific kind of pain. The main limitation of this study should be considered the lack of a standard protocol regarding RSWT and UST in scoliosis. To our knowledge, this is one of the first studies in the field, and the results appear promising. However, more studies on a larger number of patients are needed to confirm these results. Another limitation is the short-term evaluation of the outcomes assessed in the study. In our view, a follow-up examination would be the next step to be addressed in further studies. Analyzing the results, we observed that RSWT produces a statistically significant decrease in pain generated by idiopathic scoliosis versus UST. It should be noted, however, that both treatments determined an average decrease in the VAS score for pain: UST by 5.38 points, and RSWT by 6.38 points, respectively; however, RSWT proved to be statistically more effective and was further followed by a significantly higher state of emotional comfort and vitality according to the analysis of the patients’ QoL. Regarding the rest of the parameters analyzed in SF-36, we did not detect any other statistically significant differences. The mental health component of QoL SF-36 was not analyzed because the presence of mental illness was considered an exclusion criterion in our study; moreover, according to literature data, mental health factors do not influence QoL SF-36 in patients with scoliosis [ 35 ]. Radial extracorporeal shockwave therapy has been demonstrated to have a significant influence on the reduction of pain in patients with chronic low-back pain [ 36 ]. The statistically proven superior analgesic effect of RSWT may be explained by its particular mechanism of destroying unmyelinated sensory fibers [ 37 ] added to fibrinolytic properties [ 38 ] when compared to UST [ 39 ], determining the lysis of fibrin bridges in long-lasting scoliosis contractures, while also enabling effective muscle relaxation and subsequent participation in movement. Regarding the disability induced by idiopathic scoliosis in adults [ 40 ], which was investigated in our study using ODI, there were no statistically significant differences in its improvement after therapy. This finding is consistent with the fact that all patients benefited from a similar kinetic program, an element with an essential role in terms of patient functionality [ 41 ]. Both types of physiotherapy were very well-tolerated by patients, who showed no side effects; therefore, we recommend both methods in terms of safety and tolerability profile. 5. Conclusions After a short-term evaluation, radial extracorporeal shockwave therapy and ultrasound therapy were both effective in reducing pain due to muscle contraction in adult patients with idiopathic scoliosis; however, RSWT had statistically superior effects in reducing pain and increasing the quality of life, as well as bringing additional vitality and emotional comfort to the patients. In terms of disability, both treatments have similar effects when associated with kinesitherapy. Both types of physiotherapy are safe and well-tolerated by patients with idiopathic scoliosis. Further studies involving larger study groups and follow-up examinations are needed in order to extend and confirm our reported findings, as well as to develop appropriate standardized protocols for these types of patients. We would like to thank T. Spircu for his assistance with statistics and data interpretation. Additionally, we thank Toma Gheorghe for his support and dedication in aiding patient recovery in the field of physio- and kinesi-therapy. The authors declare no conflict of interest. References Trobisch, P.; Suess, O.; Schwab, F. Idiopathic scoliosis. Dtsch. Arztebl. Int. 2010 , 107 , 875–884. [ Google Scholar ] [ CrossRef ] Choudhry, M.N.; Ahmad, Z.; Verma, R. Adolescent Idiopathic Scoliosis. Open Orthop. J. 2016 , 10 , 143–154. [ Google Scholar ] [ CrossRef ][ Green Version ] Jackson, R.P.; Simmons, E.H.; Stripinis, D. Incidence and severity of back pain in adult idiopathic scoliosis. Spine 1983 , 8 , 749–756. [ Google Scholar ] [ CrossRef ] Théroux, J.; Le May, S.; Fortin, C.; Labelle, H. Prevalence and management of back pain in adolescent idiopathic scoliosis patients: A retrospective study. Pain Res. Manag. 2015 , 20 , 153–157. [ Google Scholar ] [ CrossRef ][ Green Version ] Makino, T.; Kaito, T.; Kashii, M.; Iwasaki, M.; Yoshikawa, H. Low back pain and patient-reported QOL outcomes in patients with adolescent idiopathic scoliosis without corrective surgery. Springerplus 2015 , 4 , 397. [ Google Scholar ] [ CrossRef ][ Green Version ] Alves, D.P.L.; de Araújo, B. Muscle disorders in adolescent idiopathic scoliosis: Literature review. Coluna/Columna 2016 , 15 , 73–77. [ Google Scholar ] [ CrossRef ][ Green Version ] Daia, C.; Onose, G. Towards a new indication for extracorporeal shock wave therapy: Idiopathic scoliosis. Proc. Rom. Acad. Ser. B 2017 , 19 , 171–176. [ Google Scholar ] Janicki, J.A.; Alman, B. Scoliosis: Review of diagnosis and treatment. Paediatr. Child Health 2007 , 12 , 771–776. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Miller, D.L.; Smith, N.B.; Bailey, M.R.; Czarnota, G.J.; Hynynen, K.; Makin, I.R.; Bioeffects Committee of the American Institute of Ultrasound in Medicine. Overview of therapeutic ultrasound applications and safety considerations. J. Ultrasound Med. 2012 , 31 , 623–634. [ Google Scholar ] [ CrossRef ][ Green Version ] Chapelon, J.Y.; Cathignol, D.; Cain, C.; Ebbini, E.; Kluiwstra, J.U.; Sapozhnikov, O.A.; Fleury, G.; Berriet, R.; Chupin, L.; Guey, J.L. New piezoelectric transducers for therapeutic ultrasound. Ultrasound Med. Biol. 2000 , 26 , 153–159. [ Google Scholar ] [ CrossRef ] Robertson, J.V. Dosage and treatment response in randomized clinical trials of therapeutic ultrasound. Phys. Ther. Sport 2012 , 3 , 124–133. [ Google Scholar ] [ CrossRef ] Van der Worp, H.; Van den Akker-Scheek, I.; Van Schie, H.; Zwerver, J. ESWT for tendinopathy: Technology and clinical implications. Knee Surg. Sports Traumatol. Arthrosc. 2013 , 21 , 1451–1458. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Reilly, J.M.; Bluman, E.; Tenforde, A.S. Effect of Shockwave Treatment for Management of Upper and Lower Extremity Musculoskeletal Conditions: A Narrative Review. PM R 2018 , 10 , 1385–1403. [ Google Scholar ] [ CrossRef ] [ PubMed ] Ebadi, S.; Henschke, N.; Forogh, B.; Nakhostin Ansari, N.; van Tulder, M.W.; Babaei-Ghazani, A.; Fallah, E. Therapeutic ultrasound for chronic low back pain. Cochrane Database Syst. Rev. 2020 , 7 , CD009169. [ Google Scholar ] [ CrossRef ] Zhang, L.; Fu, X.B.; Chen, S.; Zhao, Z.B.; Schmitz, C.; Weng, C.S. Efficacy and safety of extracorporeal shock wave therapy for acute and chronic soft tissue wounds: A systematic review and meta-analysis. Int. Wound J. 2018 , 15 , 590–599. [ Google Scholar ] [ CrossRef ] Izadifar, Z.; Babyn, P.; Chapman, D. Mechanical and Biological Effects of Ultrasound: A Review of Present Knowledge. Ultrasound Med. Biol. 2017 , 43 , 1085–1104. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Notarnicola, A.; Moretti, B. The biological effects of extracorporeal shock wave therapy (eswt) on tendon tissue. Muscles Ligaments Tendons J. 2012 , 2 , 33–37. [ Google Scholar ] [ PubMed ] Morishita, K.; Karasuno, H.; Yokoi, Y.; Morozumi, K.; Ogihara, H.; Ito, T.; Hanaoka, M.; Fujiwara, T.; Fujimoto, T.; Abe, K. Effects of therapeutic ultrasound on range of motion and stretch pain. J. Phys. Ther. Sci. 2014 , 26 , 711–715. [ Google Scholar ] [ CrossRef ][ Green Version ] Dedes, V.; Stergioulas, A.; Kipreos, G.; Dede, A.M.; Mitseas, A.; Panoutsopoulos, G.I. Effectiveness and Safety of Shockwave Therapy in Tendinopathies. Mater. Sociomed. 2018 , 30 , 131–146. [ Google Scholar ] [ CrossRef ] Aktürk, S.; Kaya, A.; Çetintaş, D.; Akgöl, G.; Gülkesen, A.; Kal, G.A.; Güçer, T. Comparision of the effectiveness of ESWT and ultrasound treatments in myofascial pain syndrome: Randomized, sham-controlled study. J. Phys. Ther. Sci. 2018 , 30 , 448–453. [ Google Scholar ] [ CrossRef ][ Green Version ] Dedes, V.; Tzirogiannis, K.; Polikandrioti, M.; Dede, A.M.; Nikolaidis, C.; Mitseas, A.; Panoutsopoulos, G.I. Radial Extra Corporeal Shockwave Therapy Versus Ultrasound Therapy in the Treatment of Plantar Fasciitis. Acta Inform. Med. 2019 , 27 , 45–49. [ Google Scholar ] [ CrossRef ] Greve, J.M.A.; Grecco, M.V.; Santos-Silva, P.R. Comparison of Radial Shockwaves and Conventional Physiotherapy for Treating Plantar Fasciitis. Clinics 2009 , 64 , 97–103. [ Google Scholar ] [ CrossRef ][ Green Version ] Yalvaç, B.; Mesci, N.; Külcü, D.G.; Yurdakul, O.V. Comparison of ultrasound and extracorporeal shock wave therapy in lateral epicondylosis. Acta Orthop. Traumatol. Turc. 2018 , 52 , 357–362. [ Google Scholar ] [ CrossRef ] [ PubMed ] Dedes, V.; Tzirogiannis, K.; Polikandrioti, M.; Dede, A.M.; Mitseas, A.; Panoutsopoulos, G.I. Comparison of radial extracorporeal shockwave therapy with ultrasound therapy in patients with lateral epicondylitis. J. Med. Ultrason. 2020 , 47 , 319–325. [ Google Scholar ] [ CrossRef ] [ PubMed ] De Boer, F.A.; Mocking, F.; Nelissen, E.M.; Van Kampen, P.M.; Huijsmans, P.E. Ultrasound guided Needling vs. Radial Shockwave Therapy in calcific tendinitis of the shoulder: A prospective randomized trial. J. Orthop. 2017 , 14 , 466–469. [ Google Scholar ] [ CrossRef ] Barrios, C.; Pérez-Encinas, C.; Maruenda, J.I.; Laguía, M. Significant ventilatory functional restriction in adolescents with mild or moderate scoliosis during maximal exercise tolerance test. Spine 2005 , 30 , 1610–1615. [ Google Scholar ] [ CrossRef ] [ PubMed ] Dantas, D.D.; De Assis, S.J.; Baroni, M.P.; Lopes, J.M.; Cacho, E.W.; Cacho, R.D.; Pereira, S.A. Klapp method effect on idiopathic scoliosis in adolescents: Blind ran-domized controlled clinical trial. J. Phys. Ther. Sci. 2017 , 29 , 1–7. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Onose, G. National Platform of Medical Guides in Physical Medicine and Rehabilitation: Evaluation Guide in Physical Medicine and Rehabilitation ; Onose, G., Ed.; “Carol Davila” University Publishing House: Bucharest, Romania, 2011. [ Google Scholar ] Ma, C.Z.; Ren, L.J.; Cheng, C.L.; Zheng, Y.P. Mapping of Back Muscle Stiffness along Spine during Standing and Lying in Young Adults: A Pilot Study on Spinal Stiffness Quantification with Ultrasound Imaging. Sensors 2020 , 20 , 7371. [ Google Scholar ] [ CrossRef ] Haefeli, M.; Elfering, A. Pain assessment. Eur. Spine J. 2006 , 15 (Suppl 1), S17–S24. [ Google Scholar ] [ CrossRef ] [ PubMed ] Fairbank, J.C.; Pynsent, P.B. The Oswestry Disability Index. Spine 2000 , 25 , 2940–2952, discussion 2952. [ Google Scholar ] [ CrossRef ] Lins, L.; Carvalho, F.M. SF-36 total score as a single measure of health-related quality of life: Scoping review. SAGE Open Med. 2016 , 4 , 2050312116671725. [ Google Scholar ] [ CrossRef ][ Green Version ] Lim, C.Y.; In, J. Randomization in clinical studies. Korean J. Anesthesiol. 2019 , 72 , 221–232, Erratum in 2019 , 72 , 396. [ Google Scholar ] [ CrossRef ] Karanicolas, P.J.; Farrokhyar, F.; Bhandari, M. Practical tips for surgical research: Blinding: Who, what, when, why, how? Can J Surg. 2010 , 53 , 345–348. [ Google Scholar ] Schwab, F.; Dubey, A.; Pagala, M.; Gamez, L.; Farcy, J.P. Adult scoliosis: A health assessment analysis by SF-36. Spine 2003 , 28 , 602–606. [ Google Scholar ] [ CrossRef ] [ PubMed ] Walewicz, K.; Taradaj, J.; Rajfur, K.; Ptaszkowski, K.; Kuszewski, M.T.; Sopel, M.; Dymarek, R. The Effectiveness of Radial Extracorporeal Shock Wave Therapy in Pa-tients With Chronic Low Back Pain: A Prospective, Randomized, Single-Blinded Pilot Study. Clin. Interv. Aging. 2019 , 14 , 1859–1869. [ Google Scholar ] [ CrossRef ][ Green Version ] Li, S.; Wang, K.; Sun, H.; Luo, X.; Wang, P.; Fang, S.; Chen, H.; Sun, X. Clinical effects of extracorporeal shock-wave therapy and ultrasound-guided local corticosteroid injections for plantar fasciitis in adults: A meta-analysis of randomized controlled trials. Medicine 2018 , 97 , e13687. [ Google Scholar ] [ CrossRef ] [ PubMed ] Frairia, R.; Berta, L. Biological effects of extracorporeal shock waves on fibroblasts. A Review. Muscles Ligaments Tendons J. 2012 , 1 , 138–147. [ Google Scholar ] Cherniavsky, E.A.; Strakha, I.S.; Adzerikho, I.E.; Shkumatov, V.M. Effects of low frequency ultrasound on some properties of fibrinogen and its plasminolysis. BMC Biochem. 2011 , 12 , 60. [ Google Scholar ] [ CrossRef ] [ PubMed ][ Green Version ] Kotwal, S.; Pumberger, M.; Hughes, A.; Girardi, F. Degenerative scoliosis: A review. HSS J. 2011 , 7 , 257–264. [ Google Scholar ] [ CrossRef ][ Green Version ] Shu-Yan, N.; Tsz-Ki, H.; Yin-Ling, N. Physical Rehabilitation in the Management of Symptomatic Adult Scoliosis, Different Areas of Physiotherapy, Mintaze Kerem Gunel, IntechOpen. Available online: https://www.intechopen.com/books/different-areas-of-physiotherapy/physical-rehabilitation-in-the-management-of-symptomatic-adult-scoliosis (accessed on 7 April 2021). Table 1. Patient characteristics in the three study lots regarding scoliosis type, degree, and severity. Table 1. Patient characteristics in the three study lots regarding scoliosis type, degree, and severity. RSWT 1 Lot UST 2 Lot Control Lot Patient No. Dorsal Lumbar Scoliosis Type Degree Curve Severity Type 3 Patient No. Dorsal Lumbar Scoliosis Type Degree Curve Severity Type 3 Patient No. Dorsal Lumbar Scoliosis Type Degree Curve Severity Type 3 1 left 35 moderate 17 left 32 moderate 33 left 50 severe 2 right 55 severe 18 left 20 mild 34 left 30 moderate 3 left 38 moderate 19 right 38 moderate 35 left 20 mild 4 right 38 moderate 20 right 32 moderate 36 right 35 moderate 5 right 38 moderate 21 right 55 severe 37 right 55 severe 6 left 50 severe 22 right 35 moderate 38 right 50 severe 7 right 35 moderate 23 right 55 severe 39 right 22 mild 8 right 18 mild 24 left 30 moderate 40 right 35 moderate 9 left 20 mild 25 left 48 severe 41 left 45 severe 10 left 50 severe 26 left 50 severe 42 left 50 severe 11 left 45 severe 27 right 22 mild 43 right 38 moderate 12 left 30 moderate 28 right 38 moderate 44 right 30 moderate 13 right 50 severe 29 left 36 moderate 45 left 38 moderate 14 right 35 moderate 30 left 50 severe 46 left 32 moderate 15 right 55 severe 31 right 30 moderate 47 right 38 moderate 16 right 30 moderate 32 right 50 severe 48 right 50 severe 1 RSWT = radial extracorporeal shockwave treatment. 2 UST = ultrasound treatment. 3 Severity type: mild, 10–25; moderate, 26–40; and severe, over 40 [ 26 ]. Table 2. Demographic data of age and gender. Lot Demographic Data of Age Demographic Data of Gender n Median Mean Min Max Std. Dev Female Number Male Number RSWT 1 16 46.50 51 31 78 14.43 10 6 UST 2 16 51.00 51 32 74 12.18 11 5 Control 16 47.00 50 36 74 12.21 10 6 Total 48 50.85 51 31 78 12.72 31 17 1 RSWT = radial extracorporeal shockwave treatment. 2 UST = ultrasound treatment. Table 3. Pain and disability index reported using the VAS and ODI questionnaire. Table 3. Pain and disability index reported using the VAS and ODI questionnaire. Test Parameters VAS 1 Evolution ODI 2 Evolution RSWT Lot UST Lot Control Lot RSWT 3 Lot UST 4 Lot Control Lot No. 16 16 16 16 16 16 Normal Parameters Mean −6.38 −5.38 −28.06 −29.56 −27.31 Std. Deviation 1.025 0.719 4.328 6.293 4.743 Most Extreme Differences Absolute 0.232 0.308 Positive 0.232 0.308 Negative −0.205 −0.199 Kolmogorov–Smirnov Z 0.929 1.231 Asymp. Sig. (2-tailed) 0.354 0.097 Mann–Whitney U test mean rank 9.53 11.69 21.31 Asymp. Sig. (2-tailed) 0.000 0.002 Exact Sig. (2×(1-tailed Sig.)) 0.000 0.003 ANOVA Sig. 0.465 Tamhane Sig. 0.823 0.823 0.955 1 VAS = visual analogue scale. 2 ODI = Oswestry disability index. 3 RSWT = radial shockwaves treatment. 4 UST = ultrasound treatment. Table 4. Quality of life, short form-36, subscales RSWT versus UST lots—Mann–Whitney and Wilcoxon tests. Statistic Test General Health Physical Functioning Bodily Pain Physical Role Vitality Role Emotional Social Functioning Mann–Whitney U 111.500 85.500 109.500 98.000 52.500 58.500 76.000 Wilcoxon W 247.500 221.500 245.500 234.000 188.500 194.500 212.000 Z −0.626 −1.624 −0.710 −1.154 −2.865 −2.648 −2.000 Asymp. Sig. (2-tailed) 0.531 0.104 0.478 0.249 0.004 0.008 0.046 Exact Sig. [2 × (1-tailed Sig.)] 0.539 0.110 a 0.491 0.270 0.003 0.007 0.051 a Not corrected for ties. Grouping variable: TTip. Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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(PDF) Nurturing strengths, Challenging myths: people seeking asylum and the role of support and advocacy organisations PDF | Forward: This pilot project documents ways in which support and advocacy organisations build on the strengths and capacities of people seeking... | Find, read and cite all the research you need on ResearchGate Nurturing strengths, Challenging myths: people seeking asylum and the role of support and advocacy organisations Abstract Nu rt ur in g s t re ng t hs , Cha ll en gi ng m yt hs : p eo pl e se eki ng a s yl um an d th e r o le of s up po rt a nd ad vo ca cy o r ga ni sa ti on s Paul Hodge and Faith Curtis, University of New castle, Australia (January , 201 8) 1 Nu rt ur in g st r en gt hs , Ch al le ng in g m y th s: p eo pl e se eki n g as yl u m a n d t he r o le o f su pp ort an d a dv o ca cy o rg a ni sa ti on s Foreword This pilot project documents ways in which support and advocacy organisations build on the strengths and capacities of people seeking asylum. The project represents an explicit attempt to showcase the strengths-based work of practitioners. In nurturing the strengths of people seeking asylum, practitioners challenge myths about who people seeking asylum are, and what they represent while foster ing a sense of care, belonging and hope. This report is primarily intended for those working in the sector, both non- government and government, though given its focus on strengths stories, it may be a useful resource for people working in other related sectors. Drawing on practitioner’s accounts of working with people seeking asylum, this report: (i) highlights particular aspects of programs and initiatives and the rich possibilities language and images, relationships and hope when working with people seeking asylum; and (iii) documents some of the challenges of working in the current political climate and ways that practitioners are negotiating these challenges . Our pilot project documents a wide variety of programs and initiatives currently undertaken in Newcastle, Sydney and Melbourne. In order to ‘let practitioners speak for themselves’, we have compiled direct quotes under themed headings with very brief introductions and conclusions. We chose this approach because we felt it was the best way to represent stories as they were told. We hope that by sharing these strength-based stories of care, belonging and hope, we show the difference strengths-based practices can make in this, at times, challenging context. To make explicit the strengths-based work of practitioners working with people seeking asylum is to begin to build an altogether different public discourse and policy discussion on migration in Australia. In this way, the project continues the important work of many organisations and individuals who are challenging, in various ways, Australia’s federal migration policy by practicing alternatives every day. We understand that many of the themes emerging from the pilot project will not be new and to this we would say that our hope here is to add weight to arguments for ‘spreading the net’ of strengths -based and related approaches in ways that ultimately foreground the humanity of people seeking asylum. We appreciate the time and generosity given to us by practitioners, as well as the organisations themselves, without which this pilot project would not have been possible. Geography and Environmental Studies, University of Newcastle, Australia 2 Acknowledgements We acknowledge the traditional custodians of the lands on which we live and work, the Worimi, Awabakal, Eora and Kulin Nations and pay our respects to Elders past, present and emerging and to Country itself. We acknowledge that this land was never ceded and recognise the ongoing sovereignty of Aboriginal and Torres Strait Islander peoples. We acknowledge their living culture and their unique and central role in contemporary Australia. We think it is also important to acknowledge, in light of the ongoing sovereignty of Aboriginal and Torres Strait Islander peoples, that this land’s First Nations peoples should have a key role in shaping any future alternative migration policy. These are challenging and urgent considerations that need to remain at the forefront of issues of migration in colonised lands such as Australia. We would like to acknowledge all of the participating organisations. The organisations that agreed to be identified in this pilot project include:  Asylum Seeker Resource Centre (ASRC)  Maitland Grossmann High School  NSW Service for the Treatment and Rehabilitation of Torture and Trauma Survivors (STARTTS)  Refugee Advise and Casework Services (RACS)  Refugee Council of Australia (RCOA) We would like to acknowledge the contributions of all participants who agreed to share their stories so generously. Participants who agreed to be identified include:  Sally Baker  Elise Briggs  Ron Brown  Tina Dixson  Renee Dixson  Om Dhungel  Kate Hartig For consistency, all anonymous participants are identified by their interview number i.e. Interview 1, 2, 3 etc. This pilot project was funded by the Faculty of Science, University of Newcastle. 3 About the authors Dr Paul Hodge is lecturer in the Discipline of Geography and Environmental Studies at The University of Newcastle, Australia. Paul has been involved in advocacy and support for people seeking asylum and people from refugee backgrounds since 2010, initially with Africa-Australia Alliance for Peace and Reconciliation (AAAPR) and more recently with Refugee Action Network Newcastle (RANN). Paul has published work focusing on Operation Sovereign Borders and what this has meant for people seeking asylum (Hodge, 2015). He is currently working alongside Dr Natascha Klocker (University of Wollongong) and Dr Olivia Dun (University of Melbourne) exploring farming initiatives throughout NSW and Victoria for highly skilled people from refugee backgrounds. Dr Faith Curtis lectures and tutors in the Discipline of Geography and Environmental Studies at The University of Newcastle, Australia . Faith is a community development practitioner working on collaborative arts and wellness projects in Newcastle. Faith encourages people from diverse backgrounds to express themselves through street art, in so doing, supports people to forge greater connections within their communities . Faith’s PhD research focused on the way care is performed with people from refugee backgrounds. It drew on the experiences of both care give rs and care receivers and offers a more hopeful account of the politics of caring with people from refugee backgrounds (Curtis, 2016) . Faith’s p revious honours research highlighted the everyday practices of belonging following the establishment of Inverbrackie Alternative Places of Detention, Woodside, South Australia (Curtis and Mee, 2012). 4 Table of Contents Foreword ......................................................................................................................... 1 Acknowledgements .......................................................................................................... 2 About the authors ............................................................................................................ 3 Table of Contents ............................................................................................................. 4 Background and strengths-based approaches ................................................................... 5 Why does focusing on strengths matter? .......................................................................... 6 Research background and methodology ........................................................................... 8 Practitioner’s stories ........................................................................................................ 9 Supporting what people have: creating spaces, building strengths .................................... 9 Language and images matter: shifting the narrative ........................................................ 13 Relationships and processes ........................................................................................... 16 Challenges ...................................................................................................................... 19 Hope .............................................................................................................................. 23 Conclusion ...................................................................................................................... 26 Reference list ................................................................................................................. 27 5 Background and strengths-based approaches Support and advocacy organisations for people seeking asylum in Australia are necessary and important. These organisations provide a range of material and emotional support such as food, shelter, legal assistance, spaces to meet others and socialise, health and wellbeing services, formal and informal language and support classes, advocacy, education, friendship and welcome (amongst many other things). Practitioners working with people seeking asylum in Australia are at the forefront of strengths-based work. Importantly, people from refugee backgrounds themselves are fulfilling practitioner roles across the sector adding critical insights, cultural sensitivity and guidance for how the sector might continue to be transformed to be more in tune with newly arrived communities. Strengths-based approaches (SBA) emerged out of social work and community development through the work of Kretzmann and McKnight (1993), initially in the form of assets-based community development (ABCD). Based on their research into successful community initiatives in the US in the late 1980s, Kretzmann and McKnight proposed an alternative approach that recognised that “it is the capacities of local people and their associations that build powerful communities” not the denigrating practices and pathologies that accrue from needs-based, deficiency models (in Mathie and Cunningham, 2003, p. 475-6). To tap into these capacities, SBA offer flexible and adaptable practices and processes that involve identifying and building on strengths within communities (see also Cooperrider and Whitney, 2005; McCashen, 2010 ). During the interviews, it became very clear that the types of strengths-based programs and initiatives that practitioners employ vary according to the people they are supporting. In particular, it often depends on whether a person has certainty about their future in Australia. For example, programs and initiatives designed to support people who are beginning to re-build their life in Australia (those accepted as refugees) can look very different to the types of support for people seeking asylum on various visas (who are uncertain of whether they will be able to stay in Australia). It also became apparent that people seeking asylum in these more precarious situations might need safeguarding from media attention given their uncertain status. The popularity and accessibility of social media has made this question of safeguarding people seeking asylum more acute. 6 Why does focusing on strengths matter? Migration policy is one of the most contentious issues in contemporary Australia (Australian Human Rights Commission 2013). United Nations agencies, international and national peak bodies and non-government organisations have condemned the position taken by the Liberal/National coalition and the Labor party in Australia when it comes to people seeking asylum. The dominant federal government approach is one that views people seeking asylum as potential ‘threats’ to national security, as a strain on resources, and as ‘taking local jobs’. These powerful narratives about who people seeking asylum are, and what they represent, work to shape migration and ‘border protection’ policy and public opinion all of which vilify people seeking asylum casting them in terms of deficits and deficiencies. That people seeking asylum bring with them myriad strengths and capacities, hopes and aspirations is rarely the ‘big story’. In addition, research in Australia has shown that within the support and advocacy sector, people seeking asylum are often called upon to perform a ‘refugee identity’ – a position that enables them to access services and support, but at the same time has limiting effects on their agency (Westoby & Ingamells, 2010). Moreover, presenting people seeking asylum as victims, using trauma stories to gain support from the public, which then helps to attract funding and resources, is a common, even necessary practice given current funding models. It is within this context that “refugees come to regard themselves as objects, to be acted on by others rather than as subjects and agents of their own destiny” (Westoby & Ingamells, 2010, p. 11). Advocacy t oo has tended to foreground the tragedy and despair experienced by people seeking asylum which (though a powerful advocacy tool) can, again, augment a certain ‘refugee identity’ that acts to take away a sense of agency. That people seeking asylum are more than their immigration or visa status, that they are people, with strengths, skills, capacities and hopes for their future, has been difficult to cultivate in the sector given current institutional constraints and the pervasiveness of deficit narratives . One practitioner, himself from a refugee background, reflected on needs-based analysis and what such a focus translates to in terms of thinking about where solutions lie: My own observation is that we do a lot of research to find problems. We generally find problems because we are looking for problems … Then we start looking for solutions externally … We need to look at what is community development, what are we doing now with the current approach? Are we dismantling the social capital that we [people seeking asylum and former refugees] have? I find it quite a frustrating thing [when] I meet a lot of young, fresh employees in some of the service providers and they're all excited to start … They go into doing needs analysis, ‘w e've done the needs analysis ’, ‘ we know what the community needs are now ’ . They start developing programs with little or no 7 community consultation and engagement to try to meet those needs and again we are in the same vicious cycle. ~ Om Dhungel For the reasons identified above, focusing on strengths matters. Importantly, and as reflected throughout the pilot project, many practitioners within the sector apply strength-based approaches to their work despite institutional constraints and deficit narratives. The stories shared by practitioners revealed the difference strengths-based practices make. Adopting and amplifying strengths-based practices and stories of care, belonging and hope have real, positive effects. Significantly, we argue there are important implications of this strengths-based work for an alternative migration policy and practice. This pilot project aims to amplify these alternatives. 8 Research background and methodology As academics trying to make research more engaged and impactful for those who are doing the daily work on the ground, we are convinced that research design, processes and outcomes should more willingly and openly involve research participants; their ideas, insights and hopes. With this in mind, in 2016 I (Paul) met with a friend and local practitioner in Newcastle where I live. Together we reflected on a recent Forum on people seeking asylum held at The University Newcastle run by Amnesty International. One key advocacy point coming out of the Forum was how to come up with ways to change the negative rhetoric surrounding people seeking asylum. Forum panellists, which included a former detainee, spoke about the ‘loss of truth’, referring to the federal government and its continued vilification of people seeking asylum. We talked about what research might be able to achieve in relation to ‘changing the rhetoric’. Given the power of dominant narratives, we agreed that research that compiles and shares positive programs and initiatives that make explicit the strengths and aspirations of people seeking asylum could be a very good place to start. It was this conversation, and the energy of practitioners and people from refugee backgrounds at the Forum, that inspired this pilot project. Following ethics approval through the University of Newcastle, Faith and I invited individual participants from different organisations to participate in semi-structured interviews. In total 14 interviews were conducted. The 5 sections that make up this report represent the most common themes emerging from the interviews. Often , sentiments shared in one practitioner’s story were reflected in others. We have been conscious of presenting material in a way that does not lead to identification of those organisations and ind ividuals who did not consent to be identified. For organisations (and individuals) that gave consent, the programs and initiatives may be familiar and we felt it was important to allow participants the opportunity to self-identify, something that was taken up by some interviewees. A further note on identification, given participants were not only practitioners but also advocates, educators, consultants etc., we use the term ‘practitioner’ in this report very broadly to mean those working with people seeking asylum in these various roles . As mentioned above, w e are acutely aware of not contributing to the precariousness of people seeking asylum through practitioner voices in this report, particularly if conveying criticisms of the sector. In these cases, in order to de-identify people when elaborating on a particular program or initiative, we have drawn on passages from the interviews and written them up in our own words. 9 Practitioner’s stories Supporting what people have: creating spaces, building strengt hs A number of practitioners reflected on programs and initiatives to create spaces where relationships and a sense of belonging could emerge organically. Some of these spaces were structured and intentional while others more informal and unplanned. A key characteristic in these programs and initiatives was to identify, nurture and support people’s strengths and capacities. One practitioner put it this way when describing the importance of supporting and building on existing skills as part of their Innovation Hub and having people seeking asylum lead: Having a space and a place where they can utilise their skills in a way that helps others and in a meaningful way is incredibly validating for those people [involved]. You see changes … Where we can find opportunities for their skills, for them to do a volunteer role that aligns with what their skill set is, the benefit to that individual is far greater because it just - I guess it allows them to feel like they're doing something worthwhile and they've got a core purpose. ~Interview 7 The practitioner added: One example of that is a bilingual Persian language class that's being run on a Friday by an Iranian fellow who … has come from a teaching background. He was very keen to deliver classes himself in English but as a Persian speaker. So, his classes have been really popular and well attended … So that's something that he has really taken ownership of and he's incredibly proud of and that's something that he's single-handedly running. We do have a member led initiative; there is a pool of money that's available for people if they want to get a project off the ground. ~Interview 7 Fundamental to this sense of ownership is program involvement from the outset. Elaborating on what a member led initiative looks like the prac titioner said it has to: Allow people seeking asylum to be able to be designing, delivering and evaluating the success of the programs and embedding more of a consultative approach . ~Interview 7 Another practitioner reflected on an initiative and the positive effects this has had on their members: Every fortnight we have a community space. The community comes and they take ownership and leadership in that space. Usually a group of people will cook lunch, and anywhere between 80 and 250 people will attend. They’ll cook lunch, someone will perform, someone will sing, 10 someone will share poetry. There's a tapestry artist who does tapestry, there's a sketch artist who will do your sketch. A barber, and facial hair threading … [The initiative] provided that community space where they got to do that. So , the barbers cut hair, the chefs cooked, the performers performed, the sports people - there was always a game of sports going on. It was a space where they could live out and make a contribution to community … It was there, and they wanted to contribute, and they wanted to actualise their strengths and their capabilities … This is a space where everyone is welcome. Where everyone’s strengths are realised, and it’s a place where we can all belong. ~Interview 12 Practitioners also spoke about new and unexpected things that can emerge when setting up spaces, particularly those that are more organic, inclusive and intentionally less hierarchical: So , the Innovation Hub … is a really good safe environment to test out some of these [initiatives] – yeah … supporting people to be able to hav e more autonomy and direction. ~Interview 7 Recently we had … guests [from government] come and just be a part of it [the initiative]. As I was introducing them to the space, one of the more senior people said, ‘ I don't know who is an asylum seeker, who is a volunteer, who is a staff member, who is a community leader, who’s a guest. I really don’t know’ . As they were saying it, they had that ‘ aha, I get it ’ moment … you're not meant to know because here it doesn’t matter. This is a space where everyone is welcome. Where everyone’s strengths are realised, and it’s a place where we can all belong… They [the government person] got it as they were verbalising it, but they had to go through that process of feeling really uncomfortable with [it] , when people were introduced to them on a first name basis, they didn’t know what that meant. There was a human being standing in front of them… There was no rank and title ascribed to that. So that’s why I like the [initiative] … Taking away that rank and file, and the nametag, and everything I think in the early days was really important … [and] t hat we don’ t have volunteers or staff lined up on on e side… and clients queued up on the other. For me, three and a half years later, [the initiative] is now something that the community own. It’s a space where, as people who are seeking asylum and are being granted one of these short-term visas, they're now coming back to that space and still staying engaged and involved. ~ Interview 12 Another practitioner, a teacher at Maitland Grossmann High School, spoke about a program for unaccompanied minors which started in 2011 as part of ‘community 11 detention’. Rath er than high school aged students in the area on temporary visas undertaking separate education programs, the school accepted them into mainstream classes, with great results. They loved it and in fact we had a couple who left Merrylands High to come up here. They didn't want to be in any intensive language centre. They wanted to be in amongst Australian kids … The sum of the similarities was far greater than the sum of the differences. These were boys who … loved their soccer. They liked dressing up. Some of them were interested in girls, some had no interest whatsoever. They were really wanting to do well at school. It was a hard struggle to learn English and do the HSC subjects. See, none of these boys would have been allowed - if they were in Sydney, they wouldn't be in a mainstream high school, they would be in an intensive English centre. ~ Ron Brown This has meant that the broader school community, teachers etc., have had the opportunity to connect in meaningful ways, and importantly, learn from them and become advocates themselves. I think they [Afghan students] have an integral role because when people hear from the mouths of those people who were [being] vilified, once you hear their stories and speak to them, you have a different perception of them. Look at SBS, often on their news, they'll have successful migrant stories. I'm sure some people go ‘ oh yeah, whatever, it's SBS ’, but you've got to chip away because they're facts, they're real. I think for some people in the audience - there are some people in Australia, who are never going to accept refugees or Muslims. Those people just want to look in mirrors all the time. But there is a sizeable part of the Australian population that if they are given information that they haven't received be fore about refugees, they will change their attitud e… I take the boys, whenever a new student would come, I would always make sure I introduce them to quite a number of teachers and get them to talk to the teacher. I'd say to the teacher, next time you're at a dinner party and someone starts bad mouthing refugees or badmouthing Muslims, you'll be able to say to that person, ‘ do you know any refugees? ’ ‘ Have you ever met a refugee? ’ ‘ No, I didn't think so ’ . I have and I know them and what you're saying is bullshit… ‘Y ou've never met one !’ ~ Ron Brown Again, reflecting on what personal relationships can bring about, the practitioner added: It leads to discussion in the lounge room where the father says, ‘ ooh, I don't want bloody refugees ’ and the child says, ‘ but dad, [my friend] who was here the other day, he's a refugee ’ . ~ Ron Brown 12 Another practitioner put it this way when reflecting on the impact of relating on a personal level. For me, that first-person narrative is what changes hearts and minds. That’s what I think fuels the amazingness of our community, too. I think the New South Wales and Australian community is genuinely empathetic and kind hearted. I think it’s very rare that you see people who are negative about a fellow human being. Sometimes it’s just about putting that fellow human being in front of them, and giving them the chance to connect with their own humanity as well. ~ Interview 12 There were many stories of creating spaces and building strengths shared by practitioners, too many to add in a report of this kind. The positive reaction to the invitation to participate in this pilot project signalled to us a clear understanding that creating spaces where people can flourish in their own way was a crucial part of nurturing people’s strengths and aspirations. 13 Language and images matter: shifting the narrative One of the most recurring themes to emerge from the interviews was that language and images matter. The terms and images used to label, describe or represent people seeking asylum matters in ways that can invoke victimhood or build agency. The pilot project title has itself changed through the interview process as we came to understand how our initial framing of the research could perpetuate a sense of victimhood. One practitioner, and their partner, both from a refugee background, reflected on their ‘Stories about hope’ social art project. The project explores the human dignity, identity and strengths in people from refugee backgrounds through photography and short films. We focus[ed] on the positive and what the person wants to say. People felt empowered with that. For the first time in their life they had control [of] how they were represented and they could say we… are important, not what people would impose on them. We used the strength-based approach instead of victimisation. They actually had a say in shifting the whole perception about refugees in Australia. ~ Tina Dixson [Importantly], we were not asking them to tell their story of trauma, but what they are doing now, their hopes and aspirations, issues they want to talk about. Participants were fully in control of the narrative … they were happy that they can tell their stories and be in a safe place and finally they can raise their voice to tell their stories. ~ Renee Dixson Coupled with these positive narratives illustrating people’s aspirations , multi-layered identities and experiences, the project also focused on language and its effects. There was an explicit emphasis in the project to move away from references to ‘asylum seekers’ or ‘refugees’ . In my opinion, the public wants to see people who come from a refugee background as vulnerable subjects who are apart from the white Australia. So our response to that was just to show that dignified approach to people… [and now] you could see people starting to slightly shift the language. You could see people starting to take this approach, starting to call them ‘people seeking asylum’ [rather than ‘asylum seekers’] which recognises the humanity and not just label someone by their temporal circumstance in their life. ~ Tina Dixson Recognition that language matters was evident across many of the organisations involved in the pilot project. When describing member led initiatives, another practitioner spoke about their organisational shift from ‘clients’ to ‘members’ and why the language has been chosen around that. Again, this related to the importance of pointing out similarities with the broader Australian community. 14 Another aspect of the ‘Stories about hope’ project mentioned above was gauging public sentiment through social media. They found that public engagement was much higher, in terms of donations, Facebook likes and shares, when drawing on stories of hope rather than focusing on tragedy and despair: The engagement was really high, so people would share it and people would like that stuff. If you… post articles about how horrible their situation is, the engagement with our posts will drop… Th ey [the public] stop engaging at all. It shows the hope and there is something that can be done. I could see that just in the… number of people who would write [and make] the posts… So , I could definitely say that that approach worked really well. That was something that people wanted to [be exposed to] as opposed to just having the negative stuff out there. ~ Tina Dixson A similar insight was shared by another practitioner. [W]e've tested it, the victim photo versus this [positive] photo. More people are converted over that photo than the victim [photo]. There is clearly a portion of the Australian public, which is around 25 per cent, which are really moved by seeing terrible victims. [However], there's a really big proportion of the Australian community that's like, I don't want a victim here, but who agree with and resonate with photos of people that are survivors and that are a migrant and also going to contribute. ~Interview 4 For this practitioner, it was establishing their humanity which mattered most . Similarly, another spoke about public resonance with positive stories. What also feeds my hope is the politician arranging an Ugg boot donation [for people seeking asylum] gets more Facebook likes than the negative stories. ~ Interview 12 Another practitioner reflected on their other roles as researcher/advocate. For them, research could much more effectively hone its focus for specific purposes. I would say that we have to do a better job as qualitative researchers who work with stories and are kind of crafting, constantly crafting rich narratives that tell particularly complex stories. [Also] I think we need to work out ways of translating that into something that speaks to people who don't have time; that's politicians. ~Sally Baker Practitioners are actively taking steps to challenge the negative representations of people seeking asylum and people from refugee backgrounds through the deliberate 15 use of more inclusive language and victim-less images. The approach that these practitioners communicate and embed in their programs and initiat iv es move the focus away from a p erson’s temporary circumstances and towards a sense of shared humanity. Highlighting stories that point out similarities can pl ay a role in creating and building a more inclusive Australia. How people are spoken about, and represented, matters . 16 Relationships and processes In their own way, practitioners spoke about the importance of building relationships and that this was at the core of their work. There was also a keen sense that building strong relationships needed to not only underpin and guide individual relationships but organisational processes as well, including community engagement practices. Practitioners do not just work with individuals, but with families, support networks and mainstream institutions. We observed that relationships were important through all of the themes. One practitioner described the small, informal consultation process involved as part of their initial assessment evaluations: [We] get to know each other, [we ask] ‘what’s meaningful ? What would you like to do? What would you like to do more of? What are you good at? What concerns do you have? What worries you? ’ [We] ask about friends, family, connections; what jobs they’ve had in their home country; skills and education and [what] ethnic group … [T]ypically they are multi-skilled, talented [and have] variou s jobs… these conversations [are] more informal better ones. They want to share this stuff! ‘These are the skills I have!’ ~Interview 10 For this practitioner, underlying these initial discussions are bottom-up, holistic approaches such as assets based community development (ABCD), social capital and narrative approaches. Critically, and guiding these approaches, the practitioner added, Before anything starts, we build trust and rapport before projects take place. ~Interview 10 This resonates strongly with Om Dh ungel’s reflections mentioned earlier (p. 6). Starting projects without sustained and genuine community consultation just replays “ the same vicious cycle ” of needs-based approaches and processes. Dhungel spoke about what successful consultation might look like and lead to. [I]f you're trying to get the Bhutanese community together for a briefing, you contact the relevant person in the Bhutanese community organisation [thus] supporting the existing organisational structure. ~Om Dhungel To connect to the ‘relevant person’ requires a prior relationship of trust. As D hungel describes below, an organisation that recognises and can tap into already existing community structures and relationships, will be much better placed when attempting to engage communities. So … it's about having an excuse to talk to a fellow community member and it's not external. I found it really useful. Do you want to come? I'm telling somebody else to do that. By the way, your sister might come so 17 can you tell your sister or tell your auntie… When you talk about yoga, we'll talk about so many other things as well. How is grandad doing? Oh, he's in the hospital so I should go and see him. So, all these conversations start. ~ Om Dhungel Another practitioner spoke about a project designed to provide a broad consultation process to build relationships and connections based on shared priorities. [The project] was about really just trying to create a way for community members from all different kinds of cultural backgrounds, but having the common thread of being from a refugee background … to come together and advocate for themselves about the issues that were most important to them. [We found that] there were a lot of emerging similarities in the issues that they prioritise - things like the lack of access to family reunion, issues related to employment, housing. Those were the kinds of things that came up, no matter where someone [was from] … So rather than thinking, ‘ well how can people in a particular [place] advocate for the issue for employment ?’ Thinking [instead] about how can all of the people from a refugee background that have these things that they actually have in common, get into a room together and figure out what their priorities are and how they can go about doing it and how we can support that. ~ Interview 5 Important to the process was to be conscious of not only including community leaders but also pursuing other people with capacities and interests. The practitioner added: It wasn't just about community leaders either [but rather] it was really about trying to identify individual community members who were interested in playing a role in advocacy. That meant different things to different people … We held a number of sessions with different communities and also encouraging women and young people to participate, because that was a missing link, where it was often community elders and men that were representing whole communities, whereas actually they represented a particular aspect of it. ~ Interview 5 Describing relationships developed through language classes, o ne practitioner reflected on their time working with communities in a Muslim country and the care and sense of belonging people extended to them while there. This experience left a formative impression. I guess my motivation is about extending, about giving back, but also hoping that they feel a sense of belonging [from] other people outside of their immediate community here. ~Kate Hartig 18 They added: If you don't have people of the local community accepting you for who you are, then it's probably very hard to feel that sense of belonging. ~Kate Hartig The need to build and maintain relationships was a constant theme coming out of the interviews. This was despite the persistence of institutional processes that often work against such attempts to build relationships. While many organisations aim to support programs and initiatives that nurture respect and long-term community-led engagement (and have experienced successes), it was also the case that other practitioners felt there was a gap between this rhetoric and what often happens in practice. 19 Challenges Recognising the challenges of working in the sector and the ways that practitioners are negotiating these challenges were important themes emerging from the pilot project. Some of the challenges were about the broader political climate, federal government policy, and public discourse, while other issues related to the sector itself. A number of practitioners discussed the challenge of working with people with uncertain futures, who may not be able to stay in Australia. Reflecting on this challenge, one practitioner stated: … if you're working with people who have got a guaranteed future in Australia, you work in a completely different way to the way that you would work with people who have got no idea whether they will have a future here or [if] they have to go home. ~Interview 3 Another practitioner said that: Some of them [people seeking asylum] get stuck in that all they can think about is getting their visa. Some workers get stuck in that too, building up hope towards something that is so out of anyone’s control, certainl y of any support worker… So that's something over which you can't really - you can't work towards that, because it's too much of an unknown. ~Elise Briggs This practitioner also noted that, “ with the uncertainty, belonging is very difficult ” . However, practitioners did have strategies in place to cope with this uncertainty. For example, one practitioner discussed working within short timeframes, and what this might mean for their practice: It might be next month or the month af ter that… You can't talk about long- term plans. You can't talk about enduring connections. You have to be focusing very much on building strengths that are not dependent on their ongoing presence in Australia otherwise you are building a set of expectations that are unrealistic. ~Interview 3 Another practitioner draws on the strengths of people seeking asylum as a way to negotiate and cope with this challenge. … but I must say that asylum seekers, of all people, know how to seize the day, making the most of those moments that help us to cope with whatever is around the corner. ~Elise Briggs As the following passage suggests, practitioners must also be careful not to make inappropriate comments that raise expectations : 20 So even simple comments like, ‘ you know you will have a wonderful time in Australia ’; or, ‘ you will make such a good contribution to Australia ’; or, ‘ of course you'll be able to stay in Australia ’, are totally inappropriate comments to use because you don’t know. But the person (seeking asylum) will be hearing those comments and believing because you're making them that they must be true… So that side of things, people need to understand your role [and be] absolutely truthful and honest and focus on the here and now and not making promises you can't keep. ~Interview 3 This practitioner was primarily speaking about less experienced volunteers and others who work with people seeking asylum who have no certainty about their determination. In these cases, training and preparation is key. One of the more poignant challenges raised was that there simply is not enough people with lived experience of seeking asylum working in the sector, especially in leading roles. I suppose… the biggest challenge I think is that the refugee and asylum seeker space is dominated by people with no experience of being a refugee, asylum seeker… or even migrant communities… There are barely any refugee and asylum seeker dominated organisations, or leaders that stand in the refugee space… they’re mostly white men. ~Interview 4 The practitioner spoke about how this can mean that debate is controlled by those who are not the ones most affected by the programs being put in place. Meanwhile, another spoke about complexities within groups and that appropriate ways to engage communities might not be the most obvious ones: We also have a tendency to think that, okay, if you’ve got people coming in from Syria we put on a Syrian staff person and that person will be fine to be able to work with people. Yet issues around ethnicity and class and religion are so important that quite often having somebody from somewhere totally different is much better than having somebody from the same country but the wrong class or wrong [ethnicity]. ~Interview 4 Other key challenges related to government policy: Programs are constantly changing because we're in this dynamic landscape where policies have large impacts and things come in and nothing can really be set in stone in terms of funding arrangements and things like that. So, it's a very dynamic envir onment… I think that sometimes it can lead to - not necessarily having clarity around what it is that we're trying to achieve and when we're doing it. ~Interview 7 Other challenges related to the current system for service providers: 21 As a service provider, you have a contract with the government and you have to meet the KPIs. The current system doesn’t really provide any incentive for service providers to work more collaboratively with community groups, to build social capital in the community. It's more ab out meeting those KPIs. ~Interview 14 This practitioner also suggested that “…engagement with local communities could be part of the KPIs ” offering alternatives to some of the problems they face within the system. Another challenge was about negotiating a balance between providing the public with information about the lives of people seeking asylum in Australia, while at the same time ensuring people remain safe. For example, one practitioner said: The government and the Immigration Department have very strict rules on what the refugees can say to the media, very strict rules and they … tried to silence the refugees so that mainstream Australia never heard their story. ~Ron Brown Similarly, another practitioner described the risks involved: The government continually does media monitor ing and picks up things that are going on. So, if we're going to do a media event … that actually involves any of our clients, we have to inform the department we're going to do that and tell them what it's about. ~Interview 13 There were also issues relating to sensitivities that need to be taken into account, in this case, around war and its commemoration. One practitioner spoke about an example where they invited a conversation where another person from the same community expl ained Remembrance D ay to the language class. They recalled how two women broke down and cried and how it made the practitioner cry too. I think it's quite easy within the context of a classroom to forget that these people have really suffered, members of their family - that they've lived in fear of their lives, they've lost members of their family ... I thought that was a good reminder to me that these people, just something like that … about a day we remember those who have been killed in war … could just tr igger memories of it. ~Interview 6 The issues raised here highlight just some of the challenges faced by practitioners. Issues of uncertainty around determination, training, linking communities to evaluative processes and matters relating to disclosure, anonymity and cultural sensitivity were raised often in the interviews. Likewise, issues concerning the sector make-up, as mostly people without lived experience of seeking asylum and the implications of this, 22 were ones that came through the interviews. Organisations like RISE (Refugees, Survivors and Ex-Detainees) were held up as important exemplars for how organisations could open up their workforce to better reflect organisational aims to facilitate community-led programs and initiatives. 23 Hope Despite challenges in the sector, when asked where hope lies, practitioners unanimously shared stories relating directly to people seeking asylum themselves as well as to the generosity of the Australian community as a whole. The following quotes are representative of these sentiments. Well it's two ways. I see these women that I'm working with very hopeful about their future, which is really lovely. They're very hopeful about their children's future, I mean most of them, majority, they're all mothers; they're very proud of their children and very proud that their children are going to school and can speak English and can read English. So, it's almost like they're trying to catch up with their children, they can see what their children are achieving. I think they take enormous pride, so they're hopeful and I think they're very hopeful that their children will be part of society. So, seeing their hope, I think, makes me hopeful, so it's not my personal hope so much as seeing, you know, some of their smiles, like when they've been able to do something… So , it's their hope that there is more hope. ~Interview 6 If ever I feel like I'm starting to lose that hope, I go into the community space, and I go into one of our community-based projects. I stop and I listen to the stories, because when I listen, I hear their hope. I hear their resilience, and I hear their strength, and I hear their will to want to live out something positive. This is their chance of safety. This is their chance of creating a safe life for their children, and for all the children that will follow. ~Interview 12 I mean it's in the people that I meet who are enduring just things that none of us would wish on our worst enemy and yet are still themselves, able to somehow be hopeful and that they may not be able to think about the future yet, but they can think about the small next step … So that idea that [they] can be so focused on and concerned for other people has been reflected a hundred times over in the people that I've met, that they can be going through some of the most horrible and you just see things - hopeless situations - but they not only find hope for themselves, but they're always concerned about other people and not just their family, but their community - and the community more broadly. ~Interview 5 24 There are some brilliant people who work in this sector. But the thing that gives me most hope is the people, the refugees. I mean, over decades coming across people who are extraordinarily courageous and resilient and creative and tenacious and are making such a wonderful contribution to Australia. Dr Al Muderis, the boat person who got locked up in Curtin and treated like shit and is now a world leading orthopaedic surgeon … to the ordinary people who ar e getting on with their lives… They just rolled up their sleeves and got on with it .. . I just find that remarkable. ~Interview 3 Again, I'm not approached by people who are antagonistic, but we see good will and we see support and empathy. We see people wanting to demonstrate a place of welcome, and at the same time we see people who are seeking asylum find their own place in community, where they truly belong. Where they can contribute and be a part of something bigger than just themselves. ~ Elise Briggs To be really frank. The offers of support and goodwill [from the broader community] are just tremendous, and you’ve got - I'm just trying to think of a really good example of a cross section - Girl Guides reaching out. [They’ll say], ‘We want to learn more, we want to offer, we want to do some cultural share between our girls’, who live in the inner west and may never have met someone with a forced migration experience. Also, we want to share this iconic Aussie thing with people who are new to the community. It’s a cultural share opportunity…. Honestly, in community engagement we couldn’t go a day of work without having a request for involvement [by an organisation or group] . ~ Interview 12 I think that every time the Maitland or Newcastle community have stepped up and done something extraordinary for these guys, I celebrate that hugely, because it helps to counter [the deficit stories]… Or wh en community members themselves did something amazing. That's where the hope is. It's like the guy whose friend had had his Visa refused. He got offered a job, and he didn't take the job. He got his friend to take the job, because his friend didn't have any other alternative form of income, but he would. So , he lined the job up for his friend instead. Little things like that that just - yeah, hope in humanity when you see that. ~ Interview 13 25 That gives me hope. Every day I get up in the morning to make sure that we don't miss out on some of those gems that are waiting to be polished and let them shine. That's what I see on a daily basis. Just last week we had some other wins as well because we had some engineers, young refugees they came here, finished their Year 12, - managed to get into engineering and they became engineers… We've been working together for a while now and last week one of the guys came with a funny problem. He said I've got this [job] offer in Brisbane, but I've got another offer here. So, which one do you think I should take? Now he's got a problem of which offer to take! ~ Om Dhungel 26 Conclusion So , there's a void of good news stories, and there's also this high-level negativity there. I get that. But at the community level, I don’t see it. ~ Interview 12 This quote captures what many practitioners highlighted; positive stories are everywhere and the impact of affirming stories on the Australian community and people seeking asylum themselves are real and enduring. Undermining the impact of this ‘high -level negativity ’ by emphasising the everyday experiences and relationships of care and welcome that result from the kinds of programs and initiatives described here, is a key reason for undertaking this pilot project. To make explicit the strengths, aspirations and hopes of people seeking asylum and practitioners themselves , is to begin to challenge the myths about who people seeking asylum are, and what they represent. While challenges certainly exist, practitioners, including practitioners from refugee backgrounds, are adopting SBA in ways that simultaneously embody and enact the kind of society they want to participate in. Rather than waiting for federal government policies to shift, people seeking asylum and practitioners are already ‘ living ’ the alternatives. We see our role as researchers as one of amplifying these experiences and relationships; these alternatives that challenge, confront and indeed question the veracity of the federal government’s claims that the Australian community crave ‘security’ at the cost of o ur shared sense of humanity. This is a pilot project and with that has its limitations. Further research could of course involve people seeking asylum themselves, though we would be cautious for some of the reasons raised in this report. One obvious way fo rward, would be to have people seeking asylum and people from refugee backgrounds as co-researchers in paid positions. Such community-led collaborations would, by design, build on the strengths and aspirations of those involved and potentially add to the kinds of stories shared in this report. We are currently thinking through what these kinds of projects could look like. We welcome feedback on this report and offer to present more on these findings to participating organisations and others. Dr Paul Hodge [email protected] Dr Faith Curtis [email protected] 27 Cooperrider, D.L., and Whitney, D. (2005). Appreciative Inquiry: A positive revolution in change ; San Francisco, California. Curtis, F. (2017). ‘ Performing care with people from refugee backgrounds: an Australia ’ (Unpublished doctoral thesis, University of Newcastle). Curtis, F., and Mee, K. J. (2012). Welcome to Woodside: Inverbrackie Alternative Place of Detention and performances of belonging in Woodside, South Australia, and Australia. Australian Geographer, 43 (4), 357- 375. Hodge, P. (2015). A grievable life? The criminalisation and securing of asylum seeker bodies in the ‘violent frames’ of Australia’s Operation Sovereign Borders. Geoforum, 58 (2 ), 122-131. Kretzmann, J.P., and McKnight, J.L. (1993). Building Communities from the Inside Out: A Path Toward Finding and Mobilizing a Community's Assets ; Evanston, Ill., Center for Urban Affairs and Policy Research, Northwestern Universit y. Mathie, A., and Cunningham, G. ( 2003 ). From Clients to Citizens: Asset-Based Community Development as a Strategy for Community-Driven Development , Development in Practice , 13 (5), 474 - 486 McCashen, W. ( 2010 ). The strengths approach: A strengths based resource for sharing Westoby, P., and Ingamells, A. ( 2010 ). A critically informed perspective of working with resettling refugee groups in Australia. British Journal of Social Work , 40(6), 1759-1776. Jan 2015 GEOFORUM Paul Hodge The life of those seeking asylum from persecution and other human rights abuses has become interminably precarious. As minority world governments deploy various apparatuses of security to govern the circulation of ‘unruly’ populations, the world’s most vulnerable people have been reconstituted as security threats. In this paper I trace this ‘transfer of illegitimacy’ and criminalisation of asylum seeker bodies in the context of the Australian government’s newly deployed Operation Sovereign Borders. Drawing on Foucault’s governmentality as a domain of security and Butler’s articulation of recognition, precariousness and grievability, I explore both the subjectivities formed as a function and technique of securing Australia’s borders and the way this framing produces a certain governed reality that ‘acts upon the senses’ to delimit public discourse. I argue that the range of discursive and non-discursive practises that make up Operation Sovereign Borders has dire implications for those seeking asylum in Australia. Not only do these practises constitute a social crafting where conditions for a flourishing life are diminished, but this crafting of precarity is carried out in the name of securing citizens lives. The life of the asylum seekers is a life unrecognised in the violent frames of Operation Sovereign Borders. Nov 2003 Alison Mathie Gord Cunningham Sep 2009 Faith Curtis John P. Kretzmann Jan 2017 Jan 2010 W Mccashen

https://www.researchgate.net/publication/322592400_Nurturing_strengths_Challenging_myths_people_seeking_asylum_and_the_role_of_support_and_advocacy_organisations

A Comparative Clinical Study to Evaluate the Effectiveness of ProFlor vs. Lichtenstein for Inguinal Hernia Repair - Full Text View - ClinicalTrials.gov A Comparative Clinical Study to Evaluate the Effectiveness of ProFlor vs. Lichtenstein for Inguinal Hernia Repair The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been evaluated by the U.S. Federal Government. Read our disclaimer for details. ClinicalTrials.gov Identifier: NCT02240550 Recruitment Status : Completed First Posted : September 15, 2014 Last Update Posted : January 18, 2016 Sponsor: Insightra Medical, Inc. Information provided by (Responsible Party): Insightra Medical, Inc. Study Details Study Description Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Brief Summary: This investigation will be a double-armed, randomized (Blinded patients and Blinded examiner) prospective study designed to collect perioperative and postoperative data to compare the QOL of ProFlor vs. Lichtenstein inguinal hernia repair. Condition or disease Intervention/treatment Phase Inguinal Hernia Device: ProFlor Hernia Repair System Device: Lichtenstein hernia repair Not Applicable Detailed Description: The primary objectives of this study will be to evaluate the immediate and short-term amount of post-operative pain, the quality of life (QOL), as well as return to normal activities (work) for the two procedures. Carolinas Comfort Scale (CCS) and Visual Analogue Score (VAS) assessments will be held at regular intervals to evaluate these objectives. The secondary objective of the study is to identify any short-term study related complications/adverse events. The follow up will be immediately post-operative, post-operative day 1, and 1, 2,and 4 weeks. The first and 4th week's visits are mandatory office visits (if possible). If not available for office visit, a CCS assessment will be mailed or will be conducted via phone. Up to 50 patients may be enrolled for this study (25 per arm). A report may be generated for presentation at an international meeting such as the AHS, EHS, or APHS if decided by the lead investigator. Study Design Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Resource links provided by the National Library of Medicine MedlinePlus related topics: Hernia U.S. FDA Resources Arms and Interventions Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Arm Intervention/treatment Active Comparator: ProFlor Hernia Repair System A 3-D hernia mesh Device: ProFlor Hernia Repair System The 3-D hernia mesh will be used in the repair of your inguinal hernia. Other Name: 3D mesh Active Comparator: Lichtenstein Hernia Repair Using flat polypropylene mesh Device: Lichtenstein hernia repair The standard Lichtenstein hernia repair with flat mesh will be used in the treatment of your inguinal hernia. Other Name: Polypropylene Outcome Measures Top of Page Study Description Study Design Arms and Interventions Outcome Measures Eligibility Criteria Contacts and Locations More Information Primary Outcome Measures : Evaluate the immediate and short-term amount of pain. [ Time Frame: 1 month ] We will evaluate the immediate and short-term amount of pain using the VAS and Carolinas Comfort Scale (CCS) assessments held at regular intervals. Compare the time required for return to normal activities (work). [ Time Frame: 1 month ] We will capture the time required to return to normal activities as indicated by the patient. Secondary Outcome Measures : Identify any short-term related complication/adverse events associated with the procedures. [ Time Frame: 1 month ] Layout table for eligibility information Ages Eligible for Study: 18 Years to 65 Years   (Adult, Older Adult) Sexes Eligible for Study: All Accepts Healthy Volunteers: No Criteria Inclusion Criteria: Scheduled to undergo routine inguinal hernia repair Male and Female patients between 18 and 65 years old Competent to give consent Clinically relevant inguinal hernia (classification: EHS P L/M 1/2/3) Defect size at operation is between 5mm and 35mm Diagnosed with unilateral, direct, indirect or mixed inguinal hernia Primary hernia at the operative site Exclusion Criteria: Signs of obvious local or systemic infection Any previous surgery on the hernia operative site Hernia is not in the inguinal area Hernia is not identified as indirect or direct Presenting with unstable angina or NYHA class of IV Known Pregnancy or Nursing women Active drug user Recurrence of a repair by any method Patients with giant inguinoscrotal hernia or abdominal wall defect >35 mm in diameter - e.g. large direct or combined inguinal hernia type Nyhus IIIb / EHS P L/M 1/2/3 Immunosuppression, prednisone>15 mg/day, active chemotherapy End stage renal disease Abdominal ascites Skin infection in area of surgical field BMI >35 Peritoneum cannot be closed Patient has a clinically relevant co-morbidity (antithrombic prophylaxis due to cardiovascular pathologies, diabetes requiring insulin therapy or immunodeficiency syndrome of any type) Neutropenia with absolute neutrophil count (ANC)<500 cells/mm3 Significant of life-threatening condition (e.g., endocarditis) that would confound or interfere with the procedure Patients that require anticoagulant monitoring with an activated partial thromboplastin time (aPTT) Patients unwilling to forego blood and/or blood product donation for at least 3 months from initiation of first study device Treatment with investigational medicinal product within 30 days before enrollment and for the duration of the study Investigational device present, or removed within 30 days before enrollment, or presence of device-related infection Patients who the investigator considers unlikely to adhere to the protocol or complete the clinical study Information from the National Library of Medicine Locations Layout table for location information India Healing Hands Clinic Pune, India Sponsors and Collaborators Insightra Medical, Inc. Investigators Layout table for investigator information Principal Investigator: John Murphy, MD Healing Hands Clinic Layout table for additonal information Responsible Party: Insightra Medical, Inc. ClinicalTrials.gov Identifier: NCT02240550 History of Changes Other Study ID Numbers: P0133 First Posted: September 15, 2014 Key Record Dates Last Update Posted: January 18, 2016 Last Verified: January 2016 Keywords provided by Insightra Medical, Inc.: Open Inguinal Hernia

https://clinicaltrials.gov/ct2/show/study/NCT02240550

Recommendations for examining and interpreting funnel plot asymmetry in meta-analyses of randomised controlled trials | The BMJ Research Methods & Reporting Recommendations for examining and interpreting funnel plot asymmetry in meta-analyses of randomised controlled trials BMJ 2011 ; 343 doi: https://doi.org/10.1136/bmj.d4002 (Published 22 July 2011) Cite this as: BMJ 2011;343:d4002 Article Alex J Sutton , professor 2 , John P A Ioannidis , professor and director 3 , Norma Terrin , associate professor 4 , David R Jones , professor 2 , Joseph Lau , professor 4 , James Carpenter , reader 5 , Gerta Rücker , research assistant 6 , Roger M Harbord , research associate 1 , Christopher H Schmid , professor 4 , Jennifer Tetzlaff , research coordinator 7 , Jonathan J Deeks , professor 8 , Jaime Peters , research fellow 9 , Petra Macaskill , associate professor 10 , Guido Schwarzer , research assistant 6 , Sue Duval , assistant professor 11 , Douglas G Altman , professor 12 , David Moher , senior scientist 7 , Julian P T Higgins , senior statistician 13 Author affiliations 1 School of Social and Community Medicine, University of Bristol, Bristol BS8 2PS, UK 2 Department of Health Sciences, University of Leicester, Leicester, UK 3 Stanford Prevention Research Center, Stanford University School of Medicine, Stanford, CA, USA 4 Institute for Clinical Research and Health Policy Studies, Tufts Medical Center, Boston, MA, USA 5 Medical Statistics Unit, London School of Hygiene and Tropical Medicine, London, UK 6 Institute of Medical Biometry and Medical Informatics, University Medical Center Freiburg, Germany 7 Clinical Epidemiology Program, Ottawa Hospital Research Institute, Ottawa, Ontario, Canada 8 School of Health and Population Sciences, University of Birmingham, Birmingham, UK 9 Peninsula Medical School, University of Exeter, Exeter, UK 10 School of Public Health, University of Sydney, NSW, Australia 11 University of Minnesota School of Public Health, Minneapolis, MN, USA 12 Centre for Statistics in Medicine, University of Oxford, Oxford, UK 13 MRC Biostatistics Unit, Cambridge, UK Correspondence to: J A C Sterne [email protected] Accepted 21 February 2011 Funnel plots, and tests for funnel plot asymmetry, have been widely used to examine bias in the results of meta-analyses. Funnel plot asymmetry should not be equated with publication bias, because it has a number of other possible causes. This article describes how to interpret funnel plot asymmetry, recommends appropriate tests, and explains the implications for choice of meta-analysis model The 1997 paper describing the test for funnel plot asymmetry proposed by Egger et al1is one of the most cited articles in the history of BMJ.1Despite the recommendations contained in this and subsequent papers,23funnel plot asymmetry is often, wrongly, equated with publication or other reporting biases. The use and appropriate interpretation of funnel plots and tests for funnel plot asymmetry have been controversial because of questions about statistical validity,4disputes over appropriate interpretation,356and low power of the tests.2 This article recommends how to examine and interpret funnel plot asymmetry (also known as small study effects2) in meta-analyses of randomised controlled trials. The recommendations are based on a detailed MEDLINE review of literature published up to 2007 and discussions among methodologists, who extended and adapted guidance previously summarised in the Cochrane Handbook for Systematic Reviews of Interventions.7 What is a funnel plot? A funnel plot is a scatter plot of the effect estimates from individual studies against some measure of each study’s size or precision. The standard error of the effect estimate is often chosen as the measure of study size and plotted on the vertical axis8with a reversed scale that places the larger, most powerful studies towards the top. The effect estimates from smaller studies should scatter more widely at the bottom, with the spread narrowing among larger studies.9In the absence of bias and between study heterogeneity, the scatter will be due to sampling variation alone and the plot will resemble a symmetrical inverted funnel (fig 1 ⇓). A triangle centred on a fixed effect summary estimate and extending 1.96 standard errors either side will include about 95% of studies if no bias is present and the fixed effect assumption (that the true treatment effect is the same in each study) is valid. The appendix on bmj.com discusses choice of axis in funnel plots. Fig 1Example of symmetrical funnel plot. The outer dashed lines indicate the triangular region within which 95% of studies are expected to lie in the absence of both biases and heterogeneity (fixed effect summary log odds ratio±1.96×standard error of summary log odds ratio). The solid vertical line corresponds to no intervention effect Implications of heterogeneity, reporting bias, and chance Heterogeneity, reporting bias, and chance may all lead to asymmetry or other shapes in funnel plots (box). Funnel plot asymmetry may also be an artefact of the choice of statistics being plotted (see appendix). The presence of any shape in a funnel plot is contingent on the studies having a range of standard errors, since otherwise they would lie on a horizontal line. Box 1: Possible sources of asymmetry in funnel plots (adapted from Egger et al 1 ) Reporting biases Publication bias: Delayed publication (also known as time lag or pipeline) bias Location biases (eg, language bias, citation bias, multiple publication bias) Selective outcome reporting Selective analysis reporting Poor methodological quality leading to spuriously inflated effects in smaller studies Poor methodological design Inadequate analysis Fraud True heterogeneity Size of effect differs according to study size (eg, because of differences in the intensity of interventions or in underlying risk between studies of different sizes) Artefactual In some circumstances, sampling variation can lead to an association between the intervention effect and its standard error Chance Asymmetry may occur by chance, which motivates the use of asymmetry tests Heterogeneity Statistical heterogeneity refers to differences between study results beyond those attributable to chance. It may arise because of clinical differences between studies (for example, setting, types of participants, or implementation of the intervention) or methodological differences (such as extent of control over bias). A random effects model is often used to incorporate heterogeneity in meta-analyses. If the heterogeneity fits with the assumptions of this model, a funnel plot will be symmetrical but with additional horizontal scatter. If heterogeneity is large it may overwhelm the sampling error, so that the plot appears cylindrical. Heterogeneity will lead to funnel plot asymmetry if it induces a correlation between study sizes and intervention effects.5For example, substantial benefit may be seen only in high risk patients, and these may be preferentially included in early, small studies.10Or the intervention may have been implemented less thoroughly in larger studies, resulting in smaller effect estimates compared with smaller studies.11 Figure 2 ⇓shows funnel plot asymmetry arising from heterogeneity that is due entirely to there being three distinct subgroups of studies, each with a different intervention effect.12The separate funnels for each subgroup are symmetrical. Unfortunately, in practice, important sources of heterogeneity are often unknown. Fig 2Illustration of funnel plot asymmetry due to heterogeneity, in the form of three distinct subgroups of studies. Funnel plot including all studies (top left) shows clear asymmetry (P<0.001 from Egger test for funnel plot asymmetry). P values for each subgroup are all >0.49. Differences in methodological quality may also cause heterogeneity and lead to funnel plot asymmetry. Smaller studies tend to be conducted and analysed with less methodological rigour than larger studies,13and trials of lower quality also tend to show larger intervention effects.1415 Reporting bias Reporting biases arise when the dissemination of research findings is influenced by the nature and direction of results. Statistically significant “positive” results are more likely to be published, published rapidly, published in English, published more than once, published in high impact journals, and cited by others.16171819Data that would lead to negative results may be filtered, manipulated, or presented in such a way that they become positive.1420 Reporting biases can have three types of consequence for a meta-analysis: A systematic review may fail to locate an eligible study because all information about it is suppressed or hard to find (publication bias) A located study may not provide usable data for the outcome of interest because the study authors did not consider the result sufficiently interesting (selective outcome reporting) A located study may provide biased results for some outcome—for example, by presenting the result with the smallest P value or largest effect estimate after trying several analysis methods (selective analysis reporting). These biases may cause funnel plot asymmetry if statistically significant results suggesting a beneficial effect are more likely to be published than non-significant results. Such asymmetry may be exaggerated if there is a further tendency for smaller studies to be more prone to selective suppression of results than larger studies. This is often assumed to be the case for randomised trials. For instance, it is probably more difficult to make a large study disappear without trace, while a small study can easily be lost in a file drawer.21The same may apply to specific outcomes—for example, it is difficult not to report on mortality or myocardial infarction if these are outcomes of a large study. Smaller studies have more sampling error in their effect estimates. Thus even though the risk of a false positive significant finding is the same, multiple analyses are more likely to yield a large effect estimate that may seem worth publishing. However, biases may not act this way in real life; funnel plots could be symmetrical even in the presence of publication bias or selective outcome reporting1922—for example, if the published findings point to effects in different directions but unreported results indicate neither direction. Alternatively, bias may have affected few studies and therefore not cause glaring asymmetry. Chance The role of chance is critical for interpretation of funnel plots because most meta-analyses of randomised trials in healthcare contain few studies.2Investigations of relations across studies in a meta-analysis are seriously prone to false positive findings when there is a small number of studies and heterogeneity across studies,23and this may affect funnel plot symmetry. Interpreting funnel plot asymmetry Authors of systematic reviews should distinguish between possible reasons for funnel plot asymmetry (box 1). Knowledge of the intervention, and the circumstances in which it was implemented in different studies, can help identify causes of asymmetry in funnel plots, which should also be interpreted in the context of susceptibility to biases of research in the field of interest. Potential conflicts of interest, whether outcomes and analyses have been standardised, and extent of trial registration may need to be considered. For example, studies of antidepressants generate substantial conflicts of interest because the drugs generate vast sales revenues. Furthermore, there are hundreds of outcome scales, analyses can be very flexible, and trial registration was uncommon until recently.24Conversely, in a prospective meta-analysis where all data are included and all analyses fully standardised and conducted according to a predetermined protocol, publication or reporting biases cannot exist. Reporting bias is therefore more likely to be a cause of an asymmetric plot in the first situation than in the second. Terrin et al found that researchers were poor at identifying publication bias from funnel plots.5Including contour lines corresponding to perceived milestones of statistical significance (P=0.01, 0.05, 0.1, etc) may aid visual interpretation.25If studies seem to be missing in areas of non-significance (fig 3 ⇓, top) then asymmetry may be due to reporting bias, although other explanations should still be considered. If the supposed missing studies are in areas of higher significance or in a direction likely to be considered desirable to their authors (fig 3 ⇓, bottom), asymmetry is probably due to factors other than reporting bias. Fig 3Contour enhanced funnel plots. In the top diagram there is a suggestion of missing studies in the middle and right of the plot, broadly in the white area of non-significance, making publication bias plausible. In the bottom diagram there is a suggestion of missing studies on the bottom left hand side of the plot. Since most of this area contains regions of high significance, publication bias is unlikely to be the underlying cause of asymmetry Statistical tests for funnel plot asymmetry A test for funnel plot asymmetry (sometimes referred to as a test for small study effects) examines whether the association between estimated intervention effects and a measure of study size is greater than might be expected to occur by chance. These tests typically have low power, so even when a test does not provide evidence of asymmetry, bias cannot be excluded. For outcomes measured on a continuous scale a test based on a weighted linear regression of the effect estimates on their standard errors is straightforward.1When outcomes are dichotomous and intervention effects are expressed as odds ratios, this corresponds to an inverse variance weighted linear regression of the log odds ratio on its standard error.2Unfortunately, there are statistical problems because the standard error of the log odds ratio is mathematically linked to the size of the odds ratio, even in the absence of small study effects.24Many authors have therefore proposed alternative tests (see appendix on bmj.com).4262728 Because it is impossible to know the precise mechanism(s) leading to funnel plot asymmetry, simulation studies (in which tests are evaluated on large numbers of computer generated datasets) are required to evaluate test characteristics. Most have examined a range of assumptions about the extent of reporting bias by selectively removing studies from simulated datasets.262728After reviewing the results of these studies, and based on theoretical considerations, we formulated recommendations on testing for funnel plot asymmetry (box 2). The appendix describes the proposed tests, explains the reasons that some were not recommended, and discusses funnel plots for intervention effects measured as risk ratios, risk differences, and standardised mean differences. Our recommendations imply that tests for funnel plot asymmetry should be used in only a minority of meta-analyses.29 Box 2: Recommendations on testing for funnel plot asymmetry All types of outcome As a rule of thumb, tests for funnel plot asymmetry should not be used when there are fewer than 10 studies in the meta-analysis because test power is usually too low to distinguish chance from real asymmetry. (The lower the power of a test, the higher the proportion of “statistically significant” results in which there is in reality no association between study size and intervention effects). In some situations—for example, when there is substantial heterogeneity—the minimum number of studies may be substantially more than 10 Test results should be interpreted in the context of visual inspection of funnel plots— for example, are there studies with markedly different intervention effect estimates or studies that are highly influential in the asymmetry test? Even if an asymmetry test is statistically significant, publication bias can probably be excluded if small studies tend to lead to lower estimates of benefit than larger studies or if there are no studies with significant results When there is evidence of funnel plot asymmetry, publication bias is only one possible explanation (see box 1) As far as possible, testing strategy should be specified in advance: choice of test may depend on the degree of heterogeneity observed. Applying and reporting many tests is discouraged: if more than one test is used, all test results should be reported Tests for funnel plot asymmetry should not be used if the standard errors of the intervention effect estimates are all similar (the studies are of similar sizes) Continuous outcomes with intervention effects measured as mean differences The test proposed by Egger et al may be used to test for funnel plot asymmetry.1There is no reason to prefer more recently proposed tests, although their relative advantages and disadvantages have not been formally examined. General considerations suggest that the power will be greater than for dichotomous outcomes but that use of the test with substantially fewer than 10 studies would be unwise Dichotomous outcomes with intervention effects measured as odds ratios The tests proposed by Harbord et al26and Peters et al27avoid the mathematical association between the log odds ratio and its standard error when there is a substantial intervention effect while retaining power compared with alternative tests. However, false positive results may still occur if there is substantial between study heterogeneity If there is substantial between study heterogeneity (the estimated heterogeneity variance of log odds ratios, τ 2, is >0.1) only the arcsine test including random effects, proposed by Rücker et al, has been shown to work reasonably well.28However, it is slightly conservative in the absence of heterogeneity and its interpretation is less familiar than for other tests because it is based on an arcsine transformation. When τ 2is <0.1, one of the tests proposed by Harbord et al,26Peters et al,27or Rücker et al28can be used. Test performance generally deteriorates as τ 2increases. Funnel plots and meta-analysis models Fixed and random effects models Funnel plots can help guide choice of meta-analysis method. Random effects meta-analyses weight studies relatively more equally than fixed effect analyses by incorporating the between study variance into the denominator of each weight. If effect estimates are related to standard errors (funnel plot asymmetry), the random effects estimate will be pulled more towards findings from smaller studies than the fixed effect estimate will be. Random effects models can thus have undesirable consequences and are not always conservative.30 The trials of intravenous magnesium after myocardial infarction provide an extreme example of the differences between fixed and random effects analyses that can arise in the presence of funnel plot asymmetry.31Beneficial effects on mortality, found in a meta-analysis of small studies,32were subsequently contradicted when the very large ISIS-4 study found no evidence of benefit.33A contour enhanced funnel plot (fig 4 ⇓) gives a clear visual impression of asymmetry, which is confirmed by small P values from the Harbord and Peters tests (P<0.001 and P=0.002 respectively). Fig 4Contour enhanced funnel plot for trials of the effect of intravenous magnesium on mortality after myocardial infarction Download figure Open in new tab Figure 5 ⇓shows that in a fixed effect analysis ISIS-4 receives 90% of the weight, and there is no evidence of a beneficial effect. However, there is clear evidence of between study heterogeneity (P<0.001, I 2=68%), and in a random effects analysis the small studies dominate so that intervention appears beneficial. To interpret the accumulated evidence, it is necessary to make a judgment about the validity or relevance of the combined evidence from the smaller studies compared with that from ISIS-4. The contour enhanced funnel plot suggests that publication bias does not completely explain the asymmetry, since many of the beneficial effects reported from smaller studies were not significant. Plausible explanations for these results are that methodological flaws in the smaller studies, or changes in the standard of care (widespread adoption of treatments such as aspirin, heparin, and thrombolysis), led to apparent beneficial effects of magnesium. This belief was reinforced by the subsequent publication of the MAGIC trial, in which magnesium added to these treatments which also found no evidence of benefit on mortality (odds ratio 1.0, 95% confidence interval 0.8 to 1.1).34 Fig 5Comparison of fixed and random effects meta-analytical estimates of the effect of intravenous magnesium on mortality after myocardial infarction We recommend that when review authors are concerned about funnel plot asymmetry in a meta-analysis with evidence of between study heterogeneity, they should compare the fixed and random effects estimates of the intervention effect. If the random effects estimate is more beneficial, authors should consider whether it is plausible that the intervention is more effective in smaller studies. Formal investigations of heterogeneity of effects may reveal explanations for funnel plot asymmetry, in which case presentation of results should focus on these. If larger studies tend to be methodologically superior to smaller studies, or were conducted in circumstances more typical of the use of the intervention in practice, it may be appropriate to include only larger studies in the meta-analysis. Extrapolation of a funnel plot regression line An assumed relation between susceptibility to bias and study size can be exploited by extrapolating within a funnel plot. When funnel plot asymmetry is due to bias rather than substantive heterogeneity, it is usually assumed that results from larger studies are more believable than those from smaller studies because they are less susceptible to methodological flaws or reporting biases. Extrapolating a regression line on a funnel plot to minimum bias (maximum sample size) produces a meta-analytical estimate that can be regarded as corrected for such biases.353637However, because it is difficult to distinguish between asymmetry due to bias and asymmetry due to heterogeneity or chance, the broad applicability of such approaches is uncertain. Further approaches to adjusting for publication bias are described and discussed in the appendix. Discussion Reporting biases are one of a number of possible explanations for the associations between study size and effect size that are displayed in asymmetric funnel plots. Examining and testing for funnel plot asymmetry, when appropriate, is an important means of addressing bias in meta-analyses, but the multiple causes of asymmetry and limited power of asymmetry tests mean that other ways to address reporting biases are also of importance. Searches of online trial registries can identify unpublished trials, although they do not currently guarantee access to trial protocols and results. When there are no registered but unpublished trials, and the outcome of interest is reported by all trials, restricting meta-analyses to registered trials should preclude publication bias. Recent comparisons of results of published trials with those submitted for regulatory approval have also provided clear evidence of reporting bias.3839Methods for dealing with selective reporting of outcomes have been described elsewhere.40 Our recommendations apply to meta-analyses of randomised trials, and their applicability in other contexts such as meta-analyses of epidemiological or diagnostic test studies is unclear.41The performance of tests for funnel plot asymmetry in these contexts is likely to differ from that in meta-analyses of randomised trials. Further factors, such as confounding and precision of measurements, may cause a relation between study size and effect estimates in observational studies. For example, large studies based on routinely collected data might not fully control confounding compared with smaller, purpose designed studies that collected a wide range of potential confounding variables. Alternatively, larger studies might use self reported exposure levels, which are more error prone, while smaller studies used precise measuring instruments. However, simulation studies have usually not considered such situations. An exception is for diagnostic studies, where large imbalances in group sizes and substantial odds ratios lead to poor performance of some tests: that proposed by Deeks et al was designed for use in this context.4 Summary points Inferences on the presence of bias or heterogeneity should consider different causes of funnel plot asymmetry and should not be based on visual inspection of funnel plots alone They should be informed by contextual factors, including the plausibility of publication bias as an explanation for the asymmetry Testing for funnel plot asymmetry should follow the recommendations detailed in this article The fixed and random effects estimates of the intervention effect should be compared when funnel plot asymmetry exists in a meta-analysis with between study heterogeneity Notes Cite this as: BMJ2011;342:d4002 Footnotes Contributors: All authors contributed to the drafting and editing of the manuscript. DA, JC, JD, RMH, JPTH, JPAI, DRJ, DM, JP, GR, JACS, AJS and JT contributed to the chapter in the Cochrane Handbook for Systematic Reviews of Interventionson which our recommendations on testing for funnel plot asymmetry are based. JACS will act as guarantor. Funding: Funded in part by the Cochrane Collaboration Bias Methods Group, which receives infrastructure funding as part of a commitment by the Canadian Institutes of Health Research (CIHR) and the Canadian Agency for Drugs and Technologies in Health (CADTH) to fund Canadian based Cochrane entities. This supports dissemination activities, web hosting, travel, training, workshops and a full time coordinator position. JPTH was funded by MRC Grant U.1052.00.011. DGA is supported by Cancer Research UK. GR was supported by a grant from Deutsche Forschungsgemeinschaft (FOR 534 Schw 821/2-2). Competing interests. JC, JJD, SD, RMH, JPAI, DRJ, PM, JP, GR, GS, JACS and AJS are all authors on papers proposing tests for funnel plot asymmetry, but have no commercial interests in the use of these tests. All authors have completed the ICJME unified disclosure form atwww.icmje.org/coi_disclosure.pdf(available on request from the corresponding author) and declare that they have no financial or non-financial interests that may be relevant to the submitted work. Provenance and peer review: Not commissioned; externally peer reviewed. References ↵ Egger M, Davey Smith G, Schneider M, Minder C. Bias in meta-analysis detected by a simple, graphical test. BMJ 1997 ; 315 : 629 -34. OpenUrl Abstract / FREE Full Text Google Scholar ↵ Sterne JAC, Gavaghan D, Egger M. Publication and related bias in meta-analysis: power of statistical tests and prevalence in the literature. J Clin Epidemiol 2000 ; 53 : 1119 -29. OpenUrl CrossRef PubMed Web of Science Google Scholar ↵ Lau J, Ioannidis JP, Terrin N, Schmid CH, Olkin I. The case of the misleading funnel plot. BMJ 2006 ; 333 : 597 -600. OpenUrl FREE Full Text Google Scholar ↵ Deeks JJ, Macaskill P, Irwig L. The performance of tests of publication bias and other sample size effects in systematic reviews of diagnostic test accuracy was assessed. J Clin Epidemiol 2005 ; 58 : 882 -93. OpenUrl CrossRef PubMed Web of Science Google Scholar ↵ Terrin N, Schmid CH, Lau J. In an empirical evaluation of the funnel plot, researchers could not visually identify publication bias. J Clin Epidemiol 2005 ; 58 : 894 -901. OpenUrl CrossRef PubMed Web of Science Google Scholar ↵ Ioannidis JP. Interpretation of tests of heterogeneity and bias in meta-analysis. J Eval Clin Pract 2008 ; 14 : 951 -7. OpenUrl CrossRef PubMed Web of Science Google Scholar ↵ Sterne JAC, Egger M, Moher D. Addressing reporting biases. In: Higgins JPT, Green S, eds. Cochrane handbook for systematic reviews of interventions . Wiley, 2008 . Google Scholar ↵ Sterne JAC, Egger M. Funnel plots for detecting bias in meta-analysis: guidelines on choice of axis. J Clin Epidemiol 2001 ; 54 : 1046 -55. OpenUrl CrossRef PubMed Web of Science Google Scholar ↵ Begg CB, Berlin JA. Publication bias: a problem in interpreting medical data. J R Statist Soc A 1988 ; 151 : 419 -63. OpenUrl CrossRef Google Scholar ↵ Davey Smith G, Egger M. Who benefits from medical interventions? Treating low risk patients can be a high risk strategy. BMJ 1994 ; 308 : 72 -4. OpenUrl FREE Full Text Google Scholar ↵ Stuck AE, Siu AL, Wieland GD, Adams J, Rubenstein LZ. Comprehensive geriatric assessment: a meta-analysis of controlled trials. Lancet 1993 ; 342 : 1032 -6. OpenUrl CrossRef PubMed Web of Science Google Scholar ↵ Peters JL, Sutton AJ, Jones DR, Abrams KR, Rushton L, Moreno SG. Assessing publication bias in meta-analyses in the presence of between-study heterogeneity. J R Statist Soc A 2010 ; 173 : 575 -91. OpenUrl CrossRef Google Scholar ↵ Egger M, Jüni P, Bartlett C, Holenstein F, Sterne J. How important are comprehensive literature searches and the assessment of trial quality in systematic reviews? Empirical study. Health Technol Assess 2003 ; 7 : 1 -68. OpenUrl PubMed Google Scholar ↵ Ioannidis JP. Why most discovered true associations are inflated. Epidemiology 2008 ; 19 : 640 -8. OpenUrl CrossRef PubMed Web of Science Google Scholar ↵ Wood L, Egger M, Gluud LL, Schulz KF, Jüni P, Altman DG, et al. Empirical evidence of bias in treatment effect estimates in controlled trials with different interventions and outcomes: meta-epidemiological study. BMJ 2008 ; 336 : 601 -5. Hopewell S, Clarke M, Stewart L, Tierney J. Time to publication for results of clinical trials. Cochrane Database Syst Rev 2007 ; 2 : MR000011 . Hopewell S, Loudon K, Clarke MJ, Oxman AD, Dickersin K. Publication bias in clinical trials due to statistical significance or direction of trial results. Cochrane Database Syst Rev 2009 ; 1 : MR000006 . Song F, Parekh S, Hooper L, Loke YK, Ryder J, Sutton J, et al. Dissemination and publication of research findings: an updated review of related biases. Health Technol Assess 2010 ; 14 : iii ,ix-iii,193. Dwan K, Altman DG, Arnaiz JA, Bloom J, Chan AW, Cronin E, et al. Systematic review of the empirical evidence of study publication bias and outcome reporting bias. PLoS ONE 2008 ; 3 : e3081 . Turner EH, Matthews AM, Linardatos E, Tell RA, Rosenthal R. Selective publication of antidepressant trials and its influence on apparent efficacy. N Engl J Med 2008 ; 358 : 252 -60. Rosenthal R. The “file drawer” problem and tolerance for null results. Psychol Bull 1979 ; 86 : 638 -41. Chan AW, Hrobjartsson A, Haahr MT, Gotzsche PC, Altman DG. Empirical evidence for selective reporting of outcomes in randomized trials: comparison of protocols to published articles. JAMA 2004 ; 291 : 2457 -65. Peters J, Sutton AJ, Jones DR, Abrams KR, Rushton L. Contour-enhanced meta-analysis funnel plots help distinguish publication bias from other causes of asymmetry. J Clin Epidemiol 2008 ; 61 : 991 -6. Harbord RM, Egger M, Sterne JA. A modified test for small-study effects in meta-analyses of controlled trials with binary endpoints. Stat Med 2006 ; 25 : 3443 -57. Peters JL, Sutton AJ, Jones DR, Abrams KR, Rushton L. Comparison of two methods to detect publication bias in meta-analysis. JAMA 2006 ; 295 : 676 -80. Rücker G, Schwarzer G, Carpenter J. Arcsine test for publication bias in meta-analyses with binary outcomes. Stat Med 2008 ; 27 : 746 -63. Ioannidis JP, Trikalinos TA. The appropriateness of asymmetry tests for publication bias in meta-analyses: a large survey. CMAJ 2007 ; 176 : 1091 -6. Moreno SG, Sutton AJ, Turner EH, Abrams KR, Cooper NJ, Palmer TM, et al. Novel methods to deal with publication biases: secondary analysis of antidepressant trials in the FDA trial registry database and related journal publications. BMJ 2009 ; 339 : b2981 . Eyding D, Lelgemann M, Grouven U, Härter M, Kromp M, Kaiser T, et al. Reboxetine for acute treatment of major depression: systematic review and meta-analysis of published and unpublished placebo and selective serotonin reuptake inhibitor controlled trials. BMJ 2010 ; 341 : c4737 . Kirkham JJ, Dwan KM, Altman DG, Gamble C, Dodd S, Smyth R, et al. The impact of outcome reporting bias in randomised controlled trials on a cohort of systematic reviews. BMJ 2010 ; 340 : c365 . Egger M, Schneider M, Davey Smith G. Spurious precision? Meta-analysis of observational studies. BMJ 1998 ; 316 : 140 -4.

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6102.0.55.001 - Labour Statistics: Concepts, Sources and Methods, Apr 2007 6102.0.55.001 - Labour Statistics: Concepts, Sources and Methods, Apr 2007 About this Release Contents History of Changes CHAPTER 21.10. PERSONS NOT IN THE LABOUR FORCE INTRODUCTION 21.10.1 The Persons Not in the Labour Force (PNILF) survey collects information about persons who are neither employed nor unemployed. Data from this survey are used to measure potential labour supply not reflected in employment and unemployment statistics, and in particular to measure the number and characteristics of discouraged job seekers and others marginally attached to the labour force. The policy context for the data includes labour market programs for the jobless, income support programs, retirement policies, and child care policies. The survey was first conducted in May 1975 and then in May 1977. From 1979 to 1987 the survey was collected biannually. The survey has been conducted annually since 1988. 21.10.2 This section describes only those aspects of the methodology that are unique to this survey, and hence should be used in conjunction with the overview part of this chapter, which outlines the survey methodology used in supplementary surveys. SURVEY OUTPUT 21.10.3 Data from the survey are published in Persons Not in the Labour Force, Australia (cat. no. 6220.0) and standard data service Persons Not in the Labour Force, Australia (cat. no. 6220.0.40.001) for 1994 and 1995 (available in hard copy only). More detailed estimates are available on request. 21.10.4 Data from the survey relate to persons aged 15 years and over. The main populations of interest are: persons not in the labour force: persons who are neither employed nor unemployed; persons marginally attached to the labour force: people who were not in the labour force in the reference week, want to work and were either actively looking for work but were not available to start in the survey reference week, or were not actively looking for work but were available to start work within four weeks; and discouraged job seekers: people who want work and were available to work within four weeks, but weren't actively looking for work because they believed they would not find a job. 21.10.5 Estimates are produced on an original basis only (i.e. not seasonally adjusted or trend) and include: Socio-demographic information Sex; age; marital status; relationship in household; state or territory of usual residence; country of birth; year of arrival in Australia; and full-time study status. Persons not in the labour force Whether want to work and full-time/part-time preference; whether looked for work in last 12 months; main activity when not in the labour force; and intention to enter the labour force; For persons who have previously had a job: time since last job, and reason for ceasing that job; and For persons who have held a job in the last 20 years: details of the job including occupation, status in employment, and full-time or part-time status. Persons marginally attached to the labour force Details of whether actively looking for work and whether available for work; For persons not actively looking for work: all and main reason(s) not actively looking for work; For persons not available to start work within four weeks: all and main reason not available; and For persons not actively looking for work or available to start work within four weeks for caring for children reasons: age of youngest child and number of children . 21.10.6 Data collected in the survey are compiled according to concepts and definitions outlined in Chapter 4 (Employment Measures and Classifications), Chapter 7 (Not in the Labour Force) and Chapter 16 (Other Classifications Used in Labour Statistics). SCOPE 21.10.7 The scope of this survey is restricted to persons aged 15 years and over. The standard scope restrictions for supplementary surveys outlined in the overview part of Chapter 21 also apply to this survey. DATA COMPARABILITY OVER TIME 21.10.8 In order to provide a high degree of consistency and comparability over time, changes to survey methods, survey concepts, data item definitions, frequency of collection, and analysis methods are made as infrequently as possible. Changes affecting the Labour Force Survey may also affect this survey. Such changes are outlined in Chapter 20 and are not repeated here. 1975 Initial survey conducted (November) - preliminary data published in Persons Not In the Labour Force, Preliminary (ref. no. 6.54) and final data in Persons Not In the Labour Force (ref. no. 6.59). Survey scope - persons aged 15-64. 1977 Survey conducted (May). Catalogue numbers replaced reference numbers - cat. no. 6219.0 (preliminary data) and cat. no. 6220.0 (final data). 1979 Biannual collection commenced (March and September), some definitional changes. 1980 Change to definition of persons wanting a job - broadened to include persons available to start work within 4 weeks from the end of the reference week but not actively looking for work. Preliminary publication discontinued. 1983 Concept of marginal attachment to the labour force introduced. Improvements made to measure of discouraged job seekers. Survey scope changed - persons aged 15 years and over. 1988 Survey scope changed - persons aged 15-69. Annual collection commenced (September). 1994 Standard Data Service replaced publication format (cat. no. changed to 6220.0.40.001). Sample reduced to seven-eighths of Labour Force Survey sample. 1996 Publication format replaced Standard Data Service (cat. no. changed to 6220.0). 1997 Persons living in remote and sparsely settled parts of Australia excluded from scope of survey. 2001 The introduction of the redesigned Labour Force Survey (in April 2001) allowed the identification of a new population group 'People permanently not intending to work'. These people were not asked questions about wanting to work and looking for work. As such these people were classified as 'Did not want to work' (without marginal attachment). 2005 The scope of the survey was expanded to include all persons aged 15 years and over. 2006 Estimates of the number of people who were not in the labour force because they were caring for children include people whose youngest child was aged 12 years and under. Previously questions relating to the care of children were only asked of people with children aged 11 years and under. 2007 Those 'permanently not intending to work' were further classified as either 'wanting to work' and 'did not want to work'. As a result of this change there has been a break in time series. For the data items 'All reasons for not actively looking for work', 'Main reason for not actively looking for work', and 'Main reason not available to start work within four weeks' the response category 'own ill health, physical disability or pregnancy' was changed to three separate categories; 'own short-term illness or injury', 'own long-term health condition or disability' and 'pregnancy'. For the data items 'Reason for ceasing last job', and 'Main activity when not in the labour force' the response categories 'own ill-health or injury' and 'own disability or handicap' were changed to 'own short-term illness or injury' and 'own long-term health condition or disability' respectively to ensure consistency within the survey resulting in a break in series. A new data item was collected: 'All reasons not available to start work within four weeks'. 2008 For the data item 'Main activity when not in the labour force' the category 'Home duties or caring for children' was split into 'Home duties' and 'Caring for children'. 2009 For the response categories 'All reasons for not actively looking for work' and 'Main reason for not actively looking for work' a new data item 'Believes ill-health or disability discourages employers' was collected. For the population group 'Discouraged job seekers' the response categories 'Believes no jobs in suitable hours' and 'Believes ill-health or disability discourages employers' were included resulting in a break in series. A new data item: 'Preferred number of hours' was collected. The data item 'Full-time or part-time preference' was determined using this information based on the number of hours a respondent preferred to work each week. Both data items were collected for people who intend to enter the labour force in the next 12 months, resulting in a break in series. FURTHER INFORMATION 21.10.9 For further details contact the Labour Market Statistics Section, on Canberra (02) 6252 7206 or email <[email protected]>.

https://www.abs.gov.au/ausstats/[email protected]/Latestproducts/9E077BFE0DAE708ACA2572C100244D50?opendocument

Spatial and temporal variation of dengue incidence in the island of Bali, Indonesia: An ecological study | Request PDF Request PDF | Spatial and temporal variation of dengue incidence in the island of Bali, Indonesia: An ecological study | Background: Dengue fever control in the tropical island of Bali in Indonesia carries important significance both nationally and globally, as it... | Find, read and cite all the research you need on ResearchGate Article Spatial and temporal variation of dengue incidence in the island of Bali, Indonesia: An ecological study July 2019 Travel Medicine and Infectious Disease32(7446) DOI: 10.1016/j.tmaid.2019.06.008 Authors: <here is a image 16ae2b4ebc2d8cf6-18dc12a504b4f9a6> Pandji Wibawa Dhewantara National Research and Innovation Agency <here is a image 80e3bd444df2e017-16c96f943d549e1d> Rina Marina National Instute of Health Reseach Development, INA <here is a image a6a91acd9e0bbf61-63eb73ab386af719> Ricardo J Soares Magalhães The University of Queensland <here is a image c5bd18a338822d72-104682e6f6338bdc> Wenbiao Hu Queensland University of Technology Abstract Background: Dengue fever control in the tropical island of Bali in Indonesia carries important significance both nationally and globally, as it is one of the most endemic islands in Indonesia and a worldwide popular travel destination. Despite its importance, the spatial and temporal heterogeneity in dengue risk and factors associated with its variation in risk across the island has not been not well explored. This study was aimed to analyze for the first time the geographical and temporal patterns of the incidence of dengue and to quantify the role of environmental and social factors on the spatial heterogeneity of dengue incidence in Bali. Methods: We analyzed retrospective dengue notification data at the sub-district level (Kecamatan) from January 2012 to December 2017 which obtained from the Indonesian Ministry of Health. Seasonality in notified dengue incidence was assessed by seasonal trend decomposition analysis with Loess (STL) smoothing. Crude standardized morbidity rates (SMRs) of dengue were calculated. Moran's I and local indicators of spatial autocorrelation (LISA) analysis were employed to assess spatial clustering and high-risk areas over the period studied. Bayesian spatial and temporal conditional autoregressive (CAR) modeling was performed to quantify the effects of rainfall, temperature, elevation, and population density on the spatial distribution of risk of dengue in Bali. Results: Strong seasonality of dengue incidence was observed with most cases notified during January to May. Dengue incidence was spatially clustered during the period studied with high-risk kecamatans concentrated in the south of the island, but since 2014, the high-risk areas expanded toward the eastern part of the island. The best-fitted CAR model showed increased dengue risk in kecamatans with high total annual rainfall (relative risk (RR): 1.16 for each 1-mm increase in rainfall; 95% Credible interval (CrI): 1.03-1.31) and high population density (RR: 7.90 per 1000 people/sq.km increase; 95% CrI: 3.01-20.40). The RR of dengue was decreased in kecamatans with higher elevation (RR: 0.73 for each 1-m increase in elevation; 95% CrI: 0.55-0.98). No significant association was observed between dengue RR and year except in 2014, where the dengue RR was significantly lower (RR: 0.53; 95% CrI: 0.30-0.92) relative to 2012. Conclusions: Dengue incidence was strongly seasonal and spatially clustered in Bali. High-risk areas were spread from kecamatans in Badung and Denpasar toward Karangasem and Klungkung. The spatial heterogeneity of dengue risk across Bali was influenced by rainfall, elevation, and population density. Surveillance and targeted intervention strategies should be prioritized in the high-risk kecamatans identified in this study to better control dengue transmission in this most touristic island in Indonesia. Local health authorities should recommend travelers to use personal protective measures, especially during the peak epidemic period, before visiting Bali. ... By analyzing data at a smaller scale, such as neighborhoods or districts within a city, these maps can highlight areas that are more susceptible to dengue transmission. This information is crucial for local health authorities to implement targeted interventions, allocate resources effectively, the e ciency of control measures and reduces the burden on resources [15] . Of the 45 total studies, 39 (86.67%) were applied to the administrative level, such as district, municipality, regency/city, province, and country, and 6 (13.33%) were applied to the continent and region, respectively. ... ... Retrospective descriptive is research conducted with the main aim of making a picture or description of a situation objectively by looking back. Although some studies (8, 14, 15, 21,23,24,34,36,39,45) did not use DHF data as a reference, the majority of them used secondary data on DHF cases from health surveillance system surveys. Aside from reported DHF cases, critical predictors for model generation and DHF risk maps included variables from a variety of categories, including population, demography, socioeconomic, climatology, environmental, entomological, capacity, and epidemiological data. ... ... Population density data were used in the 23 articles reviewed. Demographic data used several predictors including age, gender, skin color, occupation, and sex used in 16 studies of 45 studies reviewed (4,5,7,8,9, 15, 18,21,31,34,35,37,38,40,41,44). Meanwhile, the predictors used in social and economic data include; housing type, residential buffer, urban proportion, proximity to road/road network, toilet type, mean distance to hospital, poverty incidence, number of household rooms, number of persons per household, crowding, duration of residence, people who did not study nor worked, following any type of study, having a job, household density and gross domestic product (GDP). ... Spatial Model of Dengue Hemorrhagic Fever (DHF) Risk: Scoping Review Preprint Full-text available May 2023 <here is a image 237551263896390c-57da13de6d128c32> Ririn Pakaya D. Daniel Prima Widayani <here is a image cd9d3d704ae222bd-593429feda3915b0> Adi Utarini This review aims to provide a comprehensive overview of the important predictors, and additionally spatial modeling tools capable of producing Dengue Hemorrhagic Fever (DHF) risk maps. A literature search was conducted in PubMed, Scopus, Science Direct, and Google Scholar for studies reporting DHF risk factors. The Preferred Reporting Items for Systematic Reviews (PRISMA) 2020 statement is used to report this scoping review. It lasted from January 2011 to August of 2022. Initially 1329 articles were found, after inclusion and exclusion criteria, 45 manuscripts were selected. A variety of models and techniques were used to identify DHF risk areas with an arrangement of various multiple-criteria decision-making, statistical, and Machine Learning technique. We found that There was no pattern of predictor use associated with particular approaches; instead, a wide range of predictors was used to create DHF risk maps. Predictors are various variables or factors that are considered when assessing the likelihood or intensity of DHF outbreaks in a specific area in the context of DHF risk mapping. These predictors can include climatology factors (e.g., temperature, rainfall, humidity), socio-economic indicators (e.g., population density, urbanization level), environmental factors (land-use, elevation) and other relevant factors (e.g., mosquito abundance, previous DHF cases). The spatial model of DHF risk is a valuable tool for public health authorities, policymakers, and communities to identify areas at higher risk of dengue transmission, but its limitations underscore the importance of complementing it with other approaches and considering contextual factors for a more holistic assessment of DHF outbreaks. It enables targeted interventions, such as vector control measures and public awareness campaigns, to be implemented in high-risk areas, ultimately helping to mitigate the impact of dengue outbreaks and protect public health. ... Moran's I statistic was used at the global level and Local Indicators of Spatial Association (LISA) was for local autocorrelation (16). Positive spatial correlation meant that adjacent values in this space had similar trends while negative spatial correlation referred to the opposite (20) . The diagram of LISA was drawn in local autocorrelation analysis, which identified the distribution of "high-high, " "low-low, " "low-high" and "high-low" clusters (21) of infectious disease. ... ... Observed cases (O i ) of syphilis for the district i were assumed to have a Poisson distribution with mean µ i . The model was written as (20, 30): ... ... α 0 was an intercept term representing the baseline log of syphilis across the study region, x ij was the variate in the district i, with associated regression coefficient α j and b i was an area-specific random effect capturing the residual or unexplained log RR of disease in the area a i . b i represented the effect of latent or unobserved risk factors (20) which had spatial correlation. Winbugs14 described this model as follows: ... Spatio–temporal variation on syphilis from 2005 to 2018 in Zhejiang Province, China Article Full-text available Aug 2022 Xiaoxia Zhu Zhixin Zhu Lanfang Gu Xiuyang Li Background Syphilis has spread throughout China, especially in Zhejiang Province which endangers the health and lives of people. However, the spatial and temporal epidemiological studies of syphilis in Zhejiang are not thorough enough. The temporal and spatial variation and the relevant factors of syphilis incidence should be analyzed for more effective prevention and control in Zhejiang, China. Methods Data on confirmed cases of syphilis in Zhejiang Province from 2005 to 2018 was used and the spatio–temporal distributions were described. The spatial autocorrelation analysis and SaTScan analysis were performed to identify spatio–temporal clusters. A Bayesian spatial Conditional Autoregression (CAR) model was constructed to explore the relationships between syphilis incidence and common social and natural indicators. Results 474,980 confirmed cases of syphilis were reported between 2005 and 2018 with a large peak in 2010. Farmers and unemployed people accounted for the largest proportion of confirmed cases. And the significant spatial clusters of syphilis were concentrated in the north of Zhejiang Province, especially in more economically developed regions. Seven spatio–temporal clusters were identified and the main three high–risk areas were located in Hangzhou (RR = 1.62, P < 0.05), Zhoushan and Ningbo (RR = 1.99, P < 0.001), and Lishui (RR = 1.68, P < 0.05). The findings showed that the morbidity of syphilis was positively correlated with the Gross Domestic Product (GDP) per capita, the number of health technicians per 10,000 people, the proportion of the elderly and air temperature were negatively correlated with the proportion of the urban population, the proportion of men and precipitation. Conclusions The spatio–temporal analysis revealed that the prevalence of syphilis was still serious in Zhejiang Province. Syphilis high–risk areas were mainly located in the more developed coastal regions where more targeted intervention measures were required to be implemented. The study highlighted the need to strengthen Sexually Transmitted Diseases (STD) screening and health education for high–risk groups and improve the coverage of syphilis testing to reduce hidden syphilis cases. ... Dengue fever occurs all year in the tropics and creates epidemics in various locations (Mertha Adnyana et al., 2021). Dengue virus is transmitted through the bites of Aedes spp, Aedes aegypti, and Aedes albopictus mosquitoes (Nath et al., 2021;Widjajanti et al., 2019) which are classified as arthropod-borne viruses (arbovirus) (Dhewantara et al., 2019; Jansen et al., 2021;Ong et al., 2021). Bali Province is a dengue fever endemic location, contributing to the most cases each year (Bali Health Department, 2021a). ... ... It shows that the government's preventive efforts, such as the 3M-plus program (draining, closing and burying, and sowing powder Temephos), have not been implemented optimally. The high level of pain taken by dengue hemorrhagic fever (DHF), especially in nine districts/city in Bali Province, need to get serious attention, given this endemic disease still appears throughout the year (Dhewantara et al., 2019; Suparyatha et al., 2021). Less-controlled environmental conditions resulted in the transmission of cases due to DHF increasing rapidly. ... ... In terms of administrative areas the three districts/cities are areas with a high level of mobility. The region's proximity shows that there is spatial autocorrelation with the dispersed category in the area (Dhewantara et al., 2019) . ... PREVALENCE OF DENGUE HEMORRHAGIC FEVER IN BALI FROM 2015 TO 2020 AND DURING THE COVID-19 PANDEMIC Article Background: Dengue hemorrhagic fever cases increased significantly during the COVID-19 pandemic phase. Purpose: This study defines, describes, and evaluates the dengue hemorrhagic fever incidence in Bali Province from 2015 to 2020 and in the context of the COVID-19 pandemic. Methods: The various types of descriptive study with case series design. The study used an environmental epidemiology approach to analyze the health profile of Bali Province. The number of DHF cases in 2015-2020, DHF-related morbidity and death rates, larva-free rates, climate, population and population density, and gender were variables in this study. The study took place in May and June 2021. Results: The prevalence of dengue hemorrhagic fever in the province of Bali fluctuated between 2015 and 2020. The highest number of dengue cases in 2016 was 20,306, and they fell from 2017 to 2019. Cases doubled in 2020, up by 12,173. The endemic areas with the highest prevalence of cases are in the districts of Buleleng and Badung. The highest incidence rate was in 2016 at 483/100,000 population, and for the last five years, it has not met the national target of 49/100,000 population. The highest CFR figure in 2020 was 0.43%, an increase of 1.5 times during the COVID-19 pandemic. Conclusion: The prevalence of dengue hemorrhagic fever cases in Bali in 2015 – 2020 fluctuated significantly, and mortality due to dengue during the COVID-19 pandemic experienced a significant increase. ... The factors that contribute to risk variation within an area, as well as the spatial and temporal heterogeneity in dengue risk, have not been thoroughly investigated [3] . It implies the urgency of health problems that change based on time and place. ... ... Most of the research uses a Bayesian statistical model, such as conditional autoregressive [9], Poisson [6][10] and negative binomial [6] which contributes valuable for assessment, prediction and prevention of dengue risk. But these methods can only analyze the interaction of two factors, even though the factors that influence the risk of DHF are more complex, including the climate [3] [7] [8], socio-economic [3][11], environmental [11], and demography [11]. ... ... Most of the research uses a Bayesian statistical model, such as conditional autoregressive [9], Poisson [6][10] and negative binomial [6] which contributes valuable for assessment, prediction and prevention of dengue risk. But these methods can only analyze the interaction of two factors, even though the factors that influence the risk of DHF are more complex, including the climate [3][7] [8], socio-economic [3] [11], environmental [11], and demography [11]. ... Spatial-Temporal Visualization of Dengue Haemorrhagic Fever Vulnerability in Kediri District, Indonesia, Using K-means Algorithm Conference Paper ... [7] The transmission of Dengue Hemorrhagic Fever (DHF) is influenced by some factors, such as population mobility, climate, population density, and vector behavior . [4, 5, 6] Rice field or vegetation conversion to be residential areas exacerbates the potential DHF transmission. It components are correlated with various species in the ecosystem and the distribution patterns of viruses produced by vectors, such as dengue. ... ... Various efforts to control the prevalence of DHF incidents, especially in areas with the high or persistent transmission, are required. [,8,9,10] Climate factors, ecological changes, and socio-demographic factors can expand DHF endemic areas [5, 8] Of 9 districts and 60 sub-districts with 636 villages in Bali province, most of them are DHF endemic areas. In three consecutive years, the population in 9 districts in Bali is at risk of DHF. ... ... [10,11] Bali is ideal for Aedes aegypti breeding sites because it has an average temperature of 26- (temperature and humidity) will increase the mosquito population density, which in turn affects the transmission and spread of dengue disease. [5, 7,12] Temperature between 20-30 o C and humidity around 60 -90% allow the growth and development of the Aedes aegypti optimally. [,8,9,10] Aedes aegypti can transmit in DHF because they live in and around houses where people do activities during the day. ... Integration of Climate Factors for Dengue Fever Control Model in Bali Article Bali has a high number of dengue hemorrhagic fever (DHF) incidents, involvingfour serotypes of dengue virus. As an international and domestic tourism destination, Bali hasannually been visited by more than 4 million tourists. Aim: This study predicted the incidence ofdengue fever based on climatic factors including rainfall, humidity, temperature, and length ofexposure to sunlight, as well as control models. Setting and Design: This study was a time-series ecological study using secondary data from the Bali Provincial Health Office. These datainvolved the incidence of dengue fever (DF) and larvae free number (LFN), as well as climatedata from the Meteorology and Geophysics Agency for Region III Denpasar. Subjects andMethods: The time-series ecological research design was adopted to examine the correlation of climaticfactors. The correlation analysis of DHF and LFN was done quarterly. A simple linear regression testedthe correlation among variables. Results: Humidity was correlated with LFN, but LFN was notcorrelated with DF incidence. Dengue fever control model is predicted based on the correlationcoefficient between climatic factors and the incidence of dengue fever, control of diseasesources, transmission media, and exposure in the community. Conclusion: The DF controlmodel can be used for early prevention measures, particularly during the rainy season fromJanuary to June where standing water can facilitate mosquitos vector of Aedes Aegypti todevelop breeding sites. The increasing vector population would increase the denguetransmission. ... For high temperature studies, the ambient temperatures had been used with different temporal exposure periods, including daily, 14,23,63,64,68 bi-weekly, 69 monthly and yearly. 70 Mean temperature was frequently used as temperature metrics (n = 31). ... ... Vol 91 May, 2023and dengue incidence, 51 studies were included in the narrative synthesis. Except for a few studies (n = 13) that found insignificant associations27, 70, 84,85,[88][89][90] and negative associations67,76,86,[91][92][93][94] the rest of 90 studies reported a significant positive association between high temperature and dengue incidence. The observed associations were reported at different lag periods ranging from weeks to months. ... Effects of high temperatures and heatwaves on dengue fever: a systematic review and meta-analysis Background: Studies have shown that dengue virus transmission increases in association with ambient temperature. We performed a systematic review and meta-analysis to assess the effect of both high temperatures and heatwave events on dengue transmission in different climate zones globally. Methods: A systematic literature search was conducted in PubMed, Scopus, Embase, and Web of Science from January 1990 to September 20, 2022. We included peer reviewed original observational studies using ecological time series, case crossover, or case series study designs reporting the association of high temperatures and heatwave with dengue and comparing risks over different exposures or time periods. Studies classified as case reports, clinical trials, non-human studies, conference abstracts, editorials, reviews, books, posters, commentaries; and studies that examined only seasonal effects were excluded. Effect estimates were extracted from published literature. A random effects meta-analysis was performed to pool the relative risks (RRs) of dengue infection per 1 °C increase in temperature, and further subgroup analyses were also conducted. The quality and strength of evidence were evaluated following the Navigation Guide systematic review methodology framework. The review protocol has been registered in the International Prospective Register of Systematic Reviews (PROSPERO). Findings: The study selection process yielded 6367 studies. A total of 106 studies covering more than four million dengue cases fulfilled the inclusion criteria; of these, 54 studies were eligible for meta-analysis. The overall pooled estimate showed a 13% increase in risk of dengue infection (RR = 1.13; 95% confidence interval (CI): 1.11-1.16, I2 = 98.0%) for each 1 °C increase in high temperatures. Subgroup analyses by climate zones suggested greater effects of temperature in tropical monsoon climate zone (RR = 1.29, 95% CI: 1.11-1.51) and humid subtropical climate zone (RR = 1.20, 95% CI: 1.15-1.25). Heatwave events showed association with an increased risk of dengue infection (RR = 1.08; 95% CI: 0.95-1.23, I2 = 88.9%), despite a wide confidence interval. The overall strength of evidence was found to be "sufficient" for high temperatures but "limited" for heatwaves. Our results showed that high temperatures increased the risk of dengue infection, albeit with varying risks across climate zones and different levels of national income. Interpretation: High temperatures increased the relative risk of dengue infection. Future studies on the association between temperature and dengue infection should consider local and regional climate, socio-demographic and environmental characteristics to explore vulnerability at local and regional levels for tailored prevention. Funding: Australian Research Council Discovery Program. ... When viewed from 2018-to 2020, the number of dengue cases has increased three times from the previous year [2]. Furthermore, in the last decade, the province of Bali has been one of the top 10 contributors to the highest dengue hemorrhagic fever cases in Indonesia [3] , [4]. In the COVID-19 pandemic, Bali was recorded to occupy the second position after West Java with the highest cases, and Buleleng Regency accounted for the highest cases in Indonesia [1], [5]. ... ... The high number of cases in this region is due to population density and the dominance of tourist areas. High levels of mobilization and sanitation in various areas are poorly controlled, and climate support (weather, temperature, humidity, and high rainfall) [3] , [9], [16]. Recent research states that there are similar signs and symptoms between dengue hemorrhagic fever and COVID-19, so the COVID-19 pandemic in Bali is a high-risk factor for infection with these two diseases [17]- [19]. ... Strategy to control and eradicate dengue hemorrhagic fever vectors in Bali Dengue hemorrhagic fever cases in Bali have increased in the past decade. Control and eradication efforts must be optimized. In order to create dengue-free zones, accurate information and a comprehensive strategy for accelerating dengue vector management are required. This paper is based on empirical, field, and epidemiological studies and program evaluations guided by the health belief model approach. In this regard, Bali's lesson on dengue cases fluctuated from 2018 to 2020. The incidence and death rates are increasing, especially during the COVID-19. Many factors affect the disease agent, the host, and the environment. The SIGAP strategy is a policy brief that is studied and implemented to accelerate dengue vector control in Bali, which includes: awareness of the importance of 4M-Plus (draining, closing, reusing used goods, and monitoring) and mosquito nest control; integration of dengue information services; using natural insecticides and larvicides; observing and reporting the presence of vectors to the larva monitoring officer; and regular monitoring weekly. Implementing the SIGAP strategy can reduce dengue vectors and speed up the process of getting rid of dengue disease in Bali. ... Dengue fever (DF) is a viral disease that is transmitted to humans by mosquitoes of the Aedes genus [1]. The disease is caused by four unique dengue virus strains (DENV-1 to DENV-4) belonging to the Flaviviridae family and is transmitted by the yellow-fever and Asian tiger mosquitos [2] . Dengue fever is the most common mosquito-borne viral disease afflicting humans in the world's tropical and subtropical regions. ... ... Technological developments such as the Geographical Information System (GIS) and remote sensing (RS) have contributed significantly to public health sectors, enabling a better understanding of the transmission of vector-borne diseases, and improving the control procedures [2, 8,9]. Moreover, Machine Learning (ML) has been used in a variety of applications for the geosciences due to its ability to capture different aspects of data [10]. ... Predicting Dengue Fever Transmission Using Machine Learning Methods Conference Paper ... Overall, 10 (19.2%) studies were on chronic diseases [53][54][55][56][57][58][59][60][61][62], 8 (15.5%) on maternal and child health [63][64][65][66], 3 (5.7%) on nutrition [67][68][69] and 5 (9.6%) were applied to other health-related outcomes [58,[70][71][72][73]. Most of the studies (42, 80.7%) used an ecological study design [30,31,33,34,[37][38][39][40][41][42][43][44][45][46][47][51][52][53][54][55][56][58][59][60][61][62][64][65][66][67]69,[71][72][73][74][75] [76] [77][78][79][80][81]. Twenty-four studies (46.1%) used data from a national health survey or Demographic and Health Survey (DHS), while twenty other studies (38.4%) used data from hospital/administrative records. ... A Systematic Review of Areal Units and Adjacency Used in Bayesian Spatial and Spatio-Temporal Conditional Autoregressive Models in Health Research Article Int J Environ Res Publ Health <here is a image f0777374a727438f-40854ad898880144> Zemenu Tadesse Tessema <here is a image b5ed70a1ddc8f40c-4494fc6a80587a77> Getayeneh Antehunegn Tesema <here is a image 2636402624cc7869-1b59530c0c3c4d7b> Susannah Ahern <here is a image ee3e10ebc5a5d887-3b7400715a8a9f60> Arul Earnest Advancements in Bayesian spatial and spatio-temporal modelling have been observed in recent years. Despite this, there are unresolved issues about the choice of appropriate spatial unit and adjacency matrix in disease mapping. There is limited systematic review evidence on this topic. This review aimed to address these problems. We searched seven databases to find published articles on this topic. A modified quality assessment tool was used to assess the quality of studies. A total of 52 studies were included, of which 26 (50.0%) were on infectious diseases, 10 (19.2%) on chronic diseases, 8 (15.5%) on maternal and child health, and 8 (15.5%) on other health-related outcomes. Only 6 studies reported the reasons for using the specified spatial unit, 8 (15.3%) studies conducted sensitivity analysis for prior selection, and 39 (75%) of the studies used Queen contiguity adjacency. This review highlights existing variation and limitations in the specification of Bayesian spatial and spatio-temporal models used in health research. We found that majority of the studies failed to report the rationale for the choice of spatial units, perform sensitivity analyses on the priors, or evaluate the choice of neighbourhood adjacency, all of which can potentially affect findings in their studies. ... It is possible that the seroprevalence was overestimated in this study [44,45] as the antibodies detected were not confrmed with the recognized gold standard neutralizing test due to a lack of a biosafety level 3 laboratory. Some false positive results might be occurred due to cross-reaction with any of the JEV serogroup faviviruses, such as dengue [46] and Zika [47] viruses which are also detected in Bali. However, the use of the recombinant JEV for precoating of the ELISA plates may have reduced the occurrence of crossreactions [48]. ... Individual and Herd-Level Seroprevalence in Association with Potential Risk Factors of Japanese Encephalitis in Pigs Collected from Urban, Periurban, and Rural Areas of Bali, Indonesia Article Feb 2023 <here is a image 1b3763c998f75858-b0ff0a95548507da> Anak Agung Ayu Mirah Adi <here is a image 4189289d8af4afee-c07d15b7fc402dde> Nyoman Mantik Astawa <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Mieghan Bruce <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Made Kardena A study to assess the seroprevalence antibodies against JEV in pigs in Denpasar, Badung, and Karangasem as the representatives of urban, periurban, and rural areas in the province of Bali was conducted. Sampled pigs' blood was collected and their sera were tested for antibody detection using commercial IgG ELISA. A standard questionnaire was used to interview the pig owners or farmers to identify the determinants associated with the seropositivity of the antibodies. Overall, 96.6% (95% CI: 94.5-98.1) of 443 pig sera in individual animal-level seroprevalence were seropositive to the ELISA. Karangasem had the highest test prevalence at 97.3% (95% CI: 93.1-99.2) while Badung had a slightly lower prevalence at 96.6% (95% CI: 92.2-98.9), and Denpasar had the lowest prevalence at 96% (95% CI: 91.5-98.5) (p � 0.84). In herd-level seroprevalence, all sampled herds contained one or more seropositive pigs (overall herd-level seroprevalence 100% [95% CI: 97.7-100]). No animal-level factors were signifcantly associated with seropositivity (all p values >0.05). For the herd-level risk factors relating to pig management and husbandry practices adopted, no analysis model could be generated, as all the sampled herds were seropositive. More than 90% sero-prevalence detected in this study indicates high natural JEV infection occurred in pigs, which highlights the high public health risk of the infection in the areas. ... Gambar 1. Peta Administrasi Kota Jambi Sumber Data dan Cara Analisis Demam Berdarah merupakan salah satu penyakit di Indonesia yang kejadiannya wajib dilaporkan sejak 1968. Semua jenis infeksi Virus Dengue yang ditemukan di pelayanan kesehatan harus dilaporkan ke kantor dinas kesehatan dalam 24 jam setelah didiagnosis (Dhewantara et al., 2019) . Sumber data penelitian ini adalah data kasus DBD tahunan menurut kelurahan dari Januari 2020 sampai Desember 2020 yang berjumlah 714 kasus diperolah dari Dinas Kesehatan Kota Jambi. ... Autokorelasi Spasial Kasus Demam Berdarah di Kota Jambi Tahun 2020 Article <here is a image 24dc971fffaab97c-19919f9ff4a3c232> Muhammad Syukri Helmi Suryani Nasution Faisal Faisal Syarifah Zulfah Demam berdarah saat ini masih menjadi masalah kesehatan masyarakat yang sangat penting di Kota Jambi. Jumlah kasus DBD di Kota Jambi menempati urutan pertama dari 11 kab/kota di Provinsi Jambi. Studi ini bertujuan mengidentifikasi variasi spasial kasus DBD di Kota Jambi tahun 2020. Analisis Global dan Lokal Moran Indeks digunakan dalam penelitian ini untuk memetakan wilayah hotspots dan cold spots kasus DBD. Penelitian ini menggunakan pendekatan studi ekologi dengan unit analisis semua kelurahan di Kota Jambi yang berjumlah 62 kelurahan. Data kejadian DBD diperoleh dari laporan Dinas Kesehatan Kota Jambi dari Januari 2020 sampai Desember 2020 dengan jumlah total sebanyak 714 kasus. Studi ini menemukan adanya autokorelasi positif kasus DBD dengan 10 kelurahan yang masuk ke dalam area hotspots dan 9 kelurahan masuk ke dalam area cold spots. ... Temperature can determine the length of time that mosquitoes survive, their habitats, reproduction and maturation, as well as the infective periods [7]. Moreover, it has been shown that the land temperature influences every stage of the mosquito's life cycle, for which the ideal land temperature is 25-30°C [8] . Higher temperatures can shorten the incubation period, thereby rapidly increasing the numbers of mosquitoes and the incidence of bites. ... Spatio-temporal Modelling of Dengue Fever Cases in Saudi Arabia using Socio-economic, Climatic and Environmental Factors Article <here is a image 38287583f40c94da-691be3b7737f504a> Ali Siddiq Nagesh Shukla <here is a image 79ee96db125b8af9-9463a7f3810aa0f4> Biswajeet Pradhan Dengue Fever (DF) is a common vector-borne disease with catastrophic health implications. DF prediction modelling is a challenging task, although technologies such as Geographical Information Systems (GIS) and spatial statistics have improved our understanding of dengue dynamics. In this paper, we create a robust data analysis model to (i) provide a better understanding of confirmed dengue fever cases despite missing data, (ii) obtain better insights into risk factors associated with confirmed cases, and (iii) by means of machine learning, create clusters of patients with comparable characteristics. The last was accomplished with a self-organizing feature map (SOFM) and the density-based spatial clustering of applications with noise (DBSCAN). The approaches used to classify confirmed cases were: Decision Tree, k-nearest neighbours, Random Forest, AdaBoost, Support Vector Classification (SVC), CatBoost, and Naive Bayes. The CatBoost classifier achieved the best accuracy for the analysis of confirmed cases. Spatial analysis was conducted using the ordinary least square (OLS) and geographically weighted regression (GWR) models to identify high-risk areas. SOM can group patients with similar features into clusters, then DBSCAN detects and retrieves six clusters from this data. The clustering of confirmed cases increases CatBoost’s modelling prediction accuracy and reveals complex factors that influence prediction accuracy. Because confirmed cases in each cluster have different features, CatBoost is applied to each cluster individually to improve the prediction accuracy. Variable values in each cluster are analysed to clarify the confirmed cases of a specific subset of DF incidents. Overall, OLS outperforms GWR when identifying hotspot areas. The proposed novel, data-driven and machine-learning-based strategy facilitates the understanding and identification of patterns associated with confirmed DF cases. The study's findings can be utilized to cluster historical patient data into groups or subgroups sharing similar variables. Using identifiable patient clusters rather than raw history data improves the model accuracy provided by CatBoost. ... Flaviviruses are the main pathogens that have been facilitated by mosquitoes. The viruses reported to cause human encephalitis in this area, such as dengue [4] , Japanese encephalitis [5,6] , and Zika [7] . These disease agents are also required mosquitoes to be the vector to transmit them [8] . ... Mosquito diversity and abundance collected from paddy fields and animal farms in Bali province, Indonesia Article Full-text available Made Kardena <here is a image 24c420bffe4282a5-be1c19c0b26362bd> Anak Agung bagus Bramardipa <here is a image 1b3763c998f75858-b0ff0a95548507da> Anak Agung Ayu Mirah Adi <here is a image 64a10a34e24c5b0e-7fb22373380ed8cb> Made Dwi Dwinata This study aims were to assess the genera and number of the mosquitoes collected from rice paddy fields and animal farms in Badung and Tabanan regencies, in Bali, Indonesia. A total of eight mosquito light traps were set in the sampling areas. Only the adult female mosquitoes trapped were identified and counted in this study. The results demonstrated that four mosquito genera were found, such as Culex spp., Anopheles spp., Aedes spp., and Armigeres spp. The Culex spp. was the highest number of the mosquito trapped with total 2899 collected along the study period. Then, it was followed by Anopheles spp., Aedes spp. with 1954 and 1229 mosquitoes collected respectively. The lowest number mosquito found was Armigeres spp. with 612 mosquitoes collected along the twelve weeks collection. A significant higher number of mosquitoes Anopheles spp., Aedes spp., and Armigeres spp. collected from paddy fields found in comparison with the number of the same mosquitoes collected from animal farms in Badung regency. However, more significant number of mosquitoes Culex spp., Anopheles spp., and Aedes spp. collected from animal farms found compared to the mosquitoes collected from paddy fields in Tabanan regency. ... Meanwhile, primary clusters in Banyumas were detected from March to April. In Bali, strong seasonality of dengue incidence was observed during January to May [21] and Southeast Sulawesi Indonesia, the highest rate of dengue cases was found in January-March [22]. This result is different from the study in Pakistan which showed that the primary cluster was detected from August to October and the secondary was detected in September [23]. ... A spatiotemporal analysis of dengue hemorrhagic fever in Banyumas, Indonesia Article <here is a image 85f5f7a1788dcbf6-9ca6651c9f56f882> Dwi Sarwani Sri Rejeki Nunung Nurhayati <here is a image 4a3fc3256bd1ad65-971c41013f0fba60> Budi Aji Dengue hemorrhagic fever (DHF) is a public health problem in the world and also in Indonesia. One of the districts in Central Java that is still having problems with this disease is Banyumas, Indonesia. The incidence rate (IR) data in 2018 was 2.75 per 100,000 populations and the case fatality rate (CFR) was 3.64%. Spatiotemporal analysis was used to determine local variation, geographic determination of risk zones, and measurement of disease control interventions. Therefore, this study aimed to determine the distribution and grouping of dengue cases based on the spatiotemporal analysis. The design was observational with a cross-sectional spatial analysis. This study was conducted in Banyumas, Indonesia with the analysis unit for dengue fever patients in 2018 using as many as 57 cases. Furthermore, the data analysis used includes overlay, buffering, and clustering with SaTScan and ArcGis software. The results showed that there was a clustering of dengue cases in Banyumas, with one primary and three secondary clusters detected. The primary cluster occurred in March-April 2018, involving four sub-districts in urban areas. It was then concluded that the significantly identified clusters indicate a transmission of dengue fever in the Banyumas area with a radius of three kilometers. ... For example, dengue incidence in Bali is highly seasonal, with most cases reported during the rainy season between December and April. Evidently, the many water bodies and occurrence of lowland flooding provide excellent conditions for the vector, Aedes mosquitoes, to reproduce and undergo their larval aquatic life stage (Dhewantara et al., 2019) . Similarly, Anopheles mosquitos are required for the malaria parasite to conclude its life cycle prior to human transmission, so seasonality of the disease in humans is primarily caused by seasonality in climatic conditions required for the vector to breed (Herekar et al., 2020). ... Drivers of Infectious Disease Seasonality: Potential Implications for COVID-19 Article Full-text available Jan 2021 <here is a image 95059af722d47696-e2ad6cd844b09f11> Noga kronfeld-schor <here is a image e53deaf6e5a6a378-bcc8f19eb6af3d20> Tyler J Stevenson <here is a image d5525d3228561faf-8d3c47a1fcf5762f> Sema Nickbakhsh <here is a image 70a02b013b1e1aba-481f60ba5e49c54f> Barbara Helm Not 1 year has passed since the emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19). Since its emergence, great uncertainty has surrounded the potential for COVID-19 to establish as a seasonally recurrent disease. Many infectious diseases, including endemic human coronaviruses, vary across the year. They show a wide range of seasonal waveforms, timing (phase), and amplitudes, which differ depending on the geographical region. Drivers of such patterns are predominantly studied from an epidemiological perspective with a focus on weather and behavior, but complementary insights emerge from physiological studies of seasonality in animals, including humans. Thus, we take a multidisciplinary approach to integrate knowledge from usually distinct fields. First, we review epidemiological evidence of environmental and behavioral drivers of infectious disease seasonality. Subsequently, we take a chronobiological perspective and discuss within-host changes that may affect susceptibility, morbidity, and mortality from infectious diseases. Based on photoperiodic, circannual, and comparative human data, we not only identify promising future avenues but also highlight the need for further studies in animal models. Our preliminary assessment is that host immune seasonality warrants evaluation alongside weather and human behavior as factors that may contribute to COVID-19 seasonality, and that the relative importance of these drivers requires further investigation. A major challenge to predicting seasonality of infectious diseases are rapid, human-induced changes in the hitherto predictable seasonality of our planet, whose influence we review in a final outlook section. We conclude that a proactive multidisciplinary approach is warranted to predict, mitigate, and prevent seasonal infectious diseases in our complex, changing human-earth system. ... Access into the room in some areas through many narrow-sized doors, nonstandard stairs, and slippery roads; many mosquitoes were found in several areas Beach Access in the form of uneven limestone roads, slippery rocks, and steep paths or near cliffs; no direct access and stairs down; strong currents and high ocean waves; water activities such as bathing, surfing, canoeing, swimming, seeing mangroves using boats, diving, snorkeling, and fishing; the risk of decompression sickness in diving sites; sea snakes, monkeys and dogs; insect bites, mosquitoes, flies that affect food hygiene; lot of garbage scattered; night hours, youth gathering place, night tour; alcoholic beverages and the potential of mixed brew containing methanol; the potential for violence due to alcohol intoxication; and the existence of quite areas that are vulnerable to creating criminal action Art market Crowds and jostling; many mosquitoes and flies due to garbage; hazardous elements such as electric current due to unsafe networks Temple and place of worship Difficult entry access, ladder's reach is too high, and the stairs go down steep and slippery; winding and slippery slope; very small and cave-shaped entrance door, as well as one entry and exit access; wild animals such as monkeys and stray dogs; mosquitos can be found in the pond or water barrel; less clean place to sell food River and dam Entrance access is often in the form of damaged roads, gravelly roads, using semi-permanent and narrow bridges; danger of being dragged down, slipping and falling due to the slippery rock as a footing; when it rains heavily, the water overflows and becomes muddy; water activities such as fishing, bathing, swimming and rafting Monument, historic sites Crowd and jostling; some areas have slippery stairs when it rains; some areas are rather humid and there are many mosquitoes Amusement park Crowd in most parts of the area because there are too many visitors at certain times; the games provided in water sport run the risk of causing slipping and sinking; danger of altitude on some rides, fly board activities, fly fish and parasailing January and May [36] The increasing number of cases of acute encephalitis syndrome, a key presenting clinical sign of Japanese encephalitis infection in humans, along with increasing laboratory confirmed cases in Bali over recent years have led to the Indonesian government developing a national program of vaccination against Japanese encephalitis virus. In order to inform multidisciplinary management, a review was conducted to assess Japanese encephalitis virus-related cases in humans and animals including their determinants and detection in vectors. Along with published literature, key data from local authorized officers in Bali have been used to convey the recent situation of the disease. Related surveys detected up to 92% of the local children had antibodies against the virus with the annual incidence estimated to be 7.1 per 100,000 children. Additionally, reports on young and adult cases of infection within international travellers infected in Bali were documented with both non-fatal and fatal outcomes. Further seropreva-lence surveys detected up to 90% with antibodies to the virus in animal reservoirs. The detection of the virus in certain Culex mosquito species and high levels of seropositivity may be associated with greater risk of the virus transmission to the human population. It was also highlighted that local sociocultural practices for agriculture and livestock were potentially associated with the high density of the vector and the reservoirs, which then may lead to the risk of the disease transmission in the ecology of Bali. ARTICLE HISTORY Spatial and temporal analysis of hospitalized dengue patients in Bandung: demographics and risk Background Bandung, the fourth largest city in Indonesia and capital of West Java province, has been considered a major endemic area of dengue, and studies show that the incidence in this city could increase and spread rapidly. At the same time, estimation of incidence could be inaccurate due to a lack of reliable surveillance systems. To provide strategic information for the dengue control program in the face of limited capacity, this study used spatial pattern analysis of a possible outbreak of dengue cases, through the Geographic Information System (GIS). To further enhance the information needed for effective policymaking, we also analyzed the demographic pattern of dengue cases. Methods Monthly reports of dengue cases from January 2014 to December 2016 from 16 hospitals in Bandung were collected as the database, which consisted of address, sex, age, and code to anonymize the patients. The address was then transformed into geocoding and used to estimate the relative risk of a particular area’s developing a cluster of dengue cases. We used the kernel density estimation method to analyze the dynamics of change of dengue cases. Results The model showed that the spatial cluster of the relative risk of dengue incidence was relatively unchanged for 3 years. Dengue high-risk areas predominated in the southern and southeastern parts of Bandung, while low-risk areas were found mostly in its western and northeastern regions. The kernel density estimation showed strong cluster groups of dengue cases in the city. Conclusions This study demonstrated a strong pattern of reported cases related to specific demographic groups (males and children). Furthermore, spatial analysis using GIS also visualized the dynamic development of the aggregation of disease incidence (hotspots) for dengue cases in Bandung. These data may provide strategic information for the planning and design of dengue control programs. İtalya'da COVID-19’un Bölgeler Arası Yayılımı: Keşfedici Mekânsal Veri Analizi In this study, the spreading status of COVID-19 infectious disease among neighboring regions was investigated with the spatial dependency statistic Moran I. For this purpose, using data from 20 regions of Italy, univariate and bivariate Global and Local Moran I statistics were calculated. According to the result of the univariate Moran I statistic, it was concluded that COVID-19 infectious disease spread among neighbors before border crossings between regions were banned. The lagged effect of spatial propagation was investigated with the bivariate Moran I statistic. According to the result of the bivariate Moran I statistic, it can be said that the total number of cases occurring in neighboring regions in the first 14 days is one of the reasons for the total number of cases in the second 14 days. As a result of these findings, it can be said that by stopping the transits between provinces or regions earlier, the rapid spread of infectious diseases such as COVID-19 can be prevented. Exploring the utility of social-ecological and entomological risk factors for dengue infection as surveillance indicators in the dengue hyper-endemic city of Machala, Ecuador The management of mosquito-borne diseases is a challenge in southern coastal Ecuador, where dengue is hyper-endemic and co-circulates with other arboviral diseases. Prior work in the region has explored social-ecological factors, dengue case data, and entomological indices. In this study, we bring together entomological and epidemiological data to describe links between social-ecological factors associated with risk of dengue transmission at the household level in Machala, Ecuador. Households surveys were conducted from 2014-2017 to assess the presence of adult Aedes aegypti (collected via aspiration) and to enumerate housing conditions, demographics, and mosquito prevention behaviors. Household-level dengue infection status was determined by laboratory diagnostics in 2014-2015. Bivariate analyses and multivariate logistic regression models were used to identify social-ecological variables associated with household presence of female Ae. aegypti and household dengue infection status, respectively. Aedes aegypti presence was associated with interruptions in water service and weekly trash collection, and household air conditioning was protective against mosquito presence. Presence of female Ae. aegypti was not associated with household dengue infections. We identified shaded patios and head of household employment status as risk factors for household-level dengue infection, while window screening in good condition was identified as protective against dengue infection. These findings add to our understanding of the systems of mosquito-borne disease transmission in Machala, and in the larger region of southern Ecuador, aiding in the development of improved vector surveillance efforts, and targeted interventions. EFFICACY OF ALLIUM CEPA (AMARYLLIDACEAE) EXTRACT AGAINST DENGUE VIRUS TYPE-2 (FLAVIVIRIDAE : FLAVIVIRUS) ISOLATED FROM SURABAYA, INDONESIA Nowadays, dengue disease is still a major health problem in Indonesia. However, Surabaya, the second-largest city in the country is endemic for dengue. Dengue virus (DENV) is the causative agent of dengue fever and dengue shock syndrome or dengue haemorrhagic fever. At present, no antiviral drugs are available for treatment DENV infections. Therefore, we aimed to investigate the antiviral activity of Allium cepa extract (ACE) against dengue virus type-2 (DENV-2; NCBI accession number: KT012509) isolated from Surabaya, Indonesia in 2013. Interestingly, we revealed that ACE inhibited DENV-2 in Vero cells with IC50: 1,932.36 ìg/mL, CC50: 121.17 ìg/mL and SI: 0.06. Notably, findings presented here suggest that ACE considered as low concerning antiviral activityagainst DENV-2. In sum, ACE should be considered for re-evaluation in the development of an effective antiviral compound against DENV-2. Semi-Supervised Text Classification Framework: An Overview of Dengue Landscape Factors and Satellite Earth Observation In recent years there has been an increasing use of satellite Earth observation (EO) data in dengue research, in particular the identification of landscape factors affecting dengue transmission. Summarizing landscape factors and satellite EO data sources, and making the information public are helpful for guiding future research and improving health decision-making. In this case, a review of the literature would appear to be an appropriate tool. However, this is not an easy-to-use tool. The review process mainly includes defining the topic, searching, screening at both title/abstract and full-text levels and data extraction that needs consistent knowledge from experts and is time-consuming and labor intensive. In this context, this study integrates the review process, text scoring, active learning (AL) mechanism, and bidirectional long short-term memory (BiLSTM) networks, and proposes a semi-supervised text classification framework that enables the efficient and accurate selection of the relevant articles. Specifically, text scoring and BiLSTM-based active learning were used to replace the title/abstract screening and full-text screening, respectively, which greatly reduces the human workload. In this study, 101 relevant articles were selected from 4 bibliographic databases, and a catalogue of essential dengue landscape factors was identified and divided into four categories: land use (LU), land cover (LC), topography and continuous land surface features. Moreover, various satellite EO sensors and products used for identifying landscape factors were tabulated. Finally, possible future directions of applying satellite EO data in dengue research in terms of landscape patterns, satellite sensors and deep learning were proposed. The proposed semi-supervised text classification framework was successfully applied in research evidence synthesis that could be easily applied to other topics, particularly in an interdisciplinary context. Gambaran Kasus Demam Berdarah Dengue (DBD) Di Pulau Bali Tahun 2012-2017 Latar Belakang: Demam Berdarah Dengue (DBD) hingga saat ini masih merupakan masalah kesehatan masyarakat di Indonesia, termasuk pulau Bali. Provinsi bali yang terdiri dari 9 kabupaten/kota adalah daerah endemis DBD, padahal Provinsi Bali adalah destinasi wisata baik lokal maupun mancanegara.Penelitian ini bertujuan untuk memberikan pemaparan berupa gambaran kejadian DBD di Provinsi Bali.Metode: Penelitian ini merupakan penelitian deskriptif dengan rancang bangun caseseries. Sumberdata pada penelitian ini menggunakan data sekunder yaitu Profil Kesehatan provinsi Bali tahun2015 hingga 2017, dan data iklim di Provinsi Bali tahun 2015-2017yang diperoleh dari Badan Pusat Statistik provinsi Bali.Hasil: Hasil penelitian menunjukkan insiden DBD per 100000 penduduk di Provinsi Bali tahun 2012 hingga tahun 2017 berturut turut 65,5: 174,5: 210,2; 259,1; 483; 105. Puncak insiden tertinggi Demam Berdarah Dengue (DBD) ada pada tahun 2016.Pada tahun 2017 ada 4 kabupaten/kota yang insidennya tinggi seperti kabupaten Badung , kota Denpasar , kabupaten Buleleng dan kabupaten Gianyar. Penyebab meningkatnya insiden DBD adalah banyaknya genangan air sebagai tempat perindukan nyamuk Aedes aegypti saat musim hujan, sehingga populasi nyamuk Aedes aegypti meningkat. Insidens DBD terjadi pada bulan Januari hingga Mei, yang di pengaruhi oleh cuaca lokal, kondisi sosial ekonomi masyarakat, curah hujan, topografi maupun kepadatan,serta mobilitas penduduk.Simpulan: Insiden DBD dipengaruhi oleh pola musim hujan , di bulan Januari, Februari, Maret, April dan Mei didukung kepadatan dan mobilitas dari penduduk. ABSTRACTTitle: An Overview of Dengue Haemorrhagic Fever Cases in Bali Island 2012-2017Background: Dengue Hemorrhagic Fever (DHF)remains a public health problem in Indonesia, including the island of Bali. Bali province consists of 9 regencies / cities is a dengue endemic area, whereas the island of Bali is a tourist destination both locally and internationally. This study aims to provide an overview of the incidence rates (IR) of DHF in the island of Bali.Methods: This research is a descriptive study with case series design. Data sources in this study use secondary data obtianed from Bali Health Profile 2015 – 2017, and the climate data of Bali Province in 2015-2017 was obtained from the Central Statistics Agency (BPS) of Bali Province.Results: The results showed the Incidence Rates (IR) of Dengue Haemorrhagic Fever (DHF) in the island of Bali in 2012- 2017 were 65.5; 174.5; 210.2; 259.1; 483 and 105.7 respectively. The highest incidence rates (IR) of DHF was notified in 2016. Up to 2017 there were 4 districts that had reported high incidence of DHF, such as Badung Regency, Denpasar City, Buleleng Regency, and Gianyar Regency. Factors contributing to the increasing incidence of DHF in Bali were the existence of water container as the potential breeding places for mosquitos vector of Aedes aegypti, particularly during rainy season. This condition initiated the increased population of Aedes aegypti. The incidence of dengue fever cases intensively occurred during January – May influenced by local weather climate, socio- economic condition, rainfall, topography, as well as population density and mobilityConclusion: The incidence of DHF is significantly associated with weather seasonal patterns whereasthe highest DHF incidence rates are found in each year in January, February, March, April and May. The other significant factors are including of rainfall, population density and mobility. Paediatric dengue infection in Cirebon, Indonesia: A temporal and spatial analysis of notified dengue incidence to inform surveillance Abstract Background The recent situation of dengue infection in Cirebon district is concerning due to an upsurge trend since the year 2010. The largest dengue outbreak was reported in 2016 which has affected more than 1600 children. A study was conducted to explore the temporal variability of dengue outbreak in Cirebon’s child population in during 2011–2017, and to assess the short-term effects of climatic and environmental factor on dengue incidence. In addition, the spatial pattern of dengue incidence in children and high-risk villages were investigated. Methods A total of 4597 confirmed dengue cases in children notified from January 2011 to December 2017 were analysed. Seasonal decomposition analysis was carried out to examine the annual seasonality. A generalized linear model (GLM) was applied to assess the short-term effect of climate and normalized difference vegetation index (NDVI) on dengue incidence. The incidence rate ratio (IRR) of the final model was reported. Spatial analyses were conducted by using Moran’s I and local indicator of spatial association (LISA) analyses to explore geographical clustering in incidence and to identify high-risk villages for dengue, respectively. Results An annual dengue epidemic period was observed with peaks occurring every January/February. Based on the GLM, temperature at a lag 4 months (IRR = 1.27; 95% confidence interval, 95% CI: 1.22–1.31, P Dengue infection in international travellers visiting Bali, Indonesia Article Background Dengue, an acute febrile illness caused by infection with dengue virus (DENV), is endemic in Bali, Indonesia. As one of the world's most popular tourist destinations, Bali is regularly visited by domestic and international travellers, who are prone to infection by endemic pathogens, including DENV. Currently, limited data are available on the characteristics of dengue in travellers visiting Bali. Information on the epidemiology and virological aspects of dengue in these tourists is important to gain a better understanding of the dengue disease in international travellers. Methods We performed a prospective cross-sectional dengue study involving foreign travellers visiting Bali, Indonesia in the period of 2015–2017. Patients presenting at Kasih Ibu Hospital with fever and clinical symptoms of dengue were asked to participate in the study. Clinical and laboratory assessments were performed and sera were collected for molecular analysis, which included DENV serotyping, genome sequencing, and phylogenetic analysis. Results Among the 201 patients recruited, dengue was confirmed in 133 (66.2%) of them, based on detection of NS1 antigen and/or viral RNA. Of these, 115 (86.5%) manifested dengue fever (DF), and 18 (13.5%) dengue haemorrhagic fever (DHF). The temporal predominance of infecting DENV serotype was DENV-2 (48.7%), followed by DENV-3 (36.1%), DENV-1 (9.2%), and DENV-4 (3.4%). Phylogenetic analysis of DENV based on envelope gene sequences revealed that the source of DENVs was local endemic viruses. Conclusion Our study confirms that dengue is one of the causes of fever in travellers visiting Bali. Although it is a cause of significant morbidity, the majority of patients only experienced mild DF, with only a small proportion developing DHF. We revealed that DENVs isolated were autochthonous. Accurate diagnosis, preventive measures, and continuous disease surveillance will be useful for better management of dengue infection in travellers. DETERMINATION OF ENVIRONMENTAL FACTORS AFFECTING DENGUE INCIDENCE IN SLEMAN DISTRICT, YOGYAKARTA, INDONESIA

https://www.researchgate.net/publication/334726476_Spatial_and_temporal_variation_of_dengue_incidence_in_the_island_of_Bali_Indonesia_An_ecological_study

Wakes in Inertial Fusion Plasmas Author(s): Ellis, Ian Norman | Advisor(s): Mori, Warren B | Abstract: Plasma wave wakes, which are the collective oscillatory response near the plasma frequency to the propagation of particles or electromagnetic waves through a plasma, play a critical role in many plasma processes. New results from backwards stimulated Raman scattering (BSRS), in which wakes with phase velocities much less than the speed of light are induced by the beating of counter-propagating light waves, and from electron beam stopping, in which the wakes are produced by the motion of relativistically propagating electrons through the dense plasma, are discussed. Both processes play important roles in Inertial Confinement Fusion (ICF). In BSRS, laser light is scattered backwards out of the plasma, decreasing the energy available to compress the ICF capsule and affecting the symmetry of where the laser energy hits the hohlraum wall in indirect drive ICF. The plasma wave wake can also generate superthermal electrons that can preheat the core and/or the ablator. Electron beam stopping plays a critical role in the Fast Ignition (FI) ICF concept, in which a beam of relativistic electrons is used to heat the target core to ignition temperatures after the compression stage. The beam stopping power determines the effectiveness of the heating process. This dissertation covers new discoveries on the importance of plasma wave wakes in both BSRS and electron beam stopping.In the SRS studies, 1D particle-in-cell (PIC) simulations using OSIRIS are performed, which model a short-duration (~500/& amp;omega0 FWHM) counter-propagating scattered light seed pulse in the presence of a constant pump laser with an intensity far below the absolute instability threshold for plasma waves undergoing Landau damping. The seed undergoes linear convective Raman amplification and dominates over the amplification of fluctuations due to particle discreteness. The simulation results are in good agreement with results from a coupled-mode solver when special relativity and the effects of finite size PIC simulation particles are accounted for. Linear gain spectra including both effects are discussed. Extending the PIC simulations past when the seed exits the simulation domain reveals bursts of large-amplitude scattering in many cases, which do not occur in simulations without the seed pulse. These bursts can have amplitudes several times greater than the amplified seed pulse, and an examination of the orbits of particles trapped in the wake illustrates that the bursts are caused by a reduction of Landau damping due to particle trapping. This large-amplitude scattering is caused by the seed inducing a wake earlier in the simulation, thus modifying the distribution function. Performing simulations with longer duration seeds leads to parts of the seeds reaching amplitudes several times more than the steady-state linear theory results, similarly caused by a reduction of Landau damping. Simulations with continuous seeds demonstrate that the onset of inflation depends on the seed wavelength and incident intensity, and oscillations in the reflectivity are observed at a frequency equal to the difference between the seed frequency and the frequency at which the inflationary SRS grows.In the electron beam stopping studies, 3D PIC simulations are performed of relativistic electrons with a momentum of 10mec propagating in a cold FI core plasma. Some of the simulations use one simulation particle per real particle, and particle sizes much smaller than the interparitcle spacing. The wake made by a single electron is compared against that calculated using cold fluid theory assuming the phase velocity of the wake is near the speed of light. The results agree for the first wavelength of the wake. However, the shape of the wake changes for succeeding wavelengths and depends on the background plasma temperature, with the concavity pointing in the direction the electron is moving in cold plasmas and in the opposite direction as the plasma temperature increases. In the warm plasma the curvature is described by electrostatic Vlasov theory (for vparticle & gt;& gt; vth) and is due to the diffraction of the wave, while for cold plasmas the curvature is due to nonlinear radial oscillations of background electrons. Beams with multiple electrons exhibit correlation effects caused by electrons interacting through their wakes. Non-divergent beams are simulated, and a significant time-dependent increase in the stopping power is observed when the average electron spacing is 2c/& amp;omegape or less. This increase is caused by beam-plasma-like instabilities including self-focusing and/or filamentation and the beam-plasma-like instability. The stopping power growth rate and peak value depend on the beam size and density. For long beams with dimensions of 10c/& amp;omegape × 10c/& amp;omegape × 80c/& amp;omegape and an inter-particle separation of 0.25c/& amp;omegape (nb/n0 ≈ 4× 10-3), the peak stopping power averaged over the electrons is (1 ± 3) × 103 times that of an uncorrelated electron. These results indicate that an enhanced energy-independent or weakly dependent correlated stopping may occur for Fast Ignition scenarios, even for interparticle spacings when discreteness effects are important. The dependence of correlation effects on beam electron separation in terms of c/& amp;omegape also indicates that Fast Ignition may be possible with core densities below those designed using single-electron stopping powers. Target optimization to exploit correlated stopping in the target core may be possible once the effects of angular spread and energy spread are understood. Furthermore, this work begins to allow a connection from the discrete wakes effect to collective instabilities as the interparticle spacing is decreased relative to the size of the wake due to the use of denser beams, lower plasma densities, and the filamentation/self-focusing of the beam. Wakes in Inertial Fusion Plasmas 2014 Ellis, Ian Norman Advisor(s): Mori, Warren B ... Main Content Metrics Author & Article Info Abstract Plasma wave wakes, which are the collective oscillatory response near the plasma frequency to the propagation of particles or electromagnetic waves through a plasma, play a critical role in many plasma processes. New results from backwards stimulated Raman scattering (BSRS), in which wakes with phase velocities much less than the speed of light are induced by the beating of counter-propagating light waves, and from electron beam stopping, in which the wakes are produced by the motion of relativistically propagating electrons through the dense plasma, are discussed. Both processes play important roles in Inertial Confinement Fusion (ICF). In BSRS, laser light is scattered backwards out of the plasma, decreasing the energy available to compress the ICF capsule and affecting the symmetry of where the laser energy hits the hohlraum wall in indirect drive ICF. The plasma wave wake can also generate superthermal electrons that can preheat the core and/or the ablator. Electron beam stopping plays a critical role in the Fast Ignition (FI) ICF concept, in which a beam of relativistic electrons is used to heat the target core to ignition temperatures after the compression stage. The beam stopping power determines the effectiveness of the heating process. This dissertation covers new discoveries on the importance of plasma wave wakes in both BSRS and electron beam stopping.In the SRS studies, 1D particle-in-cell (PIC) simulations using OSIRIS are performed, which model a short-duration (~500/&omega 0 FWHM) counter-propagating scattered light seed pulse in the presence of a constant pump laser with an intensity far below the absolute instability threshold for plasma waves undergoing Landau damping. The seed undergoes linear convective Raman amplification and dominates over the amplification of fluctuations due to particle discreteness. The simulation results are in good agreement with results from a coupled-mode solver when special relativity and the effects of finite size PIC simulation particles are accounted for. Linear gain spectra including both effects are discussed. Extending the PIC simulations past when the seed exits the simulation domain reveals bursts of large-amplitude scattering in many cases, which do not occur in simulations without the seed pulse. These bursts can have amplitudes several times greater than the amplified seed pulse, and an examination of the orbits of particles trapped in the wake illustrates that the bursts are caused by a reduction of Landau damping due to particle trapping. This large-amplitude scattering is caused by the seed inducing a wake earlier in the simulation, thus modifying the distribution function. Performing simulations with longer duration seeds leads to parts of the seeds reaching amplitudes several times more than the steady-state linear theory results, similarly caused by a reduction of Landau damping. Simulations with continuous seeds demonstrate that the onset of inflation depends on the seed wavelength and incident intensity, and oscillations in the reflectivity are observed at a frequency equal to the difference between the seed frequency and the frequency at which the inflationary SRS grows.In the electron beam stopping studies, 3D PIC simulations are performed of relativistic electrons with a momentum of 10m e c propagating in a cold FI core plasma. Some of the simulations use one simulation particle per real particle, and particle sizes much smaller than the interparitcle spacing. The wake made by a single electron is compared against that calculated using cold fluid theory assuming the phase velocity of the wake is near the speed of light. The results agree for the first wavelength of the wake. However, the shape of the wake changes for succeeding wavelengths and depends on the background plasma temperature, with the concavity pointing in the direction the electron is moving in cold plasmas and in the opposite direction as the plasma temperature increases. In the warm plasma the curvature is described by electrostatic Vlasov theory (for v particle >> v th ) and is due to the diffraction of the wave, while for cold plasmas the curvature is due to nonlinear radial oscillations of background electrons. Beams with multiple electrons exhibit correlation effects caused by electrons interacting through their wakes. Non-divergent beams are simulated, and a significant time-dependent increase in the stopping power is observed when the average electron spacing is 2c/&omega pe or less. This increase is caused by beam-plasma-like instabilities including self-focusing and/or filamentation and the beam-plasma-like instability. The stopping power growth rate and peak value depend on the beam size and density. For long beams with dimensions of 10c/&omega pe × 10c/&omega pe × 80c/&omega pe and an inter-particle separation of 0.25c/&omega pe (n b /n 0 ≈ 4× 10 -3 ), the peak stopping power averaged over the electrons is (1 ± 3) × 10 3 times that of an uncorrelated electron. These results indicate that an enhanced energy-independent or weakly dependent correlated stopping may occur for Fast Ignition scenarios, even for interparticle spacings when discreteness effects are important. The dependence of correlation effects on beam electron separation in terms of c/&omega pe also indicates that Fast Ignition may be possible with core densities below those designed using single-electron stopping powers. Target optimization to exploit correlated stopping in the target core may be possible once the effects of angular spread and energy spread are understood. Furthermore, this work begins to allow a connection from the discrete wakes effect to collective instabilities as the interparticle spacing is decreased relative to the size of the wake due to the use of denser beams, lower plasma densities, and the filamentation/self-focusing of the beam. Main Content

https://escholarship.org/uc/item/5wb109v8

Intervention group and baseline child aggression plotted against Wave 2... | Download Scientific Diagram Download scientific diagram | Intervention group and baseline child aggression plotted against Wave 2 (spring 2005) teacher reports of children's academic skills. TR teacher report; BL Agg baseline child aggression index. Error bars represent standard errors. from publication: A School-Randomized Clinical Trial of an Integrated Social-Emotional Learning and Literacy Intervention: Impacts After 1 School Year | To report experimental impacts of a universal, integrated school-based intervention in social-emotional learning and literacy development on change over 1 school year in 3rd-grade children's social-emotional, behavioral, and academic outcomes. This study employed a... | School, Literacy and Achievement | ResearchGate, the professional network for scientists. Intervention group and baseline child aggression plotted against Wave 2 (spring 2005) teacher reports of children's academic skills. TR teacher report; BL Agg baseline child aggression index. Error bars represent standard errors. Source publication A School-Randomized Clinical Trial of an Integrated Social-Emotional Learning and Literacy Intervention: Impacts After 1 School Year Stephanie M Jones Joshua Brown To report experimental impacts of a universal, integrated school-based intervention in social-emotional learning and literacy development on change over 1 school year in 3rd-grade children's social-emotional, behavioral, and academic outcomes. This study employed a school-randomized, experimental design and included 942 3rd-grade children (49% boys... Context in source publication Context 1 ... also calculated effect sizes for the mean differences between the intervention and control groups at each level of the baseline child aggression index. As shown in Figure 2 , children with the highest level of baseline behavioral risk (based on teacher reports) showed the greatest positive difference in teacher report of academic skills between the intervention and control schools, with effect sizes of .40 for a score of 2 on the baseline child aggression index, .17 for a score of 1, and .05 ... Citations ... Por outro lado, a investigação existente evidenciou geralmente que a participação em programas SEL (e.g., RCCP,Caring School Community,4R,Merrell's Strong Kids,MindUp) contribuiu para uma melhoria significativa ao nível das competências socioemocionais globais e de dimensões específicas relacionadas com a negociação interpessoal, a empatia ou a tomada de perspetiva (e.g., Aber et al., 2003;Chang & Muñoz, 2006;Harlacher & Merrell, 2010; Jones et al., 2010 Jones et al., , 2011Schonert-Reichl & Lawlor, 2010;Schonert-Reichl et al., 2015). Estes resultados foram observados em amostras de crianças norte-americanas do primeiro ao sétimo ano de escolaridade, independentemente do tipo de medidas utilizadas. ... ... Apesar da heterogeneidade ao nível das amostras e dos instrumentos utilizados, a investigação existente parece enfatizar os benefícios dos programas SEL ao nível da redução da sintomatologia depressiva e na melhoria das competências socioemocionais. No entanto, a maioria dos estudos sobre os benefícios percebidos pelas crianças com a participação nos programas SEL em idade escolar, na perspetiva das crianças, que foram conduzidos até à data incidiu essencialmente em intervenções de longa duração (e.g., Aber et al., 2003;Chang & Muñoz, 2006; Jones et al., 2010 Jones et al., , 2011. Importa avaliar se os programas SEL com um menor número de sessões também se revestem de benefícios do ponto de vista da redução dos problemas emocionais (incluindo não apenas sintomas de depressão, mas também de ansiedade e stress) e das competências socioemocionais, na perspetiva das crianças. ... ... Especificamente, os nossos resultados corroboram a nossa primeira hipótese de investigação (H1), mostrando uma redução significativa de magnitude pequena do pré-teste para o pós-teste ao nível dos autorrelatos de depressão (i.e., disforia, desânimo, desvalorização da vida, auto-depreciação, falta de interesse ou de envolvimento, anedonia e inércia) que se manteve no follow-up (6 meses após o término da intervenção). Estes resultados são comparáveis às mudanças percebidas pelas crianças e jovens, após a participação em outros programas, tais como o RCCP (Aber et al., 2003), o 4R (Jones et al., 2010 ) e o MindUp (Schonert-Reichl et al., 2015, podendo ser explicados pelo facto de o programa "O Mundo dos Intergalácticos" trabalhar competências semelhantes aos programas supra, nomeadamente capacidade de autorregulação, autoreflexão e regulação e expressão de emoções, com recurso a técnicas terapêuticas de atenção plena e mindfulness. ... Benefícios de um novo programa de aprendizagem socioemocional na redução da sintomatologia psicopatológica e na promoção das competências socioemocionais globais, na perspetiva das crianças Article O presente estudo teve como objetivo examinar as perceções das crianças com idades compreendidas entre os 8 e os 12 anos, que participaram no novo programa de aprendizagem socioemocional “O Mundo dos Intergalácticos” de curta duração, acerca da evolução dos sintomas psicopatológicos (ansiedade, depressão e stresse) e das competências socioemocionais globais, do pré-teste para o pós-teste e follow-up (6 meses), considerando o sexo e a faixa etária (8-9 vs. 10-12 anos). Um total de 95 crianças, com idades compreendidas entre os 8 e os 12 anos, preencheram Escala de Depressão, Ansiedade e Stresse para Crianças e a escala. Para mim é fácil, no pré-teste, pós-teste e 6 meses após a participação no programa. Os resultados obtidos mostraram que as crianças mais novas (8-9 anos) e mais velhas (10-12 anos), de ambos os sexos, percecionaram uma redução da sintomatologia psicopatológica e uma melhoria das competências socioemocionais globais do pré-teste para o pós-teste e follow-up. ... The methodological rigor of randomized intervention studies with case-control (n = 8, 18.20%; Bavarian et al., 2013;Burnette et al., 2017;Connor et al., 2010;DiPerna et al., 2018;Dougherty, & Sharkey, 2017; Jones et al., 2010; Jones et al., 2011;Leichtentritt & Shechtman, 2010) adopted more forms of sample control and described more details in the research procedures, therefore, the research report had higher quality, which reached 79% of the analyzed criteria. ... ... Studies that reported interventions through randomized control of participants and the use of control groups described the results of different outcomes for interventions at the school level, such as the growth of mindset online (Burnette et al., 2017); the 4Rs Program (Jones et al., 2010; ; the Social Skills Improvement System Classwide Intervention Program (SSIS-CIP;DiPerna et al., 2017); the Positive Action Intervention (Bavarian et al., 2013); the individualization of teaching (Connor et al., 2010); the intervention of Reconnecting Youth (Dougherty, & Sharkey, 2017) and the Group Counseling (Leichtentritt & Shechtman;. ... ... The intervention of the 4Rs Program (i.e., 4Rs: Reading, Writing, Respect and Resolution) described in the studies by Jones et al. (2010; did not indicate strong evidence of short-term changes to the cognitive and interpersonal functioning of hostile and aggressive bias and school performance of the children evaluated. In this study, after the first year of the intervention, there was a single effect on the attributional biases of hostility and aggression in children; in self-reported symptoms of depression; and no main effect on children's literacy skills and attention span. ... Learning Motivation, Socioemotional Skills and School Achievement in Elementary School Students Ana Deyvis Santos Araújo Jesuino Abstract The motivation to learn is an element that drives student behavior and acts as a subjacent solving school demands, as well it influences the development and expression of socioemotional skills and school performance. This research constitutes a systematic review of the literature that investigated the relationship between motivation to learn, socioemotional skills and school performance in children enrolled in elementary school. A total of 9,631 abstracts from the ASSIA, EDUC, ERIC, PsycArticles, Science Direct, SCOPUS and Web of Science databases were examined, ending with 44 studies. The findings indicated a positive relationship between the three constructs, corroborating the literature that illustrates the benefits of various indicators of school and psychological well-being of children and adolescents to enhance learning and school performance. It is noteworthy that student-family and student-teacher interactions can favor student success or failure. ... Educational research and policy have shown an increasing focus on promoting the overall well-being of children and youth through the use of social-emotional learning (SEL) programs in classroom settings (Atwell & Bridgeland, 2019). The burgeoning interest in youth well-being in school contexts is underscored by a growing awareness of the internalizing and externalizing challenges that young people face, including anxiety, depression, bullying, and aggression (Fonagy et al., 2009; Jones et al., 2010) . In Canada, for instance, not only does the Mental Health Commission of Canada recognize child and youth mental health as a priority, but it also identifies schools as an important context for fostering the social-emotional skills necessary to promote positive social, emotional, mental, and academic development (Hymel et al., 2017;Mental Health Commission of Canada, 2012). ... Building elementary students’ social and emotional skills: A randomized control trial to evaluate a teacher-led intervention The Faith and Wellness: A Daily Mental Health Resource is a school-based, teacher-led social-emotional learning (SEL) intervention resource for elementary students. It is designed to address the challenges faced by existing SEL interventions, including lack of time, training, and resources. Using a randomized control trial design, this study evaluates short-term outcomes associated with the use of this resource. Participants were elementary teachers (NT1 = 201, NT2 = 129) and students (NT1 = 242, NT2 = 183; ages 4-14; 47.5% girls) from 19 Catholic school boards in [PROVINCE]. Using multi-level models, significant small to medium effect sizes indicated that intervention group teachers: taught SEL more frequently; had higher confidence in teaching SEL; and had more positive perceptions of the classroom climate, students' SEL, and students' school engagement at Time 2 than comparison group teachers. Results for students were less robust, though there was indication of dosage effects. Results highlight the role of teachers and frequent delivery in effective SEL implementation. Supplementary information: The online version contains supplementary material available at 10.1007/s12310-022-09538-x. ... These programs often involve providing teachers with training and support to promote safe and supportive learning environments that can help improve the students' self-perception, social and self-awareness, interpersonal skills, self-management and regulation of their emotions, among other skills (Denham & Brown, 2010;Tubbs et al., 2021). Numerous studies evaluating in-school and after-school SEL programs in the United States and in other Western countries have found positive impacts on student's academic performance (Jones, Brown, Hoglund, & Aber, 2010; Durlak, Weissberg, Dymnicki, Taylor, & Schellinger, 2011;Schonfeld et al., 2015) and emerging evidence suggests that offering access to SEL education programming could improve student's learning and development in crisis contexts Aber, Tubbs, et al., 2017;Tubbs et al., 2021;Brown et al., 2021; REMEDIAL SUPPORT FOR REFUGEES: IMPACT OF DOSAGE 5 2021). Nonetheless, there is limited evidence on how to implement such programs effectively (Torrente et al., 2019) under conditions that compromise children's ability to regularly attend. ... The Impact of Attending a Remedial Support Program on Syrian Children's Reading Skills: Using BART for Causal Inference Preprint Full-text available

https://www.researchgate.net/figure/Intervention-group-and-baseline-child-aggression-plotted-against-Wave-2-spring-2005_fig2_49642107

draft-ymbk-sidr-transfer-00 - Resource Transfer in the Resource Public Key Infrastructure Resource Transfer in the Resource Public Key Infrastructure Resource Transfer in the Resource Public Key Infrastructure draft-ymbk-sidr-transfer-00 Versions: Document Type This is an older version of an Internet-Draft whose latest revision state is "Expired". Authors Rob Austein , Randy Bush , Geoff Huston , George G. Michaelson Last updated 2015-05-30 RFC stream (None) Formats txt htmlized pdf bibtex Stream Stream state (No stream defined) Consensus boilerplate Unknown RFC Editor Note (None) IESG IESG state I-D Exists Telechat date (None) Responsible AD (None) Send notices to (None) draft-ymbk-sidr-transfer-00 Network Working Group R. Austein Internet-Draft Dragon Research Labs Intended status: Standards Track R. Bush Expires: December 1, 2015 IIJ Lab / Dragon Research Lab G. Huston G. Michaelson Asia Pacific Network Information Centre May 30, 2015 Resource Transfer in the Resource Public Key Infrastructure draft-ymbk-sidr-transfer-00 Abstract Transfer within the RPKI of actual address space and/or autonomous system number resources between two Internet registries (ISPs, RIRs, NIRs, etc.) is reasonably achievable for most useful operational needs. In this paper, we describe, at a high level, how this may be accomplished. Status of This Memo This Internet-Draft is submitted in full conformance with the provisions of BCP 78 and BCP 79. Internet-Drafts are working documents of the Internet Engineering Task Force (IETF). Note that other groups may also distribute working documents as Internet-Drafts. The list of current Internet- Drafts is at http://datatracker.ietf.org/drafts/current/. Internet-Drafts are draft documents valid for a maximum of six months and may be updated, replaced, or obsoleted by other documents at any time. It is inappropriate to use Internet-Drafts as reference material or to cite them other than as "work in progress." This Internet-Draft will expire on December 1, 2015. Copyright Notice Copyright (c) 2015 IETF Trust and the persons identified as the document authors. All rights reserved. This document is subject to BCP 78 and the IETF Trust's Legal Provisions Relating to IETF Documents (http://trustee.ietf.org/license-info) in effect on the date of publication of this document. Please review these documents carefully, as they describe your rights and restrictions with respect Austein, et al. Expires December 1, 2015 [Page 1] Internet-Draft Resource Transfer in the RPKI May 2015 to this document. Code Components extracted from this document must include Simplified BSD License text as described in Section 4.e of the Trust Legal Provisions and are provided without warranty as described in the Simplified BSD License. Table of Contents 1. Introduction and Terms . . . . . . . . . . . . . . . . . . . 2 2. A Simplistic Case . . . . . . . . . . . . . . . . . . . . . . 3 2.1. Steps in Simple Case . . . . . . . . . . . . . . . . . . 4 2.2. The Torn Euro Protocol . . . . . . . . . . . . . . . . . 4 3. A More Complex Case . . . . . . . . . . . . . . . . . . . . . 5 3.1. The Indirect Buyer . . . . . . . . . . . . . . . . . . . 6 3.2. The Difference Between Buyer and Seller Chain . . . . . . 7 4. Transfer in the Absense of a Common Ancestor . . . . . . . . 7 5. Transfer in process: Resources Change Forced from Above . . . 8 6. Security Considerations . . . . . . . . . . . . . . . . . . . 9 7. Acknowledgements . . . . . . . . . . . . . . . . . . . . . . 9 8. IANA Considerations . . . . . . . . . . . . . . . . . . . . . 9 9. References . . . . . . . . . . . . . . . . . . . . . . . . . 9 9.1. Normative References . . . . . . . . . . . . . . . . . . 9 9.2. Informative References . . . . . . . . . . . . . . . . . 9 Authors' Addresses . . . . . . . . . . . . . . . . . . . . . . . 9 1. Introduction and Terms To paraphrase the Introduction of [RFC6480], the "Resource Public Key Infrastructure (RPKI) represents the allocation hierarchy of IP address space and Autonomous System (AS) numbers; and is a distributed repository system for storing and disseminating the data objects that comprise the RPKI, as well as other signed objects necessary for improved routing security." An Internet Registry (IR) is the IANA, a Regional Internet Registry (RIR), a National Internet Registry (NIR), a Local Internet Registry (LIR), a Internet Service Provider (ISP), or an end site which may hold IP resources and is the subject of one or more certificates using [RFC3779] extensions in the Resource Public Key Infrastructure (RPKI), see [RFC6480]. It is desirable to transfer resources between resource-holding entities in the RPKI, and to do so without violating contracts, policies, etc., and while maintaining operational reliability and administrative accuracy with minimal administrative overhead. Austein, et al. Expires December 1, 2015 [Page 2] Internet-Draft Resource Transfer in the RPKI May 2015 +------------+ | Swing | | Point | | IR | +------------+ ^ ^ +-------+ +-------+ | | +------------+ +------------+ | Selling | | Buying | | IR | | IR | | | | | +------------+ +------------+ Fig 1. The Simplest Example of Seller, Buyer, and Swing Point Seller and Buyer are used to describe the end parties to a transfer, the selling IR transferring the resource(s) to the buying IR. For the purposes of this document, the terms seller and buyer are used, although layer nine considerations may require less commercial formal roles. Transfer is the sale and corresponding purchase of literal address space or autonomous system numbers between two parties. The seller relinquishing some amount of resource and the buyer being allocated a similar amount but not the same literal address space, is not a transfer, and is not further considered here. A Swing Point is the IR at the lowest point in the RPKI hierarchy which the seller and buyer have as a common parent and which has agreed to be used as the agent of transfer. While there is no automated method for the RPKI to assist the parties to a transaction in determining that all business and policy aspects of a transaction are satisfied, these layer eight and nine issues can be resolved using normal business practices. 2. A Simplistic Case Austein, et al. Expires December 1, 2015 [Page 3] Internet-Draft Resource Transfer in the RPKI May 2015 +------------+ | Swing | | Point | | IR | +------------+ 4 ^ ^ | ^ 4 +----------+ | | +----------+ | | | | v | | | +------------+ | | +------------+ | Selling | 2 | | 3 | Buying | | IR | | | | IR | | | | | | | +---------+--+ | | +--+---------+ | | | | 1 | | | | v | | v .---------. | | .---------. |Resources|-----+ +--->|Resources| `---------' `---------' Fig 2. Steps in a Simple Transfer 2.1. Steps in Simple Case As a formal business relationship between all parties to a transfer provides a level of trust which allows simple transactions, we first consider the simple case where the seller and the buyer are both directly known to the swing point, see Figure 1. The transfer is done in the following steps (see Figure 2): 1. The seller creates a certificate describing the subset of the seller's resources which are to be transferred. 2. The seller tells the swing point that it wishes to transfer the resources described by the certificate to the buyer. 3. The swing point issues a new expanded certificate to the buyer describing the buyer's old holdings plus the new resources. 4. When the seller and the buyer are comfortable that both the technical aspects (customers swung, routing done, etc.) and the business aspects of the transfer have been accomplished, they inform the swing point which then shrinks the seller's resource certificate, removing the transferred resources. 2.2. The Torn Euro Protocol Due to issues of cancellation, reneging, and fraud, step 4 above, where the seller and the buyer tell the swing point that the deal is done, needs to be formal in some fashion. For this purpose, we Austein, et al. Expires December 1, 2015 [Page 4] Internet-Draft Resource Transfer in the RPKI May 2015 envision a yet to be described torn Euro protocol, where the buyer and the seller each hold one half of a virtual torn Euro note, and the swing point believes the transaction to be complete when it has received both halves and they match. This protocol has yet to be described, and Steve Kent has taken on the task of looking for an existing simple example that can be borrowed for the purpose. 3. A More Complex Case What happens when the seller is not a direct customer of the swing point, see Figure 3. +------------+ | Swing | | Point | | IR | +------------+ ^ ^ +-------+ +-------+ | | +------------+ +------------+ |Intermediate| | Buying | | IR | | IR | | | | | +----.-------+ +------------+ ^ +-------+ | +------------+ | Selling | | IR | | | +------------+ Figure 3. The Case of The Indirect Seller The swing point needs to be assured that it is contractually able to move the resource given its relationship to the Other IR. As RFC 3779 extensions do not codify business issues such as PI/PA, and rights to resell, this has to be handled out of band, there is no way to automate it. But this is part of today's IR address space management process and will continue to be handled manually. Therefore the process is the same as for the simple case, except that, before issuing the expanded certificate to the buyer in step 3, the swing point must assure itself that policy and contractual issues are cleared. It might be well-advised to contact the intermediate IR Austein, et al. Expires December 1, 2015 [Page 5] Internet-Draft Resource Transfer in the RPKI May 2015 and gain its consent, possibly with the assistance of the seller. The bottom line is that the swing point does own/control the resource being transferred, and therefore has the prerogative to act within its perception of the liabilities it is incurring. This freedom allowing the seller to be indirectly related to the swing point may be induced to more levels of indirection. It is the swing point's obligation to perform diligence on the iterative financial, contractual, and policy obligations of the relationships down to the seller. Unfortunately, the RPKI can not automate this. 3.1. The Indirect Buyer The case where the buyer is not directly known to the swing point is more difficult. Among other issues, the buyer may not be an existing resource holder at all, i.e. there may be no path down from the IANA root to the buyer. In this case, the buyer must explore the graph and choose an IR with which to contract a relationship. This can be both a business issue and a policy issue, e.g. can a buyer in Asia choose a parent which is, directly or indirectly, an ARIN customer? The case where the buyer contracts directly to become a customer of the swing point has been explored above. What if the buyer becomes a grandchild of the swing point, as in figure 4? +------------+ | Swing | | Point | | IR | +------------+ ^ ^ +-------+ +-------+ | | +------------+ +------------+ | Selling | |Intermediate| | IR | | IR | | | | | +------------+ +------------+ ^ +-------+ | +------------+ | Buying | | IR | | | +------------+ Figure 3. The Case of The Indirect Buyer Austein, et al. Expires December 1, 2015 [Page 6] Internet-Draft Resource Transfer in the RPKI May 2015 Somewhat analogously to the case of the indirect seller, the swing point has to iteratively verify that the IRs between it and the buyer are all willing to contractually and technically accept the resource(s) to be allocated to the buyer. But, in the case of the indirect buyer, the iterative conditions are much stronger. In the indirect seller case, the swing point has contractual control of the chain between it and the seller. In the case of the indirect buyer, all intermediate IRs between the swing point and the buyer must give business and technical consent. The swing point can not force its child to issue a resource certificate to the buyer. Things may not be as bad as they appear at first blush. The buyer is actually contracting to its parent, and part of that contract will presumably be that the parent agrees to issue the resource certificate to the buyer when it receives the resource from it's parent. And this presumably applies to the buyer's parent's relationship to a grandparent and so forth. On the other hand, the swing point has no mechanical way to test the willingness of the IRs on the buyer's indirect chain. But the swing point can know when the buyer is happy that it has received the resources, as the buyer will give it the buyer's half of the torn Euro. 3.2. The Difference Between Buyer and Seller Chain Essentially, the difference between an indirect buyer chain and an indirect seller chain is that the swing point has the logical, though maybe not contractual, prerogative to pull address space from the seller's chain, but does not have the power to push it down the buyer's chain. All IRs on the buyer's chain must agree to certify downward toward the buyer. 4. Transfer in the Absense of a Common Ancestor For political reasons, the current RPKI structure has no single root trust anchor. There are a number of roots, e.g. the five RIRs who do not descend from the IANA. This creates considerable complexity and some risk for resource transfers between entities without a common ancestor. To work around this problem, each RIR certifies a subsidiary Certificate Authority for each other RIR to which it transfers resources, see Figure 4, and issues the transferred resources to that subsidiary CA. Austein, et al. Expires December 1, 2015 [Page 7] Internet-Draft Resource Transfer in the RPKI May 2015 +------+ | RIPE | +------+ | | +-----------+-------+-------+-----------+ | | | | | | | | | | v v | v v +-------+ +-------+ | +-------+ +-------+ | APNIC | |AfriNIC| | | ARIN | |LACNIC | +-------+ +-------+ | +-------+ +-------+ v +-----------+ | Members | +-----------+ Figure 4: The RIRs each certify proxy CAs for all of the other RIRs. But, to use the example of Figure 4, the APNIC CA to which RIPE issues resources is, in fact, run by APNIC under APNIC's Business Certificate PKI (see [RFC6492] Section 3) and uses an APNIC-provided publication point. Thus APNIC has under its control, among other things, four CAs, one with resources from each of the other CAs. And similarly for each of the other RIRs. So the swing point for a transfer from an APNIC member to a RIPE member is the APNIC CA. And an APNIC member holding resources originated by APNIC as well as resources transferred in from another RIR, e.g. RIPE, actually holds two resource certificates. This could probably be made more complicated and brittle, but it would require serious effort. 5. Transfer in process: Resources Change Forced from Above Even though both seller and buyer have agreed to a transfer, the seller might try to not relinquish the resource, hoping to sell it more than once. Therefore it may become necessary to force closure for a non-compliant seller. In this case, a resource holding would be changed, shrunk, by force from above. A 'normal' (i.e. what the RPKI design anticipated) resource shrinkage is initiated by the leaf resource holder and propagates upward toward Austein, et al. Expires December 1, 2015 [Page 8] Internet-Draft Resource Transfer in the RPKI May 2015 the root of the tree. At no point in this process does a holder claim more than their parent believes they have. When a resource is forcibly removed from 'above', the shrinkage propagates downward. Until the ultimate holder relinquishes the resource, at some point in the path down the tree a child holds more resources than its parent believes it does. As the protocol model is bottom initiated polling, see [RFC6492], the time window of exposure of this over-claiming can be relatively large. 6. Security Considerations Ghu only knows. 7. Acknowledgements Thanks Mom! 8. IANA Considerations Nothing is required of the IANA; though it would make some things a lot simpler if the IANA was the root TA/CA of the entire tree. 9. References 9.1. Normative References [RFC6492] Huston, G., Loomans, R., Ellacott, B., and R. Austein, "A Protocol for Provisioning Resource Certificates", RFC 6492, February 2012. 9.2. Informative References [RFC3779] Lynn, C., Kent, S., and K. Seo, "X.509 Extensions for IP Addresses and AS Identifiers", RFC 3779, June 2004. [RFC6480] Lepinski, M. and S. Kent, "An Infrastructure to Support Secure Internet Routing", RFC 6480, February 2012. Authors' Addresses Rob Austein Dragon Research Lab 40 Gavin Circle Reading, MA 01867 USA Email: [email protected] Austein, et al. Expires December 1, 2015 [Page 9] Internet-Draft Resource Transfer in the RPKI May 2015 Randy Bush IIJ / Dragon Research Labs 5147 Crystal Springs Bainbridge Island, Washington 98110 US Email: [email protected] Geoff Huston Asia Pacific Network Information Centre 6 Cordelia St South Brisbane, QLD 4101 AU Email: [email protected] George Michaelson Asia Pacific Network Information Centre 6 Cordelia St South Brisbane, QLD 4101 AU Email: [email protected] Austein, et al. Expires December 1, 2015 [Page 10]

https://datatracker.ietf.org/doc/draft-ymbk-sidr-transfer/00/

| Download Scientific Diagram Download scientific diagram | from publication: Interactive visual analysis of families of curves using data aggregation and derivation | Time-series data are regularly collected and analyzed in a wide range of domains. Multiple simulation runs or multiple measurements of the same physical quantity result in ensembles of curves which we call families of curves. The analysis of time-series data is extensively... | Visual, Data Model and Multiplication | ResearchGate, the professional network for scientists. Figure 1 - uploaded by Krešimir Matković Source publication Interactive visual analysis of families of curves using data aggregation and derivation Article Full-text available Zoltan Konyha Alan Lež Krešimir Matković [...] Helwig Hauser Time-series data are regularly collected and analyzed in a wide range of domains. Multiple simulation runs or multiple measurements of the same physical quantity result in ensembles of curves which we call families of curves. The analysis of time-series data is extensively studied in mathematics, statistics, and visualization; but less research is... Contexts in source publication Context 1 ... simulation run spans 500 years, represented as 500 time steps in each series. Figure 1 (a) illustrates a conventional way of storing such data. Time is an additional column in the table. ... Context 2 ... scalar di- mensions of the records need to be duplicated for each time step. If, however, we allow some columns to contain curves, we get a different model, shown in Figure 1 (b). We have now one record that contains the time series T ropicaltemperature(t) as a curve, as opposed to having 500 records with T imestep and T ropicaltemperature attributes. ... Context 3 ... values from a single simulation run are grouped into a single record. All curves populating one column in Figure 1 (b) constitute a family of curves. ... Context 4 ... while the data in levels one and two remains static throughout the analysis session, it becomes dynamic in level three. Data changes as a result of the on-demand computations-for each computation, a new dimension (a column in the table in Figure 1 (b)) is added to the data. ... View in full-text Context 5 ... example from the analysis of variable valve actuation simulation is shown in Figure 1 0. We are interested in a spe- cific shape of the curves: quick rise, a certain span of time while the valve is opened, then quick fall to a given range, and finally smooth closing. ... Context 6 ... are interested in a spe- cific shape of the curves: quick rise, a certain span of time while the valve is opened, then quick fall to a given range, and finally smooth closing. In Figure 1 0(a), the shape is defined using a similarity brush composed of four segments. The similarity thresholds can be specified per segment. ... Context 7 ... facilitates very precise control over shape. As an alterna- tive, we can compute four scalar aggregates, r1t, r2t, br, pw, shown in Figure 1 0(b). The aggregates are visualized in a parallel coordinates view in Figure 1 0(c) and we can brush desired values for all aggregates. ... Context 8 ... an alterna- tive, we can compute four scalar aggregates, r1t, r2t, br, pw, shown in Figure 1 0(b). The aggregates are visualized in a parallel coordinates view in Figure 1 0(c) and we can brush desired values for all aggregates. A similar set of curves is selected again ( Figure 1 0(d)). ... Context 9 ... aggregates are visualized in a parallel coordinates view in Figure 1 0(c) and we can brush desired values for all aggregates. A similar set of curves is selected again ( Figure 1 0(d)). We needed specific aggregates in this case, and an additional view to define the shape via aggregates. ... Context 10 ... the curve view, it supports angu- lar line brush and similarity brush which makes finding and brushing hidden relations simple. In Figure 1 1, pressure and volume are depicted in the phase diagram. The area outlined by each closed curve in the phase diagram corresponds to the energy used in one simulation run. ... Citations ... This enables us to optimize overall visibility and reduce visual clutter by displaying curves with more variability in the back and less fluctuating curves in front. However, many other properties such as visual complexity, as described by Ryan et al. [140], geometric features derived from families of curves by Konyha et al. [97] , or statistical features could be used instead. ... Visualization Hybridization with Spatialization Cues Thesis Full-text available Thomas Trautner Visualization as a tool for visual processing of any underlying data has proven to be an accepted and legitimate part of the scientific reasoning process. Many different techniques help gaining new insights from captured phenomena, support the development or evaluation of hypotheses about collected data, reveal potential misconceptions or false assumptions, simplify communicating knowledge and novel findings, and enable a multitude of additional opportunities. The reason for this effectiveness is that the human visual system is ideally suited to capture and process visually encoded data. The development of visualization from a niche to an established scientific field has made a significant contribution to this success story. A large number of journals, conferences, seminars, and workshops regularly publish new results, evaluate presented approaches, and help making knowledge globally accessible. However, this large number of contributions tailored to variable user groups, the underlying data, and the wide variety of tasks that could be performed with them, emphasizes the plethora of available techniques and the resulting difficulty in choosing the most suitable visualizations. Therefore, we investigated common data sets and analyzed typical tasks normally performed with them. Based on this, we selected well-established and most effective visualization techniques, combining them to form a hybrid representation. The goal of such a visualization hybridization was to merge advantages of individual techniques and, thereby, simultaneously eliminate their limitations. We present so-called hybrid vigors that make the underlying visualizations more widely applicable instead of either having to change required techniques sequentially, or not being able to perform certain tasks at all. Our contributions are intended to simplify the process of finding suitable visualizations for already established data sets. During our research, we focused on two-dimensional point data, depicted on the one hand as scatter plots and, on the other hand, as relationships between consecutive point such as in line charts. Our techniques can be used especially when data sets are so large, dense, and overplotted that conventional techniques reach their limits. We show that hybrid representations are well suited for combining discrete, continuous, or aggregated forms of visual representation. Our hybridizations additionally exploit spatialization cues. Such visual cues emphasize spatiality of the underlying data through shading, without having to embed the data in 3D space including its potential disadvantages. We chose this method of encoding as we consider it the most appropriate choice, given that visualization users interact naturally and preattentively with a spatial world on a daily basis. ... When x i has more than three attributes, the dataset cannot be represented in simple visualizations such as those based on the Cartesian plane. In this case, the vector produces families of curves in the m-dimensional space [19] . ... Getting over High-Dimensionality: How Multidimensional Projection Methods Can Assist Data Science Article Full-text available Evandro S. Ortigossa Fabio Felix Dias Diego Carvalho do Nascimento The exploration and analysis of multidimensional data can be pretty complex tasks, requiring sophisticated tools able to transform large amounts of data bearing multiple parameters into helpful information. Multidimensional projection techniques figure as powerful tools for transforming multidimensional data into visual information according to similarity features. Integrating this class of methods into a framework devoted to data sciences can contribute to generating more expressive means of visual analytics. Although the Principal Component Analysis (PCA) is a well-known method in this context, it is not the only one, and, sometimes, its abilities and limitations are not adequately discussed or taken into consideration by users. Therefore, knowing in-depth multidimensional projection techniques, their strengths, and the possible distortions they can create is of significant importance for researchers developing knowledge-discovery systems. This research presents a comprehensive overview of current state-of-the-art multidimensional projection techniques and shows example codes in Python and R languages, all available on the internet. The survey segment discusses the different types of techniques applied to multidimensional projection tasks from their background, application processes, capabilities, and limitations, opening the internal processes of the methods and demystifying their concepts. We also illustrate two problems, from a genetic experiment (supervised) and text mining (non-supervised), presenting solutions through multidimensional projection application. Finally, we brought elements that reverberate the competitiveness of multidimensional projection techniques towards high-dimension data visualization, commonly needed in data sciences solutions. ... Potter et al. [49] proposed a framework for visual analysis of ensemble data, consisting of linked views, which present an overview first, then guide the user towards details. Konyha et al. [35] explored methods for analyzing scalar attributes, which they called families of curves. They show different levels of data interaction through brushing, starting with single views (e.g., scatter plots), then they transition to brushing in multiple views. ... Visualization of Human Spine Biomechanics for Spinal Surgery Article Full-text available IEEE T VIS COMPUT GR Pepe Eulzer Sabine Bauer Francis Kilian Kai Lawonn We propose a visualization application, designed for the exploration of human spine simulation data. Our goal is to support research in biomechanical spine simulation and advance efforts to implement simulation-backed analysis in surgical applications. Biomechanical simulation is a state-of-the-art technique for analyzing load distributions of spinal structures. Through the inclusion of patient-specific data, such simulations may facilitate personalized treatment and customized surgical interventions. Difficulties in spine modelling and simulation can be partly attributed to poor result representation, which may also be a hindrance when introducing such techniques into a clinical environment. Comparisons of measurements across multiple similar anatomical structures and the integration of temporal data make commonly available diagrams and charts insufficient for an intuitive and systematic display of results. Therefore, we facilitate methods such as multiple coordinated views, abstraction and focus and context to display simulation outcomes in a dedicated tool. By linking the result data with patient-specific anatomy, we make relevant parameters tangible for clinicians. Furthermore, we introduce new concepts to show the directions of impact force vectors, which were not accessible before. We integrated our toolset into a spine segmentation and simulation pipeline and evaluated our methods with both surgeons and biomechanical researchers. When comparing our methods against standard representations that are currently in use, we found increases in accuracy and speed in data exploration tasks. In a qualitative review, domain experts deemed the tool highly useful when dealing with simulation result data, which typically combines time-dependent patient movement and the resulting force distributions on spinal structures. ... Similar to Azuma et al. [34] we propose to change the representation of the visualization as needed, but in our case depending on the current viewing direction. Konyha et al. [35] introduced a method to study families of curves in a multiple-view environment. The analysis of the temporal progression of attributes over time can also be interpreted as studying a set of curves. ... Popup-Plots: Warping Temporal Data Visualization Article Full-text available IEEE T VIS COMPUT GR Johanna Schmidt Dominik Fleischmann Bernhard Preim Norbert Brandle Gabriel Mistelbauer Visualizing temporal data is inherently difficult, due to the many aspects that need to be communicated to the users. This is an important topic in visualization, and a wide range of visualization techniques dealing with different tasks have already been designed. In this paper we propose popup-plots, a novel concept where the common interaction of 3D rotation is used to navigate through the data. This allows the users to view the data from different perspectives without having to learn and adapt to new interaction concepts. Popup-plots are therefore a novel method for visualizing and interacting with dependent variables over time. We extend 2D plots with the temporal information by bending the space according to the time. The bending is calculated based on a spherical coordinates approach, which is continuously influenced by the viewing direction towards the plot. Hence, the plot can be viewed from various angles with seamless transitions in between, offering the possibility to analyze different aspects of the represented data. The temporal information is encoded into the visualization itself, resembling annual rings of a tree. We demonstrate our method by applying it to data from two different domains, and we also evaluated the usability of our solution. ... Demir et al. [4] determine the most central shape from a given set, to quantify a region-wise centrality, and to compute the locally most representative shape. Konyha et al. [18] and Matkovic et al. [22] focus on the interactive visual analysis of ensembles of curves called families of curves or families of surfaces using data aggregation and attribute derivation. Piringer et al. [29] extend the work on featurepreserving downsampling of 2D functions. ... Dynamic Volume Lines: Visual Comparison of 3D Volumes through Space-filling Curves Article Full-text available IEEE T VIS COMPUT GR Johannes Weissenbock Bernhard Frohler Eduard Groller Johann Kastner Christoph Heinzl The comparison of many members of an ensemble is difficult, tedious, and error-prone, which is aggravated by often just subtle differences. In this paper, we introduce Dynamic Volume Lines for the interactive visual analysis and comparison of sets of 3D volumes. Each volume is linearized along a Hilbert space-filling curve into a 1D Hilbert line plot, which depicts the intensities over the Hilbert indices. We present a nonlinear scaling of these 1D Hilbert line plots based on the intensity variations in the ensemble of 3D volumes, which enables a more effective use of the available screen space. The nonlinear scaling builds the basis for our interactive visualization techniques. An interactive histogram heatmap of the intensity frequencies serves as overview visualization. When zooming in, the frequencies are replaced by detailed 1D Hilbert line plots and optional functional boxplots. To focus on important regions of the volume ensemble, nonlinear scaling is incorporated into the plots. An interactive scaling widget depicts the local ensemble variations. Our brushing and linking interface reveals, for example, regions with a high ensemble variation by showing the affected voxels in a 3D spatial view. We show the applicability of our concepts using two case studies on ensembles of 3D volumes resulting from tomographic reconstruction. In the first case study, we evaluate an artificial specimen from simulated industrial 3D X-ray computed tomography (XCT). In the second case study, a real-world XCT foam specimen is investigated. Our results show that Dynamic Volume Lines can identify regions with high local intensity variations, allowing the user to draw conclusions, for example, about the choice of reconstruction parameters. Furthermore, it is possible to detect ring artifacts in reconstructions volumes. ... These techniques scale to very large sets of time series but provide a limited view of the data. Konyha et al. discuss interaction techniques for filtering time series data [23] . ... Visualizing a Million Time Series with the Density Line Chart Preprint Full-text available Dominik Moritz Danyel Fisher Data analysts often need to work with multiple series of data---conventionally shown as line charts---at once. Few visual representations allow analysts to view many lines simultaneously without becoming overwhelming or cluttered. In this paper, we introduce the DenseLines technique to calculate a discrete density representation of time series. DenseLines normalizes time series by the arc length to compute accurate densities. The derived density visualization allows users both to see the aggregate trends of multiple series and to identify anomalous extrema. View Skyscraper Visualization of Multiple Time-Dependent Scalar Fields on Surfaces Article COMPUT GRAPH-UK Monique Meuschke Samuel Voß Franziska Gaidzik Bernhard Preim Kai Lawonn We present a system to visualize multiple time-dependent scalar fields defined on 3D surfaces as they arise from numerical simulations. Domain experts want to understand correlations between multiple scalar fields to verify underlying processes. However, existing visualization techniques require high exploration effort to perceive temporal changes and relations of multiple fields on 3D surfaces. This makes analysis time-consuming and carries a high risk of overlooking regions with specific correlations. Therefore, we introduce a novel concept to facilitate exploration of multiple time-dependent scalar fields simultaneously. The basic idea is to use a novel glyph-based depiction, called skyline visualization, which is inspired by the skyscraper metaphor from information visualization. The skyscrapers are represented by 2D and 3D bar charts, where their floors show the temporal behavior of scalar fields of the corresponding surface region. The user can interact with the skyline visualization to see at which surface areas the scalar fields take on striking values or whether correlations exist between them. Furthermore, the user can immediately see in which regions scalar fields change strongly over time or show strongly inverted behavior. We evaluated our system with several CFD engineers, radiologists and visualization experts. Compared to existing systems, they were able to find regions of interest quickly and with high accuracy. Line Weaver: Importance-Driven Order Enhanced Rendering of Dense Line Charts Conference Paper Full-text available Jun 2021 COMPUT GRAPH FORUM T Trautner S Bruckner Line charts are an effective and widely used technique for visualizing series of ordered two-dimensional data points. The relationship between consecutive points is indicated by connecting line segments, revealing potential trends or clusters in the underlying data. However, when dealing with an increasing number of lines, the render order substantially influences the resulting visualization. Rendering transparent lines can help but unfortunately the blending order is currently either ignored or naively used, for example, assuming it is implicitly given by the order in which the data was saved in a file. Due to the non-commutativity of classic alpha blending, this results in contradicting visualizations of the same underlying data set, so-called "hallucinators". In this paper, we therefore present line weaver, a novel visualization technique for dense line charts. Using an importance function, we developed an approach that correctly considers the blending order independently of the render order and without any prior sorting of the data. We allow for importance functions which are either explicitly given or implicitly derived from the geometric properties of the data if no external data is available. The importance can then be applied globally to entire lines, or locally per pixel which simultaneously supports various types of user interaction. Finally, we discuss the potential of our contribution based on different synthetic and real-world data sets where classic or naive approaches would fail. Task-based visual analytics for interactive multiobjective optimization Article Jun 2020 J OPER RES SOC Jussi Hakanen Kaisa Miettinen Krešimir Matković We study how visual interaction techniques considered in visual analytics can be utilized when implementing interactive multiobjective optimization methods, where a decision maker iteratively participates in the solution process. We want to benefit from previous research and avoid re-inventing ideas. Our aim is to widen awareness and increase the applicability of interactive methods for solving real-world problems. As a concrete approach, we introduce seven high-level tasks that are relevant for interactive methods. These high-level tasks are based on low-level tasks proposed in the visual analytics literature. In addition, we give an example on how the high-level tasks can be implemented and demonstrate this in the context of a real-world multiobjective optimization problem related to wastewater treatment plant operation. Finally, we make recommendations for implementations of interactive methods. We conclude that task-based visual analytics can help in implementing interaction between human decision makers and interactive multiobjective optimization methods. Challenges of Big Data Visualization in Internet-of-Things Environments Chapter Jan 2020 Doaa Mohey Eldin Aboul Ella Hassanien Ehab E. Hassanien

https://www.researchgate.net/figure/fig1_254462965

Discovery of BI 224436, a noncatalytic site integrase inhibitor (NCINI) of HIV-1 | Request PDF Request PDF | Discovery of BI 224436, a noncatalytic site integrase inhibitor (NCINI) of HIV-1 | An assay recapitulating the 3′ processing activity of HIV-1 integrase (IN) was used to screen the Boehringer Ingelheim compound collection.... | Find, read and cite all the research you need on ResearchGate Article Discovery of BI 224436, a noncatalytic site integrase inhibitor (NCINI) of HIV-1 January 2014 ACS Medicinal Chemistry Letters 5(4):422–427 DOI: 10.1021/ml500002n Authors: To read the full-text of this research, you can request a copy directly from the authors. Abstract An assay recapitulating the 3′ processing activity of HIV-1 integrase (IN) was used to screen the Boehringer Ingelheim compound collection. Hit-to-lead and lead optimization beginning with compound 1 established the importance of the C3 and C4 substituent to antiviral potency against viruses with different aa124/aa125 variants of IN. The importance of the C7 position on the serum shifted potency was established. Introduction of a quinoline substituent at the C4 position provided a balance of potency and metabolic stability. Combination of these findings ultimately led to the discovery of compound 26 (BI 224436), the first NCINI to advance into a phase Ia clinical trial. ... this novel class of HIV-1 inhibitors were initially reported in a Boehringer Ingelheim patent (9) and then by Zeger Debyser's laboratory (10). Later, several other pharmaceutical companies and academic groups reported many more lead compounds belonging to this class (11)(12)(13)(14) (15) (16)(17)(18)(19). Consequently, the molecules became known by alternative names: LEDGINs (10,19), allosteric integrase inhibitors (ALLINIs) (11,18), noncatalytic integrase inhibitors (NCINIs) (9,13,15,17), multimerization integrase inhibitors (MINIs) (11,14), and IN-LEDGF allosteric inhibitors (INLAIs) (12,16). ... ... Later, several other pharmaceutical companies and academic groups reported many more lead compounds belonging to this class (11)(12)(13)(14)(15)(16)(17)(18)(19). Consequently, the molecules became known by alternative names: LEDGINs (10,19), allosteric integrase inhibitors (ALLINIs) (11,18), noncatalytic integrase inhibitors (NCINIs) (9,13, 15, 17), multimerization integrase inhibitors (MINIs) (11,14), and IN-LEDGF allosteric inhibitors (INLAIs) (12,16). Herein, these small molecules are collectively referred to as INLAIs. ... ... Novel INLAIs based on a benzene scaffold. A range of scaffolds, including quinoline for BI-224436 (15) , thiophene for MUT-A (16), benzene for Shionogi S-I-82 (46), benzothiazole for GS-9822 (47), and pyrrolopyridine for STP0404 (21), have been used for the development of INLAIs (Fig. 1A). Importantly, all described potent INLAIs share the common chemical module comprising a carboxylic and a tert-butoxy moiety attached to a bulky hydrophobic side chain via an aromatic scaffold, which are collectively responsible for binding to the pocket on HIV-1 IN CCD dimer interface (1). ... Biological and Structural Analyses of New Potent Allosteric Inhibitors of HIV-1 Integrase Article Full-text available Jun 2023 ANTIMICROB AGENTS CH Damien Bonnard Erwann Le Rouzic Matthew R. Singer Richard Benarous HIV-1 integrase-LEDGF allosteric inhibitors (INLAIs) share the binding site on the viral protein with the host factor LEDGF/p75. These small molecules act as molecular glues promoting hyper-multimerization of HIV-1 IN protein to severely perturb maturation of viral particles. Herein, we describe a new series of INLAIs based on a benzene scaffold that display antiviral activity in the single digit nanomolar range. Akin to other compounds of this class, the INLAIs predominantly inhibit the late stages of HIV-1 replication. A series of high-resolution crystal structures revealed how these small molecules engage the catalytic core and the C-terminal domains of HIV-1 IN. No antagonism was observed between our lead INLAI compound BDM-2 and a panel of 16 clinical antiretrovirals. Moreover, we show that compounds retained high antiviral activity against HIV-1 variants resistant to IN strand transfer inhibitors and other classes of antiretroviral drugs. The virologic profile of BDM-2 and the recently completed single ascending dose phase I trial (ClinicalTrials.gov identifier: NCT03634085) warrant further clinical investigation for use in combination with other antiretroviral drugs. Moreover, our results suggest routes for further improvement of this emerging drug class. ... Structures of ALLINIs bound to the isolated CCD revealed at high resolution how small molecules could inhibit LEDGF binding via high af f inity interactions (11,(36) (37) (38)(39)47). However, the structures did not indicate how drug-induced aggregation is initiated or how other domains of IN might be involved (11,56) ( Figure 1C). ... ... We theref ore purif ied IN CCD (residues 50-212) and CTD (residues 220-271) and reconstituted the ternary complex with the ALLINI BI-224436. BI-224436 is a potent antiviral (EC50 <15 nM(60)) with f avorable solubility properties and was the f irst ALLINI tested in preclinical trials (37, 60). Its chemical structure recapitulates the most common f eatures among ALLINIs, including carboxylic and tert-butoxy groups alongside large hydrophobic moieties that optimize its f it to the hydrophobic dimer interf ace of the CCD ( Figure 1B). ... ... Instead, it directly abuts Ala-128 f rom the CCD α3 helix ( Figure 6D). In serial passage resistance studies with BI-224436, the mutations A128T and A128N arise (37, 60), which would be predicted to clash with the Tyr226 side-chain in addition to the R4 substituent noted earlier, resulting in xxx. ... Structure of a HIV-1 IN-Allosteric Inhibitor Complex at 2.93 Å Resolution: Routes to Inhibitor Optimization Preprint Jun 2022 Grant Eilers Kushol Gupta Audrey Allen Gregory D. Van Duyne HIV integrase (IN) inserts viral DNA into the host genome and is the target of the strand transfer inhibitors (STIs), a class of small molecules currently in clinical use. Another potent class of antivirals is the allosteric inhibitors of integrase, or ALLINIs. ALLINIs promote IN aggregation by stabilizing an interaction between the catalytic core domain (CCD) and carboxy -terminal domain (CTD) that undermines viral particle formation in late replication. Ongoing challenges with inhibitor potency, toxicity, and viral resistance motivate research to understand their mechanism. Here, we report a 2.93 Å X-ray crystal structure of the minimal ternary complex between CCD, CTD, and the ALLINI BI-224436. This structure reveals an asymmetric ternary complex with a prominent network of π-mediated interactions that suggest specific avenues for future ALLINI development and optimization. ... This substitution on position 3 is ideal, as it provides several hydrogen bonds with the T174 and H171 residues that mimic the LEDGF/p75 IBD binding pattern [16][17][18]. Recent derivatization studies have focused on position 4 [20, 33] . We and others have found that large aromatic substitutions at this site greatly enhance multimerization properties of the compounds by maximizing interaction with the hydrophobic pocket [20,33], including residues W132 and L102 ( Figure 1). ... ... Recent derivatization studies have focused on position 4 [20,33]. We and others have found that large aromatic substitutions at this site greatly enhance multimerization properties of the compounds by maximizing interaction with the hydrophobic pocket [20, 33] , including residues W132 and L102 ( Figure 1). We found that a para-chloro-4-phenylquinoline substitution (Figures 1 and 2) showed effective inhibition with an in vitro IN multimerization EC50 of 100 nM due to the stabilizing Cl-π interaction between the aromatic side chain of W132 and the chlorobenzene group [20]. ... ... The 4-chlorophenylquinoline (Compound 1- Figure 2 and Table 1) was utilized as reference point for relative comparison. Substitution with bromine resulted in a previously reported compound (ALLINI-2) [17, 33] showing a slight decrease in the measured EC50 value, implying increased ability to cause multimerization (Table 1). However, substitution with iodine (8a) or amino (8b) groups resulted in a two-fold decrease in the EC50 values (Table 1). ... Multi-Substituted Quinolines as HIV-1 Integrase Allosteric Inhibitors Allosteric HIV-1 integrase (IN) inhibitors, or ALLINIs, are a new class of antiviral agents that bind at the dimer interface of the IN, away from the enzymatic catalytic site and block viral replication by triggering an aberrant multimerization of the viral enzyme. To further our understanding of the important binding features of multi-substituted quinoline-based ALLINIs, we have examined the IN multimerization and antiviral properties of substitution patterns at the 6 or 8 position. We found that the binding properties of these ALLINIs are negatively impacted by the presence of bulky substitutions at these positions. In addition, we have observed that the addition of bromine at either the 6 (6-bromo) or 8 (8-bromo) position conferred better antiviral properties. Finally, we found a significant loss of potency with the 6-bromo when tested with the ALLINI-resistant IN A128T mutant virus, while the 8-bromo analog retained full effectiveness. ... A second class of drugs has been more recently developed that inhibits IN through a novel mechanism of action that is distinct from INSTIs. Using either high-throughput screening or structure-guided modeling that was intended to inhibit the interaction between IN and the cellular factor LEDGF, several groups reported on compounds that have inhibitory activity against IN [197, 198]. Similar compounds have been reported in other work and given a variety of names, including Allosteric Integrase Inhibitors (ALLINIs), Non-Catalytic Integrase Inhibitors (NCINIs), LEDGF/p75-IN interaction site (LEDGINs) and IN-LAI (IN-LEDGF allosteric inhibitors), and multimeric IN inhibitors (MINIs). Below, we will give a brief account of the main properties of these compounds. ... ... There has been an extensive literature discussing the different ALLINI compounds, chemical optimizations, mechanism of action, and their utility, and the interested reader is pointed to those works for further details [199][200][201][202]. The class of allosteric inhibitors is characterized by the presence of heterocyclic cores, such as pyridine [203,204], thiophene [205], quinoline [197, 198,[206][207][208][209], isoquinoline [210], and thienopyridine [198,211]. All compounds belonging to this class bind to the interface formed by LEDGF and IN, effectively blocking this interaction. ... ... Because IN has been implicated in RNA binding and packaging of the genomic RNA (see above), treatment with ALLINIs also leads to the improper localization of vRNA outside of the conical capsid core [51,136,138]. Early structural reports showed how ALLINI compounds bind at the CCD-CCD dimer interface, which would be normally occupied by the IBD of LEDGF/p75 [138, 197] . These, as well as more recent structural reports [51,138,197,203,210,215,[218][219][220][221], all used the CCD dimer to template different ALLINI compounds for structural studies. Several examples of ALLINI structures bound to the CCD dimer interface, and how their binding compares with that of the IBD domain of LEDGF, are shown in Fig. 13A-C. ... Retroviral integrase: Structure, mechanism, and inhibition Chapter The retroviral protein Integrase (IN) catalyzes concerted integration of viral DNA into host chromatin to establish a permanent infection in the target cell. We learned a great deal about the mechanism of catalytic integration through structure/function studies over the previous four decades of IN research. As one of three essential retroviral enzymes, IN has also been targeted by antiretroviral drugs to treat HIV-infected individuals. Inhibitors blocking the catalytic integration reaction are now state-of-the-art drugs within the antiretroviral therapy toolkit. HIV-1 IN also performs intriguing non-catalytic functions that are relevant to the late stages of the viral replication cycle, yet this aspect remains poorly understood. There are also novel allosteric inhibitors targeting non-enzymatic functions of IN that induce a block in the late stages of the viral replication cycle. In this chapter, we will discuss the function, structure, and inhibition of retroviral IN proteins, highlighting remaining challenges and outstanding questions. ... During the last two decades, there have been significant efforts to develop small molecules as clinical alternatives to INSTIs for treating HIV-1 infected individuals [20][21][22][23] [24] [25][26]. These compounds are often referred to as allosteric integrase inhibitors (ALLINIs) or LEDGF/p75-IN inhibitors (LEDGINs), IN-LEDGF/p75 allosteric inhibitors (INLAIs), noncatalytic site integrase inhibitors (NCINIs), and selective multimeric IN inhibitors (MI-NIs) [23][24][25][26]. ... ... During the last two decades, there have been significant efforts to develop small molecules as clinical alternatives to INSTIs for treating HIV-1 infected individuals [20][21][22][23][24][25][26]. These compounds are often referred to as allosteric integrase inhibitors (ALLINIs) or LEDGF/p75-IN inhibitors (LEDGINs), IN-LEDGF/p75 allosteric inhibitors (INLAIs), noncatalytic site integrase inhibitors (NCINIs), and selective multimeric IN inhibitors (MI-NIs) [23] [24] [25][26]. They bind to LEDGF/p75 binding sites on IN, leading to aberrant multimerization of HIV-1 IN, which ultimately results in defective virions [16][17][18][19][20][21][22][23][24][25]. ... ... These compounds are often referred to as allosteric integrase inhibitors (ALLINIs) or LEDGF/p75-IN inhibitors (LEDGINs), IN-LEDGF/p75 allosteric inhibitors (INLAIs), noncatalytic site integrase inhibitors (NCINIs), and selective multimeric IN inhibitors (MI-NIs) [23][24][25][26]. They bind to LEDGF/p75 binding sites on IN, leading to aberrant multimerization of HIV-1 IN, which ultimately results in defective virions [16][17][18][19][20][21][22][23] [24] [25]. Examples include BI-1001 (1), BI 224436 (2), MUT 101 (3), MUT-A (4), indole derivatives (5), 5-carbonyl-1H-imidazole-4-carboxamides (6 and 7), and N-acylhydrazone-based derivatives (8) (Figure 1) [23][24][25][26][27][28][29]. ... Studies towards the Design and Synthesis of Novel 1,5-Diaryl-1H-imidazole-4-carboxylic Acids and 1,5-Diaryl-1H-imidazole-4-carbohydrazides as Host LEDGF/p75 and HIV-1 Integrase Interaction Inhibitors Thompho Jason Rashamuse Two targeted sets of novel 1,5-diaryl-1H-imidazole-4-carboxylic acids 10 and carbohydrazides 11 were designed and synthesized from their corresponding ester intermediates 17, which were prepared via cycloaddition of ethyl isocyanoacetate 16 and diarylimidoyl chlorides 15. Evaluation of these new target scaffolds in the AlphaScreenTM HIV-1 IN-LEDGF/p75 inhibition assay identified seventeen compounds exceeding the pre-defined 50% inhibitory threshold at 100 µM concentration. Further evaluation of these compounds in the HIV-1 IN strand transfer assay at 100 μM showed that none of the compounds (with the exception of 10a, 10l, and 11k, with marginal inhibitory percentages) were actively bound to the active site, indicating that they are selectively binding to the LEDGF/p75-binding pocket. In a cell-based HIV-1 antiviral assay, compounds 11a, 11b, 11g, and 11h exhibited moderate antiviral percentage inhibition of 33–45% with cytotoxicity (CC50) values of >200 µM, 158.4 µM, >200 µM, and 50.4 µM, respectively. The antiviral inhibitory activity displayed by 11h was attributed to its toxicity. Upon further validation of their ability to induce multimerization in a Western blot gel assay, compounds 11a, 11b, and 11h appeared to increase higher-order forms of IN. ... Notably, this site had been identified as a potential inhibitor binding site before it was known to be the site of LEDGF binding (Molteni et al. 2001) . Multiple groups have synthesized and characterized ALLINIs (Fader et al. 2014; H. Wang et al. 2012;G. ... ... As with all antiretroviral classes, resistance to ALLINIs is a problem that must be addressed. Serial viral passage experiments in the presence of ALLINIs has led to the isolation of multiple resistance mutations (Christ et al. 2010;Tsiang et al. 2012;Christ et al. 2012;Le Rouzic et al. 2013;Balakrishnan et al. 2013;Sharma et al. 2014; Fader et al. 2014 (Gupta et al. 2016;Tsiang et al. 2012;Blanco et al. 2011) , meaning that ALLINIs offer a complementary approach to existing antiretrovirals targeting integrase. ... ... Importantly, ALLINIs target a site on IN distinct from the site targeted by STIs, and resistance mutations to ALLINIs do not overlap with STI resistance mutations (Gupta et al. 2016;Blanco et al. 2011;Tsiang et al. 2012) , meaning that ALLINIs offer a non-redundant approach to targeting IN. ALLINI resistance substitutions can be readily identified in serial viral passage experiments (Christ et al. 2010;Tsiang et al. 2012;Christ et al. 2012;Le Rouzic et al. 2013;Balakrishnan et al. 2013;Sharma et al. 2014; Fader et al. 2014) . The mechanisms of several resistance substitutions have been studied Slaughter et al. 2014;Hoyte et al. 2017) , albeit with a focus on the CCD side of the ALLINI binding interface. ... Structural And Biochemical Studies Of Hiv-1 Integration And Inhibition The human immunodeficiency virus (HIV) affects millions of people worldwide who rely on antiretroviral therapy to prevent the acquired immunodeficiency syndrome and halt further HIV transmission. HIV integrase (IN), one of the three retroviral enzymes, catalyzes the covalent insertion of a DNA copy of the HIV genome into host cell chromatin, an essential step in the viral replication cycle. Integration enables expression of a new generation of viral RNA genomes and subgenomic RNAs encoding viral proteins, and establishes the potential for latency, a major barrier to cure. Despite being considered initially to be “undruggable”, IN has been successfully targeted with small-molecule therapeutics, two classes of which are studied here. The strand transfer inhibitors (STIs), which block an essential step in IN catalysis, are FDA-approved and in widespread clinical use. Clinical resistance to STIs has been documented, motivating the design of new generations of STIs with high genetic barriers to resistance and novel approaches to targeting IN. The allosteric inhibitors of integrase (ALLINIs) bind to a site distinct from STIs and do not directly block catalysis, instead act by aberrantly polymerizing IN, disrupting virion maturation. The binding interface of ALLINIs and a structural explanation of their mechanism of action was recently reported by our group. Accurate structural and biochemical data are essential for ongoing drug development and efforts to understand mechanisms of resistance. Here, we report structural and biochemical advances that further our understanding of antiretrovirals targeting HIV IN. We have improved the resolution of structural models of IN·ALLINI polymers, extended these data to multiple members of this class of compounds, and revealed mechanisms of resistance. We have identified a promising clinical opportunity combining ALLINIs and STIs that exploits the hypersensitivity of STI-resistant IN to ALLINI inhibition. Through the creation of an improved in vitro model, we report structural and biochemical data that most closely recapitulates the form of IN found in vivo. Finally, we report progress toward structural and functional characterization of the catalytically active complex of IN and viral DNA, the intasome. Together, this body of work advances our understanding of a key step in the pathogenesis of HIV and provides a foundation for improvements to therapeutics. ... 6a; red, green and blue, respectively). Many ALLINIs were co-crystallized with the isolated HIV-1 IN CCD, and the structures revealed that the molecules grip onto the LEDGF/p75 IBD-binding pocket at the IN CCD dimerization interface 171, 172, 174,175,180,186,187 . The carboxylate establishes a bidentate hydrogen bond with main-chain amides of IN Glu170 and His171, perfectly recuperating the interactions made by Asp366 of LEDGF/p75 109 (fig. ... ... ALLINIs engage the LEDGF/p75-binding cleft at the IN CCD dimer interface, which is distal from the IN active site109,171 . Lead compounds in this class were discovered using two very different high-throughput approaches: a screen for inhibitors of HIV-1 IN 3′-processing activity 172 and structure-based design of small-molecule inhibitors of the HIV-1 IN-LEDGF/p75 interaction 171 . Many, though not all, of the ensuing compounds effectively inhibit IN-LEDGF/p75 binding in vitro 171,173-175 . ... ... Compounds of the last class do not effectively antagonize HIV-1 IN-LEDGF/p75 binding. Although ALLINIs have yet to advance in clinical trials, several compounds display exquisite low-nanomolar potencies to inhibit HIV-1 replication in vitro 172, 180,182,183 . ... Structure and function of retroviral integrase A hallmark of retroviral replication is establishment of the proviral state, wherein a DNA copy of the viral RNA genome is stably incorporated into a host cell chromosome. Integrase is the viral enzyme responsible for the catalytic steps involved in this process, and integrase strand transfer inhibitors are widely used to treat people living with HIV. Over the past decade, a series of X-ray crystallography and cryogenic electron microscopy studies have revealed the structural basis of retroviral DNA integration. A variable number of integrase molecules congregate on viral DNA ends to assemble a conserved intasome core machine that facilitates integration. The structures additionally informed on the modes of integrase inhibitor action and the means by which HIV acquires drug resistance. Recent years have witnessed the development of allosteric integrase inhibitors, a highly promising class of small molecules that antagonize viral morphogenesis. In this Review, we explore recent insights into the organization and mechanism of the retroviral integration machinery and highlight open questions as well as new directions in the field. ... For example, early works have shown that the 3-α-tert-butoxy acetic acid side-chain (quinoline scaffold position numbers in Fig. 2) is optimal for IN anchoring (15,16,20) because it provides several hydrogen bond interactions with T174 and H171 residues that mimic the LEDGF/p75 binding pattern. More recent derivatization efforts have focused on the position 4 (19, 29) . We and others have found that large aromatic substitutions at this site greatly enhance multimerization properties of the compounds by maximizing interaction with the hydrophobic pocket (19,29) including residues W132 and L102 (Fig. 2). ... ... More recent derivatization efforts have focused on the position 4 (19,29). We and others have found that large aromatic substitutions at this site greatly enhance multimerization properties of the compounds by maximizing interaction with the hydrophobic pocket (19, 29) including residues W132 and L102 (Fig. 2). In the present work, to further understand and quantify the specific contribution of each feature of these inhibitors toward multimerization, we have kept constant the 3α-tert-butoxy acetic acid side-chain and our previously optimized position 4 with a 1,4-benzo dioxane group (19). ... ... As previously reported with other quinoline scaffold-based ALLINIs (22, 29, 32), no measurable cytotoxicity was detected in the concentration ranges tested (Table 3). We and others have previously observed how ALLINIs interfere with proper virus particle maturation and yield noninfectious particles with eccentrically positioned RNPs. ... Optimized binding of substituted quinoline ALLINIs within the HIV-1 integrase oligomer During the integration step, HIV-1 integrase (IN) interacts with viral DNA and the cellular cofactor LEDGF/p75 to effectively integrate the reverse transcript into the host chromatin. Allosteric HIV-1 integrase inhibitors (ALLINIs) are a new class of antiviral agents that bind at the dimer interface of the IN catalytic core domain and occupy the binding site of LEDGF/p75. While originally designed to block IN-LEDGF/p75 interactions during viral integration, several of these compounds have been shown to also severely impact viral maturation through an IN multimerization mechanism. In this study, we tested the hypothesis that these dual properties of ALLINIs could be decoupled toward late stage viral replication effects by generating additional contact points between the bound ALLINI and a third subunit of IN. By sequential derivatization at position 7 of a quinoline-based ALLINI scaffold, we show that IN multimerization properties are enhanced by optimizing hydrophobic interactions between the compound and the C-terminal domain of the third IN subunit. These features not only improve the overall anti-viral potencies of these compounds but also significantly shift the ALLINIs selectivity toward the viral maturation stage. Thus, we demonstrate that in order to fully maximize the potency of ALLINIs, the interactions between the inhibitor and all three IN subunits need to be simultaneously optimized. ... In 2014, BI 224436, the first approved non-catalytic site integrase inhibitor (NCINI), was developed by Boehringer Ingelheim (Canada) Ltd. In contrast to INSTIs, BI 224436 binds a highly conserved allosteric site in CCD of IN to induce conformational changes in the catalytic site, thus disrupting interactions of IN with long-terminal repeat (LTR) DNA, and is additionally reported to inhibit 3′-processing [13, 19, 20]. NCINIs, a new class of INI drugs, can overcome the problems of INSTI-resistant viruses based on its different inhibition mechanism. ... ... BI 224436 was employed as the positive control in vitro to ascertain whether the TZM-bl system could be applied to assess the efficacy of NCINIs. Efficacy of candidates against HIV-1 AD8 was measured in TZM-bl cells (Fig. 1 [19] . The extremely low antiviral activity of BI224436 evaluated using the TZM-bl system suggests that this assay may not be suitable for assessment of NCINI efficacy. ... ... In contrast, BI 224436 showed low inhibitory activity (36% at 1000 μM and 20% at 8 μM; Fig. 3b). Our data are consistent with previous reports that BI 224436 has an EC 50 value > 50 μM and does not inhibit strand transfer [19, 20]. ... Study on suitable analysis method for HIV-1 non-catalytic integrase inhibitor Background Integrase (IN) is an essential protein for HIV replication that catalyzes insertion of the reverse-transcribed viral genome into the host chromosome during the early steps of viral infection. Highly active anti-retroviral therapy is a HIV/AIDS treatment method that combines three or more antiviral drugs often formulated from compounds that inhibit the activities of viral reverse transcriptase and protease enzymes. Early IN inhibitors (INIs) mainly serve as integrase strand transfer inhibitors (INSTI) that disrupt strand transfer by binding the catalytic core domain of IN. However, mutations of IN can confer resistance to INSTI. Therefore, non-catalytic integrase inhibitors (NCINI) have been developed as next-generation INIs. Methods In this study, we evaluated and compared the activity of INSTI and NCINI according to the analysis method. Antiviral activity was compared using p24 ELISA with MT2 cell and TZM-bl luciferase system with TZM-bl cell. Each drug was serially diluted and treated to MT2 and TZM-b1 cells, infected with HIV-1 AD8 strain and incubated for 5 and 2 days, respectively. Additionally, to analyze properties of INSTI and NCINI, transfer inhibition assay and 3′-processing inhibition assay were performed. Results During screening of INIs using the p24 ELISA and TZM-bl luciferase systems, we found an inconsistent result with INSTI and NCINI drugs. Following infection of MT2 and TZM-bl cells with T-tropic HIV-1 strain, both INSTI and NCINI treatments induced significant p24 reduction in MT2 cells. However, NCINI showed no antiviral activity in the TZM-bl luciferase system, indicating that this widely used and convenient antiretroviral assay is not suitable for screening of NCINI compounds that target the second round of HIV-1 replication. Conclusion Accordingly, we recommend application of other assay procedures, such as p24 ELISA or reverse transcription activity, in lieu of the TZM-bl luciferase system for preliminary NCINI drug screening. Utilization of appropriate analytical methods based on underlying mechanisms is necessary for accurate assessment of drug efficacy. ... In contrast to INSTIs, BI 224436 binds a highly conserved allosteric site in CCD of IN to induce conformational changes in the catalytic site, thus disrupting interactions of IN with long-terminal repeat (LTR) DNA, and is additionally reported to inhibit 3'-processing. [13, 19, 20] [13,20] We were commissioned by a domestic pharmaceutical company to evaluate the e cacy of a novel NCINI candidate but were unable to verify antiviral activity using the TZM-bl luciferase system. In view of this nding, we attempted to develop a suitable method for accurately assessing the antiviral e cacy of NCINI. ... ... And BI224436 data shows statistically signi cant differences with all INSTIs from 625 nM to 4.88 nM.In contrast to our data, BI 224436 is reported to have EC50 values in the range of 11-27 nM against HxB2, NL4.3 and a recombinant NL4.3 strain. [19] The extremely low antiviral activity of BI224436 evaluated using the TZM-bl system suggests that this assay may not be suitable for assessment of NCINI e cacy. ... ... Our data are consistent with previous reports that BI 224436 has an EC50 value >50 μM and does not inhibit strand transfer. [19, 20] ... Study on Suitable Analysis Method for HIV-1 Non-catalytic Integrase Inhibitor Preprint Background: Integrase (IN) is an essential protein for HIV replication that catalyzes insertion of the reverse-transcribed viral genome into the host chromosome during the early steps of viral infection. Highly active anti-retroviral therapy (HAART) is a HIV/AIDS treatment method that combines three or more antiviral drugs often formulated from compounds that inhibit the activities of viral reverse transcriptase and protease enzymes. Early IN inhibitors (INIs) mainly serve as integrase strand transfer inhibitors (INSTI) that disrupt strand transfer by binding the catalytic core domain (CCD) of IN. However, mutations of IN can confer resistance to INSTI. Therefore, non-catalytic integrase inhibitors (NCINI) have been developed as next-generation INIs. Methods: In this study, we evaluated and compared the activity of INSTI and NCINI according to the analysis method. Antiviral activity was compared using p24 ELISA with MT2 cell and TZM-bl luciferase system with TZM-bl cell. Each drug was serially diluted and treated to MT2 and TZM-b1 cells, infected with HIV-1 AD8 strain and incubated for 5 and 2 days, respectively. Additionally, to analyze properties of INSTI and NCINI, transfer inhibition assay and 3'-processing inhibition assay were performed. Results: During screening of INIs using the p24 ELISA and TZM-bl luciferase systems, we found an inconsistent result with INSTI and NCINI drugs. Following infection of MT2 and TZM-bl cells with T-tropic HIV-1 strain, both INSTI and NCINI treatments induced significant p24 reduction in MT2 cells. However, NCINI showed no antiviral activity in the TZM-bl luciferase system, indicating that this widely used and convenient antiretroviral assay is not suitable for screening of NCINI compounds that target the second round of HIV-1 replication. Conclusion: Accordingly, we recommend application of other assay procedures, such as p24 ELISA or reverse transcription activity, in lieu of the TZM-bl luciferase system for preliminary NCINI drug screening. Utilization of appropriate analytical methods based on underlying mechanisms is necessary for accurate assessment of drug efficacy. ... In contrast to INSTIs, BI 224436 binds a highly conserved allosteric site in CCD of IN to induce conformational changes in the catalytic site, thus disrupting interactions of IN with long-terminal repeat (LTR) DNA, and is additionally reported to inhibit 3'-processing. [13, 19, 20] NCINIs, a new class of INI drugs, can overcome the problems of INSTI-resistant viruses based on its different inhibition mechanism. Speci cally, INSTIs directly inhibit IN-LEDGF/p75 interactions while NCINIs bind CCD to inhibit IN-LTR DNA generation. ... ... BI 224436 was employed as the positive control in vitro to ascertain whether the TZM-bl system could be applied to assess the e cacy of NCINIs. E cacy of candidates against HIV-1 AD8 was measured in TZM-bl cells (Fig 1) [19] The extremely low antiviral activity of BI224436 evaluated using the TZM-bl system suggests that this assay may not be suitable for assessment of NCINI e cacy. ... ... Our data are consistent with previous reports that BI 224436 has an EC 50 value >50 μM and does not inhibit strand transfer. [19, 20] Evaluation of differences in INSTI and NCINI activities according to inhibition mechanism ... Study on Suitable Analysis Method for HIV-1 Non-catalytic Integrase Inhibitor Jul 2020 Ki Hoon Park Young Bong Kim Background: Integrase (IN) is an essential protein for HIV replication that catalyzes insertion of the reverse-transcribed viral genome into the host chromosome during the early steps of viral infection. Highly active anti-retroviral therapy (HAART) is a HIV/AIDS treatment method that combines three or more antiviral drugs often formulated from compounds that inhibit the activities of viral reverse transcriptase and protease enzymes. Early IN inhibitors (INIs) mainly serve as integrase strand transfer inhibitors (INSTI) that disrupt strand transfer by binding the catalytic core domain (CCD) of IN. However, mutations of IN can confer resistance to INSTI. Therefore, non-catalytic integrase inhibitors (NCINI) have been developed as next-generation INIs. Methods: In this study, we evaluated and compared the activity of INSTI and NCINI according to the analysis method. Antiviral activity was compared using p24 ELISA with MT2 cell and TZM-bl luciferase system with TZM-bl cell. Each drug was serially diluted and treated to MT2 and TZM-b1 cells, infected with HIV-1 AD8 strain and incubated for 5 and 2 days, respectively. Additionally, to analyze properties of INSTI and NCINI, transfer inhibition assay and 3'-processing inhibition assay were performed. Results: During screening of INIs using the p24 ELISA and TZM-bl luciferase systems, we found an inconsistent result with INSTI and NCINI drugs. Following infection of MT2 and TZM-bl cells with T-tropic HIV-1 strain, both INSTI and NCINI treatments induced significant p24 reduction in MT2 cells. However, NCINI showed no antiviral activity in the TZM-bl luciferase system, indicating that this widely used and convenient antiretroviral assay is not suitable for screening of NCINI compounds that target the second round of HIV-1 replication. Conclusion: Accordingly, we recommend application of other assay procedures, such as p24 ELISA or reverse transcription activity, in lieu of the TZM-bl luciferase system for preliminary NCINI drug screening. Utilization of appropriate analytical methods based on underlying mechanisms is necessary for accurate assessment of drug efficacy. ... In contrast to INSTIs, BI 224436 binds a highly conserved allosteric site in CCD of IN to induce conformational changes in the catalytic site, thus disrupting interactions of IN with long-terminal repeat (LTR) DNA, and is additionally reported to inhibit 3'-processing. [13, 19, 20] NCINIs, a new class of INI drugs, can overcome the problems of INSTI-resistant viruses based on its different inhibition mechanism. Speci cally, INSTIs directly inhibit IN-LEDGF/p75 interactions while NCINIs bind CCD to inhibit IN-LTR DNA generation. ... ... BI 224436 was employed as the positive control in vitro to ascertain whether the TZM-bl system could be applied to assess the e cacy of NCINIs. E cacy of candidates against HIV-1 AD8 was measured in TZM-bl cells (Fig 1) [19] The extremely low antiviral activity of BI224436 evaluated using the TZM-bl system suggests that this assay may not be suitable for assessment of NCINI e cacy. ... ... Our data are consistent with previous reports that BI 224436 has an EC 50 value >50 μM and does not inhibit strand transfer. [19, 20] ... Study on Suitable Analysis Method for HIV-1 Non-catalytic Integrase Inhibitor Seoung Eun Bae Young Bong Kim Background: Integrase (IN) is an essential protein for HIV replication that catalyzes insertion of the reverse-transcribed viral genome into the host chromosome during the early steps of viral infection. Highly active anti-retroviral therapy (HAART) is a HIV/AIDS treatment method that combines three or more antiviral drugs often formulated from compounds that inhibit the activities of viral reverse transcriptase and protease enzymes. Early IN inhibitors (INIs) mainly serve as integrase strand transfer inhibitors (INSTI) that disrupt strand transfer by binding the catalytic core domain (CCD) of IN. However, mutations of IN can confer resistance to INSTI. Therefore, non-catalytic integrase inhibitors (NCINI) have been developed as next-generation INIs. Methods: In this study, we evaluated and compared the activity of INSTI and NCINI according to the analysis method. Antiviral activity was compared using p24 ELISA with MT2 cell and TZM-bl luciferase system with TZM-bl cell. Each drug was serially diluted and treated to MT2 and TZM-b1 cells, infected with HIV-1 AD8 strain and incubated for 5 and 2 days, respectively. Additionally, to analyze properties of INSTI and NCINI, transfer inhibition assay and 3'-processing inhibition assay were performed. Results: During screening of INIs using the p24 ELISA and TZM-bl luciferase systems, we found an inconsistent result with INSTI and NCINI drugs. Following infection of MT2 and TZM-bl cells with T-tropic HIV-1 strain, both INSTI and NCINI treatments induced significant p24 reduction in MT2 cells. However, NCINI showed no antiviral activity in the TZM-bl luciferase system, indicating that this widely used and convenient antiretroviral assay is not suitable for screening of NCINI compounds that target the second round of HIV-1 replication. Conclusion: Accordingly, we recommend application of other assay procedures, such as p24 ELISA or reverse transcription activity, in lieu of the TZM-bl luciferase system for preliminary NCINI drug screening. Utilization of appropriate analytical methods based on underlying mechanisms is necessary for accurate assessment of drug efficacy. ... In contrast to INSTIs, BI 224436 binds a highly conserved allosteric site in CCD of IN to induce conformational changes in the catalytic site, thus disrupting interactions of IN with long-terminal repeat (LTR) DNA, and is additionally reported to inhibit 3'-processing. [13, 19, 20] [13,20] We were commissioned by a domestic pharmaceutical company to evaluate the e cacy of a novel NCINI candidate but were unable to verify antiviral activity using the TZM-bl luciferase system. In view of this nding, we attempted to develop a suitable method for accurately assessing the antiviral e cacy of NCINI. ... ... In contrast to our data, BI 224436 is reported to have EC 50 values in the range of 11-27 nM against HxB2, NL4.3 and a recombinant NL4.3 strain. [19] The extremely low antiviral activity of BI224436 evaluated using the TZM-bl system suggests that this assay may not be suitable for assessment of NCINI e cacy. ... ... Our data are consistent with previous reports that BI 224436 has an EC 50 value > 50 µM and does not inhibit strand transfer. [19, 20] ... Screening of Non-catalytic Integrase Inhibitors for Anti-HIV Drug Development Minjee Kim Seoung Eun Bae Young Bong Kim Background: Integrase (IN) is an essential protein for HIV replication that catalyzes insertion of the reverse-transcribed viral genome into the host chromosome during the early steps of viral infection. Highly active anti-retroviral therapy (HAART) is a HIV/AIDS treatment method that combines three or more antiviral drugs often formulated from compounds that inhibit the activities of viral reverse transcriptase and protease enzymes. Early IN inhibitors (INIs) mainly serve as integrase strand transfer inhibitors (INSTI) that disrupt strand transfer by binding the catalytic core domain (CCD) of IN. However, mutations of IN can confer resistance to INSTI. Therefore, non-catalytic integrase inhibitors (NCINI) have been developed as next-generation INIs. Methods: In this study, we evaluated and compared the activity of INSTI and NCINI according to the analysis method. Antiviral activity was compared using p24 ELISA with MT2 cell and TZM-bl luciferase system with TZM-bl cell. Each drug was serially diluted and treated to MT2 and TZM-b1 cells, infected with HIV-1 AD8 strain and incubated for 5 and 2 days, respectively. Additionally, to analyze properties of INSTI and NCINI, transfer inhibition assay and 3'-processing inhibition assay were performed. Results: During screening of INIs using the p24 ELISA and TZM-bl luciferase systems, we found an inconsistent result with INSTI and NCINI drugs. Following infection of MT2 and TZM-bl cells with T-tropic HIV-1 strain, both INSTI and NCINI treatments induced significant p24 reduction in MT2 cells. However, NCINI showed no antiviral activity in the TZM-bl luciferase system, indicating that this widely used and convenient antiretroviral assay is not suitable for screening of NCINI compounds that target the second round of HIV-1 replication. Conclusion: Accordingly, we recommend application of other assay procedures, such as p24 ELISA or reverse transcription activity, in lieu of the TZM-bl luciferase system for preliminary NCINI drug screening. Utilization of appropriate analytical methods based on underlying mechanisms is necessary for accurate assessment of drug efficacy. View Show abstract ... ALLINIs were discovered via two different high throughput approaches. One was based on a "wet bench" screen for IN 3 processing activity [211] . The other, an in silico screen, searched the co-crystal structure of the HIV-1 IN CCD dimer bound to the IN binding domain of the lentiviral IN host co-factor lens epithelium-derived growth factor (LEDGF)/p75 [212] for molecules that recapitulated molecular details of the protein-protein interaction [209]. ... ... ALLINIs stimulate IN hyper-multimerization in vitro and in virions produced from drug-treated cells [58,111,140,176,210,[222][223][224][225][226] (Figure 6B,C). Hyper-multimerized IN is defective for IN 3 processing and strand transfer activities in vitro [140,208, 211, 225,227,228] and for vRNA binding in virions [55]. Accordingly, viruses made from ALLINI-treated cells harbor a predominance of eccentric particles [7,58,176,210,223,224,226,[229][230][231] and are defective for reverse transcription [58,140,176,210,223] due to preferential degradation of IN and vRNA in infected target cells [57,111]. ... Multimodal Functionalities of HIV-1 Integrase Mamuka Kvaratskhelia Integrase is the retroviral protein responsible for integrating reverse transcripts into cellular genomes. Co-packaged with viral RNA and reverse transcriptase into capsid-encased viral cores, human immunodeficiency virus 1 (HIV-1) integrase has long been implicated in reverse transcription and virion maturation. However, the underlying mechanisms of integrase in these non-catalytic-related viral replication steps have remained elusive. Recent results have shown that integrase binds genomic RNA in virions, and that mutational or pharmacological disruption of integrase-RNA binding yields eccentric virion particles with ribonucleoprotein complexes situated outside of the capsid shell. Such viruses are defective for reverse transcription due to preferential loss of integrase and viral RNA from infected target cells. Parallel research has revealed defective integrase-RNA binding and eccentric particle formation as common features of class II integrase mutant viruses, a phenotypic grouping of viruses that display defects at steps beyond integration. In light of these new findings, we propose three new subclasses of class II mutant viruses (a, b, and c), all of which are defective for integrase-RNA binding and particle morphogenesis, but differ based on distinct underlying mechanisms exhibited by the associated integrase mutant proteins. We also assess how these findings inform the role of integrase in HIV-1 particle maturation. ... As a result, viral ribonulceoprotein complexes are mislocalized outside of the protective capsid shell in eccentric virions produced in the presence of ALLINIs [14][15][16][17][18][19][20]. While numerous attempts to discover and develop ALLINIs with various chemical scaffolds such as quinoline, benzothiazole, indole and pyridine were made [13,[21][22][23] [24] [25][26][27], none of these candidates has been successfully moved to human clinical trials. Clinical advancement of previously reported highly potent derivatives such as GS-9822 was primarily impeded by compound toxicity observed preclinically in animals [27]. ... ... Multiple chemical scaffolds such as quinoline (BI224436, Fig 1A), benzothiazole (GS-9822, Fig 1B) and pyridine (KF116, Fig 1C) were previously explored for the discovery of ALLINIs [21,23, 24, 27,28]. Here, we employed the pyrrolopyridine-based scaffold to synthesize an ALLINI candidate compound, STP0404 (Fig 1D). ... A highly potent and safe pyrrolopyridine-based allosteric HIV-1 integrase inhibitor targeting host LEDGF/p75-integrase interaction site Article Jul 2021 PLOS PATHOG Tatsuya Maehigashi Seohyun Ahn Uk-Il Kim Baek Kim Allosteric integrase inhibitors (ALLINIs) are a class of experimental anti-HIV agents that target the noncatalytic sites of the viral integrase (IN) and interfere with the IN-viral RNA interaction during viral maturation. Here, we report a highly potent and safe pyrrolopyridine-based ALLINI, STP0404, displaying picomolar IC50 in human PBMCs with a >24,000 therapeutic index against HIV-1. X-ray structural and biochemical analyses revealed that STP0404 binds to the host LEDGF/p75 protein binding pocket of the IN dimer, which induces aberrant IN oligomerization and blocks the IN-RNA interaction. Consequently, STP0404 inhibits proper localization of HIV-1 RNA genomes in viral particles during viral maturation. Y99H and A128T mutations at the LEDGF/p75 binding pocket render resistance to STP0404. Extensive in vivo pharmacological and toxicity investigations demonstrate that STP0404 harbors outstanding therapeutic and safety properties. Overall, STP0404 is a potent and first-in-class ALLINI that targets LEDGF/p75 binding site and has advanced to a human trial. ... Drug compounds that bind to inactivate the target enzyme in viruses are called inhibitors (Zimmermann et al., 2009). HIV-1 inhibitors consisting of non-catalytic site integrase inhibitor (NCINI) is an example of HIV-1 integrase inhibitors that have entered clinical trials (Fader et al., 2014) . Darunavir has bound to the amino acid residues that make up the protease's active site through hydrogen and hydrophobic bonds (Wang et al., 2011). ... ... The target compounds used in this study were catechin, catechin gallate, gallocatechin, gallocatechin gallate, (-)-epicatechin, (-)-epicatechin gallate, (-)-epigallocatechin, and (-)-epigallocatechin gallate (Wein et al., 2016;Susanto et al. 2018;Kharisma et al., 2020b). The HIV-1 antiretroviral drugs used in the study consisted of BI compound or NCINI, darunavir or protease inhibitors, and nevirapine or reverse transcriptase inhibitors (Fader et al., 2014; Bardsley-Elliot and Perry, 2000;Deek, 2014). All ligand sample preparations were retrieved on the PubChem database (pubchem.ncbi.nlm.nih.gov). ... Tea catechin as antiviral agent via apoptosis agonist and triple inhibitor mechanism against HIV-1 infection: A bioinformatics approach Full-text available Feb 2021 Muhammad Hermawan Widyananda Arif Nur Gabino Garrido Viol Dhea Kharisma Context: Human immunodeficiency virus (HIV) antiretrovirals that target the binding of viral enzyme are chosen as the lead solution in the treatment of HIV-1 infection, such as non-catalytic site integrase inhibitor (NCINI), nevirapine, and darunavir. There are natural compounds from specific plants that can be effective in treating HIV-1 infection such as tea catechin. Tea catechin administration causes a decrease in viral load and inhibition of entry mechanisms and an increased effect of apoptosis in infected cells. Aims: To identify the triple inhibitor mechanism in tea catechins against the three HIV-1 enzymes and apoptosis agonists through in silico approach as an innovation in handling HIV-1 infection. Methods: The 3D structure of tea catechin compounds from the database was examined, and then all target compounds were analyzed for drug-likeness, molecular docking, pathway prediction, and molecular interactions to determine the potential of tea catechin compounds as antiviral HIV-1 in silico. Results: Tea catechin compounds have the potential to serve as antiviral against HIV-1 through apoptosis agonist and triple inhibitor mechanisms. Apoptosis occurs due to the interaction of tea catechins with pro-apoptotic proteins in cells, and the epigallocatechin gallate (EGCG) compound is a class of tea catechins with the same binding position as control. Conclusions: The binding of the EGCG molecule complex results in low binding energy. Therefore, it allows EGCG acts as a triple inhibitor in HIV-1 infection. ... Drug compounds that bind to inactivate the target enzyme in viruses are called inhibitors (Zimmermann et al., 2009). HIV-1 inhibitors consisting of non-catalytic site integrase inhibitor (NCINI) is an example of HIV-1 integrase inhibitors that have entered clinical trials (Fader et al., 2014) . Darunavir has bound to the amino acid residues that make up the protease's active site through hydrogen and hydrophobic bonds (Wang et al., 2011). ... ... The target compounds used in this study were catechin, catechin gallate, gallocatechin, gallocatechin gallate, (-)-epicatechin, (-)-epicatechin gallate, (-)-epigallocatechin, and (-)-epigallocatechin gallate (Wein et al., 2016;Susanto et al. 2018;Kharisma et al., 2020b). The HIV-1 antiretroviral drugs used in the study consisted of BI compound or NCINI, darunavir or protease inhibitors, and nevirapine or reverse transcriptase inhibitors (Fader et al., 2014; Bardsley-Elliot and Perry, 2000;Deek, 2014). All ligand sample preparations were retrieved on the PubChem database (pubchem.ncbi.nlm.nih.gov). ... Tea catechin as antiviral agent via apoptosis agonist and triple inhibitor mechanism against HIV-1 infection: A bioinformatics approach Viol Dhea Kharisma Muhammad Widyananda Aondohemba Nege Alexander Nugraha Context: Human immunodeficiency virus (HIV) antiretrovirals that target the binding of viral enzyme are chosen as the lead solution in the treatment of HIV-1 infection, such as non-catalytic site integrase inhibitor (NCINI), nevirapine, and darunavir. There are natural compounds from specific plants that can be effective in treating HIV-1 infection such as tea catechin. Tea catechin administration causes a decrease in viral load and inhibition of entry mechanisms and an increased effect of apoptosis in infected cells. Aims: To identify the triple inhibitor mechanism in tea catechins against the three HIV-1 enzymes and apoptosis agonists through in silico approach as an innovation in handling HIV-1 infection. Methods: The 3D structure of tea catechin compounds from the database was examined, and then all target compounds were analyzed for drug-likeness, molecular docking, pathway prediction, and molecular interactions to determine the potential of tea catechin compounds as antiviral HIV-1 in silico. Results: Tea catechin compounds have the potential to serve as antiviral against HIV-1 through apoptosis agonist and triple inhibitor mechanisms. Apoptosis occurs due to the interaction of tea catechins with pro-apoptotic proteins in cells, and the epigallocatechin gallate (EGCG) compound is a class of tea catechins with the same binding position as control. Conclusions: The binding of the EGCG molecule complex results in low binding energy. Therefore, it allows EGCG acts as a triple inhibitor in HIV-1 infection. ... The allosteric inhibitors of integrase (ALLINIs) [86] [87] [88] (Fig. 4). ALLINI-induced aggregation is NaCl-dependent, so we tested aggregation at NaCl concentrations from 250 mM to 1 M. ... ... ALLINI-induced aggregation is NaCl-dependent, so we tested aggregation at NaCl concentrations from 250 mM to 1 M. At 1 M NaCl, no aggregation was observed by the ALLINIs BI-224436 [87] , BI-D [90], or CX04328 (Compound 6 from Christ et al. [86]). At NaCl concentrations where ALLINI-induced aggregation was observed, ALLINIs induced equal or greater aggregation of IN 1F as (Fig. 5a). ... Influence of the amino-terminal sequence on the structure and function of HIV integrase Background: Antiretroviral therapy (ART) can mitigate the morbidity and mortality caused by the human immunodeficiency virus (HIV). Successful development of ART can be accelerated by accurate structural and biochemical data on targets and their responses to inhibitors. One important ART target, HIV integrase (IN), has historically been studied in vitro in a modified form adapted to bacterial overexpression, with a methionine or a longer fusion protein sequence at the N-terminus. In contrast, IN present in viral particles is produced by proteolytic cleavage of the Pol polyprotein, which leaves a phenylalanine at the N-terminus (IN 1F). Inspection of available structures suggested that added residues on the N-terminus might disrupt proper protein folding and formation of multimeric complexes. Results: We purified HIV-1 IN 1F1-212 and solved its structure at 2.4 Å resolution, which showed extension of an N-terminal helix compared to the published structure of IN1-212. Full-length IN 1F showed increased in vitro catalytic activity in assays of coupled joining of the two viral DNA ends compared to two IN variants containing additional N-terminal residues. IN 1F was also altered in its sensitivity to inhibitors, showing decreased sensitivity to the strand-transfer inhibitor raltegravir and increased sensitivity to allosteric integrase inhibitors. In solution, IN 1F exists as monomers and dimers, in contrast to other IN preparations which exist as higher-order oligomers. Conclusions: The structural, biochemical, and biophysical characterization of IN 1F reveals the conformation of the native HIV-1 IN N-terminus and accompanying unique biochemical and biophysical properties. IN 1F thus represents an improved reagent for use in integration reactions in vitro and the development of antiretroviral agents. ... We retained two Mg 21 ions from the crystal structures. Hydrogen atoms were added to the HIV-1 IN, BI-D, and PIR inhibitors while retaining the (S) form chirality of the BI-D and PIR inhibitors, as the (S) form was determined to be more predominant and bioactive than was the (R) form for these compounds (17, 72) . Further, we solvated each system in a periodic box of TIP3P water molecules and ionized them with Na 1 and Cl 2 ions to a salt concentration of 150 mM (Fig. S3B and D) using VMD software (73). ... The Drug-Induced Interface That Drives HIV-1 Integrase Hypermultimerization and Loss of Function Full-text available Feb 2023 Allosteric HIV-1 integrase (IN) inhibitors (ALLINIs) are an emerging class of small molecules that disrupt viral maturation by inducing the aberrant multimerization of IN. Here, we present cocrystal structures of HIV-1 IN with two potent ALLINIs, namely, BI-D and the drug candidate Pirmitegravir. The structures reveal atomistic details of the ALLINI-induced interface between the HIV-1 IN catalytic core and carboxyl-terminal domains (CCD and CTD). Projecting from their principal binding pocket on the IN CCD dimer, the compounds act as molecular glue by engaging a triad of invariant HIV-1 IN CTD residues, namely, Tyr226, Trp235, and Lys266, to nucleate the CTD-CCD interaction. The drug-induced interface involves the CTD SH3-like fold and extends to the beginning of the IN carboxyl-terminal tail region. We show that mutations of HIV-1 IN CTD residues that participate in the interface with the CCD greatly reduce the IN-aggregation properties of Pirmitegravir. Our results explain the mechanism of the ALLINI-induced condensation of HIV-1 IN and provide a reliable template for the rational development of this series of antiretrovirals through the optimization of their key contacts with the viral target. ... Allosteric inhibitors target different sites on the integrase CCD domain, such as ALLINIs (allosteric integrase inhibitors) that bind to the dimer of HIV-1 integrase and interact with a host cell factor called lens epithelial-derived growth factor (LEDGF), essential for integration [83][84][85][86]. The peptide-type inhibitors of LEDGF interaction and multimerization as BI-224436, have also been reported in several studies and represent a new class of non-catalytic site inhibitors (NCINIs) that have entered phase 1 clinical trials for the first time [87, 88] . Later, Mitchell et al. (2017) found next generation NCINIs (GS9695 and GS9822) with higher potencies than previous NCINIs. ... Implications of Fragment-Based Drug Discovery in Tuberculosis and HIV Tuberculosis (TB) remains a global health problem and the emergence of HIV has further worsened it. Long chemotherapy and the emergence of drug-resistance strains of Mycobacterium tuberculosis as well as HIV has aggravated the problem. This demands urgent the need to develop new anti-tuberculosis and antiretrovirals to treat TB and HIV. The lack of diversity in drugs designed using traditional approaches is a major disadvantage and limits the treatment options. Therefore, new technologies and approaches are required to solve the current issues and enhance the production of drugs. Interestingly, fragment-based drug discovery (FBDD) has gained an advantage over high-throughput screenings as FBDD has enabled rapid and efficient progress to develop potent small molecule compounds that specifically bind to the target. Several potent inhibitor compounds of various targets have been developed using FBDD approach and some of them are under progression to clinical trials. In this review, we emphasize some of the important targets of mycobacteria and HIV. We also discussed about the target-based druggable molecules that are identified using the FBDD approach, use of these druggable molecules to identify novel binding sites on the target and assays used to evaluate inhibitory activities of these identified druggable molecules on the biological activity of the targets. ... When present in target cells during the early phase of infection, ALLINIs can inhibit the overall level of integration and suppress the frequency at which genes are targeted [54][55][56]82]. ALLINIs engage the HIV-1 IN CCD dimer interface at a location coincident with LEDGF/p75 and HDGFL2 binding [36,40,51,53,54,61,[82][83] [84] [85], but the roles of these virus-host interactions in ALLINImediated integration retargeting have not been systematically investigated. To address this information gap, we created HKO (for HDGFL2 knockout) cells and infected these alongside isogenic WT, LKO, and DKO HEK293T cells ( Figure 1A) [62,63]. ... Allosteric Integrase Inhibitor Influences on HIV-1 Integration and Roles of LEDGF/p75 and HDGFL2 Host Factors Allosteric integrase (IN) inhibitors (ALLINIs), which are promising preclinical compounds that engage the lens epithelium-derived growth factor (LEDGF)/p75 binding site on IN, can inhibit different aspects of human immunodeficiency virus 1 (HIV-1) replication. During the late phase of replication, ALLINIs induce aberrant IN hyper-multimerization, the consequences of which disrupt IN binding to genomic RNA and virus particle morphogenesis. During the early phase of infection, ALLINIs can suppress HIV-1 integration into host genes, which is also observed in LEDGF/p75-depelted cells. Despite this similarity, the roles of LEDGF/p75 and its paralog hepatoma-derived growth factor like 2 (HDGFL2) in ALLINI-mediated integration retargeting are untested. Herein, we mapped integration sites in cells knocked out for LEDGF/p75, HDGFL2, or both factors, which revealed that these two proteins in large part account for ALLINI-mediated integration retargeting during the early phase of infection. We also determined that ALLINI-treated viruses are defective during the subsequent round of infection for integration into genes associated with speckle-associated domains, which are naturally highly targeted for HIV-1 integration. Class II IN mutant viruses with alterations distal from the LEDGF/p75 binding site moreover shared this integration retargeting phenotype. Altogether, our findings help to inform the molecular bases and consequences of ALLINI action. ... Allosteric IN inhibitors (ALLINIs, also known as NCINIs, LEDGINs, or INLAIs) are a promising class of anti-retroviral drugs [15][16][17] [18] [19][20]. ALLINIs interact with the dimer interface of the IN catalytic core domain and induce aberrant IN multimerization [21][22][23][24][25][26]. ... HIV-1 integrase binding to genomic RNA 5′-UTR induces local structural changes in vitro and in virio Pratibha C. Koneru Wen Li Background During HIV-1 maturation, Gag and Gag-Pol polyproteins are proteolytically cleaved and the capsid protein polymerizes to form the honeycomb capsid lattice. HIV-1 integrase (IN) binds the viral genomic RNA (gRNA) and impairment of IN-gRNA binding leads to mis-localization of the nucleocapsid protein (NC)-condensed viral ribonucleoprotein complex outside the capsid core. IN and NC were previously demonstrated to bind to the gRNA in an orthogonal manner in virio; however, the effect of IN binding alone or simultaneous binding of both proteins on gRNA structure is not yet well understood. Results Using crosslinking-coupled selective 2′-hydroxyl acylation analyzed by primer extension (XL-SHAPE), we characterized the interaction of IN and NC with the HIV-1 gRNA 5′-untranslated region (5′-UTR). NC preferentially bound to the packaging signal (Psi) and a UG-rich region in U5, irrespective of the presence of IN. IN alone also bound to Psi but pre-incubation with NC largely abolished this interaction. In contrast, IN specifically bound to and affected the nucleotide (nt) dynamics of the apical loop of the transactivation response element (TAR) and the polyA hairpin even in the presence of NC. SHAPE probing of the 5′-UTR RNA in virions produced from allosteric IN inhibitor (ALLINI)-treated cells revealed that while the global secondary structure of the 5′-UTR remained unaltered, the inhibitor treatment induced local reactivity differences, including changes in the apical loop of TAR that are consistent with the in vitro results. Conclusions Overall, the binding interactions of NC and IN with the 5′-UTR are largely orthogonal in vitro . This study, together with previous probing experiments, suggests that IN and NC binding in vitro and in virio lead to only local structural changes in the regions of the 5′-UTR probed here. Accordingly, disruption of IN-gRNA binding by ALLINI treatment results in local rather than global secondary structure changes of the 5′-UTR in eccentric virus particles. Graphical Abstract ... With this new understanding of the antiviral effects of NCINIs and the continuing need for new antiviral mechanisms to treat HIV, there is increased interest in further optimization and subsequent clinical development of these compounds as a novel class of antiretrovirals with a unique, orthogonal mechanism of action. One of the first NCINIs to enter the clinic was BI-224436 (Fader et al., 2014) . ... Species-Specific Urothelial Toxicity With an Anti-HIV Noncatalytic Site Integrase Inhibitor (NCINI) Is Related to Unusual pH-Dependent Physicochemical Changes GS-9695 and GS-9822 are next generation Non-Catalytic Site Integrase Inhibitors (NCINIs) with significantly improved potency against HIV compared with previous drugs such as BI-224436. Development stopped due to vacuolation of the bladder urothelium seen in cynomolgus monkey but not in rat; this lesion was absent in equivalent preclinical studies with BI-224436 (tested in dog and rat). Lesions were unlikely to be attributable to target since NCINIs specifically target viral integrase protein and no mammalian homologue is known. Secondary pharmacology studies, mitochondrial toxicity studies, immunophenotyping and analysis of proteins implicated in cell-cell interactions and/or bladder integrity (e-cadherin, pan-cytokeratin, uroplakins) failed to offer any plausible explanation for the species-specificity of the lesion. Since it was characterized by inflammation and disruption of urothelial morphology, we investigated physicochemical changes in the bladder of cynomolgus monkey (urinary pH 5.5-7.4) that might not occur in the bladder of rats (urinary pH 7.3-8.5). In measurements of surface activity, GS-9822 showed an unusual transition from a monolayer to a bilayer at the air/water interface with decreasing pH, attributed to the strong association between drug molecules in adjacent bilayer leaflets and expected to be highly disruptive to the urothelium. Structural analysis of GS-9822 and GS-9695 showed zwitterionic characteristics over the range of pH expected in cynomolgus monkey but not rat urine. This exotic surface behaviour is unlikely with BI-224436 since it would transition from neutral to cationic (never zwitterionic) with decreasing pH. These data provide useful insights to guide discovery and development of NCINIs, related compounds and zwitterions. ... The quinoline-based compounds BI-224436 25 We have tested this hypothesis by using the thermodynamic cycle in Fig. 4 to compute the free energy changes caused by the modifications in the binding of the new compounds with the CCD dimer (denoted as CCD2), and with the CCD2-CTD multimer interface. Table 1 shows the changes in the binding free energy for the CCD dimer, which are computed using the FEP cycle that corresponds to the left half of Fig. 4: As can be seen from Table 1, the changes caused by the additions to C7 or nearby positions to the binding affinity for the CCD dimer are generally benign. ... Computational design of small molecular modulators of protein–protein interactions with a novel thermodynamic cycle: Allosteric inhibitors of HIV ‐1 integrase Article Dec 2020 Appl. WO 2009/062289 (22) Molla, A.; Vasavanonda, S.; Kumar, G.; Sham, H. L.; Johnson, M.; Grabowski, B.; Denissen, J. F.; Kohlbrenner, W.; Plattner, J. J.; (16) Parts of this work have been presented at the following conferences: (a) Fenwick J. Biol. Chem. 2003, 278, 33528−33539. (16) Parts of this work have been presented at the following conferences: (a) Fenwick, C.; Bethell, R.; Bonneau, P.; Duan, J.; http://www.who.int/mediacentre/factsheets/fs360/en/index. html (accessed REFERENCES (1) World Health Organization. Factsheet No. 360, updated June 30, 2013. http://www.who.int/mediacentre/factsheets/fs360/en/index. html (accessed July 14, 2013). (2) Palella, F. J., Jr.; Delaney, K. M.; Moorman, A. C.; Loveless, M.

https://www.researchgate.net/publication/263961996_Discovery_of_BI_224436_a_noncatalytic_site_integrase_inhibitor_NCINI_of_HIV-1

Angelucci v. Fairmont General Hospital, No. 32161 IN THE SUPREME COURT OF APPEALS OF WEST VIRGINIA January 2005 Term _____________ No. 32161 _____________ MICHAEL ANGELUCCI, Plaintiff Below, Appellant v. FAIRMONT GENERAL HOSPITAL, INC., Defendant and Third-Party Plaintiff Below, Appellee v. NATIONAL ASSOCIATION OF LETTER CARRIERS HEALTH BENEFIT PLAN, Third-Party Defendant Below _____________________________________________________ Appeal from the Circuit Court of Marion County Honorable Fred L. Fox, II, Judge Case No. 02-C-282 AFFIRMED _____________________________________________________ Filed: July 1, 2005 Brent E. Beveridge, Esq. Beveridge Law Offices Fairmont, West Virginia Attorney for the Appellant Stephen M. LaCagnin, Esq. Shannon Smith Wolfe, Esq. Parween S. Mascari, Esq. Jackson Kelly, PLLC Morgantown, West Virginia Attorneys for the Appellee The Opinion of the Court was delivered PER CURIAM. CHIEF JUSTICE ALBRIGHT and JUSTICE STARCHER dissent and reserve the right to file dissenting opinions. SYLLABUS BY THE COURT 1. “The summary judgment procedure provided by Rule 56 of the West Virginia Rules of Civil Procedure does not infringe the constitutional right of a party to a trial by jury; it is not a substitute for a trial or a trial either by a jury or by the court of an issue of fact, but is a determination that, as a matter of law, there is no issue of fact to be tried.” Syl. pt. 7, Petros v. Kellas , 146 W.Va. 619, 122 S.E.2d 177 (1961). 2. “A motion for summary judgment should be granted if the pleadings, exhibits and discovery depositions upon which the motion is submitted for decision disclose that the case involves no genuine issue as to any material fact and that the party who made the motion is entitled to a judgment as a matter of law.” Syl. pt. 5, Wilkinson v. Searls , 155 W.Va. 475, 184 S.E.2d 735 (1971). 3. “Although our standard of review for summary judgment remains de novo , a circuit court's order granting summary judgment must set out factual findings sufficient to permit meaningful appellate review. Findings of fact, by necessity, include those facts which the circuit court finds relevant, determinative of the issues and undisputed.” Syl. pt. 3, Fayette County National Bank v. Lilly , 199 W.Va. 349, 484 S.E.2d 232 (1997). Per Curiam: This action is before this Court upon the appeal of the appellant and plaintiff below, Michael Angelucci, from the March 22, 2004, order of the Circuit Court of Marion County, West Virginia, granting summary judgment in favor of the appellee and defendant below, Fairmont General Hospital, Inc. The controversy concerns the appellant's indebtedness to the Hospital in the amount of $1,663.80 for cardiac rehabilitation services. In the complaint, the appellant alleged that the Hospital breached a contractual obligation to him by failing to submit the bills for the rehabilitation services to his medical benefit provider and that the Hospital wrongfully and maliciously disclosed the debt to various credit reporting companies. In granting summary judgment for the Hospital, the Circuit Court set forth findings of fact and conclusions of law, thereby confirming the appellant's responsibility to pay the $1,663.80. This Court has before it the petition for appeal, all matters of record and the memoranda of law filed by counsel. Upon review, this Court notes that the appellant signed an agreement at the Hospital, prior to receiving treatment, which stated that he would be “directly responsible” for the payment for Hospital services not paid by his medical benefit provider. The appellant received cardiac rehabilitation services at the Hospital and does not dispute the validity or the amount of the resulting debt. Moreover, the Circuit Court determined that the Hospital, in fact, submitted invoices for the services to the appellant's medical benefit provider. The $1,663.80, however, was never paid by the provider or by the appellant. Consequently, this Court is of the opinion that the Circuit Court was warranted in granting summary judgment in favor of the Hospital. Accordingly, the March 22, 2004, order of the Circuit Court of Marion County is affirmed. I. Factual and Procedural Background From September 10, 1997 through December 19, 1997, the appellant, Michael Angelucci, received cardiac rehabilitation services from the appellee, Fairmont General Hospital, Inc., located in Marion County, West Virginia. On the first day, September 10, 1997, the appellant executed an agreement assigning the Hospital his right to benefits from his medical benefit provider for medical services. The agreement also provided that, in the event bills for such services were not paid by the benefit provider or by insurance, the appellant would be “directly responsible.” At that time, the appellant was a member of the National Association of Letter Carriers (“NALC”) and was entitled to coverage under the NALC Health Benefit Plan. Fairmont General Hospital qualified as a preferred organization under the Plan, and it is undisputed that coverage was to be provided with respect to the cardiac rehabilitation services received by the appellant. (See footnote 1) According to the Hospital, four invoices for the appellant's cardiac rehabilitation services, totaling $1,947.00, were electronically transmitted to the NALC Health Benefit Plan between October 1997 and January 1998. The invoices were in the amounts of $354.00, $708.00, $590.00 and $295.00 respectively, and copies thereof were sent to the appellant. As the record indicates, NALC received the latter invoice in the amount of $295.00, and made a partial payment of $283.20, leaving a total balance due in the amount of $1,663.80. NALC asserted, however, that it never received the earlier invoices and that, pursuant to the terms of the Health Benefit Plan, it is now too late to pay them. An on-going dispute then ensued between the appellant and the Hospital concerning whether the Hospital had, in fact, submitted the bills to the NALC Health Benefit Plan for payment. Subsequently, the Hospital placed the account with collection agencies, and the debt was disclosed to various credit reporting companies. In October 2001, the appellant completed a bank loan application to refinance his home. According to the appellant, it was during the processing of the application that he first learned he was mistaken in assuming that the $1,663.80 debt had been resolved between the Hospital and the NALC Health Benefit Plan. The appellant also learned at that time that the debt had been disclosed to the credit reporting companies. The appellant asserts that, although the bank, Branch Banking & Trust Co. (BB&T), approved the loan, he was denied the optimum interest rate because of the $1,663.80 debt. (See footnote 2) On October 23, 2002, the appellant filed an action in the Circuit Court of Marion County against the appellee, Fairmont General Hospital, Inc. The complaint set forth three counts alleging: (1) that the Hospital breached a contractual obligation to the appellant by failing to submit the bills for cardiac rehabilitation services to his medical benefit provider, the NALC Medical Benefit Plan, (2) that the Hospital wrongfully and maliciously disclosed the debt to various credit reporting companies and (3) that the appellant is thus entitled to damages and injunctive relief requiring the Hospital to facilitate the removal of the $1,663.80 debt from the records of the credit reporting companies. The Hospital filed an answer denying the allegations of the complaint and the relief requested therein. In addition, the Hospital filed a counterclaim alleging that the appellant is liable for the $1,663.80 debt. (See footnote 3) Subsequently, the Hospital filed a motion for summary judgment. Following a consideration of the appellant's response and a hearing conducted in January 2004, the Circuit Court granted the motion and entered summary judgment in favor of the Hospital. The order of March 22, 2004, setting forth findings of fact and conclusions of law, states in part as follows: The Court is of the opinion that it is highly questionable as to whether or not there was a “contract” between Mr. Angelucci and FGH [Fairmont General Hospital, Inc.] regarding the submission of his bills to his insurance carrier [NALC]. Mr. Angelucci cannot recall showing FGH staff his insurance card or completing paperwork to prove an arrangement for FGH to directly submit his medical bills to NALC. * * * On the other hand, FGH did submit Mr. Angelucci's bills to his insurance company for payment. * * * Additionally, when Mr. Angelucci received cardiac services from FGH, he signed the . . . form acknowledging responsibility for any expenses which his insurance company did not pay. Mr. Angelucci has produced no evidence of fraud, intent or malice on the part of FGH and no evidence that FGH knowingly breached any duty to him. * * * Mr. Angelucci acknowledges the debt and that FGH had every right to submit the bills to a collection agency when they remained unpaid. Thus, FGH was under no duty to remove a debt with accurate information from Mr. Angelucci's credit report. * * * Mr. Angelucci's debt was valid, and FGH had a right to take action in its collection of the debt. Furthermore, Mr. Angelucci has provided no proof his credit rating would be higher in the absence of the FGH debt. II. Discussion Pursuant to Rule 56 of the West Virginia Rules of Civil Procedure, summary judgment is proper where the record demonstrates “that there is no genuine issue as to any material fact and that the moving party is entitled to a judgment as a matter of law.” Mueller v. American Electric Power Energy Services , 214 W.Va. 390, 392-93, 589 S.E.2d 532, 534- 35 (2003); 11A M.J., Judgments and Decrees , § 217.1 (Michie 1997). As this Court explained in syllabus point 7 of Petros v. Kellas , 146 W.Va. 619, 122 S.E.2d 177 (1961): The summary judgment procedure provided by Rule 56 of the West Virginia Rules of Civil Procedure does not infringe the constitutional right of a party to a trial by jury; it is not a substitute for a trial or a trial either by a jury or by the court of an issue of fact, but is a determination that, as a matter of law, there is no issue of fact to be tried. Syl. pt. 3, Harrison v. Town of Eleanor , 191 W.Va. 611, 447 S.E.2d 546 (1994). See also , syl. pt. 7, Aetna Casualty and Surety Company v. Federal Insurance Company of New York , 148 W.Va. 160, 133 S.E.2d 770 (1963). Specifically, syllabus point 5 of Wilkinson v. Searls , 155 W.Va. 475, 184 S.E.2d 735 (1971), holds: A motion for summary judgment should be granted if the pleadings, exhibits and discovery depositions upon which the motion is submitted for decision disclose that the case involves no genuine issue as to any material fact and that the party who made the motion is entitled to a judgment as a matter of law. Syl., Redden v. Comer , 200 W.Va. 209, 488 S.E.2d 484 (1997); syl. pt. 1, Wayne County Bank v. Hodges , 175 W.Va. 723, 338 S.E.2d 202 (1985). Upon appeal, the entry of a summary judgment is reviewed by this Court de novo . Redden , supra , 200 W.Va. at 211, 488 S.E.2d at 486; syl. pt. 1, Koffler v. City of Huntington , 196 W.Va. 202, 469 S.E.2d 645 (1996); syl. pt. 1, Painter v. Peavy , 192 W.Va. 189, 451 S.E.2d 755 (1994). Nevertheless, as this Court stated in syllabus point 3 of Fayette County National Bank v. Lilly , 199 W.Va. 349, 484 S.E.2d 232 (1997): “Although our standard of review for summary judgment remains de novo , a circuit court's order granting summary judgment must set out factual findings sufficient to permit meaningful appellate review. Findings of fact, by necessity, include those facts which the circuit court finds relevant, determinative of the issues and undisputed.” Syl., Hively v. Merrifield , 212 W.Va. 804, 575 S.E.2d 414 (2002); syl. pt. 3, Glover v. St. Mary's Hospital , 209 W.Va. 695, 551 S.E.2d 31 (2001); syl. pt. 2, State ex rel. Department of Health and Human Resources v. Kaufman , 203 W.Va. 56, 506 S.E.2d 93 (1998). In the action now to be determined, the appellant executed an agreement in September 1997assigning the Hospital his right to benefits from the NALC Health Benefit Plan for medical services. The agreement also provided that, in the event bills for such services were not paid through such coverage or by insurance, the appellant would be “directly responsible.” (See footnote 4) As the agreement stated: It is understood, whether I sign as agent, patient, or as “Guarantor” that I am directly responsible and will pay for services rendered and not paid by insurance. An assignment of benefits of any insurance policy or medical reimbursement plan shall not be deemed a waiver [of] the Hospital's right to require payment directly from the undersigned or the patient. The Hospital expressly reserves the right to require such payment. The agreement concluded as follows: “I have read this form and understand its content. I have had an opportunity to ask questions which have been answered to my satisfaction.” During his subsequent deposition, the appellant acknowledged his ultimate responsibility to pay the Hospital for the cardiac rehabilitation services. (See footnote 5) Here, neither the validity of the debt nor the amount thereof are in dispute. As stated above, the original amount payable, $1,947.00, was reduced to $1,663.80 through the $283.20 payment made by the NALC Health Benefit Plan. Importantly, the Circuit Court indicated that NALC had received all the invoices relating to the debt. As the March 22, 2004, order states: “FGH did submit Mr. Angelucci's bills to his insurance company for payment.” That determination is supported by the affidavit of the Hospital's Director of Patient Accounting filed in the Circuit Court. The affidavit states in part: FGH electronically submitted invoices to NALC for services rendered to the [appellant] on a monthly basis by downloading invoices from its medical billing system, MEDITECH, to an electronic billing system, QUADAX. QUADAX then electronically forwards claims to the respective clearinghouse used by NALC. FGH tracks the submission of invoices through its MEDITECH computer system, which automatically creates a log of transactions once they are electronically forwarded to QUADAX for submission to NALC or any other insurance company. Based upon the record automatically created by MEDITECH, FGH submitted four invoices for the [appellant's] cardiac rehabilitation services to QUADAX for submission to NALC: (1) a claim dated 9/30/97, submitted 10/01/97 in the amount of $354.00; (2) a claim dated 10/31/97, submitted on 11/03/97 in the amount of $708.00; (3) a claim dated 11/30/97, submitted on 12/01/97 in the amount of $590.00; and (4) a claim dated 12/31/97, submitted on 01/01/98 in the amount of $295.00. The assertion of NALC that it did not receive the invoices, notwithstanding, an examination of the record reveals no basis for overturning the finding of the Circuit Court that the invoices were submitted to NALC for payment. The $1,663.80 never having been paid by either the NALC Health Benefit Plan or the appellant, the appellant's complaint was without justification in alleging that the Hospital wrongfully and maliciously disclosed the debt to the credit reporting companies. Consequently, the Circuit Court was warranted in granting summary judgment in favor of the Hospital. (See footnote 6) III. Conclusion Upon all of the above, the March 22, 2004, order of the Circuit Court of Marion County, West Virginia, granting summary judgment in favor of the appellee, Fairmont General Hospital, Inc., is affirmed. Affirmed Footnote: 1 The NALC Health Benefit Plan is a federal employees health benefit plan established under the Federal Employees Health Benefit Act. 5 U.S.C. § 8901 (1998). Footnote: 2 A letter dated March 7, 2003, from BB&T to appellant's counsel stated in part: We wrote a new mortgage for Mr. Angelucci [in] January, 2002. We were not able to offer him the best “secondary” rate at that time due to some unpaid collections, primarily, one placed by Fairmont General Hospital in the amount of $1,663. I do not recall the specific obstacles at that time, since I do not have a copy of his loan file. The bank had previously described the appellant's loan application as “high risk.” The appellant's credit reports revealed a number of other unpaid accounts. During his deposition, the appellant explained that many of the bills he had difficulty paying were the result of an illness suffered by his son. Footnote: 3 In June 2003, the Circuit Court granted the Hospital leave to file a third-party complaint against the NALC Health Benefit Plan to enable the Hospital to seek contribution with regard to the appellant's damage claims. By order entered on January 13, 2004, however, the Circuit Court dismissed the third-party complaint upon the ground that federal preemption barred the Hospital's claim against the NALC Health Benefit Plan. As stated in footnote 1 herein, the NALC Health Benefit Plan is a federal employees health benefit plan established under the Federal Employees Health Benefit Act. 5 U.S.C. § 8901 (1998). No issue concerning the dismissal of the third-party complaint has been raised in this appeal. Footnote: 4 Although the National Association of Letter Carriers and the NALC Health Benefit Plan do not constitute an insurance company, the coverage provided by the Plan and the term insurance are considered equivalent for purposes of this action. Footnote: 5 In his September 10, 2003, deposition, the appellant testified as follows: Q. Just so that our record is complete and accurate as I can make it. At the time you went to Fairmont General Hospital for your cardiac rehab services, you knew that there would be charges for those services? A. Yes, sir. Q. You went there because you wanted your insurance company to pay as much as possible toward those expenses? A. That's true. Q. Nonetheless, you also understood that these charges that were being incurred at Fairmont General Hospital were for your benefit and they were your responsibility to pay? A. Yes, sir. Footnote: 6 In so holding, this Court is not unmindful of the serious harm which can result to individuals from the wrongful or malicious disclosure of accounts to credit reporting companies. See , syllabus point 2 of Jones v. Credit Bureau of Huntington , 184 W.Va. 112, 399 S.E.2d 694 (1990), concerning the federal Fair Credit Reporting Act. In this action, however, the validity and the amount of the debt were not in dispute, and the appellant's “direct responsibility” to pay the debt was evidenced by the agreement he signed in September 1997.

http://www.courtswv.gov/supreme-Court/docs/spring2005/32161.htm

Brugia malayi: Acquired Resistance to Microfilariae in BALB/C Mice Correlates with Local Th2 Responses | Request PDF Request PDF | Brugia malayi: Acquired Resistance to Microfilariae in BALB/C Mice Correlates with Local Th2 Responses | The role of specific Th subsets in the regulation of acquired resistance to the filarial parasite Brugia malayi is not known. We examined... | Find, read and cite all the research you need on ResearchGate Brugia malayi: Acquired Resistance to Microfilariae in BALB/C Mice Correlates with Local Th2 Responses April 1993 Experimental Parasitology 76(2):200-8 DOI: 10.1006/expr.1993.1023 Authors: Abstract The role of specific Th subsets in the regulation of acquired resistance to the filarial parasite Brugia malayi is not known. We examined pathologic and cytokine responses in filarial antigen-sensitized BALB/c mice inoculated intraperitoneally with live microfilariae. Animals immunized three times with soluble microfilarial antigen demonstrated accelerated clearance of live parasites (12 +/- 5% of parasites recovered from the peritoneal cavity 4 days after inoculation vs 57 +/- 6% in controls, P < 0.001). Elimination of microfilariae by immunized mice was associated with local eosinophilia (1.5 x 10(7) eosinophils/ml peritoneal wash fluid compared with 2 x 10(5) eosinophils/ml in unimmunized animals), development of local eosinophil-containing granulomas, and elevated serum IgE levels (7.0 +/- 1.4 vs 2.1 +/- 0.9 micrograms/ml in controls, P < 0.01). CD4+ cells from the site of parasite challenge produced Th2-associated cytokines exclusively (IL-4 and IL-5, not IFN-gamma and IL-2) in response to Brugia antigen, whereas spleen and lymph node cells produced both Th1- and Th2-associated cytokines. Mice immunized a single time with microfilarial antigen did not clear parasites in this time, and peritoneal exudate cells from these animals produced IFN-gamma but not IL-5. These results indicate that acquired resistance to B. malayi microfilariae in mice is associated with induction of a Th2 and not a Th1 response at the site of parasite elimination. ... A variety of murine models has been used to examine the regulation of the humoral and cellular immune responses to these organisms (10). We and others have demonstrated that Th2 responses are induced selectively in mice inoculated with live Brugia mf or L3, and in animals immunized with soluble mf extracts (mf Ag) (11)(12)(13) (14) (15). These responses correlate with enhanced rates of elimination of the parasite (12,14). ... ... We and others have demonstrated that Th2 responses are induced selectively in mice inoculated with live Brugia mf or L3, and in animals immunized with soluble mf extracts (mf Ag) (11)(12)(13)(14)(15). These responses correlate with enhanced rates of elimination of the parasite (12, 14) . ... ... Serum IgE was measured by two-site ELISA by using mAb EM95 as trapping Ab and biotinylated BF-8 as detecting Ab (14, 15). ... IL-12 modulation of T helper responses to the filarial helminth, Brugia malayi Article May 1995 F P Heinzel F. E. Jr. Hazlett Eric Pearlman James Kazura The purpose of the current study was to evaluate the effects of exogenous IL-12 on: 1) development of the Th cell response to Brugia malayi microfilariae (mf); 2) established B. malayi-specific Th2 responses; and 3) resistance to mf. IL-12 given at the time of inoculation with live mf resulted in a switch in development from a dominant Th2 (IL-4, IL-5 > IFN-gamma) to a dominant Th1 response (IFN-gamma > IL-4, IL-5). Induction of Th1 activity by IL-12 was dependent on IFN-gamma in vivo. When mice were given IL-12 (0.5 microgram daily for 4 days) after mf Ag-specific Th2 responses had been established, Ag-driven IL-4 and IL-5 production by spleen and peritoneal cells were reduced respectively by approximately 75 to 90% and approximately 30 to 40%, IFN-gamma production was elevated, and 68% fewer eosinophils were recovered from the peritoneal cavity. In vivo depletion of IFN-gamma ablated the effects of IL-12 on both cytokine production and eosinophil recovery. IL-12 treatment of either naive or previously sensitized mice challenged intravenously with live B. malayi did not alter the rate of clearance of mf from the blood. These data indicate that IL-12 treatment suppresses induction of filarial driven Th2 responses and modulates recall responses of established Th2 cells, but does not alter elimination of blood-borne mf in nonimmune or immune mice. ... Mice infected with the microfilariae Brugia malayi or immunized with the parasite extract, on the other hand, develop a pronounced Th2 response [10] that can modulate the Th1 response to a mycobacterial antigen (PPD) by an interleukin (IL)-4-dependent mechanism [11]. ... [10] [11][12][13][14][15][16][17][18] control MoAb. Interleukin-10 neutralization, on the other hand, enhanced IFN-g synthesis in OA-and OA þ Asc-primed cells at the same extent (twofold). ... ... 1998 Blackwell Science Ltd, Scandinavian Journal of Immunology, 47, [10] [11][12][13][14][15][16][17][18] ... Immunomodulation Induced by Ascaris suum Extract in Mice: Effect of Anti‐Interleukin‐4 and Anti‐Interleukin‐10 Antibodies A P Ferreira Ises de Almeida Abrahamsohn Meg Macedo Eliana Faquim-Mauro Simultaneous immunization of mice with an Ascaris suum extract (Asc) and ovalbumin (OA) markedly affects the immune response to OA. The role of interleukin (IL)-4 and IL-10 induced by Asc immunization on the modulation of antigen-specific and mitogen-induced responses was investigated following single or combined cytokine-specific monoclonal antibody (MoAb) treatment of mice before immunization with OA + Asc. Immediate hypersensitivity reactions to aggregated OA and OA-specific immunoglobulin (Ig)G2a antibody production were completely restored only when both IL-4 and IL-10 were neutralized. These findings were associated with enhanced interferon (IFN)-gamma secretion by OA-stimulated lymph node (LN) cells. In addition, the Asc-specific cytokine response in anti-IL-4 plus anti-IL-10 MoAb treated mice was shifted towards a Th1 phenotype, with an increase in IFN-gamma and IL-2 levels and a decrease in IL-4, but not in IL-10, levels. Consequently, Asc-specific IgG2a antibody production increased, whereas IgE titres diminished in these animals. These results indicate that IL-4 and IL-10 act together in the Asc-induced mechanism of antigen-specific pansuppression. In contrast, modulation of Concanavalin A (Con A)-induced cytokine responses in Asc-immunized mice appears to be essentially mediated by an IL-4-dependent mechanism, since the neutralization of just IL-4 (and not of IL-10), either in vivo or in vitro, changed the cytokine profile from a Th2 towards a Th1 type. However, OA and Asc-specific cell responses were not modified by either anti-IL-4 or by anti-IL-4 + anti-IL-10 MoAbs in vitro treatments, suggesting that the induction of a Th2 response to Asc components concomitant to OA immunization has a strong suppressive effect on the priming stage of OA-specific Th1 type response. ... Therefore, determining the factors that dictate the appropriate development of a protective Th1 CD4 T cell response during such infections is essential for understanding the pathogenesis of these diseases as well as aiding in the design of more effective therapeutic interventions. While Th2 CD4 T cells are associated with exacerbated disease [68,69]; Trichinella spiralis [70][71][72][73], Heligmosomoides polygyrus [69], and Brugia malayi [74] . Moreover, the importance of a Th2 CD4 T cell response in protection against parasitic worm infections has been suggested in humans as well. ... ... T cell population in which transferred SMARTA transgenic CD4 T cells, specific to the LCMV derived epitope GP [61][62][63][64][65][66][67][68][69][70][71][72][73] [74] [75][76][77][78][79][80] , failed to persist following infection with a virulent recombinant L. monocytogenes strain expressing the cognate antigen whereas GP 61-80specific memory CD4 T cells were readily detected by tetramer staining within the endogenous T cell population [406]. Therefore, to address if the loss of antigen-specific CD4 T cells following S. typhimurium infection applies to all antigen-specific CD4 T cells or limited to a subset of the population, the expansion and persistence of endogenous CD4 T cells was assessed using MHC class II tetramers. ... CD4 T cells in the protection and pathogenesis of persistent Salmonella infection A Dissertation Tanner M Johanns ... In contrast to work done with other filaria models, almost every reported vaccine trial using B. malayi in mice has demonstrated efficacy. Irradiated larvae (46,114,129,131), dead larvae (117), the soluble fraction of larvae (117), live MF (117,132), killed MF (46), MF antigen (46, 133) , and SDS extracts of adults (134) were all protective against L3 infection, even though many of these antigen preparations contained no adjuvant. Specific antigens that have been tried in this model include abundant larval transcript (135,136), thioredoxin peroxidase (135), transglutaminase (137), Bm97 (138), HSP12.6␣c ... ... Protection from MF appears to be mediated via antibodies, as a monoclonal antibody was found to promote faster clearance in mice (132). A strong type 2 response has also been associated with protection from microfilaremia (133) . ... A Comprehensive, Model-Based Review of Vaccine and Repeat Infection Trials for Filariasis Article Jul 2013 CLIN MICROBIOL REV Christopher Paul Morris Holly Evans Sasha E Larsen Edward Mitre SUMMARY Filarial worms cause highly morbid diseases such as elephantiasis and river blindness. Since the 1940s, researchers have conducted vaccine trials in 27 different animal models of filariasis. Although no vaccine trial in a permissive model of filariasis has provided sterilizing immunity, great strides have been made toward developing vaccines that could block transmission, decrease pathological sequelae, or decrease susceptibility to infection. In this review, we have organized, to the best of our ability, all published filaria vaccine trials and reviewed them in the context of the animal models used. Additionally, we provide information on the life cycle, disease phenotype, concomitant immunity, and natural immunity during primary and secondary infections for 24 different filaria models. ... We chose to study the role of T helper cell subsets in protection against the filarial parasite, B. muluyi. Th2 cells have been implicated in protection against each of the life cycle stages (15,20, 21) , although different stages in the parasite life cycle exert opposing effects on T helper cell subsets (10). We have shown that despite profound phenotypic differences in the immune response to this infection between wild-type and IL-4 knockout mice, survival of each of the life cycle stages is unaltered. ... ... Th2 cells have also been implicated, by temporal association, in the rapid Mf clearance that occurs after repeated immunization (21) . However, in our experiments Mf, which evoke an opposite Th subset response to other stages, are eliminated over a similar time-course in intact and IL-4-knockout mice. ... Infection of IL-4 deficient mice with the parasite Brugia malayi demonstrates that host resistance is not dependent on aT helper 2-dominated immune response Article Jul 1995 William Gregory Rick M. Maizels Rachel A. Lawrence Judith E Allen Resistance of intact mice to infection with the filarial nematode, Brugia malayi is dependent on the presence of T cells. To investigate the role of Th2 cells in this protection, mice with a targeted disruption of the IL-4 gene were infected with different developmental stages of B. malayi. We examined the phenotypic changes in the immune response and the survival of each stage in these mice. In wild-type mice, adult female worms induce Th2 responses, characterized by antigen-specific IgG1 production, elevated IgE, and marked IL-4 secretion by splenocytes stimulated in vitro with Brugia extract. However, first stage larvae (microfilariae), induce Th1 responses with the appearance of antigen-specific IgG2a, IgG2b, and IgG3 and IFN-gamma secretion by splenocytes. Infection of IL-4-deficient mice revealed a dramatic change in the response to adult worms, with a severe reduction in IgG1 production and a corresponding increase in the production of IgG2a, IgG2b, IgG3, and IFN-gamma release. The switch to Th1-type responses was particularly marked in IL-4-deficient recipients of female worms, which continually release live microfilariae. In the absence of IL-4, down-regulation of the microfilarial-induced Th1 response does not occur. Despite these profound alterations to the immune response in IL-4-deficient mice, survival of infective larvae, adult worms, or microfilariae in the peritoneal cavity was unaffected. In mice, therefore, the prominent Th2-type response elicited by filarial parasites may not be an essential component of the host protective immune response. ... Microfilaremic individuals show high eosinophil counts in their blood [23, 24]. A similar finding was observed in rhesus macaques as well. ... Evaluation of a Multivalent Vaccine against Lymphatic Filariasis in Rhesus macaque Model Article Full-text available Nov 2014 PLOS ONE Gajalakshmi Dakshinamoorthy Agneta von Gegerfelt Hanne Andersen Elyard Ramaswamy Kalyanasundaram Lymphatic filariasis affects 120 million people worldwide and another 1.2 billion people are at risk of acquiring the infection. Chemotherapy with mass drug administration is substantially reducing the incidence of the infection. Nevertheless, an effective vaccine is needed to prevent the infection and eradicate the disease. Previously we reported that a multivalent fusion protein vaccine (rBmHAT) composed of small heat shock proteins 12.6 (HSP12.6), abundant larval transcript-2 (ALT-2) and large extracellular domain of tetraspanin (TSP LEL) could confer >95% protection against the challenge infection with Brugia malayi infective larvae (L3) in mouse and gerbil models. In this study we evaluated the immunogenicity and efficacy of rBmHAT fusion protein vaccine in a rhesus macaque model. Our results show that rBmHAT is highly immunogenic in rhesus macaques. All the vaccinated monkeys developed significant titers of antigen-specific IgG antibodies against each of the component antigens (16,000 for rBmHSP12.6), (24,000 for rBmALT-2) and (16,000 for rBmTSP-LEL). An in vitro antibody dependent cellular cytotoxicity (ADCC) assay performed using the sera samples from vaccinated monkeys showed that the anti-rBmHAT antibodies are functional with 35% killing of B. malayi L3s. Vaccinated monkeys also had antigen responding cells in the peripheral blood. Vaccine-induced protection was determined after challenging the monkeys with 500 B. malayi L3. Following challenge infection, 3 out of 5 vaccinated macaques failed to develop the infection. These three protected macaques had high titers of IgG1 antibodies and their PBMC secreted significantly high levels of IFN-γ in response to the vaccine antigens. The two vaccinated macaques that picked the infection had slightly low titers of antibodies and their PBMC secreted high levels of IL-10. Based on these findings we conclude that the rBmHAT vaccine is highly immunogenic and safe and can confer significant protection against challenge infections in rhesus macaques. ... Our results indicate a phenotype that consists of a spectrum of proinflammatory, alternatively activated and regulatory markers, which is in line with numerous reports stating the mixed Th1/Th2 response that is often seen after exposure to microfilariae [18,20,[35][36] [37] . The mixture of markers observed may be partly influenced by Wolbachia, the obligate endosymbiont found in all lifecycle stages of the filarial parasites W. bancrofti and B. malayi. ... Brugia malayi Microfilariae Induce a Regulatory Monocyte/Macrophage Phenotype That Suppresses Innate and Adaptive Immune Responses Article Full-text available Oct 2014 PLOS NEGLECT TROP D Noëlle Louise O'Regan Svenja Steinfelder Gopinath Venugopal Susanne Hartmann Background: Monocytes and macrophages contribute to the dysfunction of immune responses in human filariasis. During patent infection monocytes encounter microfilariae in the blood, an event that occurs in asymptomatically infected filariasis patients that are immunologically hyporeactive. Aim: To determine whether blood microfilariae directly act on blood monocytes and in vitro generated macrophages to induce a regulatory phenotype that interferes with innate and adaptive responses. Methodology and principal findings: Monocytes and in vitro generated macrophages from filaria non-endemic normal donors were stimulated in vitro with Brugia malayi microfilarial (Mf) lysate. We could show that monocytes stimulated with Mf lysate develop a defined regulatory phenotype, characterised by expression of the immunoregulatory markers IL-10 and PD-L1. Significantly, this regulatory phenotype was recapitulated in monocytes from Wuchereria bancrofti asymptomatically infected patients but not patients with pathology or endemic normals. Monocytes from non-endemic donors stimulated with Mf lysate directly inhibited CD4+ T cell proliferation and cytokine production (IFN-γ, IL-13 and IL-10). IFN-γ responses were restored by neutralising IL-10 or PD-1. Furthermore, macrophages stimulated with Mf lysate expressed high levels of IL-10 and had suppressed phagocytic abilities. Finally Mf lysate applied during the differentiation of macrophages in vitro interfered with macrophage abilities to respond to subsequent LPS stimulation in a selective manner. Conclusions and significance: Conclusively, our study demonstrates that Mf lysate stimulation of monocytes from healthy donors in vitro induces a regulatory phenotype, characterized by expression of PD-L1 and IL-10. This phenotype is directly reflected in monocytes from filarial patients with asymptomatic infection but not patients with pathology or endemic normals. We suggest that suppression of T cell functions typically seen in lymphatic filariasis is caused by microfilaria-modulated monocytes in an IL-10-dependent manner. Together with suppression of macrophage innate responses, this may contribute to the overall down-regulation of immune responses observed in asymptomatically infected patients. ... Interferon gamma (IFNc), a Th1 response has been shown to be essential for controlling T. gondii infection [40]. Th2 responses dominate in chronic filarial infections such as Brugia malayi (in a murine model) [41] and O. volvulus [42] and this tends to suppress Th1 responses. These observations suggest that individuals with chronic O. volvulus infection who are co-infected with T. gondii may be unable to mount an adequate protective response resulting in increased severity of disease e.g. ... Exposure to Multiple Parasites Is Associated with the Prevalence of Active Convulsive Epilepsy in Sub-Saharan Africa Article Full-text available May 2014 PLOS NEGLECT TROP D Gathoni Kamuyu Christian Bottomley JO Mageto Charles Newton Background Epilepsy is common in developing countries, and it is often associated with parasitic infections. We investigated the relationship between exposure to parasitic infections, particularly multiple infections and active convulsive epilepsy (ACE), in five sites across sub-Saharan Africa. Methods and Findings A case-control design that matched on age and location was used. Blood samples were collected from 986 prevalent cases and 1,313 age-matched community controls and tested for presence of antibodies to Onchocerca volvulus, Toxocara canis, Toxoplasma gondii, Plasmodium falciparum, Taenia solium and HIV. Exposure (seropositivity) to Onchocerca volvulus (OR = 1.98; 95%CI: 1.52–2.58, p<0.001), Toxocara canis (OR = 1.52; 95%CI: 1.23–1.87, p<0.001), Toxoplasma gondii (OR = 1.28; 95%CI: 1.04–1.56, p = 0.018) and higher antibody levels (top tertile) to Toxocara canis (OR = 1.70; 95%CI: 1.30–2.24, p<0.001) were associated with an increased prevalence of ACE. Exposure to multiple infections was common (73.8% of cases and 65.5% of controls had been exposed to two or more infections), and for T. gondii and O. volvulus co-infection, their combined effect on the prevalence of ACE, as determined by the relative excess risk due to interaction (RERI), was more than additive (T. gondii and O. volvulus, RERI = 1.19). The prevalence of T. solium antibodies was low (2.8% of cases and 2.2% of controls) and was not associated with ACE in the study areas. Conclusion This study investigates how the degree of exposure to parasites and multiple parasitic infections are associated with ACE and may explain conflicting results obtained when only seropositivity is considered. The findings from this study should be further validated. ... In some situations, these Th cell subsets may exacerbate autoimmune reactions, allergies, asthma, infectious disease states, such as inflammatory reactions or may promote chronic infections. For example, Th1 responses are beneficial to control many intracellular pathogens and fungi infections, but they could also exacerbate autoimmune diseases such as psoriasis, multiple sclerosis and rheumatoid arthritis [60][61][62][63][64]. Similarly, Th2 cell activation can control or eliminate extracellular pathogens such as Borrelia burgdorferi and helminthes, but these responses can worsen some viral infections, allergic responses, asthma and airway hypersensitivity reactions [65][66][67][68] [69] [70][71][72]. Th17 effector cells are important in an effective immune defense of infections caused by Klebsiella pneumoniae, Bacteroides fragilis, Candida albicans, as well as Mycobacterium tuberculosis [73][74][75]. ... The multifaceted role of T cell-mediated immunity in pathogenesis and resistance to mycoplasma respiratory disease Article Full-text available Jan 2009 Nicole Dobbs Adam N. Odeh Xiangle Sun Jerry William Simecka Mycoplasma respiratory diseases have a significant impact on the economy, health and wildlife. The hallmark of these diseases is the persistence of the mycoplasma infections and chronic inflammatory responses associated with the airways. There is still much that needs to be understood about the immune mechanisms involved in mycoplasma disease and resistance from infection. It is clear that immune responses can contribute to the generation of inflammatory lesions in mycoplasma respiratory disease, as well as provide protection from infection and extrapulmonary dissemination of the organisms. The evolution of this lung disease is under the control innate immune mechanisms and the contrasting effects of different T cell populations. The mechanisms of immunity involved in mycoplasma diseases are multifaceted, and a fascinating story of its complexity is being uncovered. Research in mycoplasma respiratory diseases have underscored the idea that immunity along the respiratory tract against infectious agents is a dynamic process and involves a network of cellular and cytokine signals that determine the type of responses generated, and ultimately, the outcome of infection. The aim of this article is to present on overview of our work on mycoplasma disease and immunity, focusing on the interactions and regulation of T cell responses that influence disease pathogenesis. We will first provide an overview of immune mechanisms involved in controlling infection and participate in the generation of T cell responses, and the role of T cell populations in generating protection and contributing to lesion development will be discussed. ... In vitro eosinophil-mediated immune serum-dependent destruction of helminths has been demonstrated with several worms, including microfilariae of Onchocerca volulus (Greene et al. 1981), schistosomula of Schistosoma mansoni (Butterworth et al. 1977), and Trichinella spiralis newborn larvae (Kazura and Aikawa 1980). Both jird and mouse models of Brugia malayi infections show increased numbers of eosinophils associated with immune-mediated killing of L3 and microfilariae (Yates and Higashi 1986; Pearlman et al. 1993) . The reduction of intracranial worms of Angiostrongylus cantonensis in mice and guinea pigs is associated with in-creased CSF eosinophilia (Sasaki et al. 1993). ... Strongyloides stercoralis:Eosinophil-Dependent Immune-Mediated Killing of Third Stage Larvae in BALB/cByJ Mice Article May 1996 H L Rotman Wiboonchai Yutanawiboonchai Richard A. Brigandi David Abraham Challenge worm survival was significantly reduced when BALB/cByJ mice were vaccinated against Strongyloides stercoralis infective third stage larvae (L3) regardless of whether the challenge infections consisted of systemically migrating L3 or L3 implanted in diffusion chambers. The only cell type that increased in number in diffusion chambers in immunized mice, 1 week after booster immunizations, was the eosinophil, and maximal levels of eosinophils were coincident with parasite killing. Mice were treated with mAb to eliminate IL-5 or granulocytes to assess the role that eosinophils play in larval killing. Treated animals showed no decrease in immunity when challenge infections consisted of systemically migrating L3 administered 3 weeks after booster immunizations. Eosinophil numbers in immunized mice decreased to control levels when measured 3 weeks post-booster immunization, both in diffusion chambers and in the peripheral blood, whereas they were elevated at 1 week after booster immunizations. Direct contact between host cells and L3 was, however, still required for larval killing in immunized hosts 3 weeks after booster immunizations. Elimination of eosinophils by treatment with mAb to IL-5 or granulocytes significantly reduced protective immunity, when L3 were implanted in diffusion chambers at 1 and 3 weeks post-booster. However, as systemically migrating L3 were still killed in immunized, eosinophil-depleted animals, other cell types may play a role in larval destruction. Two human eosinophil granule products were found to be toxic for host-adapted L3+, but had no effect on infective L3, indicating that host-adapted larvae are possible targets for eosinophil-mediated destruction of third stage larvae. These findings suggest that inactivation of eosinophils by mAb treatment abolishes protective immunity to L3 contained within diffusion chambers and that small numbers of eosinophils are sufficient for immune-mediated killing of S. stercoralis L3. ... Several reports have shown a dramatic decrease of IL-2 levels in subjects infected by O. volvulus compared with 'putatively immune' individuals [12,14,16]. The Th2-type responses are characteristic of most infections by helminths and may be involved in protective immunity [17] [18][19]. Th2-type cytokines promote humoral responses (IgG1, IgE) [20] and induce eosinophilia [21,22]. The high levels of IL-5 in Mf ¹ subjects could be associated with the strong eosinophilia (>30%, data not shown) encountered in loiasis patients. ... T helper responsiveness in human Loa loa infection; defective specific proliferation and cytokine production by CD4+ T cells from microfilaraemic subjects compared with amicrofilaraemics Article Jun 1997 Sylvain Baize Goetz Wahl P T Soboslay Alain Georges The proliferation and cytokine profiles of peripheral blood mononuclear cells (PBMC) from microfilaraemic (Mf+) subjects infected by Loa loa in response to antigens of several parasitic stages were compared with those from amicrofilaraemic (Mf-) individuals. While a strong lymphoproliferative response and consistent levels of both Th1 (IL-2, interferon-gamma (IFN-gamma)) and Th2 (IL-4, IL-5) type cytokines were observed in response to adult worm (AW) and microfilariae (Mf) antigen in Mf- individuals, Mf+ subjects were characterized by a T cell unresponsiveness, including proliferation, cytokine production and IL-2 mRNA expression. Conversely, T cell responsiveness to mitogens and non-specific antigen were similar in the two endemic populations. Depletion of lymphocyte subpopulations indicated that T CD4+ were mainly involved in the specific cellular response. In contrast to other cytokines, IL-10 was produced in response to all parasitic stages, in both Mf+ and Mf- patients. Neutralization of IL-10 did not restore cytokine production in Mf+ patients, while B7 mRNA expression was similar between Mf+ and Mf- subjects in response to Mf antigen, suggesting that IL-10 was not the only factor responsible for T cell unresponsiveness. Mf+ patients have lower Mf antigen-specific IgG levels compared with Mf-, and there is a significant correlation between Mf antigen-specific antibodies and IL-5 responses. These findings suggest that Mf- status is correlated with T helper responsiveness, including proliferation and production of both Th1- and Th2-type cytokines, whereas Mf+ status is characterized by unresponsiveness of the same cell population, induced and/or maintained by microfilariae. ... In S. mansoni infection that induces a predominant Th2 response after egg deposition, it was demonstrated that IL-10 produced by the Th2 cells down-regulate only Th1 cytokine secretion, mainly IFN-g, to sperm whale myoglobin and HIV glycoprotein, skewing the response towards Th2 [32,33]. Mice infected with the microfilariae Brugia malayi or immunized with the parasite extract develop a pronounced Th2 response [34] that can modulate the Th1 response to a mycobacterial antigen (PPD), increasing IL-4 and IL-5 secretion. This effect is mediated by an IL-4-, and not an IL-10-dependent mechanism [35]. ... The immunosuppressive activity of Ascaris suum is due to high molecular weight components Article Full-text available Dec 1998 Eliana Faquim-Mauro Meg Macedo The role of isolated components obtained by gel filtration chromatography of Ascaris suum body extract (Asc) on the modulation of the immune response to ovalbumin (OvA) was evaluated and correlated with the immunogenic properties of such components. We showed that high (PI), but not low (PIII), molecular weight components have the ability to inhibit OvA-induced immediate and DTH reactions, lymph node (LN) cell proliferation, cytokine (IL-2, interferon-gamma (IFN-gamma), IL-4 and IL-10) and antibody (IgG1, IgG2a, IgM and IgE) production in mice concomitantly immunized with OvA and these high mol. wt components. The pattern of cytokines synthesized in response to PI or PIII was totally different: the former induced more IL-4 and IL-10 and the latter more IL-2 and IFN-gamma. The levels of Asc-specific IgG1 antibodies were higher in mice immunized with OvA plus PI and IgG2a anti-Asc antibodies predominated in those immunized with PIII. IgE antibody production, however, was low in the former group of mice. These results indicate that the high mol. wt components present in the body extract from the helminth A. suum are responsible for its suppressive effect upon Th1- and Th2-dependent immune responses to an unrelated antigen. The suppression of the Th1-dependent parameters could be related to high-level expression of IL-4 and IL-10 induced by such components. ... Onchocerca volvulus L 3 induce a similar response leading to elevated serum IgE and recruitment of eosinophils to the site of the parasite inoculation, both of which are required for development of acquired immunity (Lange et al. 1994, Yutanawiboonchai et al. 1994. Furthermore, injection with live B. malayi Mf, or their antigens, induces production of significant levels of IL-4 and IL-5, in addition to elevated IgE and eosinophilia (Lawrence et al. 1993 , Pearlman et al. 1993a ,b, Subramanian et al. 1997. Microfilariae of other filarial species, such as the cattle parasite O. lienalis, also induce elevated levels of IL-4, IL-5, IgE and eosinophilia in mice, which are associated with resistance to challenge infection (Folkard & Bianco 1995, Folkard et al. 1996, 1997. ... Interleukin-12 and the host response to parasitic helminths; the paradoxical effect on protective immunity and immunopathology Article Dec 1998 Adrian Paul Mountford Eric Pearlman In general, helminth infections are associated with the development of dominant Th2-mediated immune responses which may be host protective but can also be the cause of immunopathology. Interleukin 12 (IL-12) is known to be a potent inhibitor of Th2 immune responses and as such it might be expected to have an important modulatory role in helminth-induced immune responses. In this review, we discuss the effect of IL-12 on susceptibility to infection, protective immunity and immunopathology, in the context of exposure to a range of helminths including intestinal nematodes, filariae and schistosomes. It is apparent that the effects of IL-12 are complex and can be beneficial as well as detrimental for the host. The precise role of IL-12 depends upon a number of factors including the type of helminth and the specific tissue involved in the inflammatory response. ... In the case of micro®lariae, the pro®le of response elicited is more questionable. A few authors implicate a`pure' Th1 response (Lawrence et al. 1995), whereas other report a combined Th1/Th2 or even a Th2 pro®le (Pearlman et al. 1993) . Two major factors may account for this paradox. ... Cellular responses to Loa loa experimental infection in mandrills (Mandrillus sphinx) vaccinated with irradiated infective larvae Article May 2000 Marie-Noëlle Ungeheuer Nohal Elissa A Morelli Pascal Millet In order to shed light on the mechanisms of antifilarial protective immunity, we investigated the course of experimental loaiosis after vaccination in a nonhuman primate host, Mandrillus sphinx. Six vaccinated (V) mandrills received 50 irradiated L3 while six nonvaccinated (NV) received saline solution on days -60, -30 and -15. All animals were challenged with 100 intact L3 (day 0). Parasitological and immunological status were followed for 9 months. Vaccination delayed the appearance and mean peak of microfilaraemia. Five mandrills (Mf-) were never microfilaraemic (one V mandrill) or microfilaraemic on only one occasion (2 V and 2 NV), the other seven having stable microfilaraemia (Mf+). The cytokine response of peripheral blood mononuclear cells to L3 (L3 Ag) was Th2 dominated, while microfilariae (Mf Ag) elicited a Th0-like response. During vaccination, Th2 cytokine production significantly increased in V mandrills against L3 Ag, as well as Mf Ag, whereas Th1 cytokines decreased. On day 60 postinoculation, cellular proliferation was higher in V mandrills in response to L3 and Mf Ags and PHA-L mitogen. At the end of prepatency (on day 130), mandrills with delayed appearance of microfilaraemia exhibited a high, transient IL-2 and IL-4 secretion in response to L3 Ag. Finally, high anti-Mf Th2 cytokine levels characterized Mf-mandrills not only during prepatency, but also (for IL-5) before immunization. However, the presence of a balanced Th1 anti-L3 response during prepatency in the amicrofilaraemic mandrill suggests its importance in protective immunity. Taken together, our data suggest that Th2 cells and also Th1 components of the antifilarial response, especially to larval antigen, may contribute to parasite elimination. ... The T cell responses of asymptomatic humans with microfilaraemia show a prominence of Th2 cytokines such as IL-4, while T cells from symptomatic patients of elephantiasis show a relative dominance of the Th1 cytokine IFN-␥, as do some putatively immune "endemic normal" individuals (2,12). However, Th2 responses appear to be protective in some experimental animal models (10,(25)(26) (27) . ... Macrophage Effector Functions Controlled by Bruton's Tyrosine Kinase Are More Crucial Than the Cytokine Balance of T Cell Responses for Microfilarial Clearance Article Full-text available Apr 2002 Sangita Mukhopadhyay Madhuchhanda Mohanty Anita Garg Mangla Balachandran Ravindran Macrophages from X-linked immunodeficient (xid) mice lacking functional Bruton's tyrosine kinase (Btk) show poor NO induction and enhanced IL-12 induction, and contribute to delayed clearance of injected microfilaria (mf) in vivo. We now show that Btk is involved in other macrophage effector functions, such as bactericidal activity and secretion of proinflammatory cytokines (TNF-alpha, IL-1beta), but not the T cell-directed cytokine IL-12. Induction of some transcriptional regulators of the NF-kappaB family, crucial for the expression of proinflammatory cytokines, is also poor in Btk-deficient macrophages. Thus, Btk appears to be involved in signaling for inducible effector functions, but not APC functions, in macrophages. Furthermore, adoptive transfer of T cells from mf-infected xid or wild-type mice did not alter the course of mf clearance in recipients, mf clearance was unaltered in IFN-gamma-deficient mice, and improved mf clearance was seen only if greater inducibility of IL-12 was accompanied by greater NO secretion from macrophages, as seen in Ity(r) C.D2 mice as compared with congenic Ity(s) BALB/c mice. Thus, delayed mf clearance in xid mice was correlated not with the high IL-12/Th1 phenotype but with low NO induction levels. Also, xid macrophages showed poor toxicity to mf in vitro as compared with wild-type macrophages. Inhibition of NO production blocked this mf cytotoxicity, and an NF-kappaB inhibitor blocked both NO induction and mf cytotoxicity. Thus, Btk is involved in inducing many macrophage effector functions, and delayed mf clearance seen in Btk-deficient xid mice is due to poor NO induction in macrophages, resulting in compromised microfilarial toxicity. ... Our previous studies in mice have shown that infective larvae (L3) and adult B. malayi stages induce strong Th2 cell responses (5). In contrast, microfilariae (Mf ) induce high levels of interferon (IFN)-γ throughout the course of infection, and it is only following chronic antigenic stimulation that Th2 responses to Mf are generated (5) (6) (7). In the context of cytokine responses observed in microfilaraemic humans, the induction of type 2 responses by L3 and adult worms may prevent Mf-induced type 1 cytokine production. ... Interferon-γ and nitric oxide production are not required for the immune-mediated clearance of Brugia malayi microfilariae in mice Article Jul 2002 Carolyn A. Gray Rachel A. Lawrence Previous research has shown that Brugia malayi microfilariae (Mf) primarily induce type 1 cytokine production, and that in-vitro nitric oxide (NO) can mediate Mf killing. This study addresses the role of interferon (IFN)-gamma-mediated immune responses in the clearance of Mf from fast-clearing (CBA/Ca) and slow-clearing (C57Bl/6) mouse strains. Analysis of spleen cell cytokine production at early timepoints p.i. showed that Mf-induced IFN-gamma and nitrite (NO-) levels were significantly greater in CBA/Ca mice than C57Bl/6 mice. However, in-vivo neutralization of IFN-gamma or inhibition of NO- production in CBA/Ca mice did not alter Mf survival kinetics. Similarly, the rate of Mf clearance in both C57Bl/6 mice lacking the IFN-gamma gene and (C57Bl/6 x 129) mice deficient in the receptor for IFN-gamma was similar to that of wild-type animals. Furthermore, the dramatic abrogation of NO- production in IFN-gammaR-/- mice suggests that Mf clearance in slow-clearing mouse strains is also independent of NO- production. Thus, in both rapid-clearing and slow-clearing mouse strains, IFN-gamma-mediated mechanisms are not a requirement for Mf clearance from the bloodstream. An Essential Role for Interleukin-5 and Eosinophils in Helminth-Induced Airway Hyperresponsiveness Article Sep 1998 Rajeev K. Mehlotra Laurie R. Hall Alan W. Higgins Eric Pearlman Infection with the parasitic helminth Brugia malayi can result in development of a severe asthmatic response termed tropical pulmonary eosinophilia. This disease, thought to result from a host inflammatory response to blood parasites which become trapped in the lung microvasculature, is characterized by a profound eosinophilic infiltration into the lungs. Recruitment of eosinophils also correlates with the development of airway hyperresponsiveness (AHR) to cholinergic agonists and severe asthmatic symptoms. Our studies examined the role of interleukin-5 (IL-5) in helminth-induced pulmonary eosinophilia and AHR. C57BL/6 mice immunized with killed B. malayi microfilariae and challenged intravenously with live microfilariae exhibit many of the characteristics of human disease, including peripheral and pulmonary eosinophilia. Cells recovered by bronchoalveolar lavage of sensitized mice consisted of 3.8% eosinophils on day 1 postchallenge and 84% on day 10. Extracellular major basic protein was present on the surface of airway epithelial cells as early as day 1 and continued to be evident after 8 days, indicating sustained activation and degranulation of eosinophils in the lung. These histologic changes correlated with the development of AHR to carbachol. In contrast to immunocompetent mice, immunization and challenge with B. malayi in IL-5 −/− mice did not induce peripheral or pulmonary eosinophilia, and these mice failed to show AHR in response to cholinergic agonists. Taken together, these data indicate that IL-5 and eosinophils are required for the induction of AHR by filarial helminths. Interaktion zwischen intestinalen Epithelzellen und immunkompetenten Zellen in der Mukosa unter normalen und pathologischen Bedingungen Chapter Jan 1995 A Stallmach Georg Köhne M Zeitz The intestinal immune system has specialized functions, which are characterized by protective production of immunoglobulin (Ig) A in responses to pathological antigens and a T cell-mediated suppression of the immune response against physiological antigens such as the intestinal microflora. The induction of an immune response requires a series of complex interactions between lymphocytes and antigen presenting cells (macrophages, dendritic cells). The immune response is mediated through the selective activation of two major CD4-positive T-cell subsets, which promote on one hand cellular effector response and on the other humoral immune response. Recently published data revealed that enterocytes can function adequately to macrophages or dendritic cells as antigen-presenting cells. Unlike conventional antigen-presenting cells, epithelial cells appear to stimulate selectively suppressor T cells. In contrast, in patients with inflammatory bowel disease the antigen-depending cell-surface receptor (appropriate T-cell receptor, MHC-molecules, integrins, accessory molecules) mediated interaction between intestinal enterocytes and lymphocytes, results in activation of CD4-positive cells. These data suggest that chronic inflammatory bowel disease may be a result of a pathologic enterocyte-lymphocyte interaction, which was followed by an inappropriate increased activation of T helper-cell subsets. Die Bedeutung von Interleukin-4 für den Verlauf einer Infektion mit der Filarie Litomosoides sigmodontis in Labormäusen Article Karin Haisch H Schulz-Key Manfred Kopf W H Hoffmann Einleitung Bei vielen Parasitosen wird die Immunantwort durch mindestens zwei verschiedene Subpo-pulationen von T-Helferzellen kontrolliert (20), die durch die Produktion antagonistisch wir-kender Zytokine charakterisiert sind (TH1/TH2-Paradigma) (16). Nematodeninfektionen indu-zieren eine TH2-dominierte Immunantwort, die bei Darmnematoden zu einer Abstoßung des Parasiten und zum Schutz des Wirtes vor Neuinfektionen führt (6). Interleukin-4 (IL-4) ist das Schlüsselzytokin der TH2-Antwort. Es induziert in murinen B-Zellen eine Differenzierung zu IgGl-und IgE-produzierenden Plasmazellen. Außerdem steigert es in TH2-Zellen die Sekre-tion von Interleukin-5 (IL-5) und Interleukin-10 (IL-10). IL-5 induziert eine Vermehrung der eosinophilen Granulozyten, während IL-10 zusammen mit IL-4 die Ausprägung einer TH1-Antwort hemmt. Da auch bei Humanfilariosen die klassischen Merkmale einer TH2~Immunantwort wie ho-he IgE-Titer und drastisch erhöhte Eosinophilie auftreten, sollte der Einfluß des TH2-Schlüs-selzytokins IL-4 auf den Verlauf einer experimentellen Filarieninfektion untersucht werden. Filarien sind allerdings streng wirtsspezifisch; deshalb können sich humanpathogene Arten nicht in Kleinsäugern entwickeln. Die Infektion von Labormäusen mit der Baumwollrattenfila-rie Litomosoides sigmodontis stellt daher ein bisher einzigartiges Filariosemodell dar (14). Dieser Parasit durchläuft eine vollständige Entwicklung in immunkompetenten BALB/c-Mäusen, für die ein großes Spektrum an immunologischen Reagenzien erhältlich ist. Die in unseren Versuchen verwendeten Mäuse wiesen eine gezielte Mutation des IL-4-Gens auf, die eine vererbbare IL-4-Defizienz verursacht. Um den immunmodulatorischen Einfluß der ver-schiedenen Parasitenstadien unabhängig voneinander unterscheiden zu können, wurden den Mäusen entweder dritte Larven inokuliert oder Mikrofilarien intravenös injiziert. Th1-Th2: Reliable paradigm or dangerous dogma? Sep 1997 Immunol Today Judith E Allen Rick M. Maizels The identification of crossregulating T helper (Th) cells has revolutionized current understanding of the immune response to infection. While paying tribute to this revolution, Judith Allen and Rick Maizels argue that the paradigm can be dangerously oversimplified and that the interaction between host and pathogen cannot always be addressed in the context of Th1 and Th2 cells. Roles for both CD+ and CD8+ T cells in protective immunity against Onchocerca lienalis microfilariae in the mouse Oct 1995 PARASITE IMMUNOL S G Folkard Ted Bianco Mice inoculated with microfilariae of the filarial nematode Onchocerca lienalis clear their parasites from the skin over a period of 3 to 4 months and are highly resistant to a challenge infection. The adoptive transfer of spleen cells at various time points following primary and secondary infections of mice shows that exposures of 50 days or greater are required for the generation of lymphocytes capable of transferring protection to naive recipients. This adoptive transfer of protection with spleen cells from infection-primed mice partitions with the T lymphocyte population. In contrast, the passive transfer of protection with spleen-derived B cells, or sera taken at various time points following infection was not achieved. Moreover, there was no detectable synergistic effect when B and T cells were co-administered to recipient animals. Depletion of CD4+ and CD8+ T cells with monoclonal antibodies shows that CD8 + T cells have some regulatory effect on parasite establishment early in primary infection, but this is later superseded by CD4+ T cell reactivity that is predominant both when primary infection microfilariae are cleared and also during resistance to reinfection. Measurement of cytokines in the sera of mice undergoing primary and secondary infections support a microfilariae-induced Th2 activity, with high levels of IL-5 that are sustained upon reinfection, and low levels oflFN- that are negligible at the time when mice are most strongly immune. Interleukin‐12 suppresses filaria‐induced pulmonary eosinophilia, deposition of major basic protein and airway hyperresponsiveness Oct 1998 PARASITE IMMUNOL Rajeev K Mehlotra Laurie R. Hall Alan W. Higgins Eric Pearlman Tropical Pulmonary Eosinophilia (TPE) is a severe form of allergic asthma caused by the host inflammatory response to filarial helminths in the lung microvasculature, and is characterized by pulmonary eosinophilia, increased filarial-specific IgG and IgE antibodies, and airway hyperresponsiveness. The current study examined the effect of IL-12 on pulmonary eosinophilia, deposition of eosinophil major basic protein and airway hyperresponsiveness in mice inoculated i.v. with Brugia malayi microfilariae. Injection of recombinant murine IL-12 modulated the T helper (Th) response in the lungs from Th2- to Th1-like, with elevated IFN-γ, and decreased IL-4 and IL-5 production. Consistent with this shift in cytokine response, antigen-specific IgG2a was elevated, and IgG1 and total serum IgE were decreased. In addition, eosinophils in BAL fluid from IL-12 treated mice were reduced from 56% to 11%, and there was no detectable MBP on respiratory epithelial cells. Importantly, IL-12 suppressed airway hyperresponsiveness compared with saline-injected control animals. Taken together, these data clearly demonstrate that by modulating Th associated cytokine production, IL-12 down-regulates filaria-induced lung immunopathology. Lymphatic filariasis: parallels between the immunology of infection in humans and mice Jul 2001 PARASITE IMMUNOL Eileen Devaney Rachel A. Lawrence Mouse models of Brugia infection have provided much useful quantitative and qualitative information on the immune response elicited by different life cycle stages of filarial worms. Many parallels exist between the immune response in the mouse and the infected human and in this review we highlight areas of topical interest, including the induction of specific cytokine resposes and their role in immunomodulation and protective immunity. These studies have reinforced the concept that different life cycle stages of filarial parasites each have their own mechanism of modulating responses so that potentially inflammatory IFN-γ responses are downregulated. While the precise mechanisms of protective immunity remain to be defined, studies in the mouse have suggested novel pathways, including a possible role for granulocytes. Antigens responsible for eosinophil hyporesponsiveness in Brugia pahangi microfilariae injected mice Oct 1997 Parasitol Int Md. Moazzem Hossain Setsuko Tsukidate Koichiro Fujita Nobuaki Akao In order to identify the eosinophil hyporesponsiveness factor in the microfilaremic host, stage-specific monoclonal antibodies against microfilariae (Mf) of Brugia pahangi were produced. One of these (MfG2a) was established for the first time as a monoclonal antibody of IgG2a isotype against Mf. MfG2a recognizes the eosinophil hyporesponsiveness factor, the 42 kDa excretory/secretory antigen of Mf. Treatment of MfG2a significantly (P < 0.05) induced eosinophil response with rapid reduction of microfilaremia in previously Mf injected mice which became amicrofilaremic within 2 weeks. Eosinophil hyporesponse was observed in the control microfilaremic mice and the microfilaremia persisted at high levels. Another monoclonal antibody, MfG1 of the IgG1 class, recognized the 64-kDa surface antigen of Mf, MfG1 was less effective in eosinophil response- or microfilaremia reduction. These data suggest that the 42-kDa microfilarial excretory/secretory antigen might be responsible for the eosinophil hyporesponsiveness in B. pahangi Mf injected mice. Zelluläre Immunreaktionen und immunregulatorische Zusammenhänge im Verlauf der experimentellen Lymphatischen Filariose [Elektronische Ressource] / Mareike Saunders Giessen, Univ., Diss., 2002. Computerdatei im Fernzugriff. Zelluläre Immunreaktionen und immunregulatorische Zusammenhänge im Verlauf der experimentellen Lymphatischen Filariose Mareike Saunders In der vorliegenden Arbeit wurden am Modell der Brugia malayi-Infektion des Nagers Mastomys coucha zelluläre Reaktionen und immunregulatorische Zusammenhänge im Verlauf der experimentellen Lymphatischen Filariose untersucht. Die Studie deckt bei quantitativ infizierten Tieren und nicht infizierten Kontrollen einen Zeitraum von 230 Tagen post infectionem (p.i.) ab. Zusätzliche Gruppen wurden am Tag 129 p.i. mit verschiedenen Filariziden behandelt und zum Vergleich ebenfalls bis zum Tag 230 p.i. untersucht. Infektions-und Therapieerfolge wurden durch die Bestimmung der Mikrofilariendichten im Blut und die abschließende Sektion der Versuchstiere kontrolliert. Periphere Blutlymphozyten (PBL) der Versuchstiere dienten als Grundlage für weiterführende Untersuchungen. Die Reaktion der PBL auf Stimulation mit B. malayi-Weibchen-, Männchen-, Mikrofilarien- und L3-Antigenen sowie dem T-Zell-Mitogen Con A wurde in Proliferationsassays anhand des Einbaus von 3H-Thymidin und über die m-RNA-Spiegel für IL-2, IL-4, IL-5, IL-10 und IFNg(gamma) in einer semiquantitativen RT-PCR überprüft. In den Proliferationsassays zeigte sich am 18. Tag p.i. bei infizierten Tieren nach spezifischer Stimulation eine gesteigerte Reaktion. Am Tag 35 p.i. und anhaltend bis Versuchsende trat sowohl eine unspezifische als auch in Reaktion auf Weibchen und Männchen-Antigen eine spezifische Suppression auf. Die chemotherapeutischen Behandlungen resultierten in einer generell gesteigerten Proliferationsleistung der PBL, wobei sich die Substanz mit makro- und mikrofilarizider Wirkung stärker auswirkte, als die beiden jeweils nur gegen ein Parasitenstadium wirksamen Medikamente. Bei den infizierten Tieren zeigten sich für die einzelnen Zytokine unterschiedliche Verläufe, wobei zusätzlich Abhängigkeiten von den eingesetzten Antigenen beobachtet wurden. Die chemotherapeutischen Behandlungen hatten keinen signifikanten Einfluß auf die Transkriptionsmuster von IL-4 und IL-10. Im Falle von IL-2 und IFNg(gamma), insbesondere aber bei IL-5 führte die Elimination der adulten Parasiten bis Versuchsende zur Steigerung der Transkription, wobei sich die mit einer frühen Abtötung der Mikrofilarien verbundene Applikation einer der Substanzen stärker auswirkte als der rein makrofilarizide Effekt einer anderen. Diese Effekte konnten vor allem nach Stimulation mit Adult-Antigenen beobachtet werden. In this work cellular responses and immunoregulatory interactions were examined in the course of experimental Brugia malayi-infection of Mastomys coucha as a model for lymphatic filariasis. The study covered a time span of 230 days post infection (p.i.) for quantitatively infected experimental animals and non-infected control animals. At 129 days p.i. additional groups were treated with the micro and macrofilaricidal CGP 20376, the macrofilaricide Flubendazol and the microfilaricide Bay 44-4400. Infection and response to treatment were followed using microfilarial counts from blood and all animals were autopsied at the end of the trial period. Harvested PBLs served as the basis for further investigations. The responses of harvested PBLs to stimulation with female-, male-, microfilarial- and L3-antigens as well as Con A were examined using 3H-thymidine incorporation as a measure of proliferation, and semi-quantitative RT-PCR to investigate regulation of IL-2, IL-4, IL-5, IL-10 and IFN-(gamma) gene transcription. CGP 20376 produced a rapid, longlasting and almost complete reduction of microfilaraemia and was also adulticidal. Flubendazol killed the adult parasites and thereby lead to a continual and eventually almost complete ablatement of parasitaemia. Bay 44-4400, which lacked adulticidal activity, produced complete microfilarial clearance within a few days. Proliferation assays showed increased responses after antigen-specific stimulation in infected animals at 18 days p.i.. From 35 days p.i. to the end of the study, the cells showed suppressed non-antigen-specific and male- and female-antigen specific responses. Treatment resulted in a general increase in proliferative responses. PBLs from animals treated with CGP 20376 produced stronger responses than PBLs from animals treated with stage specific medications. In infected animals the examined cytokines showed distinct patterns, which varied according to the stimulating antigens. In the case of IL-2, IFN-(gamma) and especially IL-5, clearance of the adult parasites lead to continuous elevation of transription, whereby CGP 20376 mediated simultaneous killing of adult and larval stages produced a stronger effect than the pure adulticide of Flubendazol. This was especially clear to see in responses to adult antigen. In contrast, killing and temporary clearance of microfilariae alone had no significant effect. Adult Brugia malayi mitochondrial and nuclear fractions impart Th1-associated sizeable protection against infective larval challenges in Mastomys coucha Full-text available Mar 2009

https://www.researchgate.net/publication/14745340_Brugia_malayi_Acquired_Resistance_to_Microfilariae_in_BALBC_Mice_Correlates_with_Local_Th2_Responses

3569 1151 2.8084 17/197-17:33:43.471 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1170 0.3569 1151 2.8084 17/197-17:48:43.507 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1170 0.3569 1151 2.8084 17/197-17:51:43.507 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1170 0.3569 1152 2.8109 17/197-18:03:43.538 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1170 0.3569 1151 2.8084 17/197-18:03:43.542 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1170 0.3569 1151 2.8084 17/197-18:05:13.542 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1170 0.3569 1152 2.8109 17/197-18:06:43.546 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1170 0.3569 1151 2.8084 17/197-18:17:13.570 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1171 0.3572 1151 2.8084 17/197-18:18:43.574 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1170 0.3569 1151 2.8084 17/197-18:18:43.578 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1170 0.3569 1151 2.8084 17/197-18:20:13.578 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1171 0.3572 1151 2.8084 17/197-18:21:43.582 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1170 0.3569 1151 2.8084 17/197-18:23:13.586 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1171 0.3572 1151 2.8084 17/197-18:24:43.590 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1170 0.3569 1151 2.8084 17/197-18:29:13.598 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1171 0.3572 1151 2.8084 17/197-18:33:43.618 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1171 0.3572 1151 2.8084 17/197-18:42:43.634 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1171 0.3572 1152 2.8109 17/197-18:44:13.638 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1171 0.3572 1151 2.8084 17/197-18:48:43.654 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1171 0.3572 1151 2.8084 17/197-18:56:13.666 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1172 0.3575 1151 2.8084 17/197-18:57:43.670 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1171 0.3572 1151 2.8084 17/197-18:59:13.674 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1172 0.3575 1151 2.8084 17/197-19:00:43.678 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1173 0.3578 1151 2.8084 17/197-19:03:43.686 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1174 0.3581 1150 2.8060 17/197-19:03:43.690 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1174 0.3581 1150 2.8060 17/197-19:06:43.694 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1175 0.3584 1150 2.8060 17/197-19:15:43.714 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1176 0.3587 1149 2.8036 17/197-19:17:13.718 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1175 0.3584 1149 2.8036 17/197-19:18:43.722 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1176 0.3587 1149 2.8036 17/197-19:27:43.746 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1176 0.3587 1148 2.8011 17/197-19:29:13.746 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1177 0.3590 1148 2.8011 17/197-19:33:43.766 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1177 0.3590 1148 2.8011 17/197-19:35:13.762 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1177 0.3590 1147 2.7987 17/197-19:39:43.774 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1178 0.3593 1147 2.7987 17/197-19:48:43.801 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1178 0.3593 1147 2.7987 17/197-19:50:13.798 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1179 0.3596 1148 2.8011 17/197-19:51:43.802 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1179 0.3596 1147 2.7987 17/197-19:54:43.809 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1180 0.3599 1146 2.7962 17/197-19:56:13.813 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1180 0.3599 1147 2.7987 17/197-19:57:43.817 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1181 0.3602 1147 2.7987 17/197-19:59:13.821 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1181 0.3602 1146 2.7962 17/197-20:00:43.825 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1182 0.3605 1146 2.7962 17/197-20:02:13.829 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1183 0.3608 1147 2.7987 17/197-20:03:43.837 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1183 0.3608 1147 2.7987 17/197-20:05:13.837 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1184 0.3611 1147 2.7987 17/197-20:06:43.841 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1198 0.3654 1147 2.7987 17/197-20:08:13.841 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1224 0.3733 1146 2.7962 17/197-20:09:43.849 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1241 0.3785 1145 2.7938 17/197-20:11:13.849 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1248 0.3806 1144 2.7914 17/197-20:12:43.853 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1244 0.3794 1142 2.7865 17/197-20:14:13.857 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1214 0.3703 1142 2.7865 17/197-20:15:43.861 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1196 0.3648 1143 2.7889 17/197-20:17:13.865 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1206 0.3678 1144 2.7914 17/197-20:18:43.869 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1218 0.3715 1144 2.7914 17/197-20:18:43.873 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1218 0.3715 1144 2.7914 17/197-20:20:13.873 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1233 0.3761 1145 2.7938 17/197-20:21:43.877 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1251 0.3816 1144 2.7914 17/197-20:23:13.881 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1270 0.3873 1144 2.7914 17/197-20:24:43.885 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1286 0.3922 1143 2.7889 17/197-20:26:13.889 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1296 0.3953 1142 2.7865 17/197-20:27:43.893 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1296 0.3953 1141 2.7840 17/197-20:29:13.893 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1288 0.3928 1139 2.7792 17/197-20:30:43.897 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1274 0.3886 1138 2.7767 17/197-20:32:13.901 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1265 0.3858 1137 2.7743 17/197-20:33:43.905 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1258 0.3837 1137 2.7743 17/197-20:33:43.913 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1258 0.3837 1137 2.7743 17/197-20:35:13.909 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1249 0.3809 1137 2.7743 17/197-20:36:43.913 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1234 0.3764 1137 2.7743 17/197-20:38:13.917 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1215 0.3706 1137 2.7743 17/197-20:39:43.921 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1207 0.3681 1138 2.7767 17/197-20:41:13.925 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1199 0.3657 1138 2.7767 17/197-20:42:43.929 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1192 0.3636 1138 2.7767 17/197-20:44:13.933 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1193 0.3639 1139 2.7792 17/197-20:45:43.937 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1203 0.3669 1139 2.7792 17/197-20:48:43.945 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1201 0.3663 1139 2.7792 17/197-20:48:43.949 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1201 0.3663 1139 2.7792 17/197-20:50:13.945 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1193 0.3639 1138 2.7767 17/197-20:51:43.949 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1181 0.3602 1138 2.7767 17/197-20:53:13.953 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1171 0.3572 1138 2.7767 17/197-20:54:43.957 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1171 0.3572 1139 2.7792 17/197-20:57:43.965 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1171 0.3572 1140 2.7816 17/197-20:59:13.969 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1170 0.3569 1140 2.7816 17/197-21:00:43.973 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1171 0.3572 1141 2.7840 17/197-21:02:13.977 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1170 0.3569 1141 2.7840 17/197-21:03:43.981 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1170 0.3569 1142 2.7865 17/197-21:03:43.985 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1170 0.3569 1142 2.7865 17/197-21:06:43.989 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1170 0.3569 1143 2.7889 17/197-21:11:13.997 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1169 0.3565 1144 2.7914 17/197-21:12:44.001 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1170 0.3569 1144 2.7914 17/197-21:14:14.005 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1169 0.3565 1145 2.7938 17/197-21:17:14.013 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF OFF OFF 1169 0.3565 1146 2.7962 17/197-21:18:44.021 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1169 0.3565 1146 2.7962 17/197-21:21:44.025 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1169 0.3565 1147 2.7987 17/197-21:23:14.029 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1169 0.3565 1148 2.8011 17/197-21:27:44.041 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1169 0.3565 1149 2.8036 17/197-21:33:44.060 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1169 0.3565 1149 2.8036 17/197-21:36:44.061 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1169 0.3565 1150 2.8060 17/197-21:48:44.092 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1169 0.3565 1151 2.8084 17/197-21:48:44.096 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1169 0.3565 1151 2.8084 17/197-21:51:44.100 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1169 0.3565 1152 2.8109 17/197-21:53:14.100 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1168 0.3562 1152 2.8109 17/197-21:54:44.104 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1169 0.3565 1153 2.8133 17/197-22:03:44.132 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1169 0.3565 1153 2.8133 17/197-22:09:44.144 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1169 0.3565 1154 2.8158 17/197-22:18:44.172 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1169 0.3565 1154 2.8158 17/197-22:24:44.180 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1169 0.3565 1155 2.8182 17/197-22:26:14.184 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1169 0.3565 1154 2.8158 17/197-22:27:44.188 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1169 0.3565 1155 2.8182 17/197-22:33:44.204 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1089 0.3321 1154 2.8158 17/197-22:33:44.208 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1089 0.3321 1154 2.8158 17/197-22:35:14.204 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 759 0.2315 1152 2.8109 17/197-22:36:44.208 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 249 0.0759 1138 2.7767 17/197-22:38:14.212 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 4 0.0012 996 2.4302 17/197-22:39:44.216 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1 0.0003 288 0.7027 17/197-22:41:14.220 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1 0.0003 209 0.5100 17/197-22:42:44.224 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1 0.0003 179 0.4368 17/197-22:44:14.228 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1 0.0003 178 0.4343 17/197-22:44:33.369 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1 0.0003 177 0.4319 17/197-22:45:44.232 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1 0.0003 173 0.4221 17/197-22:47:14.236 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1 0.0003 172 0.4197 17/197-22:48:44.240 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1 0.0003 207 0.5051 17/197-22:48:44.244 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 1 0.0003 207 0.5051 17/197-22:50:14.244 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 1 0.0003 338 0.8247 17/197-22:51:44.248 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 4 0.0012 1017 2.4815 17/197-22:52:33.392 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 169 0.0515 1179 2.8768 17/197-22:53:14.248 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 402 0.1226 1208 2.9475 17/197-22:54:44.256 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 873 0.2663 1220 2.9768 17/197-22:56:14.256 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 1129 0.3443 1208 2.9475 17/197-22:57:44.260 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 1163 0.3547 1195 2.9158 17/197-22:59:14.264 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 1164 0.3550 1186 2.8938 17/197-23:00:33.408 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 1164 0.3550 1180 2.8792 17/197-23:00:44.268 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 1165 0.3553 1179 2.8768 17/197-23:02:14.272 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 1165 0.3553 1175 2.8670 17/197-23:03:44.276 ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON ON 1166 0.3556 1172 2.8597 17/197-23:03:44.284 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1166 0.3556 1172 2.8597 17/197-23:05:14.280 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1166 0.3556 1169 2.8524 17/197-23:06:44.284 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1167 0.3559 1167 2.8475 17/197-23:08:14.288 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1167 0.3559 1165 2.8426 17/197-23:09:44.292 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1167 0.3559 1164 2.8402 17/197-23:11:14.296 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1168 0.3562 1163 2.8377 17/197-23:12:44.300 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1168 0.3562 1162 2.8353 17/197-23:14:14.304 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1168 0.3562 1160 2.8304 17/197-23:17:14.312 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1169 0.3565 1159 2.8280 17/197-23:18:44.319 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON 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https://lasp.colorado.edu/maven/sdc/public/data/sites/site-20171121T230149/anc/eng/sasm1/sci_anc_sasm117_196_200.drf

Inside the Government Fiasco That Nearly Closed the U.S. Air System — ProPublica The upgrade to 5G was supposed to bring a paradise of speedy wireless. But a chaotic process under the Trump administration, allowed to fester by the Biden administration, turned it into an epic disaster. The problems haven’t been solved. by Peter Elkind May 26, 2022, 5 a.m. EDT The upgrade to 5G was supposed to bring a paradise of speedy wireless. But a chaotic process under the Trump administration, allowed to fester by the Biden administration, turned it into an epic disaster. The problems haven’t been solved. by Peter Elkind May 26, 2022, 5 a.m. EDT Twitter Twitter Facebook Facebook Copy Link Link Copied! Change Appearance Auto Light Dark Contrast Sun Moon Republish ProPublica is a nonprofit newsroom that investigates abuses of power. Sign up to receive our biggest stories as soon as they’re published. The prospect sounded terrifying. A nationwide rollout of new wireless technology was set for January, but the aviation industry was warning it would cause mass calamity: 5G signals over new C-band networks could interfere with aircraft safety equipment, causing jetliners to tumble from the sky or speed off the end of runways. Aviation experts warned of “catastrophic failures leading to multiple fatalities.” To stave off potential disaster, the Federal Aviation Administration prepared drastic preventive measures that would cancel thousands of flights, stranding passengers from coast to coast and grounding cargo shipments. “The nation’s commerce will grind to a halt,” the airlines’ trade group predicted . ProPublica Get Our Top Investigations Subscribe to the Big Story newsletter. Email address: Arrow Right This site is protected by reCAPTCHA and the Google Privacy Policy and Terms of Service apply. Thanks for signing up. If you like our stories, mind sharing this with a friend? Copy link For more ways to keep up, be sure to check out the rest of our newsletters. See All Fact-based, independent journalism is needed now more than ever. Donate On Jan. 18, following nail-biting negotiations involving CEOs, a Cabinet secretary and White House aides, an eleventh-hour agreement averted these threats of aviation armageddon. Verizon and AT&T agreed not to turn on more than 600 5G transmission towers near the runways of 87 airports and to reduce the power of others. Disaster was averted. But the fact that it was such a close call was shocking nonetheless. How did a long-planned technology upgrade result in a standoff that seemed to threaten public safety and one of the nation’s largest industries? The reasons are numerous, but it’s undeniable that the new 5G deployment represents an epic debacle by multiple federal agencies, the regulatory equivalent of a series of 300-pound football players awkwardly fumbling the ball as it bounces crazily into and out of their arms. More than anything, a deep examination of the fiasco reveals profound failures in two federal agencies — the Federal Communications Commission and the FAA — that are supposed to serve the public. In the case of the FCC, the agency not only advocated for the interests of the telecommunications industry but adopted its worldview, scorning evidence of risk and making cooperation and compromise nearly impossible. In the case of the FAA, the agency inexplicably stayed silent and passively watched preparations for 5G proceed over a period of years even as the aviation industry sounded ever more dire warnings that the new networks could put air safety at risk. That’s the alarming picture that emerges, in new detail, in interviews with 51 participants and observers in the 5G rollout, along with a review of thousands of pages of documents. The problems have spanned a Republican and Democratic administration. The process first ran off the rails under President Donald Trump. It then festered under the administration of President Joe Biden — which ProPublica’s reporting shows impeded the FAA when it finally decided to act — until a crisis forced an intervention. For now, the truce between the FCC and telecom companies, on one side, and the FAA and aviation companies, on the other, is holding. The parties have mostly tempered their hostile rhetoric, sounded hopeful notes of “coexistence” and have begun to collaborate. The FAA is allowing the wireless companies to slowly turn on more 5G towers as planes mostly keep flying. (About a thousand regional jets, mostly used by JetBlue, American, Delta and United, are currently barred from landing in low-visibility conditions at many airports over fears of equipment interference.) But the underlying issues are far from resolved. Aviation companies say they need much more time — perhaps two years or more — to upgrade or replace all the equipment vulnerable to 5G interference, according to Bob Fox, a United Airlines pilot now serving as national safety coordinator for the Air Line Pilots Association, a key player in the drama. The telecom companies have no interest in such a lengthy time frame: Their agreement with the government is set to expire on July 5, and they have made no commitment to extending the restrictions on their towers past that date. The companies have been exhibiting a willingness to make short-term compromises, but they’re also showing hints of frustration that they can’t seem to bring the process to a resolution. For its part, the FCC seems aggrieved. It overwhelmingly blames the aviation agency for the problems and simultaneously says it’s cooperating with the FAA — while continuing to insist that any claims that 5G will threaten airplanes are pure fantasy. The rhetoric of the FCC chief is almost identical to that of the industry she regulates. As recently as last month, Jessica Rosenworcel, the Biden-appointed FCC chair, dismissed aviation concerns as, in effect, a shakedown — a ploy to get telecom companies to fund a nationwide upgrade of airplane equipment. Referring to the air-safety devices that the aviation industry says could be compromised by 5G, she said in an interview with ProPublica, “Has anyone spec’d the cost of altimeter replacement?” And a whole new telecom-aviation conflict could soon emerge. T-Mobile and other wireless companies are approved to roll out additional 5G service at the end of 2023, using a C-band frequency even closer to the one used by the airplane safety equipment. Like other participants in the process, T-Mobile says it’s committed to safety and to finding a reasonable solution. But if that rollout unfolds in a way that resembles the last one in the slightest way, a reasonable solution may be elusive. There was a time, a century ago, when radio was the hottest new technology in the land. Stations sprouted up everywhere, and they routinely used the same frequencies. The result was electronic bedlam: Programming was regularly interrupted by rival stations, police-radio chatter and amateur enthusiasts. Congressional legislation lamented “the present chaos of jazz bands, sermons, crop reports, sporting services, concerts and whatnot running simultaneously on the same wave lengths.” In one celebrated case, a wealthy Illinois bank president obtained a court order against a local 18-year-old whose radio transmissions had kept him from listening to broadcasts of election-night results at his home. Since its founding in 1934, the Federal Communications Commission has decided which companies would have rights to which parts of the airwaves — for television and countless other technologies. Here, an RCA engineer examines an array of ultrahigh-frequency TV antennas in 1952. Credit: Bettmann Archive/Getty Images There was a critical need for a neutral arbiter to make decisions about who could occupy which part of the airwaves. All this led to the creation of a federal agency to regulate radio, which ultimately morphed into the FCC in 1934. A big part of its mission, as the new agency told Congress, was to make “equitable distribution of the frequencies ... as congestion increases.” The march of technology over the 88 years that followed can be understood as a series of battles for the airwaves. Virtually every important communications technology, from television and satellites to cellphones and GPS, has required bandwidth. The FCC was there to dole out frequencies and referee the conflicts. There were massive financial stakes in many of the decisions. They launched entire industries, while burying or transforming others. As more and more technologies crowded into a finite set of frequencies, the opportunities for one technology to interfere with another only increased. In the mid-1990s, new digital phone technology unintentionally triggered a buzzing in some hearing aids, while interference from police radios sometimes prompted powered wheelchairs to randomly accelerate or brake, leading to serious injuries . In 2010, the shift to digital television required the replacement of wireless microphones used by actors in Broadway shows, referees at NFL games and pastors at Sunday church services. By the time 5G approached, the FCC had long ago developed an acclaimed system of selling spectrum — reserved areas of the airwaves — for commercial use, which generated large sums of money for the federal government: public auctions. The agency’s first such auction was held in 1994. Over the years that followed, the FCC successfully used the process 110 times, raising more than $233 billion. The auctions’ sophisticated format helped win a Nobel Prize for two Stanford economists who designed them . But the Trump administration didn’t initially seem inclined to leave 5G decisions to the FCC. The administration saw the fifth generation of cellular technology, with its faster speeds and automation efficiencies for industry, as its single biggest communications initiative. Top Trump officials viewed the technology through the prism of competition with China. Many in the administration also expressed fears that Huawei Technologies, a dominant maker of 5G hardware, might be a conduit for Chinese government surveillance, posing a national-security threat. (Huawei has always denied such claims.) Trump lieutenants began employing a nationalist battle cry: America needed to “ win the race to 5G ” against China. The Trump administration veered in multiple directions in pursuit of that goal. In January 2018, National Security Council officials circulated a plan to create a government-run 5G network. This idea was jettisoned almost as soon as it was proposed, amid criticism that this would constitute socialism. President Donald Trump, with AT&T’s then-CEO, Randall Stephenson, in 2017, examining a model of how 5G will be deployed in cities. Trump was a cheerleader for 5G adoption but often cast the process as part of a race against China. Credit: Olivier Douliery-Pool/Getty Images Others in the Trump orbit proposed ideas as well. Attorney General William Barr at one point suggested that the U.S. government, in the interest of developing China-free 5G networks, buy controlling interests in Nokia and Ericsson, the European telecom equipment companies. Republican insiders such as consultant Karl Rove, former House Speaker Newt Gingrich and Trump campaign manager Brad Parscale promoted a partnership in which the Defense Department would lease unused spectrum to Rivada Networks, a company backed by GOP donor Peter Thiel. This approach was embraced by a Trump campaign spokesperson and then promptly repudiated by the White House. Trump himself declared that the administration’s 5G plan should be “ private-sector driven and private-sector led .” Eventually the White House moved on to other obsessions. The FCC, and its chairman, became the driving force in the race to 5G. The agency’s brash leader, 44 when he took the role in 2017, was Ajit Pai. He had been an FCC commissioner before being elevated by Trump. Pai’s agency was legendarily friendly to the companies it regulated, with commissioners and key staff routinely moving to and from lucrative posts in the industry. Pai himself had spent two years early in his career as an in-house lawyer at Verizon and later worked at a law firm that served telecom clients. Since stepping down at the end of the Trump administration, Pai has decamped to a private-equity firm whose portfolio includes telecom and broadband companies. At the FCC, Pai joined the wireless companies in evangelizing for 5G. He would make it the central initiative of his tenure. A speedy rollout, Pai proclaimed, would “transform our economy, boost economic growth and improve our quality of life.” He regularly cited a report proclaiming that 5G could create up to 3 million new U.S. jobs and $500 billion in economic growth — without noting that those rosy figures came from a study commissioned by the wireless industry’s lobbying group. Under Pai, the path to 5G initially continued to zigzag. After the White House abandoned the central plan, the FCC turned in a new direction, one that would put a few foreign satellite companies in charge of the process. At issue was the so-called C-band, a patch of wireless real estate viewed as the sweet spot for 5G. Wireless companies coveted C-band spectrum for its ability to transmit big chunks of data rapidly over long distances; it would maximize 5G speeds while minimizing the number of expensive cell towers and transmitters the companies needed. That spectrum was owned by the federal government. But it was then being used with the government’s consent, free of charge, by four foreign satellite companies that relayed radio and TV signals around the globe. Sensing opportunity, the companies banded together and made an audacious proposal: They, the non-rent-paying users of the spectrum, would sell it to U.S. wireless companies and keep most of the expected tens of billions of dollars for themselves. (They agreed to make a voluntary contribution to the federal Treasury from the proceeds.) This “market-based solution,” the satellite companies claimed, would be the fastest way to get 5G networks up and running. Pai seriously entertained this approach for a year. The plan eventually fizzled in the face of fierce opposition led by Louisiana Republican Sen. John Kennedy , who expressed outrage that foreign satellite companies would reap most of the money from a sale of U.S. government spectrum. FCC Chairman Ajit Pai, shown in 2017, was an evangelist for 5G. He regularly cited a report proclaiming that the technology could create up to 3 million jobs, without noting that those figures came from a study commissioned by the wireless industry’s lobbying group. Credit: Chip Somodevilla/Getty Images Stymied, the FCC swerved back to its traditional approach: a public spectrum auction, in this case for a big chunk of the C-band. The agency decided that the winning bidders would pay the satellite companies up to $14.7 billion for quickly vacating those frequencies and retooling on different frequencies. That, the agency hoped, would avoid costly, time-consuming lawsuits by the satellite companies. A $14.7 billion payout was staggering, but it was an accepted FCC practice to arrange compensation for companies affected by its spectrum actions. The agency, however, would make no such provisions for another group warning of far graver consequences: the U.S. aviation industry. The battle that would threaten to ground American aviation centered on an electronic device about the size of a toaster. Called a radio altimeter, it’s used to track an aircraft’s altitude during takeoff and landing. Radio altimeters, which became standard gear in the early 1970s, work by bouncing an electronic signal off the ground, sending their readings instantly to the cockpit. This is important for low-visibility landings, at night or in bad weather. It matters at other times, too: Radio altimeters on many commercial jets feed their data into automated navigation and crash-avoidance systems, sometimes controlling the engines and braking systems. About 50,000 planes and helicopters in the U.S. carry radio altimeters. Aviation industry anxiety about the peril of radio altimeter failure inevitably cites a Turkish Airlines Boeing 737 that crashed in 2009 while attempting a landing in Amsterdam. Nine passengers and crew members died. An investigation revealed that the disaster originated with a malfunctioning altimeter , whose faulty readings triggered the jet’s auto-throttle to cut power during the final landing approach. The FCC’s plans to use the C-band for 5G rekindled these fears. The problem was that the upper part of the C-band is where radio altimeters operate, prompting concern that nearby 5G transmissions would cause them to spit out false readings or stop working. Because most altimeters had been built and installed decades earlier, when there was nothing noisy in their electronic neighborhood, they hadn’t been designed to screen out the likes of 5G. Fights over the FCC’s efforts to cram ever more users into limited spectrum became unusually common, and heated, during the Pai regime, and it wasn’t just the aviation industry that protested. Other orders granting telecom companies various 5G frequencies prompted complaints that they would disrupt networks used for satellite communications, weather forecasting, agriculture, self-driving cars, global-positioning services and military weapons systems. One of the FCC actions, still being fought, drew opposition from 14 federal agencies and departments. Again and again, the FCC, backed by the Trump White House, brushed those pleas aside. “Agencies raising concerns about the impact of 5G were just sort of being mowed down by the FCC,” according to a former high-level Trump official involved in spectrum disputes. Starting in 2018, more than a dozen aviation groups and companies told the FCC they worried that interference with radio altimeters could cause a deadly plane crash. They urged the agency to work with the FAA and delay an auction until it had identified and eliminated any risk. Aviation-industry officials also argued that the telecom companies, or the Treasury, should fund billions of dollars’ worth of altimeter upgrades to eliminate interference problems from the 5G C-band signals. But the FCC didn’t think there was a problem to solve. The agency embraced the wireless industry’s view, which was to deny that the new 5G networks posed any risk to aviation safety. “One of the things we built into the way spectrum auctions work is that various people need to be paid off,” said Blair Levin, who was FCC chief of staff when the agency first deployed its auctions. Levin said the 5G process was handled differently: “The airlines came and said, ‘We have this problem.’ Nobody asked: ‘What do you need to fix the problem? Do you need $2 billion? Do you need $4 billion? Do you need $6 billion?’ Ajit just said: ‘We don’t care; we don’t think your concerns are legitimate.’” Pai defends that position. “The FCC’s career staff did a terrific job analyzing the facts,” he told ProPublica, “and had demonstrated to all the commissioners at the FCC that there was no credible case made for the possible interference with aviation altimeters… No legitimate objective engineering work would find a legitimate case for those arguments.” The FCC was the final word on spectrum allocation. End of discussion. But that ignored a key fact: The FAA was the final word on airplane safety, and it was becoming more concerned about the risk to radio altimeters. And unlike other agencies, the FAA had sweeping authority to order dramatic steps to avoid any chance of a fatal accident. The FAA had massive power — but it didn’t seem inclined to use it. For starters, like more than one agency under Trump, it suffered from instability at the top, with an interim leader for 18 months. And the FAA was on the defensive in the wake of two Boeing 737 MAX crashes that killed a total of 346 people and tarnished the agency’s reputation. That meant that when Steve Dickson took over as chair in the summer of 2019, the FAA was distracted by the Boeing mess, according to an agency source. And in Dickson, the FAA seemed to have the temperamental opposite of the aggressive and ambitious Pai. Dickson, who was 61 when he took the agency helm, was a onetime Air Force fighter pilot who had recently retired from a three-decade career at Delta, first as a pilot and then as a company safety executive. (Along the way he also got a law degree.) Perhaps because of his background in the military or perhaps just by dint of disposition, Dickson adhered strictly to the chain of command. Methodical, cautious and measured — a top deputy said he’s never heard him raise his voice — Dickson was loath to take steps outside official procedures, and those official procedures placed a lot of emphasis on filing papers. So that’s what Dickson and his agency did. FAA Administrator Steve Dickson, shown testifying in 2021, spent a lot of his time dealing with the crisis over the Boeing 737 MAX. If Pai was aggressive and brash, Dickson tended to be rules-focused and methodical. Credit: Joshua Roberts/Getty Images Dickson did not, for example, call Pai to hash out the 5G issue. He did not march to the White House to sound the alarm. He issued no press releases to bring attention to the looming problem. The agency’s only official expression of concern in 2019 was a two-page letter from an agency engineer to a Commerce Department panel charged with resolving government spectrum disputes. It urged the FCC to delay any C-band auction until a technical study the FAA had funded had arrived and “any interference mitigations have been considered.” (Dickson declined to comment for this article.) The FAA’s passivity was particularly striking given the mounting concern in the aviation industry. A pair of reports by aviation research groups intensified those anxieties. The first was a preliminary lab study , conducted by a government-industry research cooperative at Texas A&M University. It found all seven radio altimeters it tested were susceptible to interference. It too urged further analysis. The report was submitted to the FCC in late 2019. But the FCC didn’t wait. On Feb. 28, 2020, it voted to authorize the C-band sale, and it scheduled the auction for Dec. 8. The FCC’s 258-page report and order devoted just six paragraphs to aviation safety, much of it agreeing with a T-Mobile-sponsored report that dismissed aviation concerns. The FCC contended that its precautions, including leaving a patch of spectrum vacant between 5G transmissions and the altimeter frequency, would be sufficient. The FCC order made clear whose problem this was to solve. “Well-designed equipment should not ordinarily receive any significant interference (let alone harmful interference) given these circumstances,” the order stated. “We expect the aviation industry to ... take appropriate action, if necessary, to ensure protection of such devices.” The aviation companies didn’t see it that way. Their concerns escalated in October 2020 with the issuance of a 231-page report by the RTCA, a nonprofit aviation industry research organization originally known as the Radio Technical Commission for Aeronautics. It found that 5G posed “a major risk” to altimeters with “the potential for broad impacts to aviation operations in the United States, including the possibility of catastrophic failures leading to multiple fatalities.” It too urged the FCC, FAA and industries to work together to tackle the problem. The opposite happened. The telecom and aviation industries embraced diametrically opposing views of reality. They assailed each other’s studies and methodologies. The wireless advocates declared that nearly 40 other countries had already deployed 5G on the C-band near airports, under conditions similar to those contemplated in the U.S., without incident. Aviation allies replied that power levels and other limits on 5G operations near overseas airports were meaningfully different. Each side accused the other of refusing to share technical data needed to assess the issue. The two sides also viewed risk in radically different ways. From the aviation perspective, the wireless industry simply couldn’t fathom its hypercautious safety culture, which, given the horrific consequences of an accident, demands that any critical equipment be proven to pose a probability of failure of no more than one in a billion. “If there’s the possibility of a risk to the flying public,” the FAA’s “5G and Aviation Safety” website notes , “we are obligated to restrict the relevant flight activity until we can prove it is safe.” Aviation companies were getting increasingly nervous. Yet the FAA continued to sit on its hands. Finally, with the 5G auction just a week away, in December 2020, the FAA took action of sorts: It drafted a letter. What happened next involves a tiny government agency few people have heard of. Buried deep inside the U.S. Department of Commerce, it’s called the National Telecommunications and Information Administration . It advises the president on spectrum issues and mediates fights between federal agencies. Its job is to help resolve exactly the sort of conflict that was raging over 5G. In the Trump administration, however, the NTIA was in disarray. A Government Accountability Office report would find that the agency lacked a “formalized” process for weighing in on spectrum issues. The last Senate-confirmed chief of the agency had abruptly quit in May 2019. By November 2020, it was on its third acting administrator, former Michigan State law professor Adam Candeub. Candeub had a record as a conservative legal warrior. He’d represented a white supremacist in unsuccessfully suing Twitter for permanently barring him and his organization from its platform. He was also a fervent advocate for the FCC’s aggressive 5G agenda. Right before joining the Trump administration, Candeub published a column in Forbes titled “FCC Chair Ajit Pai Must Press Forward on 5G Auctions.” The article praised Pai for cutting “bureaucratic meddling.” It was into these hands that the FAA and the Department of Transportation would deliver a four-page missive , dated Dec. 1, 2020, with a request that it be forwarded “expeditiously” to the FCC for public posting. Filing such a letter through the NTIA was the proper federal protocol. But the step hardly seemed to match the gravity of the problem. Still, what the letter was asking for was remarkable: It urged the FCC to delay its C-band auction, which was just a week away at that point. Understanding “the safety and economic ramifications” of the 5G rollout required a “comprehensive risk assessment and an analysis of potential mitigation options,” the letter stated. Its tone may have been bureaucratic, but the letter contained a dramatic warning: If 5G deployment moved forward “without addressing these safety issues,” the FAA would consider imposing flight restrictions that “would reduce access to core airports in the U.S.” This letter never made it onto the FCC’s public docket, where it would have amplified the need to resolve the dispute. Candeub never sent it. Nearly a year afterward, when news of the letter first surfaced in media reports , and some people accused him of burying the letter to help the FCC’s agenda, Candeub, back at his old job at Michigan State, denied any political motivation. His agency’s experts, Candeub told reporters, had found “serious flaws” in the RTCA report and therefore dismissed its aviation-safety warnings. In an interview with ProPublica, Candeub acknowledged discussing the FAA letter with Pai, who “wasn’t happy” about it. (Pai said he couldn’t recall whether or not he and Candeub had talked about the letter.) But Candeub said he’d made his decision based on a highly critical assessment of the RTCA report by Charles Cooper, head of NTIA’s spectrum management office and a career government employee. Candeub said Cooper thought the report had “serious errors.” Emails between Candeub and Cooper, obtained by ProPublica, reveal a different narrative. In an email on Nov. 25, 2020, Cooper wrote to Candeub that, “as requested,” he and his staff had performed an initial assessment, and it indicated “agreement” with the RTCA’s approach. “Ah ... so there is a there, there,” Candeub replied. “Do you recommend, therefore, that we work with DOT for a submission to the FCC?” “I don’t think we have a choice!” Cooper emailed back. Asked about the exchange, Candeub insisted that Cooper reversed his view after studying the matter for a few more days. “As we dug in further, the conclusion of Charles was that this did not rise to a level of concern, so the letter wasn’t sent. ... That was the final verdict I got from him.” An NTIA spokesperson, in a statement, offered a different view: “There is no record of a staff recommendation against forwarding the letter from the FAA.” (Cooper declined to comment.) The statement also noted that NTIA staff “recommended that the RTCA study be validated and offered a path forward for better understanding the issues raised. Our work evaluating those issues is ongoing.” On Dec. 8, the FCC commenced its auction for the C-band spectrum. The agency announced a few months later that the sale had raised a record $81.1 billion, roughly double what industry observers expected. Verizon bought the biggest stake, for $45.5 billion, followed by AT&T, which paid $23.4 billion. Their costs for building out 5G infrastructure, marketing and paying satellite companies to speed their exit would add tens of billions more to their tab. It left the two companies, understandably, determined to exploit their investment. “We have a license from the U.S. government saying we can proceed,” explained one executive from a wireless company. “We’re not really looking for reasons why we can’t proceed.” At that point, it appeared that the FCC had prevailed in the battle of federal agencies. Pai and Larry Kudlow, former director of Trump’s National Economic Council, would crow about how they had triumphed over Washington “swamp creatures” in a discussion on Kudlow’s Fox Business show months after both had left government. Pai insisted his agency had followed the science, and the two men denigrated concerns over aviation safety. “The FAA is bellyaching about 5G. The airlines are bellyaching about 5G. We ignored them,” Kudlow declared. “We actually fought the FAA. We won.” As the 5G auction concluded in early 2021 and winter turned into spring, the FAA resembled the bureaucratic equivalent of a turtle that had first retracted into its shell, then been flipped onto its back. It seemed helpless. After its last-minute letter seeking to halt the spectrum auction was ignored, the agency had said nothing publicly about the issue for months. The aviation industry was growing increasingly frantic at the FAA’s temporizing. In the summer of 2021, the agency told attendees at an industry forum that it was “still gathering information” on the radio altimeter problem. Aviation executives begged the agency to go public. “We wanted them to publicly state there is a huge problem, and it’s going to cause massive disruptions,” said John Shea, government affairs director for Helicopter Association International, a trade group. “We said: ‘You need to say this out loud! This can’t just be industry speculation.’” Behind the scenes, though, reality was beginning to dawn at the FAA. Top officials, who had clung to the industry’s hope that somehow the FCC could be persuaded to postpone the C-band rollout for a year or two, had finally grasped that the launch was happening. The FAA simply couldn’t wait any longer if the agency wanted to follow its methodical processes and give airlines time to prepare. Now the agency prepared to deploy its ultimate weapon: formal air-safety alerts that would pave the way for grounding commercial aircraft. By August 2021, the FAA was ready to proceed. But a new impediment had arisen: the Biden administration. The White House was discouraging any public action, as was the FCC, which was now operating with a Democratic chair but was every bit as supportive of 5G and the telecom industry’s position on it as Pai had been. They assured the FAA that the agencies and industries could somehow still work out the problem quietly. (A senior FCC official denies the agency requested any delays.) Repeatedly, the FAA deferred sending out its air warnings. Dickson privately told his staff that his agency was like Charlie Brown, with the White House and the FCC in the role of Lucy, who “keeps pulling the football out from under us.” In October 2021, the FAA finally began preparing its first airworthiness bulletin, warning of “potential adverse effects on radio altimeters” — but not until its bureaucratic opponents had gotten a chance to vet the language. The bulletin received a line-by-line review from officials at the FCC and the White House’s National Economic Council, according to an FAA staffer involved in the matter. The White House “wanted to make the problem not seem as bad as it was,” the FAA official said. “And they wanted to make sure it was worded in such a way that wireless was not seen as the villain.” (The senior FCC official said his agency “routinely” provides “technical” input on bulletins. The FAA’s parent agency, the DOT, said it supported a “collaborative approach” to “minimize any disruptions to the traveling public” but that the FAA made the final call on the language of its bulletins. As the DOT put it, “​​Part of the process failure during the last administration was a result of auctioning the spectrum without the required collaboration between stakeholders and agencies to ensure the safety of the traveling public and minimize disruptions for them — despite consistent clear requests by DOT and FAA to do so. Conversely, this Administration wanted to ensure that government agencies with technical expertise were all at the table and collaborating.”) Issued on Nov. 2, 2021, the bulletin alerted equipment manufacturers, aircraft companies and pilots to the potential for “both erroneous altimeter readings and loss of altimeter function.” This, the bulletin advised, could result in “the loss of function” of safety systems. It added that the FAA was assessing whether potential limits on flight operations were warranted. That threat instantly transformed the standoff. “That kicked in some action,” a wireless-industry executive said. “That’s the first time they said to the airlines: ‘When these guys light up on Dec. 5, we’re going to ground your planes.’” The executive added: “All of a sudden, we needed to tackle a two-year problem in 30 days.” Two days after the FAA’s airworthiness bulletin went out, Verizon and AT&T agreed to a one-month delay, pushing the 5G start date to Jan. 5, 2022. With threats of aviation shutdowns now officially in play, no one wanted to be blamed for ruining holiday travel. Now the question became how broad and long-lasting any restrictions on the 5G rollout should be. The telecom companies, backed by the White House and FCC, wanted them to be limited and temporary. The FAA and aviation interests wanted something far more sweeping and permanent. Haggling commenced in earnest. Verizon and AT&T had already offered to modestly cut power from some 5G transmitters near runways for six months. The aviation companies rejected that as “inadequate and far too narrow.” They proposed a broad swath around airports where towers would never be turned on as well as other limits. No way, countered the FCC. That would render the telecoms’ C-band spectrum “commercially unviable. ... Effectively it would cease to be 5G.” FCC officials felt that the telecom industry was being falsely cast as a villain. But momentum had turned in favor of the aviation forces. The mere specter of fatal airplane disasters was a potent message. The wireless companies were getting hammered in the press, with news reports warning that 5G could cause planes to crash. Negotiations intensified. In late December 2021, Transportation Secretary Pete Buttigieg jumped in, talking with the CEOs of Verizon and AT&T. Buttigieg announced a truce on Jan. 3. The wireless companies agreed to another two-week delay and to establish temporary modest 5G-free buffer zones around 50 airports for six months. And they’d provide the FAA with full details about their tower sites. But almost as fast as it was announced, the deal fell apart. Altimeter testing made clear that the planned buffer zones weren’t nearly big enough to resolve the FAA’s interference fears. The once-somnolent agency was now firing a fusillade of safety notices, laying out the specifics of how each airport would be affected. Thousands of planes would need to be barred from landing in low-visibility conditions at airports where 5G was present. Some big long-haul jets wouldn’t be able to fly into airports with 5G at all. One FAA notice warned that C-band interference might prevent braking systems on some Boeing 787s from kicking in during landings, causing planes to speed off runways. Buttigieg and the FAA’s Dickson went back to the CEOs of Verizon and AT&T and demanded yet more concessions. On Jan. 18, the companies capitulated, even as AT&T bitterly complained that the FAA and aviation industry had “not utilized the two years they’ve had to responsibly plan for this deployment.” When Verizon and AT&T finally switched on their networks the next day, the expanded restrictions (a 3-mile buffer zone around 87 airports) left more than 600 of their 5G towers dark — about 10% of their planned first-day service. The last-minute chaos spurred a flurry of flight cancellations. But the provisions, and continued testing of radio altimeters, allowed about 90% of planes to operate normally within days. A conspicuous exception was about a thousand Embraer regional jets, used by JetBlue, American, Delta and other airlines. Equipped with altimeters particularly susceptible to interference, they remain restricted from landing in poor-weather conditions in many cities with C-band towers. The former adversaries finally began to collaborate. The FAA built a measure of trust with Verizon and AT&T by allowing them to turn on enough towers to showcase their 5G service at the Super Bowl on Feb. 13, even though the stadium was beneath a landing approach to Los Angeles International Airport. As the weeks passed, both sides made more accommodations, shrinking the size of the buffer zones while boosting the total number of “protected” airports to 114. Most participants in the 5G process say comity and cooperation has increased among all parties. AT&T told ProPublica in a statement that it is “continuing to cooperate and collaborate with the FAA, FCC and other stakeholders to help facilitate the FAA’s technical assessments and clearance of aviation equipment. We are encouraged by the significant progress the FAA has made thus far, and we expect that progress will continue going forward.” Verizon also said it was “encouraged” by the “collaboration and pace” among the companies and agencies, adding, “We’re highly confident that the small and declining number of outstanding questions will be resolved sooner than later, without any meaningful impact to airline operations or the availability of 5G at airports.” For all the expressions of optimism, the problems aren’t yet resolved and a deadline looms: Verizon and AT&T have made no commitment to extend their “voluntary” restrictions beyond July 5. And this may not be the last such battle, either: In December 2023, T-Mobile and other wireless companies will be free to fire up a new patch of C-band, even closer to the altimeter frequency. At that point, 5G will be operating near hundreds of additional airports. In the face of this uncertainty, aviation companies are scrambling to develop the only promising short-term solution: filters designed to screen out electronic interference for the worst-performing radio altimeters. But many altimeters can’t be fitted with filters and inventing and deploying new altimeters for a 5G world will take years. Meanwhile, the industry is continuing to agitate for someone else to pay for it all. In recent months, the center of activity in the 5G saga has been the FAA, which is now led by an interim chief. (Dickson announced his resignation in February, saying it was “time to go home”; he left in March.) The FCC, more on the fringes at this stage of the process, has been talking about steps like improving its processes with the NTIA, while continuing to insist that claims of 5G risk are hooey. AT&T echoed that sentiment, saying “the physics has not changed,” in a second statement it sent to ProPublica, in late May. The company’s hope, in this statement, was beginning to sound like it was being uttered through gritted teeth. AT&T is still “working collaboratively with the FAA and the aviation industry,” it said, while noting that “we have made no additional commitments beyond July 5, but are in discussions with the FAA and aviation on a phased deployment approach that will provide the aviation industry with some additional time to complete equipment updates without stalling our C-Band deployment.” At the FAA, it seemed like one step forward, one step back as the July 5 deadline approached. On May 4, the agency convened an in-person gathering of 40 invited “stakeholders” from the wireless and aviation industries — but no FCC officials — aimed at forging a path for continued peace. Agency officials reviewed the “rapid evolution” in easing the limits on the wireless companies around airports. And they pressed aviation officials to develop a firm timetable for retrofitting the entire U.S. commercial fleet with filters and new altimeters, in short, a day when 5G can finally be unfettered. ProPublica Read More The Supreme Court Upheld the Indian Child Welfare Act. The Long Struggle to Implement the Law Continues. But barely more than two weeks later, on May 19, the follow-up meeting with the FAA sounded considerably less encouraging. One company that is preparing filters for altimeters pleaded for time, saying it needed until the end of 2023. That’s not good enough, the FAA’s acting chief responded. He told them it needs to happen by the end of this year. Efforts to reach accommodation had increased, it seemed, but so had “heartburn,” as one FAA official put it. “The wireless companies have made it very clear they’re not going to agree to an open-ended situation,” he said. “They seem willing to go past July 5, as long as they know how far past. But they’re making clear their patience is not infinite.” Do You Work for the Federal Government? ProPublica Wants to Hear From You. We’re expanding our coverage of government agencies and federal policy. With your help, we can dig deeper. Loading form... Powered by Screendoor . Expand Doris Burke contributed research. Embedded illustrations by ProPublica. Source Images: The7Dew and Peng Song/Getty Images. Filed under — Technology Regulation Peter Elkind Peter Elkind is a reporter covering government and business. Mail [email protected] Most Read Justice Samuel Alito Took Luxury Fishing Vacation With GOP Billionaire Who Later Had Cases Before the Court How a Grad Student Uncovered the Largest Known Slave Auction in the U.S. DOT Researchers Suggested a Way to Make Big Trucks Safer. After Meeting With Lobbyists, Agency Officials Rejected the Idea. 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Cutting Edge: TGF-β and Phosphatidylinositol 3-Kinase Signals Modulate Distinct Metabolism of Regulatory T Cell Subsets | The Journal of Immunology | American Association of Immunologists Abstract. Murine Foxp3+ regulatory T cells (Tregs) differentiated in vitro (induced Tregs [iTregs]) in the presence of anti-inflammatory cytokine TGF-β rely pre Cutting Edge: TGF-β and Phosphatidylinositol 3-Kinase Signals Modulate Distinct Metabolism of Regulatory T Cell Subsets * Department of Surgery and Center for Transplantation Sciences; Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129; Ryan H. Newton * Department of Surgery and Center for Transplantation Sciences; Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129; Jian-Wen Zhang 0000-0003-3787-218X ; Jian-Wen Zhang * Department of Surgery and Center for Transplantation Sciences; Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129; ‡ Department of Liver Transplantation, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong 510630, People’s Republic of China; Kelsey K. Finn Kelsey K. Finn * Department of Surgery and Center for Transplantation Sciences; Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129; Valerie A. Gerriets 0000-0002-1553-2590 ; Valerie A. Gerriets § Department of Pathology, Microbiology, and Immunology, Vanderbilt University, Nashville, TN 37232; and Alexandria Huynh Alexandria Huynh * Department of Surgery and Center for Transplantation Sciences; Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129; Jeffery C. Rathmell ; Jeffery C. Rathmell § Department of Pathology, Microbiology, and Immunology, Vanderbilt University, Nashville, TN 37232; and Bruce R. Blazar 0000-0002-9608-9841 ; Bruce R. Blazar † Division of Blood and Marrow Transplantation, Department of Pediatrics, University of Minnesota, Minneapolis, MN 55455; Laurence A. Turka Laurence A. Turka * Department of Surgery and Center for Transplantation Sciences; Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129; ¶ Rheos Medicines, Cambridge, MA 02139 Address correspondence and reprint requests to Dr. Laurence A. Turka, Rheos Medicines, 38 Sidney Street, Cambridge, MA 02139. E-mail address: [email protected] Author & Article Information Address correspondence and reprint requests to Dr. Laurence A. Turka, Rheos Medicines, 38 Sidney Street, Cambridge, MA 02139. E-mail address: [email protected] Received: March 05 2018 Accepted: August 19 2018 Published: October 15 2018 Online ISSN: 1550-6606 Print ISSN: 0022-1767 J Immunol (2018) 201 (8): 2215–2219. https://doi.org/10.4049/jimmunol.1800311 Article history Received: March 05 2018 Accepted: August 19 2018 Published: October 15 2018 Abstract Murine Foxp3 +regulatory T cells (Tregs) differentiated in vitro (induced Tregs [iTregs]) in the presence of anti-inflammatory cytokine TGF-β rely predominantly upon lipid oxidation to fuel mitochondrial oxidative phosphorylation. Foxp3 expression underlies this metabolic preference, as it suppresses glycolysis and drives oxidative phosphorylation. In this study, we show that in contrast to iTregs, thymic-derived Tregs (tTregs), engage in glycolysis and glutaminolysis at levels comparable to effector T cells despite maintained Foxp3 expression. Interestingly, exposure of tTregs to the anti-inflammatory cytokine TGF-β represses PI3K-mediated mTOR signaling, inhibits glucose transporter and Hk2expression, and reprograms their metabolism to favor oxidative phosphorylation. Conversely, replicating the effects of inflammation via elevation of PI3K signaling has minimal effects on tTregs but dramatically enhances the glycolysis of normally oxidative iTregs, resulting in reduction of Foxp3 expression. Collectively, these findings suggest both extrinsic and intrinsic factors govern the unique metabolic signature of Treg subsets. Introduction Metabolic disorders such as obesity and type 2 diabetes are associated with immune dysregulation, and immune pathological conditions themselves have been linked to metabolic maladaptation of immune cells (1). Foxp3 +regulatory T cells (Tregs) are a key component in this bidirectional link between immunity and metabolism (2). Although once considered to be a single subset, Tregs can be further subdivided into two categories. The first thymic-derived Tregs (tTregs) arise during T cell ontogeny in the thymus with TCRs that have a relatively high avidity for peptide + self-MHC, albeit not so high as to trigger negative selection (3). The second category, peripheral Tregs, develop from conventional T cells (T conv) when they undergo activation under noninflammatory conditions (e.g., low doses of Ag, presence of TGF-β, absence of IL-6, etc.) (4–6). Such cells, when generated in vitro, are referred to as induced Tregs (iTregs). Current models suggest that tTregs restrain immune responses against self-antigens, whereas peripheral Tregs play a predominant role in maintaining immune tolerance/dampening inflammation at mucosal surfaces and at the maternal–fetal interface (7–9). Immunometabolic studies have demonstrated that the metabolic preference of immune cells is not only a result of cell differentiation but can be causal as well. For instance, stimulating CD4 T cells in a manner that fosters glycolytic metabolism (i.e., strong PI3K, myc, and mTOR signals) promotes differentiation into effector T lineage (Th1, Th17, etc.), whereas prevention of glycolysis (via induction of AMPK rather than mTOR signals) and/or forced use of oxidative metabolism leads to differentiation into iTregs (10,11). This may be a self-reinforcing feature, as Foxp3 itself can drive lipid oxidation via suppression of myc (a master regulator of glycolysis) and upregulation of components of the electron transport chain required for mitochondrial oxidative phosphorylation (12–14). In contrast, available data on tTregs suggest that they can engage in glycolytic metabolism to a certain degree, which is critical for their proliferation, upregulation of suppressive molecules such as CTLA4 and ICOS, and their migration (15–17). This indicates that although Foxp3 may be a key driver of oxidative metabolism in Tregs induced from naive T cells, the same is not necessarily true in pre-existing Tregs. In this study, we perform a detailed metabolic comparison of tTregs and iTregs and show that in contrast to iTregs, tTregs can engage aerobic glycolysis comparable to differentiated effector T cells. Strikingly, exposing tTregs to iTreg-inducing conditions (i.e., TGF-β) reduces mTOR-dependent glycolytic metabolism. Interestingly, elevation of PI3K/mTOR activity via ablation of the lipid phosphatase and tensin homolog (PTEN) reconfigures the metabolism of tTregs and iTregs distinctly. Although PTEN deficiency in tTregs promotes glycolytic lipogenesis and nucleotide biosynthesis without affecting Foxp3 expression, its loss in iTregs enhances aerobic glycolysis and is associated with a significant decrease in Foxp3 expression. Together, these findings demonstrate inherent metabolic differences between Treg subsets and show that adaptations occurring because of context-dependent cues modulate Treg metabolism. Materials and Methods Mice Foxp3-YFP-Cre wild type (WT) and Pten fl/fl Foxp3-YFP-Cre (PTEN ΔTreg) mice (18) were maintained in specific pathogen-free conditions at Massachusetts General Hospital (MGH; Boston, MA). Healthy mice aged between 6 and 14 wk were used for all experiments. All animal experiments were performed in compliance with the institutional guidelines approved by the MGH Animal Care and Use Committee. Cell sorting and flow cytometry CD4 +T cells from spleens and lymph nodes (inguinal, axillary, and cervical) were enriched using the CD4 +T cell Negative Selection Kit (eBioscience) and sorted to over 95% purity on a FACSAria II (BD Biosciences) for YFP +CD4 +Tregs and YFP −CD4 +CD62L hiCD44 loCD25 −naive T cells. Phosphoflow and intracellular staining for PTEN and Glut1 was performed using the BD Phosflow protocol III with abs for PTEN (Clone A2B1; BD Biosciences), phospho Akt S473 (pAkt 473) (Cell Signaling Technology), phospho S6 (pS6) (Cell Signaling Technology), and Glut1 (Abcam). The affinity pure [F-(ab′)2] fragment donkey anti-rabbit IgG (Jackson Immunoresearch) was used as secondary Ab for staining pAkt473 and pS6. The following Abs to ICOS (clone 7E.17G9) and CTLA4 (clone UC10-4B9) were also used. In vitro cell culture and suppression assay Sorted cells were resuspended in complete Click’s media (Irvine Scientific) along with 10% FBS, 1% penicillin/streptomycin, 1% l-glut (2 mM), and 55 μM 2-ME. Forty-eight–well flat-bottom plates were coated with plate-bound αCD3 (2 μg/ml, clone 145-2C11) and αCD28 (2 μg/ml, clone 37.51) in 1× PBS overnight. For proliferation and suppression assays, cells were labeled with CellTrace Violet (Life Technologies) as indicated. The in vitro differentiation assay conditions used for 3 d were as follows: tTreg and Th0: h.rIL-2 (R&D Systems) 100 IU/ml; iTreg: IL-2 100 IU/ml, h rTGF-β 10 ng/ml (PeproTech), α-IL4 and α-IFN-γ 10 μg/ml; Th1: h.r.IL-2 100 IU/ml, mouse IL-12 5 ng/ml (PeproTech), and α-IL4 10 μg/ml. For differentiated cells analyzed at day 5, cells were split 1:2 on day 3 and rested with IL-2 100 IU/ml for 48 h before analysis as described previously (10). For suppression assays, Tregs at day 5 were cocultured with labeled CD45.1 +total T cells in the presence of soluble αCD3 at 0.25 μg/ml and irradiated APCs (5 × 10 4cells) at the indicated Treg/T cell ratios for 3 d, and proliferation of T responders was analyzed on day 8 (19). Metabolic assays The extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) were measured using the Seahorse bioflux analyzer (18). Glucose-derived lipogenesis assay was measured using radiolabeled 14C-U-glucose as described previously (15). Samples for the untargeted metabolite profiling were processed as described previously (20). The peak intensities were analyzed using MetaboAnalyst software. All the data were subject to column-wise normalization method. The media supernatants were used to analyze the changes in glucose, glutamine, and lactate and glutamate levels using the YSI 2950 metabolite analyzer. Real-time PCR and immunoblotting Quantitative PCR was performed on cDNA using the SYBR Green ROX PCR Mastermix (Qiagen) as per the manufacturer’s protocols. The relative gene expression of β-actin, Glut1, Glut3, and Hk2was calculated using the ΔΔCt method and normalized to β-actin. Immunoblotting for glucose transporters and β-actin was performed as previously described (10,21). Statistics Groups of more than three were compared using ordinary one-way ANOVA analysis with Tukey posttest analysis using GraphPad Prism software. Results and Discussion TGF-β inhibits glycolysis of tTregs For the studies below, absent an agreed-upon surface marker for tTregs, we have considered Tregs isolated directly ex vivo from spleen and peripheral nonmesenteric lymph nodes as being of thymic origin, as these cells predominantly express neuropilin-1, a marker previously associated with tTregs (22). To determine the metabolic signature of tTregs, we activated YFP +CD4 +(tTregs) in the presence of IL-2 for 3 d while simultaneously activating YFP −T conv under Th0 conditions (IL-2 alone) or Th1-differentiating conditions (in the presence of IL-2 and IL-12) and iTreg-inducing conditions (in the presence of IL-2 and the anti-inflammatory cytokine TGF-β) (Fig. 1A). By Seahorse bioenergetic flux analysis, we found tTregs exhibited essentially equivalent glycolytic function (ECAR) as Th1 cells (Fig. 1B). In contrast, iTregs had a relatively lower ECAR as well as basal OCR compared with tTregs (Fig. 1B). FIGURE 1. TGF-β inhibits glycolytic metabolism in tTregs. (A) Schematic of the 3-d activation protocol of tTregs, T effector, and iTreg differentiation and culture of tTregs in iTreg (i.e., TGF-β). (B) The ECAR ( n= 4), OCR ( n= 3) in all the cell types tested (C). Histograms of Foxp3 expression (percentage and MFI) in day 3 activated cell types as indicated, respectively ( n= 4). (D) Protein expression of Glut1, Glut3 in the indicated cell types. Result is representative of four independent experiments. (E) Relative mRNA expression of Hk2. Each point is indicative of a single experiment. All the groups were compared using one-way ANOVA analysis with Tukey posttest analysis. Error bars represent means ± SD. * p< 0.05, **** p< 0.0001. FIGURE 1. TGF-β inhibits glycolytic metabolism in tTregs. (A) Schematic of the 3-d activation protocol of tTregs, T effector, and iTreg differentiation and culture of tTregs in iTreg (i.e., TGF-β). (B) The ECAR ( n= 4), OCR ( n= 3) in all the cell types tested (C). Histograms of Foxp3 expression (percentage and MFI) in day 3 activated cell types as indicated, respectively ( n= 4). (D) Protein expression of Glut1, Glut3 in the indicated cell types. Result is representative of four independent experiments. (E) Relative mRNA expression of Hk2. Each point is indicative of a single experiment. All the groups were compared using one-way ANOVA analysis with Tukey posttest analysis. Error bars represent means ± SD. * p< 0.05, **** p< 0.0001. TGF-β, an anti-inflammatory cytokine that facilitates the development of iTregs, functions in a complementary role to Foxp3 in promoting mitochondrial oxidative metabolism (13,14). Because a key difference in the culture conditions for tTregs and iTregs is the use of TGF-β in iTreg differentiation, we stimulated tTregs in iTreg conditions (i.e., including TGF-β) to determine the effect of TGF-β on tTreg metabolism (Fig. 1A). Although inclusion of TGF-β in the cultures had no adverse effect on tTreg survival (data not shown) or Foxp3 expression (percentage and MFI) (Fig. 1C), it dramatically lowered glycolytic function of tTregs (Fig. 1B). Furthermore, TGF-β also blunted glutamine consumption, a key anaplerotic event that accompanies glycolysis in activated cells (data not shown). A key checkpoint for glycolysis is glucose uptake, which is controlled by the glucose transporter family of proteins. Among the 13 members, only four (Glut1, Glut3, Glut6, and Glut8) are detected in activated T cells, with Glut6 and Glut8 exhibiting lower copy numbers than Glut1 and Glut3. Interestingly, neither tTregs nor iTregs require Glut1 for their development or function in vivo, suggesting that other members of the Glut family may be playing a nonredundant role in these subsets (21). In accordance, tTregs in fact expressed much higher levels of Glut1 and Glut3 protein, like Th1 cells, whereas iTregs exhibited either low or undetectable levels of this protein (Fig. 1D). Most strikingly, TGF-β abrogated both mRNA (data not shown) and protein expression of Glut1 and Glut3 in tTregs (Fig. 1D). TGF-β exposure also dramatically inhibited tTreg expression of Hk2, a key enzyme in proximal glycolysis (Fig. 1E), suggesting a metabolic reprogramming of tTregs in the presence of TGF-β. TGF-β inhibits glucose metabolism and suppressive capability via inhibition of PI3K-mediated mTOR-dependent signaling in tTregs TGF-β–mediated SMAD3 activation limits CD4 T cell growth and proliferation by attenuation of CD28-mediated costimulation and PI3k/Akt/mTOR pathway activity (23). We therefore examined phosphorylation of S6 kinase and Akt, two major downstream targets in this pathway. We found that TGF-β decreased phosphorylation of S6 (an mTORC1 target) and Akt (S473) (an mTORC2 target) in tTregs (Fig. 2A). Consequently, the overall growth (data not shown) and division of tTregs stimulated in the presence of TGF-β was also diminished significantly, suggesting that TGF-β reduces Treg proliferation by inhibiting glycolytic metabolism independent of Foxp3 expression (Fig. 2B). Additionally, the mTOR-dependent glycolysis–lipogenic pathway is critical for the suppressive capability of Tregs (15). We observed that tTregs cultured in the presence of TGF-β exhibited decreased suppressive function in vitro compared with tTregs (Fig. 2C). However, they still exhibited higher levels of suppression than iTregs, suggesting that glycolysis partly contributes to the suppressive function of tTregs. Concomitantly, we also observed a significant reduction in the expression (percentage and MFI) of key Treg molecules, such as ICOS and CTLA4 to levels resembling those of iTregs (Fig. 2D). FIGURE 2. TGF-β diminishes mTOR-dependent signaling that is associated with decrease in proliferation and suppressive capability in tTregs. (A) Histogram plots with average percentages ± SD of phosphorylated S6 kinase (pS6) ( n= 5) and phosphorylated Akt S473 (pAkt473) ( n= 3). (B) Histogram plots of dilution of CellTrace Violet (cell proliferation) in the four cell types, respectively. Results are representative of three independent experiments. (C) Histograms (1:12 ratio) and percentage of inhibition of CD4 T cell proliferation on day 8 of Treg suppression assay. Data are representative of two independent experiments (D). Histogram plots of phenotypic markers such as ICOS and CTLA4. The average percentage and MFI of each marker within each cell type is indicated. Results represent four independent experiments. pvalues for all the groups were calculated using one-way ANOVA analysis with Tukey posttest. Error bars represent means ± SD. * p< 0.05, ** p< 0.01, *** p< 0.001, **** p< 0.0001. FIGURE 2. TGF-β diminishes mTOR-dependent signaling that is associated with decrease in proliferation and suppressive capability in tTregs. (A) Histogram plots with average percentages ± SD of phosphorylated S6 kinase (pS6) ( n= 5) and phosphorylated Akt S473 (pAkt473) ( n= 3). (B) Histogram plots of dilution of CellTrace Violet (cell proliferation) in the four cell types, respectively. Results are representative of three independent experiments. (C) Histograms (1:12 ratio) and percentage of inhibition of CD4 T cell proliferation on day 8 of Treg suppression assay. Data are representative of two independent experiments (D). Histogram plots of phenotypic markers such as ICOS and CTLA4. The average percentage and MFI of each marker within each cell type is indicated. Results represent four independent experiments. pvalues for all the groups were calculated using one-way ANOVA analysis with Tukey posttest. Error bars represent means ± SD. * p< 0.05, ** p< 0.01, *** p< 0.001, **** p< 0.0001. Collectively, these data suggest that TGF-β reprograms tTreg metabolism via blockade of PI3K-mediated mTOR signaling and glucose metabolism. Moreover, given the interplay between metabolic regulation and cellular function, this suggests that TGF-β may selectively modulate the metabolism of tTregs and iTregs in a context-dependent manner (i.e., TGF-β–rich environments can normalize the glycolytic differences between tTregs and iTregs, and metabolic differences observed between these subsets in vitro may be a result of their culture conditions rather than a fixed metabolic program. Effects of manipulating PI3K activity on Treg subsets Previously, others and we have shown that control of PI3K activity is critical to maintain Treg stability and function (18,24). Conditions of inflammation such as TLR activation and defective autophagy activate PI3K-Akt–mediated mTOR signaling and enhance glycolysis in Tregs, leading to impaired lineage stability and function (25,26). In T cells, the lipid phosphatase PTEN is the primary negative regulator of PI3K activity. Although PTEN is constitutively expressed in all resting T cells, it is downregulated in activated T conv, which facilitates their optimal expansion (27). Tregs, in contrast, have been reported to retain PTEN expression postactivation as a means of avoiding high PI3K activity (27), and we confirmed that this is true in both tTregs and iTregs (Supplemental Fig. 1A–C). Given that tTreg and iTreg exhibit differential requirements for glycolysis and mTOR signaling, we hypothesized that overactivation of PI3K signaling via PTEN loss would have distinct effects on glycolysis in these subsets. To establish a system to study PTEN-deficient tTregs, we used Tregs isolated from PTEN fl/fl x Foxp3-YFP-Cremice, also termed PTEN-ΔTreg mice (18). To obtain PTEN-deficient iTregs, we stimulated CD4 T conv from either Foxp3-YFP-Creor PTEN fl/fl x Foxp3 YFP-Cremice in iTreg-inducing conditions (PTENΔ−iTreg) and confirmed that PTEN expression is decreased following induction of Foxp3 (Supplemental Fig. 1D–F). Surprisingly, although others and we have previously shown increased rates of aerobic glycolysis in PTEN-deficient tTregs shortly after activation (6–24 h) (18,24), we found that by 3 d postactivation, ECAR and OCR rates of PTEN-deficient tTregs were indistinguishable from WT tTregs (Fig. 3A). This indicated that the ability of PTEN-deficient tTregs to undergo glycolysis is restored to WT levels during the late phase of activation, whereas there is no major change in mitochondrial metabolism in tTregs in the absence of PTEN. This suggested that glucose might be redirected to other metabolic pathways other than lactate production in PTEN-deficient tTregs. One of the alternate fates of glucose other than aerobic glycolysis is entry into the pentose phosphate pathway, which feeds directly into nucleotide biosynthesis. To better define this, we performed an unlabeled metabolomic analysis of freshly isolated WT and PTEN ΔTregs. Metabolite enrichment analysis revealed enhanced nucleotide biosynthetic metabolites, including ADP, ATP, GTP, and UTP in PTEN-deficient tTregs compared with WT tTregs (Supplemental Fig. 2A). These observations were consistent with our previous findings showing enhanced BrdU labeling of PTEN ΔTregs compared with WT Tregs, suggesting their rapid turnover in vivo (18). We also found that PTEN deficiency enhanced 14C glucose labeling of lipids in tTregs, suggesting enhanced lipogenesis (Supplemental Fig. 2B). FIGURE 3. Elevation of PI3K signaling specifically enhances glycolytic metabolism of iTreg subsets but not tTregs. (A) and (B) Measure of ECAR and OCR of day 3 activated tTregs and iTregs from both WT and PTEN ΔTreg mice, respectively. Results represent four independent experiments. (C) Foxp3 expression (percentage positive and MFI) in day 4 stimulated tTregs and iTregs that are either WT- or PTEN-deficient and Th1 cells (control), respectively. Results represent three mice per group. (D) tTregs and iTregs that are either WT- or PTEN-deficient were activated for 3 d in their respective conditions and rested for 2 d with IL-2. Total (both surface and intracellular) Glut1 expression (percentage positive and MFI) in these stimulated cells was analyzed on day 5. Results represent three mice per group. pvalues determined by one-way ANOVA analysis with Tukey posttest analysis. Error bars indicate means ± SD. * p< 0.05, ** p< 0.01, **** p< 0.0001. FIGURE 3. Elevation of PI3K signaling specifically enhances glycolytic metabolism of iTreg subsets but not tTregs. (A) and (B) Measure of ECAR and OCR of day 3 activated tTregs and iTregs from both WT and PTEN ΔTreg mice, respectively. Results represent four independent experiments. (C) Foxp3 expression (percentage positive and MFI) in day 4 stimulated tTregs and iTregs that are either WT- or PTEN-deficient and Th1 cells (control), respectively. Results represent three mice per group. (D) tTregs and iTregs that are either WT- or PTEN-deficient were activated for 3 d in their respective conditions and rested for 2 d with IL-2. Total (both surface and intracellular) Glut1 expression (percentage positive and MFI) in these stimulated cells was analyzed on day 5. Results represent three mice per group. pvalues determined by one-way ANOVA analysis with Tukey posttest analysis. Error bars indicate means ± SD. * p< 0.05, ** p< 0.01, **** p< 0.0001. Interestingly, PTEN loss in iTregs predominantly upregulated aerobic glycolysis with no changes in oxidative metabolism (Fig. 3B). Notably, we observed a significant reduction in Foxp3 expression in PTEN-deficient iTregs but not in tTregs by day 4 postactivation (Fig. 3C). Interestingly, by day 5 of activation post–IL-2 culturing, PTEN-deficient iTregs exhibit higher Glut1 protein expression compared with PTEN-deficient tTregs (Fig. 3D). Taken together, this indicated that although enhanced PI3K activity in tTregs led to the diversion of glucose toward nucleotide biosynthesis and the glycolytic lipogenesis pathway without substantively altering glycolysis, in iTregs, it directly enhances glycolysis (Supplemental Fig. 2C). Collectively, these findings demonstrate that upregulation of PI3K signaling has different metabolic consequences in Treg subsets. In conclusion, our study indicates that tTregs and iTregs are metabolically distinct but subject to modulation by external and context-dependent cues, such as inflammatory or anti-inflammatory stimuli (PI3K activation and TGF-β, respectively). The role of this metabolic plasticity is likely adaptation to the local environment, balancing cell division, cell survival, and regulatory function to appropriately meet the needs of the organism. Acknowledgements We thank Dr. Christian Leguern for scientific discussions, the Sanford Burnham Prebys Medical Discovery Institute for YSI analysis, and the Beth Israel Deaconess Medical Center mass spectrometry facility for processing metabolomics samples, and the MGH bioinformatics group for analyses. Footnotes This work was supported by National Institutes of Health Grants R01 HL11879 (to B.R.B.) and P01 AI056299 (to L.A.T. and B.R.B.). The online version of this article containssupplemental material. Abbreviations used in this article: ECAR extracellular acidification rate iTreg induced Treg MGH Massachusetts General Hospital OCR oxygen consumption rate PTEN phosphatase and tensin homolog T conv conventional T cell Treg regulatory T cell tTreg thymic-derived Treg WT wild type. References 1 Galgani , M. , V. De Rosa , A. La Cava , G. Matarese . 2016 . Role of metabolism in the immunobiology of regulatory T cells. J. Immunol. 197 : 2567 – 2575 . 2 Wing , K. , S. Sakaguchi . 2010 . Regulatory T cells exert checks and balances on self tolerance and autoimmunity. Nat. Immunol. 11 : 7 – 13 . 3 Curotto de Lafaille , M. A. , J. J. Lafaille . 2009 . Natural and adaptive foxp3+ regulatory T cells: more of the same or a division of labor? Immunity 30 : 626 – 635 . 4 Chen , W. , W. Jin , N. Hardegen , K.-J. Lei , L. Li , N. Marinos , G. McGrady , S. M. Wahl . 2003 . Conversion of peripheral CD4+CD25- naive T cells to CD4+CD25+ regulatory T cells by TGF-beta induction of transcription factor Foxp3. J. Exp. Med. 198 : 1875 – 1886 . PubMed Dominitzki , S. , M. C. Fantini , C. Neufert , A. Nikolaev , P. R. Galle , J. Scheller , G. Monteleone , S. Rose-John , M. F. Neurath , C. Becker Cutting edge: trans-signaling via the soluble IL-6R abrogates the induction of FoxP3 in naive CD4+CD25 T cells. Fetal regulatory T cells and peripheral immune tolerance in utero: implications for development and disease. Am. J. Reprod. Immunol. Cutting edge: distinct glycolytic and lipid oxidative metabolic programs are essential for effector and regulatory CD4+ T cell subsets. : 3299 – 3303 . 11 Gerriets , V. A. , R. J. Kishton , A. G. Nichols , A. N. Macintyre , M. Inoue , O. Ilkayeva , P. S. Winter , X. Liu , B. Priyadharshini , M. E. Slawinska , et al . 2015 . Metabolic programming and PDHK1 control CD4+ T cell subsets and inflammation. J. Clin. Invest. 125 : 194 – 207 12 Gerriets , V. A. , R. J. Kishton , M. O. Johnson , S. Cohen , P. J. Siska , A. G. Nichols , M. O. Warmoes , A. A. de Cubas , N. J. MacIver , J. W. Locasale , et al . 2016 . Foxp3 and Toll-like receptor signaling balance T reg cell anabolic metabolism for suppression. Nat. Immunol. 17 : 1459 – 1466 . 13 2017 . Foxp3 reprograms T cell metabolism to function in low-glucose, high-lactate environments. Cell Metab. 25 : 1282 – 1293.e7 . 14 Howie , D. , S. P. Cobbold , E. Adams , A. Ten Bokum , A. S. Necula , W. Zhang , H. Huang , D. J. Roberts , B. Thomas , S. S. Hester , et al . 2017 . Foxp3 drives oxidative phosphorylation and protection from lipotoxicity. JCI Insight e89160 . 15 Zeng , H. , K. Yang , C. Cloer , G. Neale , P. Vogel , H. Chi . 2013 . mTORC1 couples immune signals and metabolic programming to establish T(reg)-cell function. Nature 499 : 485 – 490 . 16 Procaccini , C. , F. Carbone , D. Di Silvestre , F. Brambilla , V. De Rosa , M. Galgani , D. Faicchia , G. Marone , D. Tramontano , M. Corona , et al . 2016 . The proteomic landscape of human ex vivo regulatory and conventional T cells reveals specific metabolic requirements. [Published erratum appears in 2016 Immunity 44: 712.] Immunity 44 : 406 – 421 . 17 Kishore , M. , K. C. P. Cheung , H. Fu , F. Bonacina , G. Wang , D. Coe , E. J. Ward , A. Colamatteo , M. Jangani , A. Baragetti , et al . 2017 . Regulatory T cell migration is dependent on glucokinase-mediated glycolysis. [Published erratum appears in 2018 Immunity 48: 831–832.] Immunity 47 : 875 – 889.e10 . 18 Huynh , A. , M. DuPage , B. Priyadharshini , P. T. Sage , J. Quiros , C. M. Borges , N. Townamchai , V. A. Gerriets , J. C. Rathmell , A. H. Sharpe , et al . 2015 . Control of PI(3) kinase in Treg cells maintains homeostasis and lineage stability. Nat. Immunol. 16 : 188 . 19 Bandukwala , H. S. , Y. Wu , M. Feuerer , Y. Chen , B. Barboza , S. Ghosh , J. C. Stroud , C. Benoist , D. Mathis , A. Rao , L. Chen . 2011 . Structure of a domain-swapped FOXP3 dimer on DNA and its function in regulatory T cells. [Published erratum appears in 2011 Immunity 34: 627.] Immunity 34 : 479 – 491 . Yuan , M. , S. B. Breitkopf , X. Yang , J. M. Asara . 2012 . A positive/negative ion-switching, targeted mass spectrometry-based metabolomics platform for bodily fluids, cells, and fresh and fixed tissue. Nat. Protoc. 7 : 872 – 881 . 21 Macintyre , A. N. , V. A. Gerriets , A. G. Nichols , R. D. Michalek , M. C. Rudolph , D. Deoliveira , S. M. Anderson , E. D. Abel , B. J. Chen , L. P. Hale , J. C. Rathmell . 2014 . The glucose transporter Glut1 is selectively essential for CD4 T cell activation and effector function. 20 : 61 – 72 . 22 Weiss , J. M. , A. M. Bilate , M. Gobert , Y. Ding , M. A. Curotto de Lafaille , C. N. Parkhurst , H. Xiong , J. Dolpady , A. B. Frey , M. G. Ruocco , et al . 2012 . Neuropilin 1 is expressed on thymus-derived natural regulatory T cells, but not mucosa-generated induced Foxp3+ T reg cells. J. Exp. Med. 209 : 1723 – 23 Delisle , J.-S. , M. Giroux , G. Boucher , J.-R. Landry , M.-P. Hardy , S. Lemieux , R. G. Jones , B. T. Wilhelm , C. Perreault . 2013 . The TGF-β-Smad3 pathway inhibits CD28-dependent cell growth and proliferation of CD4 T cells. [Published erratum appears in 2013 Genes Immun. 14: 132.] Genes Immun. 14 : 115 – 126 . 24 . 2015 . Treg cells require the phosphatase PTEN to restrain TH1 and TFH cell responses. Nat. Immunol. 16 : 178 – 187 . 25 Bailey-Bucktrout , S. L. , M. Martinez-Llordella , X. Zhou , B. Anthony , W. Rosenthal , H. Luche , H. J. Fehling , J. A. Bluestone . 2013 . Self-antigen-driven activation induces instability of regulatory T cells during an inflammatory autoimmune response. : 949 – 962 . 26 Wei , J. , L. Long , K. Yang , C. Guy , S. Shrestha , Z. Chen , C. Wu , P. Vogel , G. Neale , D. R. Green , H. Chi . 2016 . Autophagy enforces functional integrity of regulatory T cells by coupling environmental cues and metabolic homeostasis. Nat. Immunol. 17 : 277 – 285 Supplementary data Supplemental Figures 1 (PDF) - pdf file

https://journals.aai.org/jimmunol/article/201/8/2215/107025

Study of ASTX727 Plus Talazoparib in Patients With Triple Negative or Hormone Resistant/HER2-negative Metastatic Breast Cancer - Tabular View - ClinicalTrials.gov Study of ASTX727 Plus Talazoparib in Patients With Triple Negative or Hormone Resistant/HER2-negative Metastatic Breast Cancer The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been evaluated by the U.S. Federal Government. Know the risks and potential benefits of clinical studies and talk to your health care provider before participating. Read our disclaimer for details. ClinicalTrials.gov Identifier: NCT04134884 Recruitment Status : Recruiting First Posted : October 22, 2019 Last Update Posted : May 6, 2023 See Contacts and Locations Sponsor: Kathy Miller Collaborators: Pfizer Astex Pharmaceuticals, Inc. Van Andel Institute Stand Up to Cancer Team Information provided by (Responsible Party): Kathy Miller, Indiana University Study Details Tabular View Tracking Information First Submitted Date ICMJE October 18, 2019 First Posted Date ICMJE October 22, 2019 Last Update Posted Date May 6, 2023 Actual Study Start Date ICMJE April 1, 2020 Estimated Primary Completion Date December 31, 2023   (Final data collection date for primary outcome measure) Current Primary Outcome Measures ICMJE (submitted: October 18, 2019) Safety of ASTX727 plus talazoparib using National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE) v 5.0 [ Time Frame: through study completion i.e up to 1 year ] Rate of dose limiting toxicity [ Time Frame: 28 days ] rate of dose limiting toxicity will be assessed during cycle 1 (28 days) in patients enrolled during the dose escalation phase Original Primary Outcome Measures ICMJE Same as current Change History Complete list of historical versions of study NCT04134884 on ClinicalTrials.gov Archive Site Current Secondary Outcome Measures ICMJE (submitted: October 18, 2019) Overall response rate [ Time Frame: through study completion (i.e. up to 1 year) ] Clinical benefit response for triple negative disease subjects [ Time Frame: 18 weeks ] clinical benefit response defined as complete response, partial response, or stable disease Clinical benefit response for hormone receptor positive/ HER2 negative subjects [ Time Frame: 24 weeks ] clinical benefit response defined as complete response, partial response, or stable disease Progression free survival in all enrolled subjects [ Time Frame: through study completion (i.e. up to 1 year) ] Original Secondary Outcome Measures ICMJE Same as current Current Other Pre-specified Outcome Measures Not Provided Original Other Pre-specified Outcome Measures Not Provided Descriptive Information Brief Title ICMJE Study of ASTX727 Plus Talazoparib in Patients With Triple Negative or Hormone Resistant/HER2-negative Metastatic Breast Cancer Official Title ICMJE A Phase I Study of ASTX727 Plus Talazoparib in Patients With Triple Negative or Hormone Resistant/ Human Epidermal Growth Factor Receptor 2 (HER2)-Negative Metastatic Breast Cancer Brief Summary This is a Phase I study to test the safety of a combination of ASTX727 with talazoparib in patients with triple negative breast cancer or hormone resistant/HER2-negative metastatic breast cancer Detailed Description The phase I portion will use a traditional 3 + 3 design and standard definitions of DLT based on toxicity experienced during the first cycle of therapy. Patients with triple negative breast cancer (TNBC) and hormone resistant/HER2 negative (HRBC) metastatic disease will be enrolled and analyzed together during the dose escalation cohorts. Once the maximum tolerated dose is determined, we will enroll a small expansion cohort to further characterize safety and provide preliminary efficacy estimates.The expansion cohort will be limited to 14 patients; 7 with TNBC and 7 with HRBC. The dose level selected for expansion will be based on the totality of the data available including toxicity during the DLT evaluation period, toxicity during subsequent cycles, and correlative results. Study Type ICMJE Interventional Study Phase ICMJE Phase 1 Study Design ICMJE Allocation: N/A Intervention Model: Sequential Assignment Intervention Model Description:The dose escalation phase will begin by enrolling 3 patients to the cohort 1 dose level. Patients will be observed for 28 days or one cycle of therapy for dose limiting toxicity (DLT).If 0 of 3 patients experience a DLT, the study will proceed to cohort 2. If 1 patient experiences a DLT, 3 additional patients will be enrolled into cohort 1. If 1 of 6 patients experience a DLT, the study will proceed to the cohort 2 dose level. If > 2/3 or 2/6 patients in cohort 1 experience DLT, we will de-escalate to cohort -1. Identical DLT evaluation and dose escalation/de-escalation decision rules will be used in subsequent cohorts. A total of 6 patients will be treated at the highest dose level achieved to ensure that 6 patients have been treated at the proposed maximum tolerated dose before proceeding to the expansion cohorts. Masking: None (Open Label) Primary Purpose: Treatment Condition ICMJE Metastatic Breast Cancer Triple Negative Breast Cancer Hormone Receptor Positive Tumor Intervention ICMJE Drug: Talazoparib Talazoparib will be taken on days 4-21, 6-21 or 1-21 at a dose level of 0.25, 0.5, 0.75 or 1.0 mg depending on cohort assignment Drug: ASTX727 ASTX727 will be taken on days 1 and 3, days 1,3,5 or days 1 through 5 of the 28 day cycle at doses of 10 mg:100 mg or 15 mg:100 mg depending on cohort assignment Study Arms ICMJE Experimental: ASTX727 + Talazoparib Interventions: Drug: Talazoparib Drug: ASTX727 Publications * Not Provided *   Includes publications given by the data provider as well as publications identified by ClinicalTrials.gov Identifier (NCT Number) in Medline. Recruitment Information Recruitment Status ICMJE Recruiting Estimated Enrollment ICMJE (submitted: August 7, 2021) 38 Original Estimated Enrollment ICMJE (submitted: October 18, 2019) 32 Estimated Study Completion Date ICMJE July 1, 2024 Estimated Primary Completion Date December 31, 2023   (Final data collection date for primary outcome measure) Eligibility Criteria ICMJE Inclusion Criteria: ≥ 18 years old at the time of informed consent Ability to provide written informed consent and HIPAA authorizationLocally recurrent (not amenable to local therapy with curative intent) or metastatic breast cancer Patients with triple negative breast cancer must have received at least one prior chemotherapy regimen for metastatic disease. Patients with hormone-positive, HER2-negative disease must have received treatment with and progressed on at least one prior endocrine therapy including a CDK4/6 inhibitor in the metastatic setting. Measurable or evaluable disease based on RECIST 1.1 criteria.Only subjects who have disease amenable to biopsy will be asked to consent to serial tumor biopsies. Consent for biopsy is not required for participation.a. NOTE: If no amendable disease is present at the time of biopsy, subjects may continue participation in the study and further study specific biopsies will not be required. Eastern Cooperative Oncology Group Performance Status 0 or 1 Patients with treated, asymptomatic central nervous system (CNS) disease may participate if the patient is > 4 weeks from completion of CNS therapy (radiation and/or surgery), is clinically stable at the time of study entry, and is receiving a stable or decreasing dose of corticosteroid therapy. Brain MRI or head CT is required at screening for patients with known brain metastases.Adequate organ function as indicated by: Total bilirubin </= ULN (upper limit of normal) (except in patients with documented Gilbert's disease, who must have a total bilirubin </= 3.0 mg/dL) Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) </= 3.0 x ULN (</= 1.5-3.0 x baseline if baseline is abnormal) Calculated creatinine clearance of >/= 60 mL/min using the Cockcroft-Gault formula Absolute neutrophil count (ANC) >/= 1.5 K/mm3 Platelets >/= 100 K/mm3 Hemoglobin (Hgb) >/= 9.0 g/dLWomen of childbearing potential must have a negative pregnancy test within 14 days of protocol registration. Women are considered to have childbearing potential (regardless of sexual orientation, having undergone a tubal ligation, or remaining celibate by choice) unless they meet one of the following criteria: Has undergone a hysterectomy or bilateral oophorectomy; or Has been naturally amenorrheic for at least 24 consecutive months. Women of childbearing potential and men must agree to use effective contraception throughout the study and for 7 months after the last study treatment. Note: Acceptable methods of birth control include abstinence, partner with previous vasectomy, placement of an intrauterine device (IUD), condom with spermicidal foam/gel/film/cream/suppository, diaphragm or cervical vault cap, or hormonal birth control (pills or injections).Exclusion Criteria: Prior treatment with decitabine, guadecitabine or other known DNA Methyltransferase inhibitors (DNMTis) Prior treatment with talazoparib or other known PARPi (poly(ADP-ribose polymeras inhibitor) Known deleterious breast cancer susceptibility gene (BRCA) mutation. Patients with BRCA variants of unknown significance (VUS) or who have not had germline genetic testing may participate. Active or symptomatic CNS diseasePatients with HER2+ disease HER2 will be considered positive if scored 3+ by immunohistochemistry (IHC) or 2+ by IHC associated with a fluorescence in situ hybridization (FISH) ratio of > 2.0 or > 6 total HER2 gene copies per cell. Patients with active malignancy other than breast cancer. Patients with prior malignancies without recurrence after standard treatment will not be excluded Chemotherapy within 3 weeks of registration Radiation therapy within 2 weeks of registration Hormone therapy within 2 weeks of registration Patients requiring ongoing therapy with strong P-gp inhibitors

https://clinicaltrials.gov/ct2/show/record/NCT04134884

The Association of Haplotype at the Lumican Gene with High Myopia Susceptibility in Taiwanese Patients | Request PDF Request PDF | The Association of Haplotype at the Lumican Gene with High Myopia Susceptibility in Taiwanese Patients | To investigate the association of genetic polymorphisms in the dermatan sulfate proteoglycan 3 (DSPG3), lumican (LUM), and decorin (DCN) genes... | Find, read and cite all the research you need on ResearchGate Article The Association of Haplotype at the Lumican Gene with High Myopia Susceptibility in Taiwanese Patients August 2009 Ophthalmology116(10):1920-7 DOI: 10.1016/j.ophtha.2009.03.023 Source PubMed Authors: <here is a image f2f62089ad6ff61c-d19572ff1996375a> Zoe Tzu-Yi Chen Zoe Tzu-Yi Chen <here is a image 27a66ac73e324777-d630edeec7414892> I-Jong Wang National Taiwan University Hospital <here is a image f2f62089ad6ff61c-d19572ff1996375a> Yung-Feng Shih Yung-Feng Shih <here is a image f2f62089ad6ff61c-d19572ff1996375a> Luke Long-Kung Lin Luke Long-Kung Lin Abstract To investigate the association of genetic polymorphisms in the dermatan sulfate proteoglycan 3 (DSPG3), lumican (LUM), and decorin (DCN) genes (component genes of the sclera) with high myopia susceptibility in Taiwanese people. Prospective case-control study. Hospital clinic-based samples of 120 unrelated patients with extremely high myopia were studied. One hundred thirty-seven unrelated emmetropic individuals served as controls. Four, 8, and 4 single nucleotide polymorphism (SNPs) were genotyped within the DSPG3, lumican, and decorin genes, respectively, using direct DNA sequencing. Pairwise linkage disequilibrium, haplotype analysis, adjusted logistic regression, and multifactor dimensionality reduction (MDR) methods were used to determine significant associations. The association of haplotypes at the lumican gene with high myopia development. The lumican gene SNP rs3759223:T-->C demonstrated a significant association with high myopia (P = 2.83 x 10(-4)). Four lumican SNPs showed significant linkage disequilibrium and formed a haplotype block. Sliding window haplotype analyses revealed that the block consisting of rs3759223 and rs3741834 showed significant goodness of fit (global P = 1.0725 x 10(-6)). Haplotype-specific tests showed that the C-C and T-C haplotypes were associated significantly with high myopia, with odds ratios (95% confidence intervals) of 19.32 (2.55-146.54) and 0.69 (0.46-1.04), respectively. rs3759223 and rs3741834 are in a putative regulatory element of the lumican gene, which influences fibrillogenesis of scleral collagen fibers and the development of myopia. The results of an MDR analysis corroborated the single-locus association and suggested a significant 2-locus interaction model composed of SNPs rs2300588 and rs3741834 in the lumican gene. Genetic variation in the regulatory domains of the lumican gene, where both rs3759223 and rs3741834 are located, are associated with high myopia susceptibility among the Han Chinese, making this region worthy of further investigation. <here is a image 175d1bd4b7439d95-754927124da715c1> ... Another group explored four DCN and four EPYC polymorphisms, with only four (rs2070985 of DCN and rs1920748, rs1920751 and rs1920752 of EPYC) being polymorphic in the Chinese population under study, and these four polymorphic markers were found not to be associated with high myopia in a study of 120 cases (SE Յ Ϫ10.0 D) and 137 controls. 31 Two DCN and three EPYC tSNPs were screened in the present study and were also found not to be associated with high myopia (Table 2). We did not examine rs2070985 of DCN and rs1920748 of EPYC, because their MAFs (0.089 and 0.081, respectively) documented in the HapMap database for Han Chinese are less than the selection threshold (MAF Ն 0.10) for our study. ... ... Another Taiwanese group investigated eight SNPs in the LUM gene in 120 cases of high myopia (Յ Ϫ10.0 D) and 137 controls (Ϫ1.5 to ϩ0.5 D) and found that rs3759223 showed a significant association with high myopia (P ϭ 2.83 ϫ 10 Ϫ4 ). 31 Nonetheless, this association was not substantiated in two other studies of Chinese subjects. 32, 33 We did not examine rs3759223. ... ... Note that rs2300588 is in strong LD with rs3759223 (r 2 ϭ 0.773). This discrepancy may be due to the use of different thresholds for defining high myopia in different studies: Ϫ10.0 D in the positive study, 31 Ϫ8.0 D in the present study, and Ϫ6.0 D in the other two negative studies. 32,33 We focused on common polymorphisms of these six candidate genes, but did not explore the role of their rare variants in high myopia. ... Evaluation of Proteoglycan Gene Polymorphisms as Risk Factors in the Genetic Susceptibility to High Myopia Article Full-text available Sep 2011 <here is a image 9d1180a55ee9944e-e33a3285750cabba> Shea Ping Yip <here is a image f8fff7014566481a-a06f0303a869bd14> kim hung Leung Po Wah Ng Maurice K H Yap PURPOSE. To investigate the relationship between high myopia and single nucleotide polymorphisms (SNPs) in six proteoglycan genes: aggrecan (ACAN), fibromodulin (FMOD), decorin (DCN), lumican (LUM), keratocan (KERA), and epiphycan (EPYC). These genes were selected for study because they are involved in induced myopia in animals and/or are within the human MYP3 locus identified by linkage analysis of families with high myopia. METHODS. Two groups of Chinese subjects were studied: group 1 (300 cases and 300 controls) and group 2 (356 cases and 354 controls). Cases were high myopes with spherical equivalent (SE) ≤ -8.00 D, and controls had SE between +1.0 and -1.0 D. From these candidate genes, 60 tagging SNPs were selected. First, 12 DNA pools were each constructed from 50 samples of the same phenotype from group 1 subjects and were tested for association with the SNPs. Second, putatively positive SNPs were confirmed by individual genotyping of group 1 subjects. Finally, positive results were replicated in group 2 subjects. RESULTS. Of the 58 SNPs successfully screened by DNA pooling, 8 ACAN SNPs passed the threshold of P ≤ 0.10 (nested ANOVA) and were then genotyped in the individual samples. Haplotypes rs3784757 and rs1516794 showed significant association with high myopia. However, the positive result could not be replicated in the second subject group. CONCLUSIONS. These six proteoglycan genes were not associated with high myopia in these Chinese subjects and hence are unlikely to be important in the genetic predisposition to high myopia. ... Scleral thinning is a feature of the development of high myopia. Scleral remodeling affecting the scleral extracellular matrix could occur during myopia progression due to axial length elongation [77] . Another candidate gene in the MYP3 locus is IGF-1. ... Myopia Genetics and Heredity Article Full-text available Mar 2022 <here is a image 278c62ddcafb9a73-3b5110c90390834a> Yumeng Wang <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Shi Yao Lu <here is a image aea647a06b862fd7-dc209bbbdce9cb0a> Xiu Juan Zhang <here is a image 1585ec4dfe5162c7-68bb6bdf495557af> Jason C S Yam Myopia is the most common eye condition leading to visual impairment and is greatly influenced by genetics. Over the last two decades, more than 400 associated gene loci have been mapped for myopia and refractive errors via family linkage analyses, candidate gene studies, genome-wide association studies (GWAS), and next-generation sequencing (NGS). Lifestyle factors, such as excessive near work and short outdoor time, are the primary external factors affecting myopia onset and progression. Notably, besides becoming a global health issue, myopia is more prevalent and severe among East Asians than among Caucasians, especially individuals of Chinese, Japanese, and Korean ancestry. Myopia, especially high myopia, can be serious in consequences. The etiology of high myopia is complex. Prediction for progression of myopia to high myopia can help with prevention and early interventions. Prediction models are thus warranted for risk stratification. There have been vigorous investigations on molecular genetics and lifestyle factors to establish polygenic risk estimations for myopia. However, genes causing myopia have to be identified in order to shed light on pathogenesis and pathway mechanisms. This report aims to examine current evidence regarding (1) the genetic architecture of myopia; (2) currently associated myopia loci identified from the OMIM database, genetic association studies, and NGS studies; (3) gene-environment interactions; and (4) the prediction of myopia via polygenic risk scores (PRSs). The report also discusses various perspectives on myopia genetics and heredity. ... Most of the cells were vimentin + , indicating the successful isolation of scleral fibroblasts. Lumican expression has been demonstrated to alter collagen synthesis, leading to scleral remodeling, lengthening of the ocular axis and the development of myopia (29) . Lumican binds to integrin on the cell surface to regulate cell proliferation and apoptosis (30). ... Effects of lumican expression on the apoptosis of scleral fibroblasts: In vivo and in vitro experiments Article Full-text available Mar 2021 Jinsong Wu Yanzhi Zhao Yanmei Fu Xu Zhang Lumican serves an important role in the maintenance of sclera biomechanical properties. However, whether lumican expression is altered in myopia and the mechanisms of action involved are unknown. In the present study, the expression of lumican in cultured scleral fibroblasts and in the scleral tissue of a rat model of form-deprivation myopia was assessed. It was confirmed that diopter was decreased, whereas axial length was increased in modeled eyes relative to normal control eyes, indicating that the model of myopia was successfully established. These pathologic changes were accompanied by the upregulation of lumican and tissue inhibitor of metalloproteinases (TIMP)-2, as well as the downregulation of matrix metalloproteinase (MMP)-2 and MMP-14. The same trends in TIMP-2, MMP-2 and MMP-14 expression were observed when lumican was overexpressed in cultured scleral fibroblasts. Additionally, cell proliferation decreased whereas apoptosis increased compared with those of control cells. Inhibiting lumican expression had no effect on cell proliferation or apoptosis, but stimulated the expression of MMP-2 and MMP-14 while decreasing that of TIMP-2. The results suggested that lumican overexpression contributed to myopia by promoting apoptosis in scleral fibroblasts via the modulation of TIMP-2, MMP-2 and MMP-14 expression. ... In addition to collagen, the scleral extracellular matrix contains small leucine-rich proteoglycans such as lumican, decorin, and keratocan, which can regulate collagen fibrillogenesis; polymorphisms in the genes encoding these substances have been associated with myopia development in human genetics studies [13, 14]. In mice with both lumican and fibromodulin knockout, the collagen fibril diameter distribution was severely disrupted, and the phenotype resembled that of high myopia in humans; that is, increased AL, thin sclera, and retinal detachment [15]. ... Stepwise candidate drug screening for myopia control by using zebrafish, mouse, and Golden Syrian Hamster myopia models Background We developed a preclinical protocol for the screening of candidate drugs able to control myopia and prevent its progression. The protocol uses zebrafish, C57BL/6 mice, and golden Syrian hamster models of myopia. Methods A morpholino (MO) targeting the zebrafish lumican gene (zlum) was injected into single-cell zebrafish embryos, causing excessive expansion of the sclera. A library of 640 compounds with 2 matrix metalloproteinase (MMP) inhibitors (marimastat and batimastat), which have the potential to modulate scleral remodelling, was screened to identify candidates for mitigating scleral diameter expansion in zlum-MO-injected embryos. The myopia-prevention ability of compounds discovered to have superior potency to inhibit scleral expansion was validated over 4 weeks in 4-week-old C57BL/6 mice and 3-week-old golden Syrian hamsters with form-deprivation myopia (FDM). Changes in the refractive error and axial length were investigated. Scleral thickness, morphology of collagen fibrils in the posterior sclera, messenger RNA (mRNA) expressions, and protein levels of transforming growth factor-β2 (TGF-β2), tissue inhibitor of metalloproteinase-2 (TIMP-2), MMP-2, MMP-7, MMP-9, and collagen, type I, alpha 1 (collagen Iα1) were investigated in C57BL/6 mice, and MMP-2, MMP-9, and MMP activity assays were conducted in these mice. Findings In the zebrafish experiment, atropine, marimastat, batimastat, doxycycline, and minocycline were the drugs that most effectively reduced expansion of scleral equatorial diameter. After 28-day treatment in diffuser-wearing mice and 21-day treatment in lid-sutured hamsters, myopic shift and axial elongation were significantly mitigated by eye drops containing 1% atropine, 50 µM marimastat, 5 µM batimastat, or 200 µM doxycycline. MMP-2 mRNA expression in mouse sclera was lower after treatment with atropine, marimastat, batimastat, or doxycycline. The protein levels and activity of MMP-2 and MMP-7 were significantly reduced after treatment with atropine, marimastat, batimastat, doxycycline, and minocycline. Furthermore, scleral thickness and collagen fibril diameter were not lower after treatment with atropine, marimastat, batimastat, or doxycycline than those of occluded eyes. Interpretation Stepwise drug screening in a range of models from zlum-MO-injected zebrafish to rodent FDM models identified effective compounds for preclinical myopia control or prevention. On the basis of the 640 compounds that were screened, MMP inhibitors may offer alternatives for clinical trials. Funding This research was supported by grants from Taiwan's Ministry of Science and Technology and Ministry of Health and Welfare. ... Chakravarti et al. reported that fibromodulin-null and double lumican-and fibromodulin-null mice displayed an increased axial length, scleral thinning, and retinal detachment [84]. Hospital-based studies of Asian and European populations reported single nucleotide polymorphisms (SNPs) in the lumican promoter as a risk factor for myopia [190,192, 193] . While gene mutations or altered SLRP expression, such as that of lumican, potentially influences myopia pathogenesis through the disruption of scleral collagen organization, the roles of SLRPs and other proteoglycans should be further investigated in their regulation of the scleral ECM. ... Pharmacotherapeutic candidates for myopia: A review BIOMED PHARMACOTHER Wen-Yi Wang Camille Chen <here is a image 47e4db0642ef85ea-54c1d830470435d1> Justine Chang <here is a image 27a66ac73e324777-d630edeec7414892> I-Jong Wang This review provides insights into the mechanism underlying the pathogenesis of myopia and potential targets for clinical intervention. Although the etiology of myopia involves both environmental and genetic factors, recent evidence has suggested that the prevalence and severity of myopia appears to be affected more by environmental factors. Current pharmacotherapeutics are aimed at inhibiting environmentally induced changes in visual input and subsequent changes in signaling pathways during myopia pathogenesis and progression. Recent studies on animal models of myopia have revealed specific molecules potentially involved in the regulation of eye development. Among them, the dopamine receptor plays a critical role in controlling myopia. Subsequent studies have reported pharmacotherapeutic treatments to control myopia progression. In particular, atropine treatment yielded favorable outcomes and has been extensively used; however, current studies are aimed at optimizing its efficacy and confirming its safety. Furthermore, future studies are required to assess the efficacy of combinatorial use of low-dose atropine and contact lenses or orthokeratology. ... Decorin is detected in human sclera which may contribute to scleral disorders, such as high myopia [31]. However, Chen et al. found that a decorin gene polymorphism has no significant correlation with high myopia [32] . At the same time, Yip et al. also indicated that proteoglycan genes are not associated with high myopia in these subjects with single nucleotide polymorphisms, and therefore, they may be controversial in the genetic predisposition to high myopia [33]. ... The expression of cytokines in aqueous humor of high myopic patients with cataracts Purpose: To analyze the expression of 440 human cytokines in aqueous humor of high myopic patients with cataracts. Methods: Eighty-five patients with cataracts were recruited in this study. In the screening stage, the RayBio G-Series Human Cytokine Antibody Array 440 was used to assay the aqueous humor samples collected from nine high myopic patients with cataracts and eight non-myopic patients with cataracts right before the surgery. The array was further used for verification of the screened cytokines, with aqueous humor samples obtained from 34 eyes of high myopic patients with cataracts and 34 eyes of non-myopic patients with cataracts. Results: Compared with the non-myopic patients with cataracts, the expression levels of decorin, receptor activator of NF-kB (RANK), angiopoietin-1 (ANG-1), C-X-C motif ligand 16 (CXCL16), β-inducible gene-h3 (bIG-H3), insulin-like growth factor-binding protein 2 (IGFBP-2), and interleukin-17B (IL-17B) were statistically significantly higher in high myopic patients with cataracts (all p<0.000114). The matrix metalloproteinase-2 (MMP-2) level also increased in the aqueous humor of high myopic patients with cataracts (p = 0.0034). The concentrations of ANG-1 and MMP-2 were also increased in the aqueous humor of the confirmatory stage (all p<0.05). Conclusions: In this study, numerous cytokines in aqueous humor were detected in high myopic patients with cataracts and non-myopic patients with cataracts, and it was confirmed that the MMP-2 level in the aqueous humor of patients with high myopia was statistically significantly increased. Further verification also revealed the elevation of ANG-1 in the aqueous humor of high myopic patients with cataracts, which suggests that ANG-1 may be related to the pathogenesis of high myopia. ... 36 An even higher prevalence of myopia was reported in an urban sample in Taiwan: 19% in first graders and as high as 76% in sixth graders. 37 A high prevalence of myopia in Taiwan may be related to susceptible genetic backgrounds shared by East Asian ethnicities, [38] [39][40] while some lifestyle factors may also play a role. Visual problems are usually diagnosed before entrance to junior high school, thus making this problem indifferently found in adolescents from all SEP strata. ... Poverty and the prediction of health status in adolescents from low-income families in Taiwan Background: A comparison of the performance between monetary and an asset-based socioeconomic position (SEP) index in predicting adolescents' health outcomes was conducted only in low-income countries. Our study aims to compare these two indices among adolescents from low-income families in a high-income country. Methods: Data of adolescents aged 12-18 years was used from the Taiwan Database of Children and Youth in Poverty (n = 2529). The asset-based index was based on a set of weighted self-reported household conditions using principal components analysis. Multivariable logistic regression was conducted to examine the association between these two indices and the mental and physical health outcomes among adolescents. Results: Health outcomes were significantly associated with the asset-based SEP index but not with the monetary index. An increased trend was shown in mental illness symptoms, infections, injuries, allergies and dental problems when the regression was performed in the asset-based SEP index (P < 0.05) but not in the monetary SEP index. Conclusions: An asset-based SEP index serves as a better index associated with a social gradient in health inequality. To prioritize giving care to people in need, healthcare policymakers should consider including an assessment of household assets and resources, supplementary to the conventional monetary index. ... Термин «очень высокая/экстремальная/сверхвысокая миопия» используют не столь часто и также дают ему различные определения. Наиболее распространенный критерий: <(-)10 дптр [4][5] [6] [7][8][9] или ≤(-)10 дптр [10]. Реже границей «экстремальной» миопии служат (-)12 дптр [11,12], (-)15 дптр [13] и др. ... Axial ametropias of very high degree To characterize a complex of disorders developing in patients with extreme values of anteroposterior axis of the eye, it is necessary to create an appropriate nomenclature. As a topic for discussion, authors suggest to supplement the existing classifications of myopia and hyperopia with 'character of ametropia' section (axial, refractive, mixed) and the definition of ametropias of very high (ultra-high/extreme) degree (a criterion for myopia is: <-10 D). In prospect, classification of such ametropias should be based on the length of the anteroposterior axis of the eye. ... It's suggested that altered expression of LUM may contribute to myopia. Increasing studies have showed that single nucleotide polymorphisms (SNPs) in the LUM gene were associated with high myopia [10][11] [12] [13] . Majava et al [14] found that a novel SNP c.893-105G>A may contribute to pathogenesis of high myopia. ... The association of lumican polymorphisms and high myopia in a Southern Chinese population Qing-Shan Ji Gui-Fang Wang Bing Qi Jing-Xiang Zhong Aim: To investigate the correlation between lumican (LUM) gene and high myopia in a Southern Chinese population. Methods: The study comprised of 95 high myopia patients with a spherical equivalent ≤-6.5 diopters (D). The control group recruited 95 individuals with a spherical equivalent ranging from -0.5 D to +0.5 D. Direct sequencing was used to detect the single nucleotide polymorphisms (SNPs) of LUM gene in coding region. Genotype distributions were tested for Hardy-Weinberg disequilibrium. Genotypic and allelic frequencies were analyzed through Chi-square test or Fisher's exact test. Results: We identified 3 SNPs of the LUM gene: LUM c.32 (rs577456426), LUM c.507 (rs17853500) and LUM c.849 (rs181915277). Among the three SNPs, the genotype and allele frequencies of rs17853500 showed a significant difference between patients and control subjects (P<0.05). However, there were no significant differences in rs181915277 and rs577456426 between the two groups (P>0.05). Conclusion: LUM c.507 polymorphism may be a risk factor for the pathogenesis of high myopia in the Southern Chinese population. ... In previous studies, some genetic loci associated with high myopia were mapped to 12q21-q22, including the locus for LUM. LUM is a candidate gene associated with high myopia, especially in Asian populations [25][26][27] [28] . In our previous study, we verified that LUM is susceptible to pathological myopia in the northern Han ethnic Chinese people [29]. ... Enlargement of the Axial Length and Altered Ultrastructural Features of the Sclera in a Mutant Lumican Transgenic Mouse Model PLOS ONE Yanzheng Song Fengju Zhang Yanyan Zhao Junfeng Hao Lumican (LUM) is a candidate gene for myopia in the MYP3 locus. In this study, a mutant lumican (L199P) transgenic mouse model was established to investigate the axial length changes and ultrastructural features of the sclera. The mouse model was established by pronuclear microinjection. Transgenic mice and wild-type B6 mice were killed at eight weeks of age. Gene expression levels of LUM and collagen type I (COL1) in the sclera were analyzed by quantitative real-time polymerase chain reaction (qPCR), and the protein levels were assessed by Western blot analysis. Ocular axial lengths were measured on the enucleated whole eye under a dissecting microscope. Ultrastructural features of collagen fibrils in the sclera were examined with transmission electron microscopy (TEM). Lumican and collagen type I were both elevated at the transcriptional and protein levels. The mean axial length of eyes in the transgenic mice was significantly longer than that in the wild-type mice (3,231.0 ± 11.2 μm (transgenic group) vs 3,199.7 ± 11.1 μm (controls), p<0.05 =). Some ultrastructural changes were observed in the sclera of the transgenic mice under TEM, such as evident lamellar disorganizations and abnormal inter-fibril spacing. The average collagen fibril diameter was smaller than that in their wild-type counterparts. These results indicate that the ectopic mutant lumican (L199P) may induce enlargement of axial lengths and abnormal structures and distributions of collagen fibrils in mouse sclera. This transgenic mouse model can be used for the mechanistic study of myopia. ... To date, there are very few human genetic mutations that are linked to SLRP genes. The only collagen-related disorders attributed to mutations in SLRP genes within the human populations are primarily associated with corneal pathologies (Bredup et al, 2005;Majava et al, 2007;Wang et al, 2006; Chen et al, 2009; Pellegata et al, 2000). Note that the double-deficient Fmod 2/2 ; Lum 2/2 mice had a completely penetrant bicuspid AV phenotype, and the PV also had an increase in bicuspid malformations. ... Development of Myotendinous-like Junctions that Anchor Cardiac Valves Requires Fibromodulin and Lumican Article Aug 2016 DEV DYNAM Loren E. Dupuis <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Lorna Doucette A. Kittrell Rice <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Christine B Kern Background: There are many patients that exhibit connective tissue related cardiac malformations but do not have mutations in collagen genes. The Small Leucine Rich Proteoglycans (SLRP) fibromodulin (FMOD) and lumican (LUM) bind collagen and regulate fibril assembly in other biological contexts. Results: FMOD deficient mice and double deficient FMOD;LUM mice exhibited anomalies in regions where cardiac valve tissue interdigitates with adjacent muscle for support. Ectopic connective and/or myocardial tissue(s) was associated with the more severe cardiac valve anomalies in FMOD;LUM deficient mice. At postnatal day 0 (P0) there was an increase in the mesenchymal cell number in the regions where valve cusps anchor in FMOD;LUM deficient mice compared to WT. The cardiac valve anomalies correlated with the highest levels of FMOD expression in the heart and also where myotendinous junctions (MTJ) components biglycan, collagen type I alpha 1, and collagen type VI, are also localized. Conclusion: The postnatal assembly of the collagen-rich ECM in regions where cardiac valves anchor, that we have designated 'myotendinous-like junctions' (MTLJ) requires the SLRPs FMOD and LUM. Moreover, FMOD and LUM may facilitate mesenchymal cell differentiation in late stages of cardiac valve development. This article is protected by copyright. All rights reserved. ... To date, many case-control studies have been carried out to investigate the role of LUM gene polymorphism in the development of high myopia [10][11][12][13][14] [15] [16][17][18]. However, these studies have yielded inconsistent results, especially concerning the SNP loci rs3759223 and rs3759222, which are the focus of significant current research interest. ... Meta-Analysis of the Association between Lumican Gene Polymorphisms and Susceptibility to High Myopia Article Full-text available Jun 2014 PLOS ONE Miao He <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Wei Wang Dina Ragoonundun <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Wenyong Huang Backgrounds Many studies have evaluated the association between lumican (LUM) gene polymorphisms and high myopia. However, the results remain controversial. This meta-analysis aims to comprehensively evaluate the relationship between two common LUM polymorphisms (rs3759223 and rs3759222) and the risk of high myopia. Methods A comprehensive literature search for studies published up until September of 2013 was performed. Data were extracted independently by two investigators, and the weighted Odds Ratios (ORs) and 95% Confidence Intervals (CIs) for the associations were obtained by using a random-effects model. Results Eight studies (1425cases and 1271 controls) were identified for the analysis of the association between rs3759223 polymorphism and high myopia. The results indicated that rs3759223 polymorphism was associated with high myopia under a recessive model (OR = 1.71, 95%CI 1.04–2.81). Further subgroup analysis indicated that this polymorphism was associated with high myopia among Chinese people in the additive model (OR = 1.17, 95%CI 1.06–1.29) and a recessive model (OR = 1.75, 95%CI 1.00–3.06) with control group coming from hospital based population. Four studies (1024 cases and 1163 controls) were identified for the analysis of the association between rs3759222 polymorphism and high myopia. The results indicated that rs3759222 polymorphism was not associated with high myopia in all genetic models, even the subgroup analysis couldn't provide relative proof to assure the outcome. Conclusion This meta-analysis suggests that LUM polymorphisms are associated with the risk of high myopia. However, well-designed studies with larger sample sizes and more ethnic groups are required to further validate this association. ... Three SNPs were associated with pathological myopia in a study looking at Northern Han ethnic Chinese [25]. In a similar study looking at Taiwanese Han Chinese, two SNPs were observed to be significantly associated with high myopia, including one that was also present in the previous Northern Chinese study [26] , as well as in other studies [27,28], in which the polymorphism was rs3759223 (C → T). These results have not been confirmed in other studies looking at Southeastern Han Chinese [29], Northeastern Han Chinese [30], and Koreans [31]. ... The Role of Lumican in Ocular Disease Article Full-text available Jul 2013 <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Shahriar Amjadi <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Kelly Mai Peter McCluskey <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Denis Wakefield Lumican is keratan sulfate proteoglycan of the small leucine rich proteoglycan family. Through studies in animal models lumican has been found to be critical in maintaining corneal clarity. It maintains ordered collagen fibrils which are vital in keeping the cornea transparent. It may also be important in primary open angle glaucoma influencing aqueous outflow. Lumican deficiency in mice results in increased axial length with fibromodulin deficiency and thinner sclerae. There is evidence suggesting that this characteristic may be pertinent in humans and lumican gene polymorphisms could be related to high myopia. Lumican plays a fundamental role in inflammation and wound healing. It localises macrophages to the site of corneal injury and recruits neutrophils in lipopolysaccharide-induced keratitis in mice. It has also been shown to bind lipopolysaccharide which may be critical in inflammatory diseases such as uveitis. Lumican is also important in wound healing revealing decreased synthesis in scar tissue and mediating Fas-Fas ligand interactions. It is present in human placenta and amniotic membrane suggesting that it may ensure viable amniotic membrane grafts. Lumican may also be involved in the formation of posterior capsular opacification following cataract surgery. Research into the pivotal role of lumican in the pathogenesis of ocular disease has resulted in greater understanding of the key role which proteoglycans play in human disease. ... respectively, have previously been associated with several multifactorial conditions. [15][16][17][18] [19] [20] The genes encoding biglycan (BGN) on chromosome Xq28, decorin (DCN) on chromosome 12q21. 33 and fibromodulin (FMOD) on chromosome 1q32, have, however, not been associated with any multifactorial conditions until today. ... Genes encoding proteoglycans are associated with the risk of anterior cruciate ligament ruptures Article Full-text available Feb 2014 BRIT J SPORT MED <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Sasha Mannion <here is a image 569c69cf69e6e547-6e257d48ab64e10a> Asanda Mtintsilana <here is a image 68b6f9148e239be5-43140cb00af1ab93> Michael Posthumus <here is a image c9f669b79c1f6b13-6c684a6c273885bb> Alison V September Genetic variants within genes involved in fibrillogenesis have previously been implicated in anterior cruciate ligament (ACL) injury susceptibility. Proteoglycans also have important functions in fibrillogenesis and maintaining the structural integrity of ligaments. Genes encoding proteoglycans are plausible candidates to be investigated for associations with ACL injury susceptibility; polymorphisms within genes encoding the proteoglycans aggrecan (ACAN), biglycan (BGN), decorin (DCN), fibromodulin (FMOD) and lumican (LUM) were examined. A case-control genetic association study was conducted. 227 participants with surgically diagnosed ACL ruptures (ACL group) and 234 controls without any history of ACL injury were genotyped for 10 polymorphisms in 5 proteoglycan genes. Inferred haplotypes were constructed for specific regions. The G allele of ACAN rs1516797 was significantly under-represented in the controls (p=0.024; OR=0.72; 95% CI 0.55 to 0.96) compared with the ACL group. For DCN rs516115, the GG genotype was significantly over-represented in female controls (p=0.015; OR=9.231; 95%CI 1.16 to 73.01) compared with the ACL group and the AA genotype was significantly under-represented in controls (p=0.013; OR=0.33; 95% CI 0.14 to 0.78) compared with the female non-contact ACL injury subgroup. Haplotype analyses implicated regions overlapping ACAN (rs2351491 C>T-rs1042631 T>C-rs1516797 T>G), BGN (rs1126499 C>T-rs1042103 G>A) and LUM-DCN (rs2268578 T>C-rs13312816 A>T-rs516115 A>G) in ACL injury susceptibility. These independent associations and haplotype analyses suggest that regions within ACAN, BGN, DCN and a region spanning LUM-DCN are associated with ACL injury susceptibility. Taking into account the functions of these genes, it is reasonable to propose that genetic sequence variability within the genes encoding proteoglycans may potentially modulate the ligament fibril properties. ... Myopia is a complex disease caused by environmental and genetic factors. To date, although many studies have evaluated various candidate genes and susceptible loci of high myopia [6][7] [8] [9][10][11], no single gene has been consistently responsible for the condition. In addition to candidate gene studies, a genome-wide approach has also been performed by several groups. ... Insulin-like growth factor 1 is not associated with high myopia in a large Japanese cohort Article Full-text available May 2013 MOL VIS <here is a image 7ddb4127d4f506af-e1d4f54dad1e9ebc> Masahiro Miyake <here is a image 9a2a7b7b42519a2e-5a6c1197fdd219c4> Kenji Yamashiro Hideo Nakanishi <here is a image 18773c20e13d9eaa-cf0fc54a48436da3> Nagahisa Yoshimura To investigate whether genetic variations in the insulin-like growth factor 1 (IGF-1) gene are associated with high myopia in Japanese. A total of 1,339 unrelated Japanese patients with high myopia (axial length ≥26 mm in both eyes) and two independent control groups were evaluated (334 cataract patients without high myopia and 1,194 healthy Japanese individuals). The mean axial length (mm±SD) in the case group was 29.18±1.85 mm, and the mean spherical equivalent (D±SD) of the phakic eyes was -12.69±4.54 D. We genotyped five tagging single nucleotide polymorphisms (SNPs) in IGF-1: rs6214, rs978458, rs5742632, rs12423791, and rs2162679. Chi-square tests for trend, multivariable logistic regression, and haplotype regression analysis were conducted. We found no significant association between the IGF-1 SNPs and high or extreme myopia (axial length ≥28 mm in both eyes, 837 subjects) in the additive model, even when compared with the cataract and general population controls, with or without adjustments for age and sex. The evaluation using dominant and recessive models also did not reveal any significant associations. The haplotype analysis with a variable-sized sliding-window strategy also showed a lack of association of IGF-1 SNPs with high or extreme myopia. The results of the present study using a Japanese subset do not support the proposal that the IGF-1 gene determines susceptibility to high or extreme myopia in Caucasians and Chinese. Further studies are needed to confirm our reports in other populations and to identify the underlying genetic determinants of these ocular pathological conditions. ... Several common high myopia loci[26,34,35,41], as well as Mendelian loci[23,28,42,43], have been identified in Han Chinese. SNPs in genes including PAX6[44][45][46], LAMA1[47], LUM[48] [49] [50], TGFB1[51,52], and CTNND2[33,41], and mutations in ZNF44[53]also have been identified in Chinese families with Mendelian high myopia. However, some regions have not been consistently identified as risk loci[54,55]. ... Association of markers at chromosome 15q14 in Chinese patients with moderate to high myopia Article Full-text available Oct 2012 MOL VIS <here is a image e99b2d597fe78f18-033ee81c6a081adc> Xiaodong Jiao <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Panfeng Wang Shiqiang Li <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> J. Fielding Hejtmancik To investigate the association of two reported regions on chromosome 15 with moderate to high myopia in two Chinese cohorts from southern China. Two candidate regions on 15q14 and 15q25 were selected based on reported association with refractive error in the literature. Five single nucleotide polymorphisms (SNPs) were genotyped in 300 university students with high myopia at Guangzhou and 308 without refractive error, and 96 university students of Chaoshan Chinese origin with moderate to high myopia and 96 without refractive error. Genotypes were evaluated using direct sequencing and analyzed with chi-square, Armitage trend, and Mantel-Haenszel tests, and regression analysis. Of the five SNPs screened, alleles of rs634990 and rs524952 in the 15q14 region showed evidence of allelic association with moderate to high myopia (p<8.81×10(-7) and p<1.57×10(-6), respectively) in the Guangzhou group, but not in the Chaoshan group. The SNPs at 15q25 did not show significant association in any group. Association of rs634990 and rs524952 were still significant when both groups were combined into a single analysis (p<1.66×10(-6) and p<2.72×10(-6), respectively), and for genotypic, additive, and dominant models. This study confirms the significant association of rs634990 and rs524952 on chromosome 15q14 previously reported in European and Japanese populations with high myopia in the Guangzhou but not the Chaoshan Chinese populations, suggesting that genetic contributors to high myopia in the Chaoshan population might be different from other Chinese populations. ... Of these candidate loci, 11 have been associated with high myopia: MYP1-5 [2,[6][7][8][9], MYP11 [10], MYP12 [11], MYP13 [12], MYP15 [13], MYP16 [14], and MYP18 [15]. In the same chromosomal regions, many candidate genes associated with high myopia have been studied, such as collagen, type I, alpha 1 (COL1A1) [16], transforming growth factor, beta 1 (TGF beta1) [17,18], transforming growth factor-beta-induced factor (TGIF) [19], lumican (LUM) [20, 21], hepatocyte growth factor (HGF) [22], myocilin (MYOC) [23], paired box 6 (PAX6) [24,25], and uromodulin-like 1 (UMODL1) [26], but none of these are now thought to be responsible for or associated with high myopia. ... Evaluation of BLID and LOC399959 as candidate genes for high myopia in the Chinese Han population Article Full-text available Oct 2010 MOL VIS <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Fuxin Zhao <here is a image 80c2fed84628b644-6aee4e66f5046228> Jian Bai <here is a image 036fae21c18444b6-2f740ebd259bebc6> Wei Chen Xiangtian Zhou BH3-like motif containing, cell death inducer (BLID) and LOC399959 are two genes associated with the single nucleotide polymorphism (SNP) rs577948, which is a susceptibility locus for high myopia in Japanese subjects. The purpose of this study was to determine if BLID and LOC399959 are associated with high myopia in Chinese Han subjects. High myopia subjects (n=476) had a spherical refractive error of less than -6.00 D in at least one eye and/or an axial length greater than 26 mm. Genomic DNA was extracted and genotyped from peripheral blood leukocytes of high myopes and controls (n=275). Using a case-control association study of candidate regions, linkage disequilibrium blocks for 19 tag SNPs (tSNPs), including rs577948, harbored within and surrounding the BLID and LOC399959 genes were analyzed on a MassArray platform using iPlex chemistry. Each of the tSNPs had an r(2)>0.8 and minor allele frequency >10% in the Chinese Han population. Haplotype association analysis was performed on Haploview 4.1 using Chi-square (χ(2)) tests. None of the 19 tSNPs were statistically associated with high myopia. While rs577948 may be associated with high myopia in Japanese subjects, it and the other tSNPs near the BLID and LOC399959 genes are not susceptibility loci for high myopia in the Chinese Han population. Thus, associations of SNPs with high myopia as determined by Genome-Wide Association Study (GWAS) may be restricted to certain ethnic or genetically distinct populations. Without systematic replication in other populations, the results of GWAS associations should be interpreted with great caution. The versatile roles of lumican in eye diseases: A review Article Jun 2023 OCUL SURF <here is a image 67ddb8ed3870d7e1-e81473dc68e60d47> Mei-Chi Tsui Hsin-Yu Liu <here is a image c7c7a1cd4a1c75f8-af9ec56e8a160105> Hsiao-Sang Chu <here is a image 27a66ac73e324777-d630edeec7414892> I-Jong Wang Lumican is a keratan sulfate proteoglycan (KSPG) that belongs to the small leucine-rich proteoglycan family. Research has lifted the veil on the versatile roles of lumican in the pathogenesis of eye diseases. Lumican has pivotal roles in the maintenance of physiological tissue homogenesis and is often upregulated in pathological conditions, e.g., fibrosis, scar tissue formation in injured tissues, persistent inflammatory responses and immune anomaly, etc. Herein, we will review literature regarding the role of lumican in pathogenesis of inherited congenital and acquired eye diseases, e.g., cornea dystrophy, cataract, glaucoma and chorioretinal diseases, etc. Association of MTOR and PDGFRA gene polymorphisms with different degrees of myopia severity Article Jan 2022 EXP EYE RES Xuan Li Jianxiong Long Yue Liu Chunling Li This study aims to investigate the associations of SNPs in the mammalian target of rapamycin (MTOR) and the platelet derived growth factor receptor alpha (PDGFRA) genes with different degrees of myopia severity in Han and Zhuang populations. The SNPs of MTOR (rs1057079, rs1064261, and rs2536) and PDGFRA (rs1800812, rs35597368, rs4358459, rs6554162, and rs7677751) were analyzed among 1347 patients with myopia (849 patients with high myopia and 498 patients with mild to moderate myopia) and 453 controls without myopia in Guangxi, China (collected 2016–2018). Genetic model association analysis was performed on each SNP in different myopia subgroups. The associations of rs1057079 and rs1064261 with mild to moderate myopia were observed under the dominant models (rs1057079: OR = 1.324, 95%CI: 1.005–1.744, P = 0.046; rs1064261: OR = 1.597, 95%CI: 1.099–2.319, P = 0.014). However, the association of SNP rs1057079 could not withstand multiple correction. The number of adverse genotypes in each sample was counted. Results showed that in the high myopia group, the levels of risk of myopia in patients carrying three to four and five to eight adverse genotypes were 1.734 and 2.062 times the level of risk in patients carrying two or lower genotypes, respectively. After the stratified analyses of Han and Zhuang populations, the Zhuang populations consistently had high frequencies of myopia. This study provides evidence suggesting that the MTOR and PDGFRA genes are associated with different degrees of myopia severity and have gene–gene interactions. In addition, this study discovered a new SNP of MTOR (rs1064261) that is associated with myopia. Thus, further longitudinal studies are warranted. Consortium for Refractive Error and Myopia (CREAM): Vision, Mission, and Accomplishments Chapter Full-text available Feb 2021 <here is a image 26d0d971b9f0d046-f28c69941115faea> Virginie Johanna Maria Verhoeven <here is a image 28dc959bcbf509e7-d198b6f3fb78f001> A.E.G. Haarman <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Milly Tedja <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Magda A Meester-Smoor The Consortium for Refractive Error and Myopia (CREAM) is an international collaboration founded to increase knowledge on the genetic background of refractive error and myopia. The consortium was established in 2011 and consists of >50 studies from all over the world with epidemiological and genetic data on myopia endophenotypes. Due to these efforts, almost 200 genetic loci for refractive error and myopia have been identified. These genetic risk variants mostly carry low risk but are highly prevalent in the general population. The genetic loci are expressed in all retinal cell layers and play a role in different processes, e.g., in phototransduction or extracellular matrix remodeling. The work of CREAM over the years has implicated the major pathways in conferring susceptibility to myopia and supports the notion that myopia is caused by a light-dependent retina-to-sclera signaling cascade. The current genetic findings offer a world of new molecules involved in myopiagenesis. However, as the currently identified genetic loci explain only a fraction of the high heritability, further genetic advances are needed. It is recommended to expand large-scale, in-depth genetic studies using complementary big data analytics, to consider gene-environment effects by thorough measurements of environmental exposures, and to focus on subgroups with extreme phenotypes and high familial occurrence. Functional characterization of associated variants is simultaneously needed to bridge the knowledge gap between sequence variance and consequence for eye growth. The CREAM consortium will endeavor to play a pivotal role in these future developments. The Genetics of Myopia Chapter Full-text available Oct 2019 Stuart MacGregor <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Milly Tedja <here is a image 28dc959bcbf509e7-d198b6f3fb78f001> A.E.G. Haarman <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Magda A Meester-Smoor Myopia is the most common eye condition worldwide and its prevalence is increasing. While changes in environment, such as time spent outdoors, have driven myopia rates, within populations myopia is highly heritable. Genes are estimated to explain up to 80% of the variance in refractive error. Initial attempts to identify myopia genes relied on family studies using linkage analysis or candidate gene approaches with limited progress. More genome-wide association study (GWAS) approaches have taken over, ultimately resulting in the identification of hundreds of genes for refractive error and myopia, providing new insights into its molecular machinery. These studies showed myopia is a complex trait, with many genetic variants of small effect influencing retinal signaling, eye growth and the normal process of emmetropization. The genetic architecture and its molecular mechanisms are still to be clarified and while genetic risk score prediction models are improving, this knowledge must be expanded to have impact on clinical practice. IMI - Myopia Genetics Report Article Full-text available Feb 2019 INVEST OPHTH VIS SCI <here is a image 28dc959bcbf509e7-d198b6f3fb78f001> A.E.G. Haarman MA Meester-Smoor X Zhou <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Milly Tedja The knowledge on the genetic background of refractive error and myopia has expanded dramatically in the past few years. This white paper aims to provide a concise summary of current genetic findings and defines the direction where development is needed. We performed an extensive literature search and conducted informal discussions with key stakeholders. Specific topics reviewed included common refractive error, any and high myopia, and myopia related to syndromes. To date, almost 200 genetic loci have been identified for refractive error and myopia, and risk variants mostly carry low risk but are highly prevalent in the general population. Several genes for secondary syndromic myopia overlap with those for common myopia. Polygenic risk scores show overrepresentation of high myopia in the higher deciles of risk. Annotated genes have a wide variety of functions, and all retinal layers appear to be sites of expression. The current genetic findings offer a world of new molecules involved in myopiagenesis. As the missing heritability is still large, further genetic advances are needed. This Committee recommends expanding large-scale, in-depth genetic studies using complementary big data analytics, consideration of gene-environment effects by thorough measurement of environmental exposures, and focus on subgroups with extreme phenotypes and high familial occurrence. Functional characterization of associated variants is simultaneously needed to bridge the knowledge gap between sequence variance and consequence for eye growth. IMI – Myopia genetics report Article Full-text available Feb 2019 <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Milly Tedja <here is a image 28dc959bcbf509e7-d198b6f3fb78f001> A.E.G. Haarman <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Magda A Meester-Smoor <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Caroline C W Klaver The knowledge on the genetic background of refractive error and myopia has expanded dramatically in the past few years. This white paper aims to provide a concise summary of current genetic findings and defines the direction where development is needed. We performed an extensive literature search and conducted informal discussions with key stakeholders. Specific topics reviewed included common refractive error, any and high myopia, and myopia related to syndromes. To date, almost 200 genetic loci have been identified for refractive error and myopia, and risk variants mostly carry low risk but are highly prevalent in the general population. Several genes for secondary syndromic myopia overlap with those for common myopia. Polygenic risk scores show overrepresentation of high myopia in the higher deciles of risk. Annotated genes have a wide variety of functions, and all retinal layers appear to be sites of expression. The current genetic findings offer a world of new molecules involved in myopiagenesis. As the missing heritability is still large, further genetic advances are needed. This Committee recommends expanding large-scale, in-depth genetic studies using complementary big data analytics, consideration of gene-environment effects by thorough measurement of environmental exposures, and focus on subgroups with extreme phenotypes and high familial occurrence. Functional characterization of associated variants is simultaneously needed to bridge the knowledge gap between sequence variance and consequence for eye growth. Insight into the molecular genetics of myopia Article Full-text available Dec 2017 MOL VIS Jiali Li Myopia is the most common cause of visual impairment worldwide. Genetic and environmental factors contribute to the development of myopia. Studies on the molecular genetics of myopia are well established and have implicated the important role of genetic factors. With linkage analysis, association studies, sequencing analysis, and experimental myopia studies, many of the loci and genes associated with myopia have been identified. Thus far, there has been no systemic review of the loci and genes related to non-syndromic and syndromic myopia based on the different approaches. Such a systemic review of the molecular genetics of myopia will provide clues to identify additional plausible genes for myopia and help us to understand the molecular mechanisms underlying myopia. This paper reviews recent genetic studies on myopia, summarizes all possible reported genes and loci related to myopia, and suggests implications for future studies on the molecular genetics of myopia. Construction of adenovirus vectors encoding the lumican gene by gateway recombinant cloning technology Aim: To construct adenovirus vectors of lumican gene by gateway recombinant cloning technology to further understand the role of lumican gene in myopia. Methods: Gateway recombinant cloning technology was used to construct adenovirus vectors. The wild-type (wt) and mutant (mut) forms of the lumican gene were synthesized and amplified by polymerase chain reaction (PCR). The lumican cDNA fragments were purified and ligated into the adenovirus shuttle vector pDown-multiple cloning site (MCS)-/internal ribozyme entry site (IRES)/enhanced green fluorescent protein (EGFP). Then the desired DNA fragments were integrated into the destination vector pAV.Des1d yielding the final expression constructs pAV.Ex1d-cytomegalovirus (CMV)>wt-lumican/IRES/EGFP and pAV.Ex1d-CMV>mut-lumican/IRES /EGFP, respectively. Results: The adenovirus plasmids pAV.Ex1d-CMV>wt-lumican/IRES/EGFP and pAV.Ex1d-CMV>mut-lumican/IRES/EGFP were successfully constructed by gateway recombinant cloning technology. Positive clones identified by PCR and sequencing were selected and packaged into recombinant adenovirus in HEK293 cells. Conclusion: We construct adenovirus vectors containing the lumican gene by gateway recombinant cloning technology, which provides a basis for investigating the role of lumican gene in the pathogenesis of high myopia. Genes Involved in the Development of Myopia Both environmental factors and genetic factors have been implicated in the development of myopia. Family studies and twin studies have revealed the heritability of myopia since the 1960s. To identify the causative genes for myopia, common myopia (>−6D) and high myopia (≤−6D) have been investigated in familial studies, twin studies, case–control studies, and cohort studies. In familial studies and twin studies, linkage analysis using microsatellite markers has identified 19 loci for myopia: MYP1 to MYP19 (Table 2.1). Although many genes in these loci were evaluated as candidate genes for myopia or high myopia, most genes were found not to be involved in the pathogenesis of myopia or high myopia. After the completion of the Human Genome Project, many researchers performed genome-wide association studies (GWAS) in case–control cohorts and population-based cohorts. The allele frequency in single nucleotide polymorphisms (SNPs) was compared between cases of myopia or high myopia and controls in case–control studies, and the association of the allele to refractive error or axial length was evaluated in quantitative trait locus (QTL) analysis. In spite of these intensive studies, genes for myopia have not been determined until recently. Proteoglycans of the Intervertebral Disc Proteoglycans are present within the extracellular matrix (ECM) of the intervertebral disc and on the surface of its cells. The disc possesses many matrix proteoglycans, with most also being present in hyaline cartilages. The best studied of these are aggrecan and versican (members of the hyalectan/lectican family) and decorin, biglycan, fibromodulin, lumican, PRELP, and chondroadherin (members of the small leucine-rich repeat protein/SLRP family). More recently, the disc has also been shown to contain perlecan and lubricin, which were previously thought to be characteristic of basement membranes and the surface of articular cartilage, respectively. Much less is known about the disc cell-associated proteoglycans, though it is likely that several members of the syndecan (Tkachenko et al. 2005) and glypican (Fransson et al. 2004) families will be present, together with other unrelated proteoglycans such as NG2 (Akeda et al. 2007; Stallcup 2002). While little is known concerning the specific function of the cell-associated proteoglycans within the disc, it has been shown that syndecan-4 expression is increased when elevated levels of interleukin-1 (IL-1) or tumor necrosis factor-α (TNF-α) are present (Wang et al. 2011), and that it may play a role in promoting aggrecanase-mediated proteolysis within the disc. However, due to the scarcity of information on disc cell-associated proteoglycans, this chapter will focus on the matrix proteoglycans, in particular aggrecan. Lack of Association Between LUM rs3759223 Polymorphism and High Myopia Purpose: Previous evidence has indicated that the lumican (LUM) gene is a candidate susceptibility gene of high myopia; however, the association between LUM promoter regions rs3759223 polymorphism and high myopia remains controversial and ambiguous. This study performed a meta-analysis to clarify the association between the rs3759223 polymorphism and high myopia risk. Methods: Eligible studies were identified by comprehensive search of PubMed, EMBASE, Web of Science, and Chinese Biomedical Literature database. The crude odds ratios (ORs) and their corresponding 95% confidence intervals (CIs) were used to estimate the association between the rs3759223 polymorphism and high myopia susceptibility. Meta-regression and subgroup analyses were performed to identify the source of heterogeneity. Results: Finally, six studies including 1238 cases and 1059 healthy controls were included. Meta-analyses showed no association between rs3759223 polymorphism and high myopia susceptibility in all genetic models (CC vs. TT, OR = 1.089; 95% CI, 0.690 to 1.718; CT vs. TT, OR = 0.865; 95% CI, 0.646 to 1.157; CC + CT vs. TT, OR = 1.202; 95% CI, 0.730 to 1.980; CC vs. CT + TT, OR = 0.914; 95% CI, 0.771 to 1.083) and no significance in subgroup analyses according to the definition of high myopia (based on more myopic than -6.00 diopters vs. not based on more myopic than -6.00 diopters). Publication bias was not evident in this study. Conclusions: This meta-analysis has suggested that there is a lack of association of the rs3759223 polymorphism with high myopia risk. However, further large and well-designed studies with the consideration of LUM gene locus interactions and gene-gene and gene-environment interactions are still required to further evaluate high myopia risk. Update on the epidemiology and genetics of myopic refractive error While myopia is an increasingly common refractive error worldwide, its prevalence is greatest in urbanized regions in east Asia. Myopia is a complex multifactorial ocular disorder governed by both genetic and environmental factors and possibly their interplay. Evidence for a genetic role in myopia has been derived from studies of syndromal myopia, familial correlation studies and linkage analyses. More recently, candidate gene and genome-wide association studies have been utilized. However, a high proportion of the heritability of myopia remains unexplained. Most genetic discoveries have been for high myopia, with the search for genes underpinning myopia of lesser severity yielding fewer positive associations. This may soon change with the use of next-generation sequencing, as well as the use of epigenetics and proteomic approaches. [Analysis of COL9A2 gene mutations in a Chinese Han population with pathological myopia] To analyze the mutation of COL9A2 gene and investigate the molecular pathogenesis of pathological myopia in a Han Chinese population. Mutation in the coding region of the COL9A2 gene was screened by Sanger sequencing in 200 subjects with pathological myopia and 200 normal controls. The detected variants were genotyped by SNaPshot method in another 200 myopic cases and 200 normal controls. Sanger sequencing has failed to detect the reported D281fs frameshift mutation in the 200 cases. A novel variant, c.143G>C heterozygous missense mutation in exon 2, was identified in a myopic subject, and another novel variant, c.884G>A heterozygous missense mutation in exon 17, was found in another case. Neither was found in normal controls. One SNP (rs2228564) was detected in the coding region of the COL9A2 gene, but there was no significant difference in its allelic frequencies between the two groups (P> 0.05). Genotyping of the remainder 200 cases and 200 controls by SNaPshot method has found a c.143G>C in 1 case and c.884G>A in 2 cases, though no significant difference between the two groups was detected (P> 0.05). The D281fs frameshift mutation in the COL9A2 gene is not associated with pathological myopia in the studied Han Chinese population. Two novel mutations, c.143G>C in exon 2 and c.884G>A in exon 17 of the COL9A2 gene, may contribute to the development of pathological myopia. Association between a Lumican Promoter Polymorphism and High Myopia in the Chinese Population: A Meta-Analysis of Case-Control Studies Purpose: To evaluate the relationship between lumican polymorphisms and high myopia in Chinese populations. Methods: An electronic search was conducted in Pubmed, Embase, Cochrane Library and the China Biological Medicine Database for articles published prior to September 30, 2012. A meta-analysis was performed to assess heterogeneity, combine results and determine publication bias. Results: This meta-analysis, including 1,545 subjects from 5 studies, indicated that Chinese lumican rs3759223 C allele carriers had a decreased risk of high myopia in comparison to T allele carriers (odds ratio: 0.531; 95% confidence interval, CI: 0.304-0.925; p = 0.025). There was some heterogeneity between studies. A metaregression showed that the mean axial length of controls weakens the effect of rs3759223 on high myopia (slope: -0.914; 95% CI: -1.490 to 0.337; p = 0.002). Sensitivity analysis confirmed the reliability and stability of this meta-analysis. Conclusion: Chinese lumican rs3759223 C allele carriers may be at reduced risk of high myopia. Association between lumican gene-1554 T/C polymorphism and high myopia in Asian population: A meta-analysis To investigate the association between lumican gene -1554 T/C polymorphism and high myopia susceptibility. We searched the published literature in the Medline, Embase, and CBM databases from inception to July 2013. A meta-analysis was performed by the programs RevMan 5.1 and Stata 12.0, and the odds ratio (OR) with 95% confidence interval (CI) was calculated in fixed or random effect model based on heterogeneity test among studies. Seven case-control studies with a total of 1 233 cases and 936 controls were included. A statistical significant association with high myopia was observed in the recessive model (TT vs CT+CC: OR=1.92; 95%CI=1.14-3.23) and codominant model (TT vs CT: OR=1.81, 95%CI=1.19-2.75). The present meta-analysis suggested that lumican -1554 T/C polymorphism might be moderately associated with high myopia susceptibility. This conclusion warrants confirmation by further studies. Association of Lumican Gene Polymorphism With High Myopia: A Meta-Analysis. Numerous studies have evaluated the association between the single-nucleotide polymorphism (SNP) (rs3759223, C→T) in Lumican gene and high myopia risk in the Chinese population. However, the results have been inconsistent. We therefore here examined whether the rs3759223 polymorphism confers high myopia risk by conducting a meta-analysis. PubMed, EMBASE, Science Citation Index, and Chinese National Knowledge Infrastructure (up to November 30, 2012) were searched by two investigators independently. Pooled relative ratios (RRs) and 95% confidence interval (CI) were used to assess the strength of the associations between SNP rs3759223 and myopia. Statistical analysis was undertaken using the program STATA 11.0 software (Stata Corporation, College Station, TX). Five case-control studies involving 923 patients with high myopia and 622 controls were included in this meta-analysis. A significant relationship between SNP rs3759223 and high myopia in the Chinese population was found under the homozygote (RR = 1.47, 95% CI 1.01-2.12, p = 0.04) and recessive (RR = 1.69, 95% CI 1.09-2.62, p = 0.02) genetic models. However, no significant association was found under the heterozygote (RR = 1.01, 95% CI 0.76-1.35, p = 0.93) and dominant (RR = 1.06, 95%CI 0.90-1.26, p = 0.48) genetic models. This meta-analysis showed the evidence that SNP rs3759223 may affect individual susceptibility to high myopia in the Chinese population. Given the limited sample size, further investigations are needed to validate the association. Genetic Susceptibility to Refractive Error: Association of Vasoactive Intestinal Peptide Receptor 2 (VIPR2) with High Myopia in Chinese Myopia is the most common ocular disease worldwide. We investigated the association of high myopia with the common single nucleotide polymorphisms (SNPs) of five candidate genes - early growth response 1 (EGR1), v-fos FBJ murine osteosarcoma viral oncogene homolog (FOS), jun oncogene (JUN), vasoactive intestinal peptide (VIP), and vasoactive intestinal peptide receptor 2 (VIPR2). We recruited 1200 unrelated Chinese subjects with 600 cases (spherical equivalent ≤-8.00 diopters) and 600 controls (spherical equivalent within ±1.00 diopter). A discovery sample set was formed from 300 cases and 300 controls, and a replication sample set from the remaining samples. Tag SNPs were genotyped for the discovery sample set, and the most significant haplotypes and their constituent SNPs were followed up with the replication sample set. The allele and haplotype frequencies in cases and controls were compared by logistic regression adjusted for sex and age to give P a values, and multiple comparisons were corrected by permutation test to give P aemp values. Odd ratios (OR) were calculated accordingly. In the discovery phase, EGR1, JUN and VIP did not show any significant association while FOS and VIPR2 demonstrated significant haplotype association with high myopia. In the replication phase, the haplotype association for VIPR2 was successfully replicated, but not FOS. In analysis combining both sample sets, the most significant association signals of VIPR2 were the single marker rs2071625 (P a = 0.0008, P aemp = 0.0046 and OR = 0.75) and the 4-SNP haplotype window rs2071623-rs2071625-rs2730220-rs885863 (omnibus test, P a = 9.10e-10 and P aemp = 0.0001) with one protective haplotype (GGGG: P aemp = 0.0002 and OR = 0.52) and one high-risk haplotype (GAGA: P aemp = 0.0027 and OR = 4.68). This 4-SNP haplotype window was the most significant in all sample sets examined. This is the first study to suggest a role of VIPR2 in the genetic susceptibility to high myopia. EGR1, JUN, FOS and VIP are unlikely to be important in predisposing humans to high myopia. Purpose: To determine the association of single nucleotide polymorphisms (SNPs) in the promoter region of the lumican (LUM) gene with high myopic Korean patients. Methods: Genomic DNA samples were obtained from 128 unrelated Korean patients with high myopia who had refractive errors ≤ -9.25 and axial lengths ≥ 26.5 mm in both eyes, and 235 control subjects. We investigated two promoter SNPs of the LUM gene. Results: For the rs3759222, the C/C genotype was less prevalent in the high myopia group compared to the control group (46.1% vs. 53.2%); however, there was no statistical significance (p = 0.068, OR = 0.754, 95% CI: 0.491-1.159). The "C" allele frequency in the high myopia group (68.0%) was slightly lower than the control group (72.6%), but this difference was not statistically significant (p = 0.061, OR = 0.810, 95% CI:0.582-1.126). For the rs3759223, the genotype frequencies of T/T, T/C, and C/C were 67.2%, 26.6%, and 6.2%, respectively, in the high myopia group and 64.7%, 30.6%, and 4.7 %, respectively, in the control group. The allele frequency of T was 80.5% in the high myopia group and 80.0% in the control group (p = 0.077, OR = 1.03, 95% CI: 0.703-1.508). There were no significant differences in the distribution of genotype and allele frequencies for the two promoter SNPs tested. Conclusions: The current study did not support an association between the promoter SNPs of the LUM gene with high myopia in the Korean population. Extracellular matrix proteins have been implicated in protein remodelling of the sclera in refractive error. The matrix metalloproteinases (MMPs) falling into the collagenase (MMP1, MMP8, MMP13), gelatinase (MMP2, MMP9) and stromelysin (MMP3, MMP10, MMP11) functional groups are particularly important. We wished to assess their association with myopia, refractive error and ocular biometric measures in an Australian cohort. A total of 543 unrelated individuals of Caucasian ethnicity were genotyped including 269 myopes (≤-1.0D) and 274 controls (>-1.0D). Tag single nucleotide polymorphisms (SNPs) (n = 53) were chosen to encompass these eight MMPs. Association tests were performed using linear and logistic regression analysis with age and gender as covariates. Spherical equivalent, myopia, axial length, anterior chamber depth and corneal curvature were the phenotypes of interest. Initial findings indicated that the best p values for each trait were 0.02 for myopia at rs2274755 (MMP9), 0.02 for SE at both rs3740938 (MMP8) and rs131451 (MMP11), 0.01 for axial length at rs11225395 (MMP8), 0.01 for anterior chamber depth at rs498186 (MMP1) and 0.02 at rs10488 (MMP1). However, following correction for multiple testing, none of these SNPs remained statistically significant. Our data suggests that the MMPs in the collagenase, gelatinase and stromelysin categories do not appear to be associated with myopia, refractive error or ocular biometric measures in this cohort. To investigate whether the PAX6, Lumican, and MYOC genes are related to high myopia in Han Chinese since the association between these genes and high myopia is unclear in this patient population. Peripheral venous blood samples were collected for DNA extraction from 220 subjects with high myopia (refractive error ≤-10.00 D) vs. normal controls among the Han Chinese of Northeastern China. Mass spectrometry was applied to detect 10 SNP loci of the PAX6, Lumican, and MYOC genes. The candidate region was analyzed using case-control correlation analysis. The χ(2) test was used to analyze the allele and genotype frequencies in the myopic group vs. the control group. Haploview software was used for haplotype analysis. The χ(2) test was used to compare the allele and genotype frequencies of SNPs in patients and control subjects and the results showed that ten SNPs of the PAX6, Lumican, and MYOC genes were not significantly associated with high myopia. Our results confirm that the PAX6, Lumican, and MYOC genes were not associated with high myopia in the Han Chinese in Northeastern China. The small leucine-rich proteoglycans (SLRPs) comprise an expanding family of proteoglycans and glycoproteins that now encompass five distinct groups including three canonical and two noncanonical classes based on shared structural and functional parameters. SLRPs are tissue organizers by orienting and ordering various collagenous matrices during ontogeny, wound repair, and cancer and interact with a number of surface receptors and growth factors, thereby regulating cell behavior. The focus of this chapter is on novel conceptual and functional advances in our understanding of SLRP biology with special emphasis on genetic diseases, cancer growth, fibrosis, osteoporosis, and other biological processes where these proteoglycans play a central role. Wojciechowski R. Nature and nurture: the complex genetics of myopia and refractive error. The refractive errors, myopia and hyperopia, are optical defects of the visual system that can cause blurred vision. Uncorrected refractive errors are the most common causes of visual impairment worldwide. It is estimated that 2.5 billion people will be affected by myopia alone within the next decade. Experimental, epidemiological and clinical research has shown that refractive development is influenced by both environmental and genetic factors. Animal models have showed that eye growth and refractive maturation during infancy are tightly regulated by visually guided mechanisms. Observational data in human populations provide compelling evidence that environmental influences and individual behavioral factors play crucial roles in myopia susceptibility. Nevertheless, the majority of the variance of refractive error within populations is thought to be because of hereditary factors. Genetic linkage studies have mapped two dozen loci, while association studies have implicated more than 25 different genes in refractive variation. Many of these genes are involved in common biological pathways known to mediate extracellular matrix (ECM) composition and regulate connective tissue remodeling. Other associated genomic regions suggest novel mechanisms in the etiology of human myopia, such as mitochondrial-mediated cell death or photoreceptor-mediated visual signal transmission. Taken together, observational and experimental studies have revealed the complex nature of human refractive variation, which likely involves variants in several genes and functional pathways. Multiway interactions between genes and/or environmental factors may also be important in determining individual risks of myopia, and may help explain the complex pattern of refractive error in human populations. The GEnes in Myopia (GEM) study in understanding the aetiology of refractive errors Refractive errors represent the leading cause of correctable vision impairment and blindness in the world with an estimated 2 billion people affected. Refractive error refers to a group of refractive conditions including hypermetropia, myopia, astigmatism and presbyopia but relatively little is known about their aetiology. In order to explore the potential role of genetic determinants in refractive error the "GEnes in Myopia (GEM) study" was established in 2004. The findings that have resulted from this study have not only provided greater insight into the role of genes and other factors involved in myopia but have also gone some way to uncovering the aetiology of other refractive errors. This review will describe some of the major findings of the GEM study and their relative contribution to the literature, illuminate where the deficiencies are in our understanding of the development of refractive errors and how we will advance this field in the future. The lumican gene (lum), which encodes one of the major keratan sulfate proteoglycans (KSPGs) in the vertebrate cornea and sclera, has been linked to axial myopia in humans. In this study, we chose zebrafish (Danio rerio) as an animal model to elucidate the role of lumican in the development of axial myopia. The zebrafish lumican gene (zlum) spans ∼4.6 kb of the zebrafish genome. Like human (hLUM) and mouse (mlum), zlum consists of three exons, two introns, and a TATA box-less promoter at the 5′-flanking region of the transcription initiation site. Sequence analysis of the cDNA predicts that zLum encodes 344 amino acids. zLum shares 51% amino acid sequence identity with human lumican. Similar to hLUM and mlum, zlum mRNA is expressed in the eye and many other tissues, such as brain, muscle, and liver as well. Transgenic zebrafish harboring an enhanced GFP reporter gene construct downstream of a 1.7-kb zlum 5′-flanking region displayed enhanced GFP expression in the cornea and sclera, as well as throughout the body. Down-regulation of zlum expression by antisense zlum morpholinos manifested ocular enlargement resembling axial myopia due to disruption of the collagen fibril arrangement in the sclera and resulted in scleral thinning. Administration of muscarinic receptor antagonists, e.g. atropine and pirenzepine, effectively subdued the ocular enlargement caused by morpholinos in in vivo zebrafish larvae assays. The observation suggests that zebrafish can be used as an in vivo model for screening compounds in treating myopia.

https://www.researchgate.net/publication/26680692_The_Association_of_Haplotype_at_the_Lumican_Gene_with_High_Myopia_Susceptibility_in_Taiwanese_Patients

Abdorrahman Boroumand Center :: Shahie Setrakian: One Person’s Story One Person’s Story Shahie Setrakian About Age: 31 Civil Status: Unknown Case Date of Killing: April 18, 1983 Location of Killing: Beirut, Lebanon Mode of Killing: Bombing Charges: Unknown charge About this Case Ms. Shahie Setrakian was one of 63 victims of the bombing of the American Embassy in Beirut, Lebanon, which took place on April 18, 1983. At approximately 1:05 PM, a truck loaded with nearly 2,000 pounds of explosives careened through the driveway of the American embassy and crashed into the building. A massive explosion ripped through all seven levels of the embassy, sending debris flying hundreds of feet into the air and causing the burning building to collapse on itself. In addition to those who lost their lives, at least 120 people were injured. At the time, it was the deadliest attack on an American diplomatic mission since World War II. Ms. Setrakian was one of 38 local employees of the American Embassy (36 presumed Lebanese, one Tunisian, and one Egyptian) who were killed in the explosion. She was 31 years old at the time of her death. Background Ms. Setrakian is in Omid because of the overwhelming evidence connecting the Islamic Republic of Iran to the 1983 bombing at the American embassy in Beirut. This attack, claimed by a group called “Islamic Jihad,” was the first in a series of related bombings carried out in Lebanon between 1983 and 1984, which claimed at least 396 lives. In the early 1980s, Lebanon was a highly volatile country, home to conflicting military and political agendas and where the Islamic Republic exercised growing influence with the stated aim of spreading the Islamic Revolution. Already hostile to American influence in the Middle East and further angered by the United States’ military support of Iraq, Iran’s foe in the Iran-Iraq war, the Islamic Republic’s leaders became exasperated when, following the Israeli invasion of Lebanon in the summer of 1982, the United States brokered a Lebanese-Israeli truce and deployed a large contingent of American servicemen in the framework of a Multinational Peacekeeping Force. “Islamic Jihad” was a heretofore little-noticed group known only for a string of grenade attacks launched on French, Italian, and American members of the Multinational Force in March of 1983. When it claimed responsibility for the embassy bombing, “Islamic Jihad” declared that the attack was “part of the Iranian revolution’s campaign against imperialist targets throughout the world.” The group promised that “We shall keep striking at any imperialist presence in Lebanon, including the Multinational Force.” (Two massive bomb attacks against American and French servicemen indeed followed on October 23, 1983). “Islamic Jihad” was a shadowy subset of Hezbollah (Party of God), a radical, armed Shi’ite organization with deep ideological and organizational ties to the Islamic Republic of Iran. As early as August of 1979, Iran’s Supreme Leader, Ayatollah Khomeini, had alluded to the establishment of a “Party of God” that would unite Muslims around the goal of resisting American and Israeli designs for the Middle East. “So far, the oppressed have been disunited and nothing happens through disunity. Now that an example of the unity of the downtrodden has materialized in the land of Muslims [the Islamic Revolution], this example will have to be applied in a wider context, to encompass all humanity, through the establishment of a ‘Party of the Downtrodden,’ which is the same as the ‘Party of God’ [ Hezbollah], and which is in line with the exalted will of God … Past mistakes must be rectified through Muslim unity and the establishment of the ‘Party of the Downtrodden’ against the great powers, led by the criminal America and its very corrupt stooge, Israel.”* Punishing those who supported Iraq in the Iran-Iraq war was another publicly discussed subject. The Islamic Republic’s leaders were blunt about the potential consequences of the United States’ support for Iraq. As Khomeini stated in a speech of July 25, 1982, “If we see that … governments are giving financial and military assistance to the Iraqi government, we consider them guilty … we will deal with them as we do with guilty people, and we will enforce the Islamic sentences concerning them.” Ayatollah Khomeini repeatedly demonized the United States and expressed concern about its influence in the Muslim world. He emphasized that Muslims had a duty to rise up against “autocratic” leaders and that Iran would help them drive America out. In his speeches, he routinely called on Muslims, both Sunni and Shia, to follow the Iranian line on matters of foreign policy. “Muslims’ problems are all caused by great powers and their insinuations and suggestions to their stooges in the region,” he said in January of 1983. “All problems are caused by them, and these problems will not be resolved unless we rid ourselves of them.” Khomeini was particularly harsh when it came to Egypt and Lebanon: “Is it not shameful for Muslims that a country … which is considered the enemy of Islam and the enemy of humanity, reaches out from the other side of the world to determine the fate of Muslim countries? ... America pursues its corrupt goals through [Islamic and Arab] governments dependent on it, through wicked writers and speakers, and Muslims are just sitting [and doing nothing]. Is it not our duty? Do Muslims have no duties any longer in this day and age?” Days after the Israeli invasion of south Lebanon, in mid-June 1982, Iran openly stated through the Speaker of the Parliament, Hashemi Rafsanjani, that it planned to use a proxy to expel the United States from Lebanon. “We must not be afraid of confronting America,” he said. “If America becomes embroiled in this war [in Lebanon] with Muslims, it would be in our interest, and it would end America’s intervention in the region.” Once the American peacekeepers had arrived in Lebanon, Rafsanjani again warned that “the occupation of Lebanon, this small, sectarian and war-stricken country,” would inevitably bring “revenge.” As early as 1982, the Islamic Republic’s leadership, responding to Ayatollah Khomeini’s call for an organized effort to drive the United States out of Lebanon, had begun the process of transforming Hezbollah from loose, uncoordinated groups of Islamist militants into an organized and trained fighting force. Following the Israeli invasion, Iran deployed a contingent of 1,500 Revolutionary Guards to Baalbek, a city in eastern Lebanon and a hub of Shi'ite activism. In a speech delivered June 13, 1982, the Speaker of Parliament alluded to the training mission of the Revolutionary Guards when he stated that a confrontation with the United States and Israel would be undertaken “with the assistance of Islamic states, in particular the resistance front that we are creating.” He went on to say, “We must resist, stay and fight in a bid to deal with Israel once and for all.” In Baalbek, the Revolutionary Guards, in coordination with the Iranian Ministry of Intelligence and Security (MOIS), began training militants, providing them with weapons, paying them salaries and healthcare subsidies, and cultivating their ideological allegiance to the Islamic Republic by sending pro-Iranian clerics into the city’s mosques. Hezbollah pledged loyalty to the Islamic Republic’s Supreme Leader and depended on Iran for its funding, military training, and matériel. Iranian officials were also entrenched in Hezbollah’s highest governing body, the “consultative” ( shoura) council, which handed down policy, strategy, and orders for Hezbollah operations. Two (sometimes one) Iranian officials sat on the Consultative Council, including, in the early 1980s, Iran’s then-ambassador to Syria, Ali Akbar Mohtashami; the chargé d’affairesin Lebanon; and diplomatic staff at the Iranian embassy in Beirut. There were additional ties between Iran and Hezbollah officials.** Further, according to investigators, the attack on the embassy required not only training but financial backing; in Hezbollah’s early history these two dimensions were almost entirely under Iranian control. A plethora of evidence corroborating Iranian involvement in this and other “Islamic Jihad” bombings was collected by American intelligence agencies.*** Forensic evidence also pointed towards the intervention of a state actor. The U.S. Federal Bureau of Investigation, sent in to investigate the embassy bombing, identified the explosive used in the attack as the “bulk form” of PETN (pentaerythritol tetranitrate). The FBI came to this conclusion because PETN of the variety available commercially is wholly consumed in an explosion; however, in this case, there remained unconsumed particles of PETN at the blast site. This indicated that the explosive was the “bulk form” of PETN, a form generally not available commercially and which could only be procured from the manufacturer. According to the FBI’s on-scene forensic explosive investigator, in the Middle East, the bulk form of PETN was produced for military purposes by state-sponsored manufacturers. Such factories did not exist in Lebanon, but bulk form PETN was being manufactured in Iran. Incidentally, the same explosive was used in the October 21, 1983, bombing that killed 241 American servicemen.**** Investigations by the American media also revealed the existence of intelligence, not necessarily verifiable, on Iran’s involvement in the attack. For example, Washington Postinvestigative journalist Jack Anderson reported on May 10, 1983 that the National Security Agency had intercepted “communications … [that] gave a clear indication that a pro-Iranian Shiite Moslem group, fanatically loyal to Ayatollah Ruhollah Khomeini, was planning to bomb the embassy in Beirut.” The same article noted that analysis of the available intelligence led to the conclusion that “preparations for the bomb attack were supervised by a high official in the Iranian Foreign Ministry, who also gave final approval.” Further, Iran’s role became apparent in another attack against the U.S. Embassy in Beirut in 1984. As David Martin of CBS News and John Walcott of the Wall Street Journal reported in Best Laid Plans, their investigation of American anti-terrorism policy in the 1980s, satellite imagery obtained by the Central Intelligence Agency showed that the Revolutionary Guards were involved in the preparation of the bombing of the American Embassy Annex on September 20, 1984. Images of the Shaykh Abdallah barracks, which were controlled by the Guards and housed both American hostages held by Hezbollah and Hezbollah fighters, showed an exact replica of the concrete barriers guarding the U.S. Embassy Annex, in the vicinity of the barracks. Upon closer inspection, the images were found to show tire tracks swerving between the mock-up concrete blocks, suggesting that this Iranian-controlled location was where the suicide bomber had trained for the operation against the American Embassy. Finally, Iranian leaders boasted, directly and indirectly, of their responsibility for the string of bombings on American targets in 1983-1984. One such example is the statement by Mohsen Rafiqdust, who at the time of the bombing served as commander of the Iranian Revolutionary Guard Corps. In a speech delivered July 20, 1987, Rafiqdust boasted that the explosive in the Multinational Forces bombing of October 23, 1983, had indeed come from Iran: “In the victory of the revolution in Lebanon … the United States has felt our power on its ugly body; and it knows that both the TNT and the ideology that, in one blast, sent to hell 400 officers, NCOs, and soldiers at the Marine Headquarters, were provided by Iran.” Taken together, Iran’s activities in Lebanon, the stated motives of punishing the United States and driving it out of Lebanon, and other statements of the Islamic Republic’s leaders make the case for their involvement in the bombing of the American Embassy. The April 18, 1983, bombing, the first in a chain of bombings throughout the 1980s and in 1994, also fits the Islamic Republic’s pattern of using violence against civilians to further its foreign policy agenda. Notes for the “About this case” section: Information about the life of Ms. Setrakian has been gathered from the online memorial to victims of the Embassy bombing maintained by the American Embassy in Beirut, Lebanon. Notes for the “Background” section: *Sources for the statements presented (in order of appearance):1.Ruhollah Khomeini, Speech of August 7, 1979 on the occasion of Qods day in Qom (Iran), Sahife-ye Emam( Collected works and speeches of Imam Khomeini), Vol. 9, p.280.2.Ruhollah Khomeini, Speech of July 25, 1982 before the Hoseiniyeh Jamaran (Tehran), Sahife-ye Emam, Vol. 16, p. 393.3.Ruhollah Khomeini, Speech of January 2, 1983 before the Hoseyniyeh Jamaran (Tehran), Sahife-ye Emam, Vol. 17, pp. 208-209.4.Hashemi Rafsanjani, Speech of June 13, 1982 before Parliament, Notqha-ye Qabl Az Dastur( Addresses of Hashemi Rafsanjani before Parliament), p.130.5.Hashemi Rafsanjani, Speech of September 7, 1982 before Parliament, Notqha-ye Qabl Az Dastur, pp. 132-133. The full quote reads: “We say to America and Israel: do not become arrogant over this limited victory of occupation of Lebanon, this small, sectarian and war-stricken country. No matter how big a force you deploy there, you will not be as strong as the Shah was in Iran; sooner or later, nations lose patience and it is then that they will take revenge.”6.Hashemi Rafsanjani, Speech of June 13, 1982 before Parliament, Notqha-ye Qabl Az Dastur( Addresses of Hashemi Rafsanjani before Parliament), p.130. The full quote reads: “In Lebanon today, these Sa‘’ad Haddads [creator of the South Lebanon Army, a faction allied with Israel], Phalangists [a right-wing Christian faction in Lebanon], America and several other forces are on the side of Israel. With the assistance of Islamic states, in particular the resistance front that we are creating; we must resist, stay and fight in a bid to deal with Israel once and for all. Moreover, we must not be afraid of confronting America. If America becomes embroiled in this war with Muslims it would be in our interest and it would end America's intervention in the region.”7.Mohsen Rafiqdust, speech reported in Iran’s state-run newspaper, Resalat, July 20, 1987, p. 8. **Hussein Musawi, the leader of Islamic Amal, a group closely tied to Hezbollah, was a protégé of Mostafa Chamran, an important public figure in the Islamic Revolution. Before the 1979 Iranian revolution, Chamran had helped establish Amal, the main Shi‘ite political party in Lebanon; he later served as Iran’s Defense Minister from 1979 to 1981. The Lebanese cleric Ibrahim al-Amin was, in the early 1980s, Amal’s representative in Tehran. He defected to “Islamic Amal” when this group (eventually incorporated into Hezbollah) was established by Hussein Musawi following the Israeli invasion of June 1982; Al-Amin returned to Beirut in August of 1983 and shortly thereafter became a Hezbollah spokesperson, and issued Hezbollah’s manifesto in 1985. Sheikh Muhammad Ismail Khaliq, another senior Hezbollah official, was the personal representative in Lebanon of Iran’s Ayatollah Montazeri, whose son Mohammad founded and at the time supervised the Office of Liberation Movements, a department run by the Revolutionary Guards which was responsible for supporting opposition movements abroad. Ali Akbar Mohtashami, the Iranian Ambassador to Syria who served on Hezbollah’s shouracouncil in the early 1980s, meanwhile, had cultivated close ties to Hezbollah’s future leaders during his studies under Ayatollah Khomeini at a theological seminary in Najaf (Iraq). ***Some of this information was officially made public or declassified, some was leaked to the press or disclosed in memoirs, and some came to light in the course of civil suits filed against the Islamic Republic of Iran in the early 2000s. To cite just a few examples: Regarding the1983 American Embassy bombing, the Washington Post’s Jack Anderson reported in the May 10, 1983 article cited above that the National Security Intercepts reported to the United States government in the National Intelligence Daily (classified above top-secret) of April 20, 1983, and which pointed to Iranian designs to strike at the American Embassy, had been corroborated by human intelligence gathered by the CIA. Six days after Anderson published his article, on May 16, 1983, CBS News reported similar information – that unspecified “U.S. intelligence agencies” had intercepted, among other transmissions, a cable from the Iranian Foreign Ministry to the Iranian Embassy in Damascus authorizing “a $25,000 payment for a terrorist attack against an unidentified installation in Beirut.” In his Washington Postarticle, Anderson noted that the Intelligence Daily was “hedged with suggestions that the intelligence information could not be verified, and in at least one case came from an untested source.” Nonetheless, further information divulged to the Miami Heraldand published December 7, 1986 indicated that a distinct, $1 million transfer was made “from the government of Iran to the Iranian Embassy in Lebanon, where it was disbursed for the bombings of the U.S. Embassy and the Marine barracks.” Whereas the White House had never publicly confirmed the information leaks in 1983, the Miami Herald’s report was corroborated by an unnamed White House official. Regarding the1983 Marine barracks bombing, in addition to the forensic evidence mentioned above, Bob Woodward reported in his 1987 book Veil: The Secret Wars of the CIA, 1981-1987(p.275) that according to Israeli intelligence that was passed on to the United States, a “shadowy Lebanese financial emissary” named Hassan Hamiz had cashed a $50,000 voucher at the Iranian Embassy in Damascus, Syria; the emissary had close ties to Ali Akbar Mohtashami, Iran’s then-Ambassador to Syria, who had close connections to Hezbollah and served on its governing council. Further, a Hezbollah member testifying under the cover of anonymity in the civil suit Peterson v. Islamic Republic of Iranfiled in the U.S. District Court in the District of Columbia stated that the suicide bomber who had driven the truck into the Marine Barracks was Iranian (testimony cited in the Memorandum Opinion by the Honorable Royce C. Lamberth, May 20, 2003, 264 F.Supp.2d at 54-56). Evidence introduced in the same suit ( Peterson v. Islamic Republic of Iran) showed that on October 25, 1983, American naval intelligence was notified of the intercept of a message sent from Iran’s Ministry of Intelligence and Security to Ambassador Mohtashami in Damascus. The message instructed the Ambassador to contact Hussein Musawi, the leader of Islamic Amal (a group with close ties to Hezbollah) and instruct him to carry out attacks against the Multinational Forces, and specifically to “take a spectacular action against the United States Marines.” (See testimony of Admiral James A. Lyons, Deputy Chief of Naval Operations for Plans, Policy and Operation from 1983-85, in Peterson v. Islamic Republic of Iran, cited in the Memorandum Opinion by the Honorable Royce C. Lamberth, May 20, 2003, 264 F.Supp.2d at 54. Reports as to whom Ambassador Mohtashami in fact contacted vary, but all reports concerning the telephone conversation report the instruction to carry out a “spectacular action against the United States Marines”). According to the New York Timesof October 5, 1984 (p. A1), the Central Intelligence Agency, prior to the1984 Embassy Annex bombing, had indications that Iran was assisting the transfer of explosives from Syria into Lebanon. This information was confirmed by the U.S. Senate report on the September bombing, which stated that before the bombing, “US intelligence was aware of certain patterns of activity involving the transport of explosives by Iranians operating in Damascus under the shield of diplomatic immunity.” ( The security of American personnel in Lebanon: A staff report prepared for the committee on foreign relations United States Senate, October 1984, Government Printing Office). ****Mr. Danny Defenbaugh, the FBI’s lead explosives investigator for the two Embassy bombings and the Marine barracks bombing, gave this testimony before the U.S. District Court in the District of Columbia in the course of the civil suit Peterson v. Islamic Republic of Iran. The suit was filed by relatives of the 214 American servicemen who died in the October 23, 1983 bombing of the Marine barracks. For the relevant citations from Mr. Defenbaugh’s testimony, see the Memorandum Opinion by the Honorable Royce C. Lamberth, May 20, 2003, 264 F.Supp.2d at 56-58. *****The bombing at the American Embassy was not the last act of terror of this kind, nor were Americans the only ones targeted. A number of bombings in the 1980s appear to have connections to the Islamic Republic, including at least four in Lebanon, six in Kuwait, and eleven in France. Together with related assassinations and hostage-takings, these acts of terror killed over 400 men, women, and children (262 Americans, 76 Lebanese, 71 Frenchmen, 6 Iranians, and 11 persons of other nationalities have so far been identified). References: Background research was conducted through interviews of persons familiar with the events or involved with the investigations, and through consultation of sources including, but not limited to: 1. Monographs (in chronological order): David C. Martin and John Walcott, Best Laid Plans: The Inside Story of America's War Against Terrorism (1988); Robert “Bob” Woodward, Veil: The Secret Wars of the CIA, 1981-1987 (1988); Robert Fisk, Pity the Nation: Lebanon at War (first published 1990); Howard Teicher and Gayle Radley Teicher, Twin Pillars to Desert Storm: America's Flawed Vision in the Middle East From Nixon to Bush (1993); Roger Faligot and Rémi Kauffer, Histoire mondiale du renseignement. Tome 2, Les maîtres espions : de la guerre froide à nos jours (1994); Magnus Ranstorp, Hizb'allah in Lebanon: The Politics of the Western Hostage Crisis (1997); Robert Baer, See No Evil: The True Story of a Ground Soldier in the CIA's War Against Terrorism (2003); and Augustus Richard Norton, Hezbollah: A Short History (2007). 2. Scholarly articles such as: Marius Deeb, “Militant Islamic Movements in Lebanon: Origins, Social Basis, and Ideology,” Georgetown University Center for Contemporary Arab Studies Occasional Papers (November 1986), 27 pages; Simon Shapira, “The Origins of Hizballah,” Jerusalem Quarterly , No. 46 (Spring 1988), pp. 115-130; Augustus Richard Norton, “Hizballah and the Israeli Withdrawal from Southern Lebanon,” Journal of Palestine Studies , Vol. 30, No. 1 (Autumn 2000), pp. 22-35; and Magnus Ranstorp, “The Hizballah Training Camps of Lebanon,” in James Forest, ed., The Making of a Terrorist: Recruitment, Training, and Root Causes, Vol. 2 (2006), Chapter 15, pp.243-262, notes pp. 364-367. 3. Newspaper and magazine articles including: Mohammed Selhami, “J’ai rencontré les ‘hommes suicide’,” Jeune Afrique , January 25, 1984, pp. 41-51; Bob Woodward, Richard Harwood, and Christian Williams, “The Terror Factor,” a six-part series published in The Washington Post between February 1 and 12, 1984; Robert Fisk, “Iranian radical looks to ‘life’ after Madrid,” The Independent , October 23, 1991; Mark Bowden, “The Dark Art of Interrogation,” The Atlantic , October 2003; as well as a large number of reports covering topics such Iranian foreign policy, American-Iranian relations, and Iranian connections to acts of terror. In addition to articles cited in the footnotes above, these reports were published between 1983 and 1988 in news sources including but not limited to the Associated Press , United Press International , the Washington Post , the New York Times , and the Los Angeles Times , as well as the Lebanese newspapers Al-Nahar , Al-Nida , Al-Watan Al-Arabi , Monday Morning , and L'Orient-Le Jour . 4. Reports of the Foreign Broadcast Information Service dated May 17, 1982; July 27, 1982; July 28, 1982; May 6, 1983; May 19, 1983; January 1, 1984; March 25, 1984; March 26, 1984; March 30, 1984; and June 26, 1984. 5. Speeches and statements of Ayatollah Ruhollah Khomeini and Hojatoleslam Hashemi Rafsanjani (published, respectively, as Sahife-ye Emam , meaning “ Collected works and speeches of Imam Khomeini ,” and Notqha-ye Qabl Az Dastur , meaning “ Addresses of Hashemi Rafsanjani before Parliament ”). 6. Court opinions and exhibits associated with the following civil suits brought against the Islamic Republic of Iran in the U.S. District Court in the District of Columbia: (a) “Decision and Order” by the Honorable Thomas P. Jackson, Anderson v. Islamic Republic of Iran, et al. (No. CIV.A.99-0698, a suit filed by journalist Terry Anderson, held hostage by Hezbollah for seven years), March 24, 2000, 90 F.Supp.2d 107, particularly at 112; (b) “Opinion” by the Honorable Paul L. Friedman, Surette v. Islamic Republic of Iran, et al. (No. CIV.A.01-0570, a suit filed by the longtime partner of CIA officer Robert Buckley, who died in Hezbollah captivity), November 1, 2002 as amended November 4, 2002, 231 F.Supp.2d 260, particularly at 266; (c) “Memorandum Opinion” by the Honorable Royce C. Lamberth, Peterson v. Islamic Republic of Iran, et al. (Nos. CIV.A. 01-2094 and CIV.A.01-2684, suits filed by relatives of the 241 American servicemen killed in the Marine Barracks bombing on October 23, 1983), May 30, 2003, 264 F.Supp.2d 46, particularly at 51-58 and Footnote 10; (d) “Memorandum Opinion (Findings and Conclusions)” by the Honorable John D. Bates, Dammarell v. Islamic Republic of Iran, et al. (No. CIV.A.01-2224, a suit filed by relatives of the American victims of the bombing of the American Embassy on April 18, 1983), September 8, 2003, 281 F.Supp.2d 105, particularly Exhibits 8, 9, 10, 17, 19, 22, 27, 28, 30, 31, and 34; (e) “Report and Recommendation” by the Honorable Alan C. Kay, Welch v. Islamic Republic of Iran, et al. (No. CIV.A.01-863, a suit filed by relatives of Chief Warrant Officer Kenneth Welch (U.S. Army), killed in the bombing of the American Embassy Annex on September 20, 1984), September 20, 2007, 2007 WL 7688043 (D.D.C.), particularly WL page 6 (testimony of Dr. Bruce Tefft); (f) “Findings of Fact and Conclusion of Law” by the Honorable Ricardo M. Urbina, Kilburn v. Islamic Republic of Iran, et al. (No. CIV.A.01-1301, a suit filed by relatives of Peter Kilburn, an librarian at the American University in Beirut who died in Hezbollah captivity), March 30, 2010, 699 F.Supp.2d 136, particularly at 141-145 and 148; and (g) “Decision and Order” by the Honorable Thomas P. Jackson, Wagner v. Islamic Republic of Iran, et al. , (No. Civ.A.00-1799, a suit filed by relatives of Petty Officer First Class Michael R. Wagner (U.S. Navy), killed in the bombing of the American Embassy Annex on September 20, 1984), November 6, 2011, 172 F.Supp.2d 128, particularly at 132. What we do Who we are Contact us

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(PDF) Square Kilometer Array Telescope - Precision Reference Frequency Synchronisation via 1f-2f Dissemination PDF | The Square Kilometer Array (SKA) is an international effort to build the world's largest radio telescope, with one square kilometer collecting... | Find, read and cite all the research you need on ResearchGate Square Kilometer Array Telescope - Precision Reference Frequency Synchronisation via 1f-2f Dissemination DOI: 10.1038/srep13851 Authors: Abstract and Figures The Square Kilometer Array (SKA) is an international effort to build the world's largest radio telescope, with one square kilometer collecting area. Besides its ambitious scientific objectives, such as probing the cosmic dawn and cradle of life, SKA also demands several revolutionary technological breakthroughs, with ultra-high precision synchronisation of the frequency references for thousands of antennas being one of them. In this report, aimed at applications to SKA, we demonstrate a frequency reference synchronization and dissemination scheme with the phase noise compensation function placed at the client site. Hence, one central hub can be linked to a large number of client sites, forming a star-shaped topology. As a performance test, the 100 MHz reference signal from a Hydrogen maser clock is disseminated and recovered at two remote sites. Phase noise characteristics of the recovered reference frequency signal coincides with that of the hydrogen-maser source and satisfies SKA requirement. Schematic diagram of the client-side, 1f-2f actively compensated frequency dissemination system.: PDRO: phase-locked dielectric resonant oscillator. OCXO: oven-controlled crystal oscillator. WDM: wavelength-division multiplexer. FPD: fast photodiode. … Results of relative frequency stability measurements.: (a) Relative frequency stability between the free-running OCXO and H-maser clock. (b) Measured frequency stability of the dissemination system without compensation. (c) Relative frequency stability between the recovered 100-MHz signal at RX and Vref at TX with the PLL closed. (d) Relative frequency stability of the two recovered 100-MHz signals at the two RX sites with both PLLs closed. Curves (c,d) were measured simultaneously. … Results of phase-noise measurements for various 100-MHz frequency signals.: (a) SSB phase noise of the H-maser at TX. (b) SSB phase noise of the free-running OCXO used in the experiment. (c) SSB phase noise of the phase-locked OCXO in wideband locking mode. (d) SSB phase noise of the phase-locked OCXO in narrowband locking mode. … Figures - uploaded by Bo Wang www.nature.com/scientificreports Square Kilometre Array T elescope—Precision Reference Frequency Synchronisation via 1f-2f Dissemination B. Wang 1,2 , X. Zhu 1,3 , C. Gao 1,2 , Y . Bai 1,3 , J. W. Dong 1,2 & L. J. Wang 1,2,3,4 The Square Kilometre Array (SKA) project is an international eort to build the world’s largest radio telescope, with a one-square-kilometre collecting area. In addition to its ambitious scientic objectives, such as probing cosmic dawn and the cradle of life, the SKA demands several revolutionary technological breakthroughs, such as ultra-high precision synchronisation of the frequency references for thousands of antennas. In this report, with the purpose of application to the SKA, we demonstrate a frequency reference dissemination and synchronisation scheme in which the phase-noise compensation function is applied at the client site. Hence, one central hub can be linked to a large number of client sites, thus forming a star-shaped topology. As a performance test, a 100-MHz reference frequency signal from a hydrogen maser (H-maser) clock is disseminated and recovered at two remote sites. The phase-noise characteristics of the recovered reference frequency signal coincide with those of the H-maser source and satisfy the SKA requirements. Beneting from innovations in modern at omic clocks, time and frequency have become the physical quantities that can be most accurate ly and stably measured and con trolled 1–4 . e powerful tools for precision time and frequency dissemination a nd synchronisation that ha ve been developed 5–12 , although initially intended for metrology 13 , can be utilised in an increasing n umber of elds by converting meas- urements of target para meters into measurements o f time and frequency. F or example, in radio tel- escope array application s 14–16 , the image delity is related to the accuracy and stabili ty of the phase measurements. High-precisio n, short-term (1–100 s) phase (frequency) synchronisation among di erent telescope dishes is required. is requir ement typically cannot be satised by traditional synchr onisation methods via satellite links, which can achieve only long-term ( > 1000 s) frequency synchronisatio n 17,18 . Furthermore, satellite-based time and frequen cy synchronisation cannot meet the radio silence requir e- ment of radio astrono my observations. In this regard, r ecently developed bre-based frequency syn- chronisation methods are mo re suitable because they can achieve ultra-high-precision p hase (frequency) synchronisation for in tegration times from 1 to 10 6 s 19 . To incr ease the availability and accessibility o f this approach, several bre-based mul tiple-access frequency dissemination schemes have been pr oposed and demonstrated 20–25 . Ho wever, there a re still many unsolved pro blems related to these methods. ere are three major r equirements for br e-based reference frequency synchronisation syst ems that are specic to the SKA 14 . First, in the rst phase of the S KA (SKA1), the synchronisatio n system must distribute the reference frequen cy signal to hundreds of receivers in a star -shaped topology. is r equire- ment will be extended to thousands of receivers in the second phase (SKA2). Second, in accordance with 1 Joint Institute for Measurement Science, Tsinghua University , Beijing 100084, China. 2 State Key Laboratory of Precision Measurement T echnology and Instruments, Department of Precision Instruments, Tsinghua University, Beijing 100084, China. 3 Department of Physics, Tsinghua University , Beijing 100084, China. 4 National Institute of Metrology, Beijing 100013, China. Correspondence and requests for materials should be addressed to L.J.W . (email: [email protected]) R eceived: 09 April 2015 P ublished: 09 September 2015 www.nature.com/scientificreports/ 2 SCIENTIFIC RepoRts | 5:13851 | DOI: 10.1038/srep1 3851 the coherence requiremen ts for the reference frequency signals, which must su pport astronomical obser- vations at frequencies of u p to 20 GHz with a maximum coher ence loss of 2% over periods of 1–100 s, the frequency dissemination stability should be better than 1 × 10 − 12 /s, 1 × 10 − 13 /10 s and 1 × 10 − 14 /100 s, respectively . Finally, in acco rdance with the jitter requir ements for the recover ed sampling clock at each receptor , t he phase noise of the r eference frequency signal should not degrade through dissemination. Simply speaking, the frequency stability and phase-noi se specications of the H-maser clock at the SKA central station should be recover ed at each dish site. ese requiremen ts cannot be well satised by current bre-based frequency dissemination schemes with active co mpensation. In all presen t schemes, the phase-noise detection and compensation function s are typically applied at the tran smitting site. In the case of SKA1, for which hundr eds of telescope dishes are planned, a corres ponding number of com- pensation modules would need to be placed at the same central statio n. is requirement would incur extraordinary space requirements and ca use unnecessary complexities and diculties for future expan- sion. Furthermore, the phase-noise com p ensation bandwid th is related to the propaga tion delay of light in the bre, which would cause so-called delay-unsup pressed bre noise. In this report, we propose and demon strate a new frequency dissemination and synchro nisation method that features phase-noise co mpensation performed at the client (dis h) site. One central trans- mitting module can thus be linked to multi ple client sites, and futur e expansion to additional receiv- ing sites will not disrupt the structure of the central tran smitting station. As a performance tes t, using two 50-km bre spools, we recover 100-MHz di sseminated reference frequency signals at two separa te remote sites. Relativ e frequency stabilities between the two recovered frequency signals of 3.7 × 10 − 14 /s and 3.0 × 10 − 17 /10 5 s are ob tained; these values far exceed the SKA requirements. F ur thermore, the phase noise of the recovered 100-MHz signal is measur ed and found to coincide with that o f the H-maser clock’ s source signal at low frequency osets ( < 60 Hz) and to be better at higher frequency osets. e proposed scheme can well satisfy all frequency dissemination and synchro nisation requirements o f the SKA. Results Client-side, 1f-2f actively compensated frequency dissemination method. Figure1 presents a schematic diagram of the client-side actively co mpensated frequency dissemination system. One tran s- mitting site (TX), which represents the cen tral station, is connected to two receiving sites (RX). As a performance test and for con venience of phase-dierence measurements, TX is linked to RX-I and RX-II via two 50-km bre spools. In our case, unlike in current active pha se compensation schemes, the func- tions of TX are very simple—modulating and br oadcasting. T o increase the signal-to-noise ratio for compensation, the 100-MHz ref erence frequency (V ref from an H-maser clock) is boosted to 2 GHz via a phase-locked dielectric resonant oscillator (PDRO); this signal can be expr essed as V 0 = cos( ω 0 t + φ 0 ) (without considering its am plitude). V 0 is used to modulate the am plitude of a 1547.72-nm diode laser . As an example, in the dissemina tion channel from TX to RX-I, aer passing through a “2 to 1” bre coupler and an optical circulat or, the modula ted laser light is coupled to the 50-km bre link. e struc- ture of the RX site is relative ly complex. A 1-GHz PDRO , which can be expressed as V 1 = cos( ω 1 t + φ 1 ), Figure 1. Schematic diagram of the client-side, 1f-2f actively compensated frequency dissemination system. PDRO: phase-locked dielectric resonant oscillator. OCXO: o ven-controlled crystal oscillator . WDM: wavelength-division multi plexer. FPD: fast p hotodiode. www.nature.com/scientificreports/ 3 SCIENTIFIC RepoRts | 5:13851 | DOI: 10.1038/srep1 3851 is phase locked to a 100-MHz oven-con trolled crystal oscillator (OCXO). e phase of the OCXO can be controlled by app lying an external voltage. V 1 is used to modulate the a mplitude of a 1548.53-nm diode laser. W ith the aid of two optical circulator s at the RX-I and TX sites, the modulated 1548.53-nm laser light propagates thro ugh the same 50-km bre spool via the route from RX-I to TX and back. Aer the round-trip propaga tion, the 1548.53-nm laser light can be separated from the received one-wa y 1547.72-nm laser light by a wavelength-division m ultiplexer (WDM). e modulated 1547.72-nm laser is then detected by a fast photodiode (FPD1). e directly received 2-GHz frequen cy signal from TX can be expressed as V 2 = cos( ω 0 t + φ 0 + φ p ), where φ p is the phase uctuation induced during the 50-km bre dissemination. e modulated 1548.53-nm laser carrier is detected by FPD2, and the r ecovered 1-GHz frequency signal can be expressed as () ωφ φ =+ + ′ Vt co s p 31 1 , where φ ′ p is the phase uctuation induced by the 100-km (round-trip) br e dissemination. W e then down mix the signals V 2 and V 3 to obtain ωω φφ φφ =    (− )+ +− − ′    V co st pp 40 1 01 . Furthermore, down-mixing the signals V 1 and V 4 yields an error signal ωω φφ φφ =    (− )+ +− − ′    V co s2 t2 pp 50 1 01 . Finally, V 5 is u sed to close the phase-locked loop (PLL) to control the phase o f the OCXO. In this case, the rela tion ω 0 = 2 ω 1 is satised, and under a bre dispersion con dition (discussed later), the one-way phase uctua tion of the 2-GHz frequency signal is the same as the round-trip accumulated phase uctua tion of the 1-GHz frequency signal, φ φ = ′ pp . Consequently , t he error signal can be expressed as V 5 = cos( φ 0 − 2 φ 1 ). When the PLL is closed, the phase of the OCXO at RX-I is locked to the phase of V ref at TX, with φ 0 = 2 φ 1 , thus providing stable-frequency dissemination from TX to RX-I. As previously men tioned, the light propaga tion delay in the bre link limits the feedback contro l time and cause delay-unsupp ressed bre noise. T o ensure that the phase noise of the 100-MHz OCXO at the clien t site is not worsened by this uns uppressed bre noise, we use a custom-designed narrow-bandwidth PLL. I ts bandwidth can be tuned and set based on the phase-noise characteristics of the free-running OCXO and H-maser clock. I n our case, the PLL ’s band- width is set to 10 Hz. e same structure can be applied to RX-II and all other future si tes. e method is referred to as the “1f-2f ” method because of the factor-of-2 r elationship between the TX and RX frequencies. Measurements of the frequency dissemination stability. T o test the dissemination stability of the proposed client-site active co mpensation method, we performed a series of measurements.  e results are shown in Fig.2. Curve (a) repr esents the measured relative stab ility of the 100-MHz frequency signals between the H-maser at TX and the OCXO at RX-I with the PLL open. e relative frequency Figure 2. Results of relative frequency stability measurements. ( a ) Rela tive frequency stability between the free-running OCXO and H-maser clock. ( b ) Measured frequency stability o f the dissemination system without compensation. ( c ) Rela tive frequency stability between the recovered 100-MHz signal a t RX and V ref at TX with the PLL closed. ( d ) Relative frequency stability of the two reco vered 100-MHz signals at the two RX sites with both PLLs closed. Curves ( c , d ) were measured simultaneously . www.nature.com/scientificreports/ 4 SCIENTIFIC RepoRts | 5:13851 | DOI: 10.1038/srep1 3851 stabilities of 1.6 × 10 − 11 /s and 1.2 × 10 − 9 /10 5 s reect the frequency stability of the OCXO that we used. Curve (b) represents the relativ e frequency stability between V 0 and V 2 . e relative frequency stabili- ties of 2.0 × 10 − 12 /s and 3.4 × 10 − 14 /10 5 s reect the dissemination stability of the 50-km br e without compensation. Curve (c) repr esents the measured relative stabili ty of the 100-MHz frequency signals between the H-maser clock at TX and the OCXO at RX-I with the PLL closed at a 10-Hz eective band- width. Relative frequency stabilities of 3.1 × 10 − 14 /s and 2.7 × 10 − 17 /10 5 s were obtained, thus indicatin g that the OCXO at RX-I had become phase locked to the H-maser at TX. W e also measured the relative frequency stability between the two OCXOs at RX-I and RX-II when the phase-locked loops at both dissemination links were closed, as repr esented by curve (d). Relative frequency stabilities of 3.7 × 10 − 14 /s and 3.0 × 10 − 17 /10 5 s were achieved, thereby indicating tha t the OCXOs at the two RX sites were phase coherent with each other within the com pensation bandwidth of the dissemination sys tem. Phase-noise measurements. In the above measuremen ts, we tested the frequency dissemination stability for integra tion times from 1 to 10 5 s, i.e., the long-term stability (longer than 1 s) of the recovered 100-MHz frequency signal. However , outstanding short-term stability (shorter than 1 s) for the recovered 100-MHz frequency signal is also required by the SKA. In the frequen cy domain, the short-term phase instability can be dened in terms of the one-sided spectral density () = () φ Lf Sf 2 , which is half of the double-sided spectral density of phase uctuations, S φ ( f ) (inc luding the uctuations in both the upper and lower sidebands of the carrier) 26 . T ypically, L(f) is expressed in decibels (dB) as 10 Lg [ L ( f )] and is referred to as the single-sideband (SSB) phase noi se. is quantity was directly measured using a com- mercial signal source analyser (Agilent E5052B). Figure 3 shows the SSB phase-noise spectra of the 100-MHz signals measured in various cases. Curves (a) and (b) represen t the measured SSB phase-noise spectra of the H-maser clock and free-running OCXO, respectively , that we used. Obviously , t he long-term phase stability of the free-running OCXO is wo rse than that of the H-maser clock, whereas its high-frequency-oset ( > 60 Hz) phase-no ise characteristics are better than those of the H-maser clock. e main crossing point o f these two spectra occurs at approximatel y 10 Hz. Curve (c) represents the SSB phase noise of the OCXO when the PLL was closed in a wideband locking mode. In this case, the loop bandwidth is determined by the light pro pagation delay and can be calculated as 1/4 τ ~ 500 Hz 27 , where τ is the round-trip propagation time for ligh t in the 50-km bre link. Consequently , there is a large bump in curve (c) at a frequency oset of ap proximately 500 Hz. Furthermore, the unsuppressed high-frequency phase noise ( > 500 Hz) induced during bre dissemination worsens the phase-locked OCXO’ s phase noise. Curve (d) represents the SSB phase noise of the OCX O when the eective band- width of the PLL is set to 10 Hz. In this case, the SSB phase noise of the OCXO coincides with that of the H-maser at frequency osets of less than 60 Hz and is even better at frequency osets higher than 60 Hz. Based on these measurements, we observed that the long-term stability of the recover ed 100 MHz frequency signal was locked to the H-maser at the TX site and that its short-term phase-noi se charac- teristics can be improved b y carefully choosing the locking bandwidth. Figure 3. Results of phase-noise measurements for various 100-MH z frequency signals. ( a ) SSB phase noise of the H-maser at TX. ( b ) SSB phase noise of the free-running OCXO used in the experiment. ( c ) SSB phase noise of the phase-locked OCXO in wideband locking mode. ( d ) SSB phase no ise of the phase-locked OCXO in narrowband locking mode. Discussion In the proposed scheme, the wavelengths o f the two laser carriers are dierent. Consequen tly, the b re dispersion may cause a phase-delay di erence between the two laser carriers, which may limit the fre- quency dissemination stability . Here, we discuss this phenomenon and analyse its e ects in a real-world situation. e phase time delay τ ( λ , T ) of the disseminated frequency signal is τλ λ (, )= (, )⋅ () , () T nT LT c 1 where c is the speed of light in vacuum, n ( λ , T ) is the refractive index of the bre at optical wavelen gth λ and temperature T , and L ( T ) is the bre length. In practice, tem perature uctuation s in the bre link are the dominant factor tha t will degrade the frequency dissemination stability . We calculate the second-order partial derivative of the phase time delay as f ollows: τλ λ λ λ λ λ κλ α ∂( ,) ∂∂ =⋅ ∂    ⋅( )+ ⋅( ,)    ∂ = ∂( ,) ⋅( ) ∂ =( )⋅ ∂( ,) ∂ +( ,) ⋅ ∂( ) ∂ =( )⋅ +( ,) ⋅, () λ λλ ∂( ,) ∂ ∂( ) ∂ T Tc LT nT T DT LT T LT DT T DT LT T LT DT 1 [] [] 2 nT LT 2 where the bre dispersion is λ λ λ (, )= ⋅ (, ) , () DT c dn T d 1 3 the thermal coecient of chromatic dispersion is κ λ = (, ) , () dD T dT 4 and the thermal expansion coecient for the br e length is α = () ⋅ () . () LT dL T dT 1 5 For a commercial G652 br e, α = 5.6 × 10 − 7 /° C 28 , D = 17 ps /( nm · km ) and κ = − 1.45 × 10 − 3 ps /( km · n- m ·° C ) 29 at appro ximately 1550 nm. Obviously , the thermal coecient of chromatic dispersion i s the dom- inant factor that con trols the phase time delay dierence. F or dissemination over a 100-km br e, the phase time delay dierence is δτ = − 0.14 ps /( nm ·° C ). Supposing a diurnal tem perature uctuation of 30 ° C , the dierence in the phase time delays of the frequency signals carried by 1547.72- and 1548.53-nm carriers (a 0.81-nm dierence) will be 3.5 ps, which corresponds to a dissemination stability of 8.1 × 10 − 17 over an integration time of half a da y. is frequency dissemina tion stability can satisfy the requirements o f almost all current practical applications. U sing the dense wavelength-division multiplexing (D WDM) technique, the wavelengths of the two laser carriers can be within 0.4 nm of each other, and this will further reduce the impact of chromatic dispersio n on the dissemination stability . Conclusions Unlike all current br e-based frequency dissemination schemes, in this proposed and demonstrat ed 1f-2f scheme, the phase-noise compensation function is a pplied at the receiving site. is key change makes this method suitable for use in any star -shaped topology f requency dissemination a pplications. e space requirements and co mplexity of the frequency dissemination system a t the central station can be dra- matically reduced. Furthermore, in this scalable scheme, fu ture expansions in terms of the n umber of receiving sites will not disrupt the structure of the cen t ral transmitting sta tion. is approach can sa tisfy all three basic requirements for SK A frequency reference synchronisa tion as mentioned earlier . Further verication on a telescope array will be performed in the near future . Methods Dissemination stability measurements. As noted above, curves (c) and (d) in Fig. 2 were meas- ured simultaneously . However, o nly one commercial phase-noise measuremen t de vice (model 5125A, Microsemi Corporation) was available. Consequen tly, w e used the “voltmeter” method to measure the dissemination stability with active com pensation. During the measurement, two 100-MHz signals wer e mixed into a direct current (DC) signal. e voltage  uctuation of this DC signal corresponded to the www.nature.com/scientificreports/ 6 SCIENTIFIC RepoRts | 5:13851 | DOI: 10.1038/srep1 3851 relative phase uctua tion between the two 100-MHz signals. e voltage uctuation was m easured using a precise voltmeter and reco rded by a computer . e relative phase time uctuation was calculated using () () =⋅ π () / yt ar csin f V V 1 2 t 2 pp . In this formula, f is the frequency of the compa rison signal, with the relation ω = 2 π f (here, f = 100 MHz); V (t) is the measured real-time voltage uctuation o f the DC signal; and V pp is the peak-to-peak voltage of the DC signal when the relative phase change between the two com parison signals is greater than 2 π , which can be measured when the PLL is open. U sing the commercial soware package “Stable 32” , we calculate the overlapping Allan deviation from the measur ed phase time uctu- ation. In practice, to enhance the sensitivi ty of phase-uctuation measuremen ts, V (t) is always initially set to approxima tely 0. is can be achieved by choosing a suitable len gth for the radio frequency (RF) cable between the OCXO and phase comparison mixer . Here, the result measured using the vol tmeter method was veried using a model 5125A commercial device. e two results were f ound to be identical. Compared with the 5125A, the voltmeter method oer s certain advantages. e frequency measurement range of the 5125A is 1–400 MHz, whereas the voltmeter method can be used to measure frequencies higher than 400 MHz. e disadvantages of the voltmeter method are the follo wing: (1) it cannot be used to measure signals with dierent frequencies, a nd (2) it cannot be used to measure unstable signals whose relative phase uctuation is gr eater than 2 π , e.g., curves (a) and (b) in Fig.2. References 1. T aamoto, M., Hong, F . L., Higashi, . & atori, H. An optical lattice cloc. Nature 435, 321–324 (2005). 2. Chou, C. W ., Hume, D. B ., oelemeij, J. C. J ., Wineland, D . J. & osenband, T . Frequency comparison of two high-accuracy Al + optical clocs. Phys. ev . Lett. 104, 070802 (2010). 3. Hinley , N. et al. An atomic cloc with 10 − 18 instability. Science 341, 1215–1218 (2013). 4. Bloom, B. J . et al. An optical lattice cloc with accuracy and stability at the 10 − 18 level. Nature 506, 71–75 (2014). 5. Fujieda, M.,  umagai, M., Gotoh, T. & Hosoama, M. Ul trastable frequency dissemination via optical ber at NICT . IEEE Trans. Instrum. Meas. 58, 1223–1228 (2009). 6. Marra, G. et al. H igh-resolution microwave frequency transf er over an 86-m-long optical ber networ using a mode-loced laser. Opt. Lett. 36, 511–513 (2011). 7. Predehl, . et al. A 920-ilom eter optical ber lin for frequency metrology at the 19th decimal place. Science 336, 441–444 (2012). 8. W ang, B. et al. Precise and continuous time and frequency synchro nisation at the 5 × 10 − 19 accuracy level. Sci. ep . 2, 556 (2012). 9. Lopez, O. et a l. Simultaneous remote transfer o f accurate timing and optical frequency over a public ber networ. A ppl. Phys. B. 110, 3–6 (2013) 10. Ning, B. e t al. High-precision distribution of highly stable o ptical pulse trains with 8.8 × 10 − 19 instability. Sci. ep . 4, 5109 (2014). 11. Giorget ta, F. . et al. Op tical two-way time and frequency transfer over free space. Nat. Photon . 7, 435–439 (2013). 12. Miao, J . et al. Ultra-stable radio frequency dissemination in free space. ev. Sci. Instru m . 84, 104703 (2013). 13. Fujieda, M. et al. All-op tical lin for direct comparison of distant op tical clocs. Opt. Express 19, 16498–16507 (2011). 14. T urner, W . SA phase 1 system (level 1) requirements specication. (2014) Ava ilable at: https://www .satelescope.org/ey- documents/. (Date of access: 31/03/2015). 15. Cliche, J . F . & Shillue, B. Precision timing contro l for radioastronomy: Maintaining f emtosecond synchronisation in the A tacama Large Millimeter Array . IEEE Contr . Syst. Mag . 26, 19–26 (2006). 16. Shillue, W . Fiber distribution of local oscillator for Atacama Larg e Millimeter Array. P aper presented at OFC/NFOEC 2008 , San Diego, CA USA, OSA (2008, February , 24-28). 17. Bauch, A. et al. Com parison between frequency standards in Europe and the USA at the 10 − 15 uncertainty level . Metrologia 43, 109–120 (2006). 18. Levine, J. A review of time an d frequency transfer methods. Metrologia . 45, S162–S174 (2008). 19. W ang, B. et al. Fiber based time and frequency synchronization system. Paper presen ted at China Satell ite Navigat ion Conference (CSNC) 2013 Proceedings , S pringer (doi: 10.1007/978-3-642-37407-4_32) (2013, May , 15-17). 20. Grosche, G. V erfahren zum bereitstellen einer referenz-frequenz. DPMA Paten t application DE 10 2008 062 139 ( 2010 ) . 21. Gao, C. et a l. Fiber-based multiple-access ultrastable frequency dissemination. Opt. Lett. 37, 4690–4692 (2012). 22. Bai, Y . et al. Fiber-based multiple-access optical frequency dissemination. Opt. Lett. 38, 3333–3335 (2013). 23. Grosche, G. Eavesdr opping time and frequency: phase noise cancellation along a time-varying path, such as an op tical ber. Opt. Lett. 39, 2545–2548 (2014). 24. Bercy , A. In-line extraction of an ultrastable frequency signal over an optical ber lin. J. Op t. Soc. Am. B 31, 678–685 (2014). 25. rehli, P ., Śliwczyńsi, Ł., Bucze, Ł. & Lipińsi, M. Mul tipoint dissemination of F frequency in ber optic lin with stabilized propagation delay . IEEE Trans. Ultrason. F er roelet r. Fr eq. Control . 60, 1804–1810 (2013). 26. Ferre-Pial, E. S. et a l. Dra revision of IEEE STD 1139-1988 standard denitions o f physical quantities for fundamen tal frequency and time metrology-random instabilities. Pa per presented at Proceedi ngs of Interna tional Freq uency Control Sympos ium , Orlando, FL USA, IEEE (doi: 10.1109/FEQ.1997.638567) (1997, M ay, 28-30). 27. Williams, P . A., Swann, W . C. & Newbury, N. . High-stability transfer of an op tical frequency over long ber-optic lins. J. Opt. Soc. Am. B 25, 1284–1293 (2008). 28. Bousonville, M. & ausch, J . Ve locity of signal delay changes in bre optic cab les. Paper presented at Pr oceedings of DIP AC09 , Basel, Switzerland, PSI (2009, May , 25-27). 29. Zhong, . et al. Experimental st udy on temperature dependence of dispersion o f G. 652 ber and its eect on high speed optical communication system and com pensation. Paper presented a t Optoelectronic Devices a nd Integra tion III , Beijing, China, SPIE (doi: 10.1117/12.870355) (2010, October, 18-20). Acknowledgements W e acknowledge funding support from the National Key Scientic Instrumen t and Equipment ... Apart from metrology laboratories and those involved directly in time, frequency or metrology activities [1]- [3], the applications include a broad range of scientific cases, like e.g. spectroscopy [4], astronomy (VLBI) [5] - [7] or geodesy [8]. Due to the high stability and accuracy possible by fiber optic transfer, there is also a growing number of sensitive commercial applications, like in telecommunication [9], [10], navigation and positioning [11], smart grids [12] or in the financial sector [13], where a certain amount of independency from global navigation satellite systems (GNSS), ubiquitously used for T&F transfer, is important [14]. ... ... . This feature is shown in Fig. 5a where the scale at the right side was drawn assuming 5 = 2550 MHz (in this case the abovementioned scaling factor equals to about 0.05). In addition, (5) and (6) suggest that the discussed systematic frequency error can be expected to be independent on the specific shape of the noise spectrum , # %, as the only noise-related parameter appears to be ( . ... ... This parameter reflects the fact that the sensitivity of the prescaler to the change of its input frequency decreases when the SNR decreases. It can be calculated using (6) as Q = 0 ( # 5 % 0 5 ⁄ . The relevant curves are shown in Fig. 7b. ... Systematic Frequency Error in Laser Synchronization Circuits for Fiber-Optic Time Transfer Systems The paper addresses the problem of a systematic frequency error occurring in semiconductor-laser frequency-synchronization circuits based on counting the beat note between the two lasers in a reference time interval using a high-frequency prescaler. Such synchronization circuits are suitable for operation in ultra-precise fiber-optic time-transfer links, used e.g. in time/frequency metrology. The error occurs when the power of the light coming from the reference laser, to which the second laser is synchronized, is below about -50 dBm to -40 dBm, depending on the details of particular circuit implementation. The error can reach tens of MHz if left out of consideration and does not depend on the frequency difference between the synchronized lasers. Its sign can be positive or negative, depending on the spectrum of the noise at the prescaler input and the frequency of the measured signal. In the paper we present the background of the systematic frequency error, discuss important parameters allowing for predicting the error value, and describe the simulation and theoretical models being helpful for designing and understanding operation of discussed circuits. The theoretical models presented here show good agreement with the experimental data, which demonstrates the usefulness of proposed methods. Implementing polarization scrambling to mitigate the effect of polarization misalignment of the lights of the lasers used was considered and the resulting penalty was determined. View Show abstract ... In order to achieve the high-precision optical fiber-based frequency transfer, it is usually necessary to adopt a phase compensation technology, and the compensation methods have two types, one is active approach and the other is passive. In these reported works, due to the advantage of low timing fluctuation, most frequency transfer schemes used remote active phase compensation to suppress the fluctuation [21][22] [23] [24]. Radio frequency (RF) signal is loaded onto a laser and transmitted over a round-trip fiber link, where the timing fluctuation between the reference and returning signals is measured with a phase discriminator. ... Femtosecond-Level Frequency Transfer at 10 GHz over Long Fiber Link with Optical–Electronic Joint Compensation We report a fiber-optic 10 GHz frequency transfer technique based on an optical–electronic joint phase compensator. A highly stable frequency signal at 10 GHz was transferred in a 50-km long fiber link by using this technique. Two key parameters of the frequency dissemination, the timing fluctuation and frequency stability were both measured. The experimental results show the root-mean-square timing fluctuation of the transferred microwave is about 103 fs within 10,000 s, and the frequency stability for the transmission link is 2.2 × 10−14 at 1 s and 8.5 × 10−17 at 2000 s. The technique proposed in this paper provides a powerful tool which can be used to transfer atomic clocks (e.g., commercial H-master and Cs clocks) in a long fiber link. ... Atomic clock synchronisation is a key component in a number of emerging areas of physics including the square kilometer array [1] , atomic clock geodesy [2,3,4,5,6,7,8,9,10,11] and searching for physics beyond the standard model [12]. Tracking the frequency differences between two atomic clocks over a long distance is now a well established concept. ... A novel method of synchronising two atomic clocks Adam Walton Ben Varcoe In this paper we present a novel method of atomic clock synchronisation combining digital error correction and phase tracking. Microwave broadcasts are used to measure the difference in frequency between a pair of atomic clocks. We use the fact that desynchronised atomic clocks give rise to a phase error in a QPSK broadcast, which can be measured with high precision. The rate of change of this phase error is used to find the relative error in frequency of reference signals provided by two clocks, with an uncertainty in the region of 1e-15 Hz over a measurement time of 10 minutes. We find that the base stability that can be measured with this technique is 2.66e-17 Hz, making it potentially applicable to applications like measuring geodetic height to the centimeter level. ... H IGH-precision frequency standards have been playing an important role in various research fields such as: fundamental physics [1], [2] (e.g., search for dark matter and general relativity effect verification [3]- [5], optical clock comparison [6]- [8]), radio astronomy [9] , positioning and navigation timing [10], [11]. The low-noise optical frequency transmission has the best performance because of its high carrier frequency and no fiber chromatic dispersion effect [12]- [18], but most practical applications still require the transmission of stable radio or microwave frequency to interface with electronic system [19], [20]. ... Simultaneous Optical and Radio Frequency Transmission Using Optical Phase Stabilization Combined With Phase Conjugation Technology We propose and demonstrate a new simultaneous optical and radio frequency (RF) transmission scheme using optical phase stabilization combined with phase conjugate technology. The acousto-optical modulator (AOM) is used as a frequency actuator to compensate the optical frequency phase noise introduced by the optical fiber link, and the electro-optical modulator (EOM) is used to modulate two optical carrier sidebands simultaneously to stabilize the RF phase by phase conjugate technology. The novelty is that forward and backward RFs are distinguished by the AOMs used in standard optical carrier transfer to avoid the backscattering without WDM, so that the optical link is more symmetric, reducing the degradation of frequency stability due to polarization fluctuation and chromatic dispersion thermal sensitivity. We experimentally demonstrate 193.4 THz optical frequency transfer with a frequency stability of 1.2 $ \times $ 10 <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">−17</sup> at the integral time of 1 s and 3.3 $ \times $ 10 <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">−19</sup> at 10000 s, 1 GHz RF with a frequency stability of 7 $ \times $ 10 <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">−13</sup> at 1 s and 3 $ \times $ 10 <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">−16</sup> at 10000 s over a 50.4 km fiber spools. This novel scheme demonstrates the compatibility of two contrasting fiber noise cancellation techniques, and is superior to the current optical frequency and RF simultaneous transmission schemes in terms of cost and complexity. ... In this system, the asymmetry of the fiber link is derived from the group velocity dispersion (GVD). Based on the phase noise cancellation scheme, we assumed that the forward transmission signal and back transmission signal would go through the same fiber Photonics 2022, 9, 399 9 of 10 link with the same delay τ; however, τ is closely related to the wavelength of the optical carrier and the temperature fluctuation [27] . The chromatic propagation delay uncertainty can be calculated as: ∆τ = (λ 1 − λ 2 )D(T)L. ... Microwave Frequency Dissemination over a 212 km Cascaded Urban Fiber Link with Stability at the 10−18 Level Article Full-text available Honglei Quan Wenxiang Xue Wenyu Zhao Shougang Zhang To synchronize standard frequency signals between long-distance laboratories, we carried out a frequency dissemination experiment over a 212 km cascaded urban fiber link. This cascaded link was composed of two 106 km fiber links, in which the fiber noise was compensated by two microwave frequency dissemination systems. The two adjacent frequency dissemination systems used different frequency transmitted signals, preventing the influence of signal crosstalk between the received signal of the previous stage and the transmitted signal of the second stage caused by microwave signal leakage. The frequency dissemination over the cascaded link showed a dissemination fractional frequency instability of 6.2 × 10−15 at 1 s and 6.4 × 10−18 at 40,000 s, which is better than the transfer stability over the same 212 km single-stage link. Quantum two-way time synchronization over a 103 km urban fiber Huibo Hong Runai Quan Xiao Xiang As a new approach to realizing high-precision time synchronization between remote time scales, quantum two-way time synchronization via laboratory fiber link has shown significant enhancement of the synchronization stability to several tens of femtoseconds. To verify its great potential in practical systems, the field test in long-haul installed fiber optic infrastructure is required to be demonstrated. In this paper, we implement the two-way quantum time synchronization over a 103 km urban fiber link. A time synchronization stability of 3.67 ps at 10 s and 0.28 ps at 40000 s has been achieved, despite the large attenuation of 38 dB leading to fewer than 40 correlated events per second. This achievement marks the first successful step of quantum two-way time synchronization in the task of high-precision long-distance field synchronization systems. Phase-Modulation-Based Stable Radio Frequency Transmission Via 125 Km Fiber Optic Link Jun 2023 Chenxia Liu Tianwei Jiang Hanyue Wang Song Yu This paper proposes a stable radio frequency (RF) transfer scheme based on phase modulation. The passive compensation method is used to compensate the phase variations. The use of only one mixer reduces the power consumption during the frequency mixing process. By utilizing a phase modulator, the issue of bias drifting, encountered in RF transmission systems with intensity modulation method, is eliminated. A single-mode fiber (SMF) serves as a dispersive medium for phase modulation-to-intensity modulation (PM-to-IM) conversion. Our passive compensation system with phase modulation has the characteristics of robustness and low insertion loss at the modulation module. The RF signal with frequency of 2.4 GHz is transferred via a 125 km fiber optic link. The measured standard Allan deviation (ADEV) of our transmission system is 3.66× $10^{-13}$ at 1 s and 2.26× $10^{-16}$ at 10000 s, which is better than that of the reference atomic clock. The proposed system will be useful for further applications such as in square kilometer arrays and remote clock comparison. Kierunki optymalizacji konstrukcji w aspekcie nowoczesnych rozwiązań podwyższających własności osiągowe śmigłowców Chapter Jakub Kocjan Stanisław Kachel Robert Rogólski Praca podejmuje zagadnienia rozwijane w ramach uczelnianego projektu badawczego realizowanego w Wojskowej Akademii Technicznej w r. 2022 pn. "Metody optymalnego projektowania statków powietrznych w zastosowaniu do poprawy ich własności konstrukcyjnych i aerodynamicznych". Potrzeba prowadzenia badań jest konsekwencją analizy aktualnie eksploatowanych konstrukcji w wojskach NATO, która wykazała, że większość płatowców zakończy swój resurs docelowy w latach 2030-2050. W wyniku czego NSPA (NATO Support and Procurement Agency) opublikowało wymagania dla śmigłowca nowej generacji NGRC (Next Generation Rotorcraft Capability). Praca przedstawia analizę rozwiązań konstrukcyjnych ostatnich piętnastu lat w dziedzinie konstrukcji wiropłatów, a w szczególności śmigłowców o przeznaczeniu militarnym. Celem prowadzonych badań była analiza możliwości rozwoju potencjału operacyjnego wiropłatów stosowanych na nowoczesnym polu walki. W ramach prowadzonych badań zrealizowano przegląd nowoczesnych rozwiązań aerodynamicznych i konstrukcyjnych implementowanych we współczesnych śmigłowcach. Analizą objęto zarówno materiały udostępnianie przez producentów statków powietrznych jak i najnowsze publikacje naukowe w dziedzinie rozwoju wiropłatów. Wzięto również pod uwagę doświadczenia eksploatatora jakim są Siły Zbrojne Rzeczypospolitej Polskiej. Następstwem przeprowadzonej analizy jest matematyczna parametryzacja określonych podzespołów. W ramach analizy wskazano parametry projektowe i ich wpływ na osiągi śmigłowca. Korzystając z opracowanych danych opracowano parametryczny model łopaty jako przykładowy kierunku rozwoju projektowania wiropłatów z wykorzystaniem pętli optymalizacji. W podsumowaniu pracy dokonano oceny aktualnie wdrażanych nowoczesnych rozwiązań technicznych. Bazując na przeglądzie najnowszych informacji i raportów technicznych wskazano również potencjalne kierunki rozwoju i prognozowane rozwiązania dedykowane dla bojowych śmigłowców przyszłości. High-precision optical frequency transfer over 96km urban fiber link Dec 2022 Lei Liu Nan Cheng Jialiang Wang You-Zhen Gui We demonstrated the optical frequency transfer over a 96 km urban business network in Shanghai. The key factors affecting the optical frequency transmission system, such as fiber link quality, feedback compensation strength, and out-of-loop fiber temperature variation, are studied for the urban fiber link characteristics. The effective suppression technique of complex urban fiber link noise with different feedback compensation parameters is studied. With active phase noise suppression, the optical frequency stability can reach 1.9 × 10 − 16 at 1 s and 2.2 × 10 − 18 at 10,000 s over a 96 km urban fiber link. This work potentially plays an important role in optical clock frequency comparison, and it makes a good foundation for future research on long-distance optical frequency transfer over an urban business network. Generation of a coherent distributed RF array with a strong positive correlation OPT EXPRESS Yiwen Lu Zhongze Jiang Zhengyi Wan Yitang Dai In this work, we present a coherent distributed radio frequency (RF) array, discover and quantitatively describe the strong positive correlation between reconstructed signals for the first time. Eight replicable parallel receivers are connected to the phase-locked common trunk link via eight optical couplers spaced 1 km apart. The forward and backward signals at each receiver, extracted from two ports of optical couplers, are recovered to RF signals separately and then mixed to achieve upward frequency conversion. The link delay jitter is counteracted by wavelength-tuning of the optical carrier. With the long-term stability of point-to-multipoint fiber-optic RF dissemination effectively improved, the coherent distributed array is generated, and further the relative frequency stability between signals at different receivers is studied. The proposed correlation coefficient at 10 ³ s is ∼0.8 and shows a slight downward trend with the increase of averaging time based on our experimental results. High-Precision Distribution of Highly Stable Optical Pulse Trains with 8.8 × 10 instability Bo Ning Shuangyou Zhang Dong Hou Jianye Zhao The high-precision distribution of optical pulse trains via fibre links has had a considerable impact in many fields. In most published work, the accuracy is still fundamentally limited by unavoidable noise sources, such as thermal and shot noise from conventional photodiodes and thermal noise from mixers. Here, we demonstrate a new high-precision timing distribution system that uses a highly precise phase detector to obviously reduce the effect of these limitations. Instead of using photodiodes and microwave mixers, we use several fibre Sagnac-loop-based optical-microwave phase detectors (OM-PDs) to achieve optical-electrical conversion and phase measurements, thereby suppressing the sources of noise and achieving ultra-high accuracy. The results of a distribution experiment using a 10-km fibre link indicate that our system exhibits a residual instability of 2.0 × 10(-15) at1 s and8.8 × 10(-19) at 40,000 s and an integrated timing jitter as low as 3.8 fs in a bandwidth of 1 Hz to 100 kHz. This low instability and timing jitter make it possible for our system to be used in the distribution of optical-clock signals or in applications that require extremely accurate frequency/time synchronisation.

https://www.researchgate.net/publication/275364140_Square_Kilometer_Array_Telescope_-_Precision_Reference_Frequency_Synchronisation_via_1f-2f_Dissemination

DELANEY v. E.P.A. | 898 F.2d 687 (1990) | f2d68711444 | Leagle.com WIGGINS Circuit Judge Residents of Maricopa and Pima counties Arizona petition this court to vacate as arbitrary and capricious...f2d68711444 DELANEY v. E.P.A. No. 88-7368. View Case Cited Cases Citing Case 898 F.2d 687 (1990) Roberta DELANEY; Gayle Hartmann, et al., Petitioners, v. ENVIRONMENTAL PROTECTION AGENCY, Respondent. United States Court of Appeals, Ninth Circuit. https://leagle.com/images/logo.png Argued and Submitted June 26, 1989. Decided March 1, 1990. As Amended April 11, 1990. Attorney(s) appearing for the Case David S. Baron, Arizona Center for Law in the Public Interest, Tucson, Ariz., for petitioners. Alan W. Eckert, Associate General Counsel, U.S. Environmental Protection Agency, Washington, D.C., for respondent. Before TANG, REINHARDT and WIGGINS, Circuit Judges. United States Court of Appeals, Ninth Circuit. WIGGINS, Circuit Judge: Residents of Maricopa and Pima counties, Arizona, petition this court to vacate as arbitrary and capricious the Environmental Protection Agency's approvals of the counties' Clean Air Act implementation plans. We have jurisdiction under 42 U.S.C. § 7607(b)(1) (1982). We vacate the EPA's approvals of the plans, and direct the EPA to take further action consistent with this opinion. BACKGROUND The Clean Air Act, as amended in 1970, mandated that states as expeditiously as practicable, but not later than December 31, 1975, reduce the levels of certain ambient pollutants to comply with the National Ambient Air Quality Standards set by the EPA. 42 U.S.C. § 7409(a), (b) (1982). Preliminarily, each state had to develop a state implementation plan to achieve this goal. 42 U.S.C. § 7410(a) (1982). [898 F.2d 689] After many areas of the country failed to attain some or all of the national ambient air quality standards on time, Congress amended the Act in 1977, providing new deadlines for nonattainment areas. 42 U.S.C. §§ 7501-7508 (1982). The 1977 amendments required states to submit by January 1, 1979, for each of their nonattainment areas, a revised implementation plan that provided for implementation of all reasonably available control measures to attain the relevant national ambient air quality standards as expeditiously as practicable, but not later than December 31, 1982. 42 U.S.C. § 7502. If a state did not submit an adequate revised state implementation plan by the 1979 deadline, the EPA had to promulgate its own federal implementation plan. 42 U.S.C. §§ 7410(c)(1), 7502(b)(1). The 1977 amendments provided only one exception: If a state demonstrated in its revised implementation plan that the carbon monoxide or ozone ambient air quality standard could not be attained by the close of 1982 despite implementation of all reasonably available control measures, the state could have until the end of 1987 to attain the relevant air quality standard in that nonattainment area. 42 U.S.C. § 7502(a)(2). In that case, the state had to submit a second revised implementation plan to the EPA by July 1, 1982. 42 U.S.C. § 7502(c). In 1978, the EPA designated large areas of both Maricopa and Pima counties as nonattainment areas for carbon monoxide. In 1979, Arizona submitted revised implementation plans for both areas. In 1983, the EPA approved the plans subject to certain conditions. Neither area, however, satisfied the EPA's conditions or attained the carbon monoxide standard by the 1982 deadline. Arizona failed in its attempt to extend both areas' attainment deadlines to 1987, but submitted additional revised plans for the areas to attain by the 1987 deadline anyway. The EPA rejected these additional revised plans and we upheld that decision. Arizona v. Thomas,824 F.2d 745(9th Cir.1987), later proceeding829 F.2d 834(9th Cir.1987). When Arizona thereafter failed to submit adequate plans for Maricopa and Pima counties, petitioners filed suit in the Arizona district court. That court ordered the EPA to promulgate implementation plans for both counties by March 30, 1988 (later extended to August 10, 1988), unless before this date Arizona submitted, and the EPA approved, adequate state plans. McCarthy v. Thomas,No. 85-344 (D.Ariz. August 10, 1987). Arizona submitted, and the EPA ultimately approved (with revisions), state implementation plans for both counties on August 10, 1988. Petitioners now challenge these approvals. DISCUSSION We will not set aside the EPA's approval of a state implementation plan unless it is arbitrary, capricious, or otherwise not in accordance with law. 5 U.S.C. § 706(2)(A); Arizona v. Thomas,824 F.2d at 748. We will not set aside an agency's construction of a statute it is implementing unless that construction conflicts with clear congressional intent or is unreasonable. Chevron USA, Inc. v. Natural Resources Defense Counsel, Inc.,467 U.S. 837, 842-45, 104 S.Ct. 2778, 2781-83, 81 L.Ed.2d 694 (1984). I. The EPA's Decision That Delinquent Nonattainment Areas May Take Three Years From Approval of Their State Implementation Plan to Attain the National Ambient Air Quality Standards Because the Clean Air Act amendments of 1977 made the 1982 compliance deadline absolute (with the one exception noted above), the amendments did not specify an additional deadline for nonattainment areas that failed to obtain revised implementation plan approval and national ambient air quality standard compliance by the 1982 deadline. In this circumstance, the EPA adopted the policy that it should evaluate the adequacy of the Maricopa and Pima county plans based on whether they provide for attainment by three years from the date it approved those plans, August 10, 1991. Petitioners contend that the EPA's policy is arbitrary and capricious. Now that the 1977 amendments' deadline [898 F.2d 690] for compliance has passed, petitioners assert, Pima and Maricopa Counties must attain the carbon monoxide ambient air quality standard as soon as possible utilizing every available control measure. The EPA contends that Congress expressed no clear intent on the attainment deadline that the EPA should apply in evaluating the Maricopa and Pima county plans because the 1977 amendments did not specify a deadline for nonattainment areas that failed to obtain revised state implementation plan approval and national ambient air quality standard compliance by the 1982 deadline. The EPA argues that this amounts to a "statutory gap." As a result, the EPA believes, we must uphold its policy of allowing compliance within three years of state implementation plan approval because it is a reasonable construction of the 1977 amendments. The EPA argues that if Congress had considered that some nonattainment areas would not meet the deadline specified in the 1977 Amendments, Congress would have intended that the EPA administratively establish a new three year attainment period from the date of implementation plan approval deadline, like those provided in the Clean Air Act amendments of 1970. The EPA contends that this result is consistent with the language, history, and overall purposes of the Act. The EPA contends that Congress knew some states would not attain by the 1982 deadline and did not intend that states implement draconian measures. The EPA explains the seemingly absolute nature of the 1977 amendments' 1982 deadline by stating that some legislators felt the need to keep fixed deadlines, even as "somewhat of a legal myth," because they "provide a basis for obtaining maximum progress towards clean air." Transcript, Mark-up, Clean Air Act Amendments, at 13 (May 4, 1977) (Statement of Sen. Domenici), reprinted inState Implementation Plans for Nonattainment Areas for Ozone and Carbon Monoxide: General Preamble and Notice of Future Actions, 52 Fed.Reg. 26,408 (July 14, 1987) [hereinafter "Preamble and Notice"]. If the deadlines turned out not to be realistic, there would be "plenty of time for legislative relief." 123 Cong.Rec. 18,038 (June 8, 1977) (Statement of Sen. Stafford), reprinted inPreamble and Notice, 52 Fed.Reg. 26,408 (July 14, 1987). The EPA contends that petitioners' position would require the draconian measures that Congress allegedly intended to avoid. The EPA argues that, in effect, it is just doing what Congress did in 1977 when, faced with many states' failure to meet attainment deadlines, Congress revised the Clean Air Act, extending those deadlines at least three years. Although we recognize the EPA's predicament, we cannot accept the EPA's position. As the EPA itself recognizes, Congress explicitly declined to allow extensions of the deadline because it believed an absolute deadline to be necessary. Other circuits have characterized the 1982 deadline as the "`heart' of the 1977 Amendments," City of Seabrook v. EPA,659 F.2d 1349, 1357 (5th Cir.1981), cert. denied,459 U.S. 822, 103 S.Ct. 51, 74 L.Ed.2d 57 (1982), implemented according to a precise schedule to avoid a repetition of the failure of many states to attain the national ambient air quality standards by the deadline specified in the 1970 amendments. Connecticut Fund for the Environment v. EPA,672 F.2d 998, 1001-02 (2d Cir.), cert. denied,459 U.S. 1035, 103 S.Ct. 445, 74 L.Ed.2d 601 (1982). This court has already found the deadlines of the 1977 amendments to be "clear and unambiguous." Abramowitz v. EPA,832 F.2d 1071, 1079 (9th Cir.1987). We stated: Because we find the language of the Act clear and unambiguous, we do not believe that EPA has the discretion to ignore the statutory deadline. We are informed by counsel for both sides of their expectation that Congress will extend the deadline once again in the near future, but we must apply the law as it now stands, not as it may become .... Until the Clean Air Act is further considered ... the Agency must give effect to the unambiguously expressed intent of Congress. Id.(citations omitted). We, and the EPA, are bound by the statutory scheme until Congress alters that scheme. As the EPA states in one of its national guidance documents: [T]he legislative history shows that Congress set up the Part D system in order to force communities and industry to do their utmost to bring about attainment as rapidly as possible and expected that a future Congress would change the course it had set, if necessary, to avoid any unacceptable consequences. . . . . . ... Plainly, the original intention of the committee, which evolved ultimately into the current Part D system, was to generate [state implementation plans] that would provide for attainment by a specific date, even if they contained draconian measures, and to leave the task of making adjustments to a future Congress. Preamble and Notice, 52 Fed.Reg. 26,407-08 (July 14, 1987). When Congress has explicitly set an absolute deadline, congressional intent is clear. It is a semantic game to claim that once a state fails to meet an absolute deadline, a statutory gap is created because Congress has not provided a back-up deadline for its explicitly absolute deadline. If Congress had been more lenient and allowed a one-year fall back extension, the EPA would be bound by that directive. The EPA cannot extract leeway from a statute that Congress explicitly intended to be strict. In any case, the EPA's national guidance documents state that for those areas that qualified for an extension from 1982 to 1987, if their control measures "are not adequate to demonstrate attainment by 1987, additional measures which can be implemented after 1987 must be identified and adopted and attainment must be demonstrated by the earliest possible date." Environmental Protection Agency Criteria for Approving 1982 Ozone and Carbon Monoxide Plan Revisions, 46 Fed.Reg. 7186 (January 22, 1981) [hereinafter "Criteria"]. "The 1982 [state implementation plan] revision ... must demonstrate that all possible measures will be implemented...." Id.at 7188 (listing four examples of additional control measures). We believe that it is unreasonable for the EPA to treat more leniently areas that did not qualify for the 1987 extension than areas that did qualify. We believe that the only reasonable interpretation of the 1977 amendments is that if the 1982 deadline that Congress specified is not met, the national ambient air quality standards must be attained as soon as possible with every available control measure, including those that the EPA identified in its criteria for approving 1982 plans. II. The EPA's Decision that the Maricopa and Pima Plans Provide for Sufficient Control Measures The plans that the EPA approved for Maricopa and Pima Counties are similar. Both require that: (1) gas stations sell only high oxygen content fuel during winter months (when carbon monoxide levels are highest); (2) large employers reduce the amount of single car commuting by their employees; (3) automobiles be tested for emission levels while under load as well as while idling. Petitioners point out that neither the Maricopa plan nor the Pima plan adopts most of the forty-five measures recommended in a study of Maricopa County conducted by the Maricopa Association of Governments, a regional planning organization. Similarly, neither plan adopts most of the twelve measures recommended by Cambridge Systematics, a group that conducted an EPA-Sponsored study of Maricopa County. The excluded measures include major expansion of mass transit, imposition of parking controls, significant use of bus and carpool lanes, reductions in bus fares, restrictions on truck travel during peak periods, application of the Maricopa and Pima county oxygenated fuel program and automobile emission testing program to the entire state, elimination of waivers to the automobile [898 F.2d 692] emission testing program, and adoption of year-round daylight savings time. The Maricopa Association of Governments noted that immediate implementation of all the measures it identified could have produced attainment within one year. Petitioners contend that it was arbitrary and capricious for the EPA to approve the plans without more control measures. The EPA contends that it properly approved the Maricopa and Pima plans with just the three control measures that they contain, because additional control measures "could not be demonstrated to further accelerate the projected attainment date." The EPA has, however, arbitrarily shifted from Arizona the burden of demonstrating that control measures would not accelerate the projected attainment date. An EPA guidance document explicitly provides that each of the eighteen measures listed in 42 U.S.C. § 7408 is presumed reasonably available; a state can reject one of these measures only by showing that the measure either would not advance attainment, would cause substantial widespread and long-term adverse impact, or would take too long to implement. The document states: [I]f a state adopts less than all [reasonably available control measures] and demonstrates (a) that reasonable further progress and attainment of the [national ambient air quality standards] are assured, and (b) that application of all [reasonably available control measures] would not result in attainment any faster, then a plan with less than all [reasonably available control measures] may be approved. State Implementation Plans; General preamble for Proposed Rulemaking on Approval of Plan Revisions for Nonattainment Areas, 44 Fed.Reg. 20,375 (April 4, 1979) (emphasis added). Neither the Maricopa plan nor the Pima plan contains serious commitment to many of the measures listed in section 7408, including: 1) limiting portions of roads to common carriers; 2) improving transit systems with major changes in existing facilities; 3) controlling on-street parking; 4) establishing auto-free zones; 5) instituting road user fees that discourage single occupant automobile trips; or 6) retrofitting older vehicles with emission control devices. See42 U.S.C. § 7408(f)(1)(A)(v), (vi), (vii), (ix), (xiii), (xvii). Yet Arizona has made no claim that any of the control measures that section 7408 or the Maricopa Association of Governments identified is impracticable or unreasonable in either Maricopa County or Pima County. Furthermore, as noted above, for a nonattainment area that qualified for deadline extension to 1987, the 1977 amendments require implementation of not only all reasonably available control measures, but also any additional measures necessary to ensure timely attainment. 42 U.S.C. § 7502(b)(11)(C). If such an area failed to attain the relevant ambient air quality standard by 1987, the EPA required implementation of "all possible measures" and more extensive evidence to justify failure to adopt any of the measures listed in section 7408 in order to ensure "the most expeditious [attainment] date beyond 1987." Criteria, 46 Fed.Reg. 7188 (January 22, 1981). The EPA expressly applied these requirements to nonattainment areas that failed to meet the 1982 statutory deadline and did not qualify for deadline extensions to 1987. Guidance Document for Correction of Part D SIP's For Nonattainment Areas, January 27, 1984, at 32-33 [hereinafter "Part D SIP's for Nonattainment Areas"]. We, therefore, conclude that the EPA arbitrarily and capriciously found that the Maricopa and Pima plans provide for sufficient control measures. III. The EPA's Decision that the Maricopa Plan Provides Adequate Contingency and Conformity Provision The EPA's published guidelines require post-1982 carbon monoxide implementation plan revisions to contain a contingency plan to be implemented if short-falls in emission reductions occur during the period covered by the plan. See Criteria, 46 Fed.Reg. 7188 (January 22, 1981) (prescribing the policy for nonattainment extension areas). The contingency plan [898 F.2d 693] must provide: (1) a list of planned transportation measures that may adversely affect air quality and that will be delayed while the state implementation plan is being revised, if expected emission reductions or air quality improvements do not occur; and (2) a description of the process that will be used to determine and implement additional transportation measures beneficial to air quality that will compensate for unanticipated shortfalls in emission reductions. Id. The Maricopa plan contains neither of these. Petitioners contend that the EPA arbitrarily and capriciously waived its contingency plan requirement. The EPA contends that its contingency plan provision applies only to nonattainment extension areas. But as we note above, the EPA expressly applied its nonattainment extension area requirements to nonattainment nonextension areas. SeePart D SIP's for Nonattainment Areas, at 32-33. Indeed, it would be anomalous for the EPA not to require as stringent measures for delinquent areas as for areas that qualified for extensions. The EPA alternatively contends that each plan contains two traffic control measures beyond those necessary to demonstrate attainment by December 31, 1991, and that these additional measures are the functional equivalent of a contingency plan. We disagree. First, we held above that the EPA erred in not requiring compliance before December 31, 1991. Second, the EPA concedes that the two traffic control measures in question — a voluntary no-drive day program and a minimum market share for alcohol fuels — are of such speculative value that the EPA could not assign them any emission reduction value. SeeApproval and Promulgation of Implementation Plans; Arizona State Implementation Plan Revision; Maricopa County Carbon Monoxide Plan, 53 Fed.Reg. 30,232 (August 10, 1988) [hereinafter "Approval of Maricopa Plan"]. Programs of such speculative benefit are not the functional equivalent of a list of specific projects that will be delayed and a description of the specific process that will be used to determine and implement additional transportation control measures if they become necessary. We find that the EPA has arbitrarily and capriciously waived its requirement of a contingency plan. The Clean Air Act explicitly requires that all federal activities conform to Clean Air Act Implementation Plans. 42 U.S.C. § 7506(c). The EPA guidelines direct that to assure compliance with this mandate, each implementation plan "should identify, to the extent possible, the direct and indirect emissions associated with major federal actions. . . ." Criteria, 46 Fed.Reg. 7188 (January 22, 1981). The state implementation plan must contain "[a]dministrative and technical procedures and agency responsibilities for ensuring ... that transportation plans, programs and projects" will conform with the state implementation plan. Id.The EPA does not dispute that the Maricopa plan fails to meet either requirement. Petitioners argue that the EPA arbitrarily and capriciously waived these requirements. The EPA argues, however, that we should sustain as within its discretion, its finding that the plan sufficiently assures conformity "given the flexibility which should be accorded [the EPA's] practical implementation of the [Clean Air Act]'s statutory provisions." Again, we cannot agree. The EPA must adhere to its own guidelines. Morton v. Ruiz,415 U.S. 199, 235, 94 S.Ct. 1055, 1074, 39 L.Ed.2d 270 (1974); Service v. Dulles,354 U.S. 363, 388, 77 S.Ct. 1152, 1165, 1 L.Ed.2d 1403 (1957). Because it is undisputed that the Maricopa plan fails to meet either of the EPA's own requirements and the EPA offers no reason for waiving those requirements, we can conclude only that the EPA arbitrarily and capriciously approved the plan. See Illinois State Chamber of Commerce v. EPA,775 F.2d 1141, 1147 (7th Cir.1985). IV. The EPA's Decision that the Maricopa Plan Provides Adequate Maintenance of the National Ambient Air Quality Standards The Clean Air Act requires that implementation plans provide for maintenance of national ambient air quality standards after attainment. 42 U.S.C. [898 F.2d 694] § 7410(a)(2)(B). Petitioners contend that the Maricopa provisions for maintenance are inadequate because the plan considers air quality only ten years into the future. The EPA required projections only ten years into the future because it found that projections beyond ten years are unreliable. Of course, this finding would not have justified the EPA's disregard for its rules without a formal revision of the rules in accordance with the Administrative Procedure Act. But even assuming that the EPA's own rule generally requires maintenance projections twenty years into the future applies to delinquent nonattainment nonextension areas (which the EPA disputes), that rule allows plans to provide maintenance projections only ten years into the future. The rule states: The time period analyzed shall extend at least 20 years or, if modified under § 51.63, not less than 10 years from the date on which the Administrator identified the area. 40 C.F.R. § 451.42 (1987). Section 51.63(a) adds: At the request of a State, or under his own initiative, the Administrator, where he determines it appropriate, may approve alternative [air quality maintenance area] analysis and plan development procedures as allowed under §§ 51.42, 51.44, 51.45, 51.46, 51.48(b), and 51.60(a). He may consider all relevant factors including but not limited to air quality problems, financial and manpower limitations, administrative feasibility, and existing commitments by the State. 40 C.F.R. § 51.63(a) (1987). The preamble to these regulations specifically states that one of Section 51.63(a)'s "relevant factors" is the reliability of the maintenance projections: The time periods will be selected by the Administrator, through his Regional Administrators, considering such factors as State resources, other planning programs that may significantly affect air quality, the reliability of projections, and the extent of present and potential air quality problems. Maintenance of National Air Quality Standards — Summary, 41 Fed.Reg. 18,386 (May 3, 1976) (emphasis added). In this case, the EPA specifically found that projections beyond ten years into the future "become too speculative to be reliable." Approval of Maricopa Plan, 53 Fed. Reg. 30,234 (August 10, 1988). Petitioners' own contention that traffic projections changed dramatically from February 1987 to October 1987 supports the EPA's conclusion that projections are quite speculative. We, therefore, cannot say that the EPA arbitrarily or capriciously required a maintenance projection only ten years into the future. V. The EPA's Decision not to Consider Revised, Increased Vehicle Traffic Projections in Evaluating the Maricopa Plan The Maricopa plan based its projection that Maricopa County would attain the carbon monoxide ambient air quality standard by 1991 on traffic projections made in 1984 and in February 1987. The EPA refused to consider revised, increased vehicle traffic projections for Maricopa County that petitioners presented during the public comment period for the Maricopa plan. The EPA argues that petitioners submitted this new data only four weeks before the approval deadline and thus the EPA lacked sufficient time to incorporate it into the plan. Petitioners contend that the EPA acted arbitrarily and capriciously in failing to consider the revised, increased vehicle traffic projections. They also contend that the EPA's decision that the Maricopa plan provides adequate maintenance of the national ambient air quality standards was arbitrary and capricious because it failed to rely on the revised vehicle traffic projections. We do not determine whether the EPA acted arbitrarily and capriciously in its decision not to consider the revised traffic projections. Our holdings on the other issues in this case require the EPA to develop a new Maricopa county implementation plan. To develop that plan properly the EPA and Arizona will, of course, need to consider [898 F.2d 695] the most recent vehicle traffic projections currently available. Whether, due to time constraints, the EPA in 1988 justifiably refused to consider the revised, increased traffic projections is now an issue of only academic significance. CONCLUSION We grant the petition for review. We vacate the EPA's approvals of Maricopa and Pima counties' Clean Air Act implementation plans. We direct the EPA to disapprove these plans and to promulgate federal implementation plans consistent with this opinion within six months. 1To summarize, the new plans must utilize all available control measures to attain the carbon monoxide ambient air quality standard as soon as possible. The new plans must contain contingency and conformity plans in accordance with EPA guidelines and must be based on the most recent traffic projections currently available. We grant petitioners' motion for fees and costs and direct petitioners to submit appropriate documentation to this court. FootNotes 1. Arizona may, of course, submit proposals to the EPA for its consideration in developing the plans.

https://www.leagle.com/decision/19901585898f2d68711444

Mr Joseph Maclise and the Epistemology of the Anatomical Closet, by Michael Sappol | Article index | Articles | British Art Studies An online, open-access and peer-reviewed academic journal for research on all aspects of British art, architecture and visual culture. Mr Joseph Maclise and the Epistemology of the Anatomical Closet One ObjectbyMichael Sappol Joseph Maclise, Superior Mesenteric Artery (detail), from Richard Quain, The Anatomy of the Arteries of the Human Body(London: Taylor & Walton, 1841–1844), Plate 51, circa 1841–1844, coloured lithograph, 67.5 × 55 cm. Wellcome Collection. 5. Joseph Maclise, Taylor & Walton, and Publishing on Gower Street in the 1840s, by William Schupbach 6. “It Should Be on Every Surgeon’s Table”: The Reception and Adoption of Joseph Maclise’s Surgical Anatomy (1851) in the United States, by Naomi Slipp Abstract This article takes up the case of Joseph Maclise (1815–1891), a talented and truculent surgeon, anatomist, and medical illustrator of mid-nineteenth-century Britain. Maclise left behind a corpus of brilliant, idiosyncratic anatomical images, and opinionated commentaries, but almost no evidence of his social interactions or affective relations. Homoerotic desire was heavily policed in Maclise’s time. Given the conditions under which the archive was created (or suppressed, or lost, or shamed into reticence), we can never know with certainty what he intended or felt, or what his readers received—but we do have a rich evidentiary base of visual materials. Using narrative history, close readings of images and texts, detailed comparisons with other illustrated anatomies, and open-ended theoretical and methodological approaches (a mash-up of queer theory, Foucault, gaze theory, genre analysis, and contextualization)—an argument is joined: a book can be a closet and a queer space. Maclise’s drawings, ostensibly designed to contribute to the improvement of medical knowledge, theory, and practice, show good-looking young men and cadaveric bodies in various states of dissection. Penises, testicles, anuses, faces, sensuous hands on skin are crisply rendered in illusionistic perspective, with a highly cultivated aestheticism—often without any relevance to the anatomical topic discussed—and little attention is paid to the female body. In historical context, and from our twenty-first-century vantage point, the hypothesis of homoerotic investment leads to productive interpretations. This article poses more questions than answers but comes to rest with this: it is plausible and meaningful to take Maclise’s anatomical illustrations, and the figures depicted therein, as queer objects of queer desire. The Mystery of Mr Joseph Maclise The closet : a condition of smothered homosexuality, suppressed desire, longing. A claustrophobic space of isolation, where shameful feelings are incarcerated, expression is secret, or entirely thwarted. 1 2 The closet is not a grave where desire comes to die, but a private and provisionally safe queer space—shielded from public spectatorship and policing—where desire is imaginatively free to take shape, can be self-nurtured and articulated, in reverie as in linguistic and visual and gestural code, and in meaning-laden silence, arousal, and “the telling secret”. 3 But when same-sex desire and its objects are unavowed, and evidence is limited, how should we proceed? What can we know when “closeted-ness itself is a performance initiated … by the speech act of a silence”? 4 This is the paradox so cogently addressed in Eve Kosofsky Sedgwick’s Epistemology of the Closet (1990). Sedgwick acknowledges that there can be no epistemological certainty when it comes to a system of meaning that is covered over. But, she argues, if we only limit ourselves to what is epistemologically secure, we miss everything important. 5 But, leaving those specific claims aside, and following Sedgwick’s lead, let’s entertain the possibility that Maclise’s renderings, in some queer and occluded fashion—maybe not even fully apparent to Maclise himself—instantiated other principles. Maybe they were a space in which he unveiled himself, sent out flares of homoerotic desire. To our twenty-first-century eyes, Maclise’s “pictures of form” are open-ended, offer many propositions from which many possible solutions can be derived. But they do have tendencies: a deeply felt mimetic sensuality, an overabundant pleasure in bodies, line, and texture, an argumentative assertion of principles and positions, and a queer intensity. And so we’ll take the case of Maclise and his images in three contrapuntal registers: the historical method (chronological, biographical, contextual, narrative); close reading and analysis of images (and some texts); and the comparative method (comparing Maclise’s pictures to those of his predecessors and contemporaries). (Which is only fitting: Maclise was a passionate exponent of the comparative method.) Over its long history, anatomical illustration has often been a multivalent conveyor of meanings, open to idiosyncratic clowning, horror shows, allegorical play-acting, memento mori , and erotic expression. And often enough—given the centrality of the figure of Man as the emblem of the universal anatomical human—homoerotic expression. So, to set the scene before Maclise enters stage left, let’s entertain the possibility that this spectacular life-size colored engraving from Paolo Mascagni’s Anatomia universa (or Grande anatomia del corpo umano ) (1823–1830) performs something that exceeds the instrumental purpose of demonstrating the anatomy of the back of the arm, lower back, buttocks, and rectum ( Fig. 1 ). 6 7 Figure 1.Antonio Serrantoni, Back, Stratum Primum, from Paolo Mascagni, Anatomia universale … rappresentata con tavole … ridotte a minori forme di quelle della grande edizione pisana per Antonio Serrantoni(Firenze: V. Batelli, 1833), Plate 2, 1833, hand-coloured copper-plate engraving. Collection of Hagströmer Library. Digital image courtesy of Hagströmer Library (public domain). The erotics of anatomical illustration went mostly unmentioned in eighteenth- and nineteenth-century medical discourse. (Anatomists William Hunter and John Bell never explicitly discussed flirtation or sensuality in the essays that prefaced their anatomical atlases, but signaled their disdain for all forms of representational play.) 8 And, again, that’s a problem for historians: should we bracket erotic valences as a kind of unknowable semiological dark matter—disqualify ourselves on the grounds that we will be tempted to fill up the epistemological void with our own aisthesis Sharing Medical Eyes and Hands: Jacob and Bourgery’s Traité complet Between Men also happens to be the title of another seminal work by Sedgwick, an extended riff on masculine sociality and its troubled ramifications. 9 viewing and owning of those things. Anatomical objects occupied a place of privilege among the collectables: beautifully wrought sculptures, paintings, drawings, and prints of beautiful nude and semi-nude men and women; and authoritative, expensive, artfully illustrated and printed, often large-sized, books dealing with the subject of anatomy, atlases which stood atop the hierarchy of medical publication. 10 So it makes sense that a reviewer of the first two installments of Richard Quain’s lithographic “imperial” folio, Anatomy of the Arteries of the Human Body (inaugurated in 1840, completed in 1844), praises the “delineations of Mr. Maclise”, for evincing “artistical talent of the very highest order”, and makes a connoisseurial assessment: Maclise’s plates “will bear comparison with any of those splendid specimens of anatomical drawing, so abundant on the continent, but heretofore so very rare in our own country …”. 11 There was, it seems, something Continental Traité complet de l’anatomie de l’homme (1832–1854) of surgeon-anatomist Jean-Baptiste-Marc Bourgery (1797–1849) and his gifted collaborator, “professor-draughtsman-anatomist” Nicolas Henri Jacob (1781–1871). 12 Take, for example, this lithographic demonstration of an operation for the surgical removal of a breast cancer, which shows disembodied masculine hands working on the body of a supine and beautiful woman ( Fig. 2 ). The scene is a brilliant composition brilliantly executed. Jacob was a skilled practitioner of Jacques-Louis David’s “corporal aestheticism” (Jacob studied under David)—but was just as much under the influence of Jean-Auguste-Dominique Ingres, who famously used color and shading to show body volume, mass, texture, and silky, creamy, perfect skin. 13 So, a sumptuous, perfectly polished illustration of a desirable young woman with a fashionable hairdo, undergoing a terrible operation (before anesthesia comes into common usage) without any expression of terror. Like many other images in the Traité complet , it is offered as a how-to guide to a difficult surgical operation, a horror, and also a visual delight, an incitement. Figure 2.Nicolas Henri Jacob, Removal of the Breast, from J.B.M. Bourgery and Claude Bernard, Traité Complet de l’Anatomie de l’Homme(Paris: 1832–1854), Vol. 7 (1866–1871), Plate 27, coloured lithograph. Wellcome Collection. Digital image courtesy of Wellcome Collection (CC BY 4.0). Two sets of almost identical hands share the surgical work, mimetically evoking both the haptic experience of surgery and the touch-sensation of masculine hands on the naked flesh of a female beauty and her anatomical subsurface. 14 A shared set of touchings, which in turn are offered for sharing among those with privileged access to the plate in a fine medical library, or who can afford to subscribe to the installments (expensive), or purchase volumes as they were completed (very expensive), or purchase (from 1854 onward) the full set of eight volumes (obscenely expensive). 15 The Traité complet Traité complet , that asymmetrical relation, between undepicted gazer and depicted object of the gaze, inscribes a hierarchical relation—between spirit (the sight-unseen surgeon and reader/viewer, the subject) and matter (the pictured patient and cadaver, the object). 16 17 But, as we shall see, some constructions are more sexualized than others. Traité complet was on dissected bodies and body parts. But the showstoppers were the atlas’s nude and semi-nude figures, some female, but most male ( Fig. 3 ). Beautiful faces and bodies—objects of desire—subjected to horrific or just peculiar scenes of surgical-anatomical intervention, in a work that was itself an object of desire, arrayed in libraries alongside other works of bibliophilic desire. (And not far away: collections of desired specimens, models, instruments, objets d’art .) DOI Figure 3.Nicolas Henri Jacob, Ligature des arteres iliaque primitive, externe et interne, from Traité complet anatomie de l’homme, 8 vols. (Paris: 1832–1854), Vol. 6 (1841), Plate 48, coloured lithograph. Wellcome Collection.Digital image courtesy of Wellcome Collection (CC BY 4.0). 18 But even Jacob had to negotiate boundaries. Compare his preparatory drawing of the ligature of the iliac arteries with the finished 1841 print. The preparatory drawing shows a hand inside a surgical cut made in the body of a well-proportioned, almost classical, masculine torso ( Fig. 4 DOI Figure 4.Nicolas Henri Jacob, Study for Ligature des arteres iliaque primitive, externe et interne, prepared for Traité complet anatomie de l’homme, 8 vols. (Paris: 1832–1854), Vol. 6 (1841), Plate 48, drawing. Collection of Bibliothèque Nationale de France.Digital image courtesy of gallica.bnf.fr / Bibliothèque Nationale de France (all rights reserved). écorché > nude > dressed—that was a foundational part of the curriculum of drawing pedagogy. 19 And the nude stage of that hierarchy (often connected to the study of live nude models) provided the opportunity for a certain kind of private erotic pleasuring, followed by a more modest covered public presentation. 20 DOIWhat makes all of this germane to our discussion is that Joseph Maclise—who followed in the footsteps of Bourgery and Jacob, and situated himself in the tradition of perfectionist, sensual, volumetric figure drawing—departed from the conventions of medical illustration and, in his finished publications, didn’t cover up his irrelevant penises … DOI A Bit of Puppetry: Maclise’s Superior Mesenteric Artery 21 22 Consider, then, this plate, from Richard Quain’s The Anatomy of the Arteries of the Human Body (1844), featuring “drawings from nature and on stone”—Joseph Maclise’s first major publication as medical illustrator ( Fig. 5 ). 23 Figure 5.Joseph Maclise, Superior Mesenteric Artery, from Richard Quain, The Anatomy of the Arteries of the Human Body(London: Taylor & Walton, 1841–1844), Plate 51, c. 1841–1844, coloured lithograph, 67.5 x 55 cm. Wellcome Collection. Digital image courtesy of Wellcome Collection (CC BY 4.0). The wealth of extraneous detail adds to the verisimilitude of the scene, attests to Maclise’s commitment to truth telling, his skill at staging a persuasive view, what Roland Barthes famously termed “the reality effect”. 24 Maclise’s aphorism speaks of the “picture of form”, but what of the form of this picture? 25 The mesenteric artery is framed by intestines, which are framed by the anatomist’s cut, which is framed by the skin of the cadaver, which is framed by the non finito outline drawing of the sheet with the left hand completing the circle, and then framed by the negative space around the whole. Frames around frames. Almost a rosette. The plate demonstrates a bravura dissection no doubt, flaunts the anatomist’s erudition and skill with the scalpel. 26 27 28 Said to be “strictly conservative”, and an advocate for education in the liberal arts for the public and medical profession (as he contended in his Hunterian Oration of 1869), Quain argued that a surgeon should be a man of culture and discrimination, a gentleman and not just a crude mechanic of the body. 29 His “friend and former pupil” Joseph Maclise came from humbler stock. 30 31 And established himself as a working surgeon and medical illustrator of uncommon ability. Early on, Quain was his mentor and protector, a bond likely forged from their County Cork connection. After the accolades accorded The Anatomy of the Arteries of the Human Body , Maclise was raised up from Member to Fellow at the Royal College of Surgeons. From there, he went on to produce his own atlases, most notably Surgical Anatomy (1st ed.: London: John Churchill, 1851; 2nd ed.: London: John Churchill, 1856; also in large format, though smaller than Quain’s atlas) ( Fig. 6 ). While Surgical Anatomy was a critical and commercial success, and perhaps earned him some income (we have no records of book sales, finances, or arrangements with publishers), it failed to win him a hospital or university appointment. His career ambitions, and likely his social aspirations, were thwarted. Figure 6.Joseph Maclise, Surgical Anatomy, (London: John Churchill, 1851), Plate XVIII. Wellcome Collection. Internet Archive (CC BY 4.0). DOI Queer Scenes: Maclise and the Bells Compared Plate 60 of The Anatomy of the Arteries of the Human Body is a tour de force , a cascade of visual rhymes in deep-focus, high-definition, illusionistic perspective ( Fig. 7 32 Not with eyes (it has none). With hands? There is no haptic equivalent for staring back, yet Maclise’s cadaver seems to feel back … touch … grope … Figure 7.Joseph Maclise, The Arteries of the Pelvis of the Male, from Richard Quain, The Anatomy of the Arteries of the Human Body(London: Taylor & Walton, 1841–1844), Plate 60, c. 1841–1844, coloured lithograph, 67.5 x 55 cm. Wellcome Collection. Digital image courtesy of Wellcome Collection (CC BY 4.0). Maclise keeps his hands to himself. Unlike Bourgery and Jacob’s Traité complet 33 But the hands in Maclise’s Plate 60 (and Plate 51) do nothing of the sort. They are irrelevant, unpedagogical, unindexical. If anything, they distract from surgical and anatomical business (and go unmentioned in captions and commentaries). Distraction, of course, is a key technique of stage magic and mesmerism. Hypnotists rely on our habit of following the indexical hand to induce a trance that allows them to take over the consciousness of a hypnotized subject. Are Maclise’s cadaveric hands doing something similar? Is he entrancing us? Or indulging in a bit of auto-suggestion? If the hands in Plate 60 aren’t a proxy for the surgical hand, then for what? The desirous self? The scene of Plate 60 is oddly private, personal. Recruited to empathically feel the feeling hands of an anatomical subject, the reader is ushered into a space where Maclise’s flashing desire can, momentarily, become palpable. and artistic powers, which are not entirely separate matters. The beauty, precision, and detail of the artist’s rendering are distinguishing marks of anatomical excellence, as they had been for the grand productions of Maclise’s eminent anatomical forerunners: Govard Bidloo, Albrecht von Haller, Antonio Scarpa, William Hunter, Friedrich Tiedemann (all cited in Quain’s preface to the folio), and the Traité complet of Bourgery and Jacob (uncited, but arguably a lurking contextual presence). Remember also the super-size format: The Anatomy of the Arteries of the Human Body is an “imperial” folio, life-size, 1:1 scale, printed on expensive paper—of a measure and quality that bespeaks the aspiration to create a masterpiece. 34 All of which redounded to the credit of author Richard Quain (who occupied the professorial chair once held by the venerated Charles Bell), credit shared to some degree by his protégé Maclise. The Anatomy of the Arteries of the Human Body Fig. 8 ). Art! Which, given the centrality of the male nude in figure drawing, was full of homoerotic potentials. Maclise aimed to satisfy the highest scientific standards of medical publication and the highest aesthetic, connoisseurial standards so prized by formally trained fine artists and collectors. If Maclise displayed his ability to perform beautiful dissections, and skillfully render beautiful men, beautiful bodies, beautiful body parts, he also displayed his gift for making beautiful pictures . DOI Figure 8.Annibale Carracci, Nude Study of a Young Man Lying on his Back with his Head Thrown Back, Eyes Closed, circa 1584, red chalk on cream paper, 35.5 × 39.7 cm. Collection of National Museum Sweden (NMH 769/1863).Digital image courtesy of Cecilia Heisser / National Museum Sweden (all rights reserved). DOITo get a fuller sense of Maclise’s aesthetic maximalism, consider Plate 62, depicting the operation for lithotomy (Fig. 9). It’s all a bit much. Ostentatious virtuosity. Provocative, fulsome representation of skin surfaces. An approach, which Maclise justifies, in later writings, with a surgical rationale: “The surface of the living body is […] a map explanatory of the relative position of the organs beneath …” The depiction of skin surfaces—even those that aren’t adjacent to his dissections—is “an aid”, which furnishes “the memory with as clear an account of the structures” as if they were “perfectly translucent”. 35 DOI Figure 9.Joseph Maclise, A Deeper Dissection of the Perineum, the Pudic Artery and the Artery of the Bulb are Shown, from Richard Quain, The Anatomy of the Arteries of the Human Body(London: Taylor & Walton, 1841–1844), Plate 62, circa 1841–1844, coloured lithograph, 67.5 × 55 cm. Wellcome Collection.Digital image courtesy of Wellcome Collection (CC BY 4.0). DOI 36 A corpse obviously cannot resist, but the body in death becomes noncompliant (“dead weight”, rigor mortis But the logic of the pose becomes apparent when you consider the illustration’s iconographical origins and purpose—to show the anatomical structures and positioning that a surgeon would need to know to “cut for stone”. Maclise’s plate is perhaps inspired by, or in dialogue with, the “posture for lithotomy” illustration in John Bell’s Principles of Surgery (1801–1808), an awkward engraving of a living patient subjected to the cutting for stone operation ( Fig. 10 ). 37 DOI Figure 10.Charles Bell, Posture for Lithotomy, from John Bell, Principles of Surgery, 3 Vols. (Edinburgh: Cadel & Davies & Longman, 1801-1808), Vol. 2, Fig. 1, 1801, etching. Wellcome Collection.Digital image courtesy of Wellcome Collection (CC BY 4.0). That fits with John Bell’s persona as a gruff enemy of medical gentility and prettification. In his 1794 Engravings of the Bones, Muscles and Joints Engravings of the Bones, Muscles and Joints , as just that.) If the “posture for lithotomy” in Principles of Surgery Fig. 11 ). 38 In the preface to his Engravings of the Bones, Muscles, and Joints , John Bell deplores the fact that most anatomists have to depend on professional artists whose “striving for elegance of form” introduces inaccuracies, falsifies the image. 39 DOI Figure 11.John Bell, Engravings of the Bones, Muscles and Joints, (London: Printed for Longman and Rees, 1804), Plate 11, 1804, etching. Wellcome Collection.Digital image courtesy of Wellcome Collection (CC BY 4.0). Yet, Maclise’s lithotomy images, contravening Bell, combine great “elegance of form” with meticulous rendering, verisimilitude. The plates of The Anatomy of the Arteries of the Human Body counterpoint anatomical rupture, violation, and destruction with sensuality and beauty, and make it look real , which is to say true. But they also perform a bit of magic: make the cadavers seem subtly alive. The contrast between Bell’s 1794 engraving and the work of Maclise is even more pronounced in Maclise’s Plate 61, the first of the pair dealing with the anatomy of the perineum and rectum ( Fig. 12 40 The feeling, again, is both intimate and grand—the artist’s immersive private view (Maclise’s view) is also a printed monument. Maclise aims for a heedless transcendence, entirely sequestered from the growing debate over whether beauty was antagonistic to the goal of accuracy in science (an issue he never broached in his commentaries). 41 DOI Figure 12.Joseph Maclise, The Anatomy of the Perineum and Rectum, from Richard Quain, The Anatomy of the Arteries of the Human Body(London: Taylor & Walton, 1841–1844), Plate 61, circa 1841–1844, coloured lithograph, 67.5 × 55 cm. Collection of Hagströmer Library.Digital image courtesy of Hagströmer Library (public domain). A “technique of echoing bodily forms in drapery and upholstery”, art historian Mechthild Fend notes, was “often used in 18th-century erotic art, in paintings by Fragonard” and in “Boucher’s 1745 Odalisque where the forms of the reclining nude […] are multiplied in the plush fabrics”. 42 In both his dissection and his arrangement of reclining nudes (in Plates 61 and 62), Maclise plays with the difference between surface and underlying layers. Hardness, in the hardness of bone and muscle and the wooden table. Softness, in the smooth textures of skin and cloth. The approach is sculptural and tactile. 43 Maclise exquisitely renders skin contours and surfaces, especially around the area of the anatomical opening, which softens (feminizes?) the undressed masculine subject. His pencil is a caress, almost an invitation to touch (what cannot be touched). (And what of his scalpel?) It’s as though “the alluring surface … ‘solicits touch … as it prohibits it’”. 44 If, as Fend suggests, anatomy is an unveiling and the skin a veil, “the symbolic removal of which results in the revelation of an ‘inner truth’ or essence”, Maclise reverses that. His anatomy is an opportunity to dress the body in sumptuous, silky, hairless skin and display its utter nakedness. 45 Irrelevant Penises (a Gallery) The Anatomy of the Arteries of the Human Body , often in scenes where it is irrelevant to the anatomical region demonstrated. Maclise and Quain focus almost entirely on male bodies and body parts ( Figs. 13 – 20 Figure 13. Joseph Maclise, Plate 49 from Richard Quain, The Anatomy of the Arteries of the Human Body, (London: Taylor & Walton, 1841–1844), circa 1841–1844, coloured lithograph, 67.5 × 55 cm. Wellcome Collection. Digital image courtesy of Wellcome Collection (CC BY 4.0). Figure 14. Joseph Maclise, Plate 52 from Richard Quain, The Anatomy of the Arteries of the Human Body, (London: Taylor & Walton, 1841–1844), circa 1841–1844, coloured lithograph, 67.5 × 55 cm. Wellcome Collection. Digital image courtesy of Wellcome Collection (CC BY 4.0). Figure 15. Figure 16. Joseph Maclise, Plate 54 from Richard Quain, The Anatomy of the Arteries of the Human Body, (London: Taylor & Walton, 1841–1844), circa 1841–1844, coloured lithograph, 67.5 × 55 cm. Wellcome Collection. Figure 17. Joseph Maclise, Plate 53 from Richard Quain, The Anatomy of the Arteries of the Human Body, (London: Taylor & Walton, 1841–1844), circa 1841–1844, coloured lithograph, 67.5 × 55 cm. Wellcome Collection. Figure 18. Joseph Maclise, Plate 52 (detail) from Richard Quain, The Anatomy of the Arteries of the Human Body, (London: Taylor & Walton, 1841–1844), circa 1841–1844, coloured lithograph, 67.5 × 55 cm. Wellcome Collection. Figure 19. Joseph Maclise, Plate 55 (detail) from Richard Quain, The Anatomy of the Arteries of the Human Body, London: Taylor & Walton, 1841–1844), circa 1841–1844, coloured lithograph, 67.5 × 55 cm. Wellcome Collection. Figure 20. Joseph Maclise, Plate 53 (detail) from Richard Quain, The Anatomy of the Arteries of the Human Body, (London: Taylor & Walton, 1841–1844), circa 1841–1844, coloured lithograph, 67.5 × 55 cm. Wellcome Collection. That special interest didn’t wane in Maclise’s subsequent work. The 1856 edition of Surgical Anatomy Surgical Anatomy and only briefly discusses operations on women. Other contemporary British anatomies—the large format chromolithographic atlases of George Viner Ellis  (a former student of Quain) ( Fig. 21 ) and Francis Sibson ( Fig. 22 ), and the smaller chromolithographic manuals of Thomas Morton (another former student of Quain) ( Fig. 23 Figure 21. George Henry Ford, Dissection of a Portion of the Face and Neck, with the Muscles, Blood Vessels and Trachea Indicated: Shown Resting on a Block, from George Viner Ellis, Illustrations of Dissections in a Series of Original Coloured Plates, the Size of Life, Representing the Dissection of the Human Body(London, 1865), Plate 25, 1865, coloured lithograph. Wellcome Collection. Figure 22. William Fairland, The body of a man lying down with the trunk dissected: two figures showing the lungs after breathing out (above) and after breathing in (below, simulated by inflating the lungs), from Francis Sibson, Medical Anatomy(London, John Churchill, 1869): Plate 17, 1866, coloured lithograph. Wellcome Collection (no. 642390i). Figure 23. Andrew Morton, Plate 1 from Thomas Morton, The Surgical Anatomy of Inguinal Hernia, the Testis and its Coverings, (London: Taylor, Walton, & Moberly, 1844), 1844, coloured lithograph. Collection of Hagströmer Library. DOI “Transcendental Anatomy” and the Semblance of a Crucifixion We’ve looked at headless cadavers, with sensate hands and exposed genitals. Now consider Plate 47 of The Anatomy of the Arteries of the Human Body , another figure study and another queer scene: the nearly bodiless head of a handsome young man with flawless skin, flowing hair—but no hands, no arms, no genitalia ( Fig. 24 ). Seemingly asleep, his head rests on a horizontal block of wood, artfully lit (slightly from below, the left side). But that beautiful face is irrelevant (unnecessary, like the penises) to the demonstration of “the thoracic or descending aorta with the intercostal arteries”. 46 As in so many of his eye-catching illustrations, Maclise’s hunger to depict masculine beauty (or just Beauty) converges with his eagerness to flaunt his artistic gifts and aesthetic sensibility. DOI Figure 24.Joseph Maclise, The Thoracic Aorta and the Intercostal Arteries, with the Intercostal Veins, Vena Azygos, the Thoracic Duct, Seen in Connexion with the Parietes of the Thorax, from Richard Quain, The Anatomy of the Arteries of the Human Body(London: Taylor & Walton, 1841–1844), Plate 47, circa 1841–1844, coloured lithograph, 67.5 × 55 cm. Wellcome Collection.Digital image courtesy of Wellcome Collection (CC BY 4.0). There is, of course, also anatomy to attend to: an inverted V-cut incision at the neck leads down to the heavily dissected aortic cavity. Maclise leaves off at the bottom with a non finito sketch of the kidneys. What’s left of the body, after the radical paring away of parts, is framed by a non finito wooden armchair, the staves of the chair echoing the ribs of the thoracic cage and their architectural/structural function. Maclise is doubling and redoubling, embellishing his image with a kind of platonic ghost, in step with the dualism that was then coursing through British and Continental fiction, philosophy, and fine art, with its elevation of capital “B” Beauty and refined abstraction and getting-it-right detail. (Think of the dualism of real and ideal beauty in the art criticism of William Hazlitt and John Ruskin.) That open-ended, ramifying aesthetic, in turn, neatly connects Maclise’s love of beauty in realistic depiction to French naturalist Geoffroy Saint-Hilaire’s preachings on the organic “unity of plan”. 47 Quain did all the talking in the text of The Anatomy of the Arteries of the Human 48 In the prefaces and commentaries to Comparative Osteology (1847), Surgical Anatomy (1851 and 1856), and On Dislocations and Fractures 49 Quain’s commentary for Plate 47 states simply that “[t]he body was in the sitting posture”, probably in a wooden armchair, “while the drawing was being made”. 50 But Maclise takes the liberty here (and elsewhere) to play with aesthetic elements and visually encode “transcendental anatomical” principles that Quain might not have fully endorsed, maybe wasn’t even fully cognizant of. One reviewer commented that “Mr. Quain has allowed himself to be completely overshadowed by the artist”. 51 You get the feeling that Maclise hijacked the project. While from Maclise’s later account, the two men agreed on “the Law of a ‘unity of organization’” (Quain appears on a list of the scientific men Maclise says he learned it from), Quain’s text abstains from discussion of transcendental anatomy. 52 If that disappointed Maclise, maybe he compensated by speaking loudly through the artwork. 53 Maclise dedicated Surgical Anatomy to Liston. (Quain’s name only appears in a few passing references.) 54 Quain can’t have liked that. Did the two have a falling out? Was Quain jealous, Maclise resentful? From the peevish tone of his writings on matters anatomical, it’s evident that Maclise carried a sizeable chip on his shoulder. It’s not hard to imagine the two men clashing on philosophy, money, deference, or something else. DOI Surgical Anatomy (1851 and 1856), Maclise asserts that there are two kinds of facts, “ideal and real”, which he takes to be elements of a visual dialogue. 55 The placement of tools then could be signaling his double commitment: to the truthful depiction of “real facts” (the pin); and to “ideal facts” (the compass); the anatomized body as actually dissected; larger anatomical principles as thereby revealed. 56 This packing of signals, gestures, allusions, metaphors, details into what-the-eye-sees anatomical representation wasn’t unprecedented—historians of anatomy will be reminded of Gerard de Lairesse’s drawings for Govard Bidloo’s Anatomia humani corporis (1685). However, over 150 years separate Lairesse from Maclise. The plates of The Anatomy of the Arteries of the Human Body and the two London editions of Surgical Anatomy are utterly unlike anything to be found in the atlases of Maclise’s Victorian medical contemporaries and immediate predecessors. To fully get the nature of that difference, compare The Anatomy of the Arteries of the Human Body with the canonical work Quain aims to supersede, Friedrich Tiedemann’s Tabulae arteriarum corporis humani (1822) ( Figs. 25 – 27 ). 57 Tiedemann doesn’t bring readers into the anatomy room. There are no “bedroom scenes”, echolalic crucifixions, or non finito Touching Hands But gaze doesn’t capture the intensity of Maclise’s immersive representational tactics. For that, we need to return to touch, and the representation of hands on skin and skin-to-skin contact. In Plate 26, the sensual hand that gently grazes the thigh ( Fig. 28 ) … Is that caress merely another cadaveric proxy for the surgeon-artist’s desirous touch, special feelings for his beautifully proportioned anatomical subject? Or does it also double for the artist’s touche with crayon, pencil, and brush? 58 In the 1851 first edition, Maclise shows a bit more ( Fig. 29 ). Below the genital region, the subject’s left leg appears to be touching the right leg, just in back of the knee, as if scratching an itch or performing a Scottish country dance or balletic cou-de-pied . See also Plate 27, which initiates a small series of side-by-side dissections ( Fig. 30 ). The penis on the right appears to brush the thigh of the figure on the left. It’s rare to see two cadavers touch in anatomical illustration. This kind of touching is rarer still. DOI Figure 28.Joseph Maclise, The Surgical Dissection of the Superficial and Deep Structures of the Inguino-Femoral Region, from Joseph Maclise, Surgical Anatomy(London: John Churchill, 1856), Plate 26, 1856, coloured lithograph. Wellcome Collection.Digital image courtesy of Wellcome Collection (CC BY 4.0). DOI Figure 29.Joseph Maclise, The Surgical Dissection of the Superficial and Deep Structures of the Inguino-Femoral Region, from Joseph Maclise, Surgical Anatomy(London: John Churchill, 1851), Plate 16, 1851, coloured lithograph. Wellcome Collection.Digital image courtesy of Wellcome Collection (CC BY 4.0). DOI Figure 30.Joseph Maclise, The Surgical Dissection of the Superficial and Deep Structures of the Inguino-Femoral Region, Hernial Protrusions are Very Liable to Occur at the Groin, from Joseph Maclise, Surgical Anatomy(London: John Churchill, 1856), Plate 27, 1856, coloured lithograph. Wellcome Collection.Digital image courtesy of Wellcome Collection (CC BY 4.0). “Images of skin”, Mechthild Fend comments, “recall a double sensation of touching and being touched.” 59 Fig. 31 ). A caress within a caress. Exquisite sensation. Reference to the visible brush stroke, as the record of a caress of the surface of the paper or canvas—and, by analogy, a caress of the body of the subject—was a part of the rhetoric of eighteenth- and nineteenth-century art discourse. 60 61 Maclise orchestrates a haptic imaginary in words, image, and line. (And if hand is “autograph of mind”, what is on Maclise’s mind when he autographs with his hand?) Critics mostly loved Surgical Anatomy . A reviewer in the Edinburgh Medical & Surgical Journal , commenting on the first three installments, singled out Maclise’s “anatomical knowledge”, “talents as a draughtsman”, “great fidelity and accuracy”, his “very correct and beautiful view” and arrangement of dissected subjects in “the best and most effective positions … before the eye of the beholder” so as to stage them in an “impressive light”. 62 But images like Plate 27 push the limits. One reviewer deplored “the crowding of two anatomical figures on the same page”. Was that a displaced expression of discomfort with the erotic implications of such “crowding”? With less space on the page, the smaller format 1851 Philadelphia edition places the figures on separate pages. 63 In a larger format, it’s easier to put more than one figure on the same page, and perhaps more pedagogically effective: the proximity of the two figures makes the comparison easier for readers to see. But it also provides an opportunity for Maclise to stage a teasing contact between anatomical bodies. The convergence of artist’s touche and anatomist’s touch was an oft-recited topos of art criticism and commentary. According to Fend, the eighteenth-century French portraitist Louis Tocqué, in an essay on painting, described how “the brush penetrates the body like an anatomist’s knife”: 64 Figs. 32 and 33 ). Intoxicated by the metaphorology and the haptics of his double pleasure as artist and anatomist, in some plates of The Anatomy of the Arteries of the Human Body Maclise includes the tools of the artist’s trade within the image: inkwell, paper, and pen ( Fig. 34 65 Figure 34.Joseph Maclise, The Anterior Tibial Artery, the Circulatory System: Dissection of the Front of the Lower Leg and the Ankle, with Arteries Indicated in Red and Blue, from Richard Quain, The Anatomy of the Arteries(London: Taylor & Walton, 1844), Plate 82, 1841–1844, coloured lithograph, 64.2 x 49.1 cm. Wellcome Collection (no. 579465i). Digital image courtesy of Wellcome Collection (CC BY 4.0). Maclise’s Men, an Imaginary Confraternity? Maclise’s focus on the male genitalia and rectum, and the erotics of skin and touch are not the whole story. While he largely refrained from classicizing gestures and conventions, Maclise paid heed to contemporary ideals of masculine beauty in the selection of his cadavers. And seems to have had, at some junctures, a large pool of specimens to choose from. According to Quain, for the plates of The Anatomy of the Arteries of the Human Body , “the bodies … received during a series of years for the study of anatomy into the School of Medicine in University College” amounted to 1,040 or 930. (Quain’s two prefaces differ on the exact number; either way, it’s a lot.) 66 For the plates of Surgical Anatomy , Maclise doesn’t total up his cadavers, but states, “in guarantee of their anatomical accuracy”: natural specimens [Maclise’s emphasis], recent and preserved, which I had the opportunity of examining at the Hospitals and Museums in Paris, London, and elsewhere. 67 68 Even there, his choices of cadavers to illustrate were undoubtedly limited: some plates feature older men; others are likely composites (incorporating those “natural specimens”). But, whether true-to-nature or composites, one doesn’t have to search hard to assemble a gallery of handsome young men from Maclise’s drawings. In fact, Maclise himself assembled such a gallery in Surgical Anatomy (Plates 11–15, 1851 edition; Plates 21–25, 1856 edition) ( Figs. 35 – 39 DOI Figure 35. Joseph Maclise, The Relations of the Principal Blood Vessels to the Viscera of the Thoracico-Abdominal Cavity, from Joseph Maclise, Surgical Anatomy(London: John Churchill, 1851), Plate 13, 1851, coloured lithograph. Wellcome Collection. Figure 36. Joseph Maclise, The Relation of the Internal Parts to the External Surface of the Body, from Joseph Maclise, Surgical Anatomy(London: John Churchill, 1851), Plate 15, 1851, coloured lithograph. Wellcome Collection. Figure 37. Joseph Maclise, Illustrating the Action of the Thoracico-Abdominal Apparatus as Effecting the Motions of the Contained Viscera, from Joseph Maclise, Surgical Anatomy(London: John Churchill, 1856), Plate 25, 1856, lithograph. Wellcome Collection. Figure 38. Joseph Maclise, The Form of the Abdomen and the Relative Position of its Contained Organs in Health and in Sickness, from Joseph Maclise, Surgical Anatomy(London: John Churchill, 1851), Plate 11, 1851, coloured lithograph. Wellcome Collection. Figure 39. Joseph Maclise, The Relation of the Internal Parts to the External Surface of the Body, from Joseph Maclise, Surgical Anatomy(London: John Churchill, 1851), Plate 14, 1851, coloured lithograph. Wellcome Collection. DOI Maclise’s nudes were, of course, anatomized corpses. Was he thereby participating in the mid-century Romantic valorization and eroticization of the dead body? The kind of thing that Paul Hippolyte Delaroche was performing in his over-the-top painting, The Wife of the Artist, Louise Vernet, on her Deathbed (1845/46) and that Edgar Allan Poe adverted to in his famous remark: “The death of a beautiful woman is, unquestionably, the most poetical topic in the world” (also from 1846). 69 70 Maclise lived in a social environment that disapproved of, punished, homoerotic expression and relations. If his erotic attachments were principally directed to men, as we have some reason to suspect, perhaps he took his gallery of men as an imaginary confraternity. Not exactly pin-ups, but a playful projection of desired objects of affection, imaginary friends. A representational consolation for relations and attachments never realized, or (another possibility) figurations of the affective life he did enjoy. Maybe Maclise imagined them as he sketched: transported from the dissecting tables back to life, congregating in student rooms, the medical school lecture hall, or pub … Coda: Maclise’s Fate and the Queer Anatomical Figure Study 71 72 Yet Maclise, uniquely licensed and committed to illustrating the male genitalia—with a sense of himself as an artist in the grand tradition, but located in the domain of medicine, not gallery art—was able to make figure studies of what was scanted in the genre of illusionistic perspectival figure study. And what was scanted in British anatomical and surgical illustration: naturalistic, sensual, proportionate renderings of nude and semi-nude men with genitalia. Images which could be privately archived between the covers of the anatomical book, on the shelves of a medical library, or displayed on the walls of a dissecting room or classroom or lecture hall, openly exhibited to groups of privileged viewers. Shared among men in the homosocial clubs of professional medicine. While hidden from view. The anatomical closet. Maclise was in, or adjacent to, those clubs. If we can’t say anything definite about his social networks or relations, here’s what we do know. In the 1840s and most of the 1850s, he receives laudatory mentions in the Lancet (then still an oppositional journal, a champion of political and medical radicalism), and attracts notice as a firebrand in the transformism debates. 73 As he settles into a career as a practicing surgeon, medical author, and illustrator, his atlases are acclaimed and he is made a Fellow of the Royal College of Surgeons. But, in spring 1852, he is passed over for an appointment at a London hospital. A less distinguished man is named. ( The Lancet protests on Maclise’s behalf.) 74 Then, in 1858, at the height of his authorial success, Maclise enters into a pointless two-year-long controversy, which he brings upon himself by penning an off-kilter diatribe against William Harvey’s account of the action of the heart, oddly inserted into On Dislocations and Fractures , a book on an entirely different topic. It was a peculiar provocation—Harvey was a beloved, celebrated icon of British medical science. The convoluted literary tone of Maclise’s diatribe, and convoluted defense of his position in letters to the Lancet , work to discredit him. 75 (Was he losing his mind?) In 1861, Maclise turns up as candidate for the position of conservator at the Hunterian Museum. The five applicants are ranked by the committee; Maclise comes in last among them. 76 As the 1860s unfold, he produces no new book or article and drops out of the pages of the Lancet DOI During this period, at some intervals, Joseph lives with brother Daniel in Bloomsbury and Chelsea—the two men were said to be close. (An unmarried sister, Isabella, “keeps house” for them until her untimely death in 1865.) 77 Perhaps Maclise decides to concentrate on his surgical practice. Or retires to live off his savings and royalties (if they amounted to anything) or his brother’s fortune (we don’t know anything about the size or disposition of Daniel’s estate). Or takes ill. Or becomes morbidly despondent over the loss of his beloved siblings: Daniel dies in 1870. DOI Or stakes a place as a defiant outsider. Maclise was on the far edge and losing side of the debates on evolution, an evolutionist but not a Darwinian. He also courted disfavor by condemning empirical research—in an age of progress, when research programs were abundantly productive, new findings were piling up and receiving acclaim in scientific journals and the press. In the preface to Surgical Anatomy , Maclise fulminates: corpus mortuum … ? How long are they to lead us on this dull road, weed-gathering, tracking out bloodvessels, tricking out nerves, and slicing the brain into still more delicate atoms … in order to coin new names and swell the dictionary? 78 No subtleties there. It’s not the only passage in which Maclise bludgeons the opposition, lays out grievances, and expounds on disagreements amounting almost to hatred. If he lays it on thick in his drawings, he lays it on thicker in his prose. To reviewers who called him on his textual excesses, he replies in the “Notice to The Second Edition” of Surgical Anatomy : “I have at times left the beaten march but to seek recreation”, then obscurely breaks off into four not very revelatory sentences on science and nature, in Latin, from Bacon’s Novum Organum . And finishes with this: “I have throughout … recorded ideal and real facts … for the seemingly exhausted subject of Anthropotomy [human anatomy] which have not hitherto been either written, spoken, or known …” 79 DOI Even with the brilliance of his illustrations, rhetoric like that could derail a career. (One imagines readers skipping the introduction and going directly to the plates.) But let’s also entertain the possibility that Maclise might be alluding (consciously or unconsciously) to other things when musing over recreation-seeking off “the beaten march” and “facts … not hitherto … written, spoken, or known …”. DOI Was there anything? We know that Maclise was in Paris in the 1830s and later traveled with Daniel “over the continent” (Paris, Lyon, Florence, Munich, Naples, Milan, Rome). It is tempting to speculate about what went on in those places: the “continent” was notoriously an occluded zone where British men, especially artists, went to let their hair down, explore aspects of hetero- and homosexuality that were more rigorously policed in their native habitat. 80 If that was the case with the brothers Maclise, we might have gotten clues from Daniel’s letters to Charles Dickens. But Dickens, who delivered the eulogy at Daniel’s funeral, destroyed that “immense correspondence, … because I could not answer for its privacy being respected when I should be dead”. 81 No letters from Joseph to Daniel or anyone else have thus far turned up, and no letters to Joseph either. Likely also destroyed, as so often happened in nineteenth-century Britain. So a blind spot. DOI Then darkness. Quain dies in 1887 and receives chatty (though not entirely laudatory) obituaries in the Lancet and the British Medical Journal. 82 In contrast, Maclise, once the Lancet ’s pet, receives no obituary, not in the Lancet or any other journal. So what happened? We haven’t a clue. His name drops out of the Medical Directory in 1879 (the year most catalogs and references give as his death date). Maybe he retired in 1879, but there was no announcement. And no announcement or remembrance upon his death from “bronchitis” and “syncope” in 1891. Which also seems odd, considering his distinguished record of publication and distinguished brother. 83 The death certificate describes him as a “retired army surgeon”. (That was likely mistaken, but if he did at some point enter the service, that would add another twist to Maclise’s life story.) DOI * * * DOI By the century’s end—a generation after Maclise’s zenith—the old medical order was coming undone. Works like The Anatomy of the Arteries of the Human Body and Surgical Anatomy , while still valued by collectors, no longer seemed modern, and the arty bibliophilic super-sized folio no longer stood securely atop the hierarchy of medical publications. Improvements in surgical techniques and photography bypassed Maclise’s illustrated plates. As enthusiasm shifted to embryology, microbiology, and neuroanatomy, Gray’s Anatomy and similar works rationalized and democratized anatomical publication for a new generation ( Fig. 40 ). DOI Figure 40.Henry Gray, detail from page 632,, Henry Gray, Anatomy Descriptive and Surgical(London: Longmans, 1897), 1897, coloured lithograph. Collection of Francis A. Countway Library of Medicine, Harvard University.Digital image courtesy of Open Knowledge Commons and Harvard Medical School (public domain). DOI Even if Surgical Anatomy retained some usefulness as a practical guide, its visual holism, high aesthetics, cranky politics, and glowing sensuality began to look deficient, old fashioned—maybe a little queer . The tour de force DOI 84 And if beauty was an enemy, sexy beauty was even more so. DOI In this new world, fine art’s truth claims were increasingly siphoned off from some order of independently verifiable objectivity (realist aesthetics and reportage) to a hodgepodge of hard-to-define oppositional essences (symbolism, impressionism, primitivism, cubism, etc.). 85 As anatomist and 86 Did he ever pause to consider how his own drawings might complicate that precept? About the author Footnotes This article is taken and adapted from a book-length work-in-progress by the author: Queer Anatomies: Aesthetics and Perverse Desire in 18th- and 19th-century Medical Illustration; or The Epistemology of the Anatomical Closet. 1 Michel Foucault, The History of Sexuality, Vol. 1, An Introduction, trans. Robert Hurley (New York: Vintage, 1990), Parts I and II, especially 44–49. 2 “The closet is not a grave” is an off-hand allusion to Leo Bersani, “Is the Rectum a Grave?” October43 (Winter 1987): 197–222. “The telling secret” is from Eve Kosofsky Sedgwick, Epistemology of the Closet(Berkeley, CA: University of California Press, 2008 [1990]), 67. 3 Sedgwick, Epistemology of the Closet, 3. 4 Joseph Maclise, Preface to Surgical Anatomy, 2nd ed. (London: John Churchill, 1856). 5 Paolo Mascagni, Anatomia universa or Grande anatomia del corpo umano (Pisa: Presso Niccolò Capurro, 1823–1830), Plate 2. Art: Antonio Serrantoni; 98.5 x 71.5 cm. 6 Michael Sappol, Dream Anatomy(Washington, DC: GPO, 2003). For an influential late eighteenth-century art anatomy full of fantastical macabre scenes, see Jacques Gamelin, Nouveau recueil d'ostéologie et de myologie, dessin‚ d’après nature … pour l’utilit‚ des sciences et des arts(Toulouse: J.F. Desclassan, 1779). But over the course of the nineteenth century, art anatomy pedagogy was also winnowed down to basics: figures posed in the nude, écorché, or as skeleton, without grand gesture, affect, or scenography. 7 William Hunter, The Anatomy of the Human Gravid Uterus(Birmingham: John Baskerville, 1774); and John Bell, Engravings of the Bones, Muscles, and Joints, Illustrating the First Volume of the Anatomy of the Human Body, 2nd ed. (London: Printed for Longman and Rees, 1804 [1794]). Yet, despite their endorsement of anatomical sobriety, Hunter and Bell choreographed and orchestrated provocative image play. 8 Eve Kosofsky Sedgwick, Between Men: English Literature and Male Homosocial Desire(New York: Columbia University Press, 1985). For erotics as the boundary of aesthetic pleasure, see also Abigail Zitin, “Wantonness: Milton, Hogarth, and The Analysis of Beauty”, Differences27, no. 1 (2016): 25–46. 9 For the social aims of high medical publication, and the way in which knowledge production affirmed claims of authority and the search for patronage and prestige, see Carin Berkowitz, “The Illustrious Anatomist: Authorship, Patronage and Illustrative Style in Anatomy Folios, 1700–1840”, Bulletin of the History of Medicine(2015): 171–208. 10 Richard Quain, Anatomy of the Arteries of the Human Body, with its Applications to Pathology and Operative Surgery: In Lithographic Drawings with Practical Commentaries[“drawings from nature and on stone by Joseph Maclise”] (London: Printed for Taylor and Walton, 1844); “imperial folio”; 67 cm. Quote from “Review of New Books: The Anatomy of the Arteries of the Human Body, with its Applications to Pathology and Operative Surgery, in Lithographic Drawings, with Practical Commentaries. By Richard Quain, &c. &c. The Delineationsby Joseph Maclise, Esq. Surgeon. Parts I and II. London: Taylor and Walton. 1840”, Provincial Medical & Surgical Journal1–1, no. 12 (19 December 1840), 203, doi:10.1136/bmj.s1-1.12.203. 11 Jean-Baptiste-Marc Bourgery and Nicholas Henri Jacob, Traité complet de l’anatomie de l’homme: Comprenant la médecine opératoire, avec planches lithographiées d’après nature, 8 vols (Paris: C.A. Delaunay, 1831–1854; 2nd ed., 1866–1871), 44 cm; art: Nicolas Henri Jacob. Later volumes also list Claude Bernard as co-author. Jacob was the primary artist, but some plates are by his students or associates: Ludovic Hirschfeld, Gerbe, Lévéillé, Roussion, Leroux, Dumoutier, and others. 12 The phrase comes from Dorothy Johnson, Jacques-Louis David: The Art of Metamorphosis(Princeton, NJ: Princeton University Press, 1993). See also Melissa Lo, “The Wounded Ideal Goes Anatomical”, in Hidden Treasure, ed. Michael Sappol (New York: Blast Books, 2011), 44–49. 13 For the trope of disembodied medical hands on a passive subject, indexicality, elided tactility, and their accompanying erotic frisson, see Elizabeth Stephens, “Touching Bodies: Tact/ility in 19th-Century Medical Photographs and Models”, in Bodies, Sex and Desire from the Renaissance to the Present, ed. Kate Fisher and Sarah Toulalan (London: Palgrave Macmillan, 2011), 87–101. Stephens discusses the use of disembodied medical hands to exhibit living subjects in pathological photographs, illustrations, and live demonstrations. She doesn’t discuss the atlases of Maclise or Bourgery and Jacob, but much of her study is also relevant to surgical anatomies and handbooks. See also Kathryn Hoffmann, “Sleeping Beauties in the Fairground: The Spitzner, Pedley, and Chemisé Exhibits”, Early Popular Visual Culture4, no. 2 (July 2006): 139–159. 14 Bourgery and Jacob’s Traité completwas initially sold to subscribers in installments. From 1831 to 1840, seventy installments were issued, “each containing eight panels of illustrations and eight sheets accompanying the text”. A monochrome set of installments cost about 800 francs, a color set 1600 francs, and the “high price obviously was an obstacle for the spread of the work”; quoted from “Jean-Baptiste Marc Bourgery”, Wikipedia,https://en.wikipedia.org/wiki/Jean-Baptiste_Marc_Bourgery. See also Naomi Slipp, “International Anatomies: Teaching Visual Literacy in the Harvard Lecture Hall”, in Bodies Beyond Borders: Moving Anatomies, 1750–1950, ed. Kaat Wils, Raf de Bont, and Sokhieng Au (Leuven: Leuven University Press, 2017), 228, n. 40. 15 Stephens, “Touching Bodies”, 89, 96, and 98–99. 16 Stephens, “Touching Bodies”, 89. 17 Vanessa R. Schwartz, Spectacular Realities: Early Mass Culture in Fin-de-Siècle Paris(Berkeley, CA: University of California Press, 1999), 45–88; and John Edmondson, “Death and the Tourist: Dark Encounters in Mid-Nineteenth-Century London via the Paris Morgue”, in The Palgrave Handbook of Dark Tourism Studies, ed. Philip R. Stone, Rudi Hartmann, Tony Seaton, Richard Sharpley, and Leanne White (London: Palgrave, 2017), 77–101. 18 See, for example, Helen McCormack, William Hunter and his Eighteenth-Century Cultural Worlds: The Anatomist and the Fine Arts(London: Routledge, 2017), 146–149. 19 There were, of course, erotically pleasurable scenes, lots of nudity, in painting and sculpture, sometimes even uncovered male genitalia (but never female), all legitimated by ostensibly religious, historical, moral, purposes. 20 Michel Foucault, The Birth of the Clinic: An Archaeology of Medical Perception, trans. Alan M. Sheridan Smith (1973) (New York: Routledge, 2003 [1963]), 204. Smith, translating Foucault’s le regardas “the gaze” ( le regard medicale= the medical gaze), thereby bequeathed the stronger word to Anglophone scholarship. 21 James Elkins, The Object Stares Back: On the Nature of Seeing(New York: Harcourt Brace, 1996). Walter Benjamin’s notion of “aura”, in “The Work of Art in the Age of Mechanical Reproduction” (1935–1939), also describes investment in objects and the effects such objects have on the viewer, not just in viewing but in structuring perception and making hierarchies; see Walter Benjamin, “The Work of Art in the Age of Mechanical Reproduction”, trans. Harry Zohn, in Illuminations, ed. Hannah Arendt (New York: Schocken/Random House, 1969),https://www.marxists.org/reference/subject/philosophy/works/ge/benjamin.htm. See also John Berger, Ways of Seeing(London: Penguin, 1973). 22 Richard Quain, The Anatomy of the Arteries of the Human Body(London: Taylor & Walton, 1844); originally published in installments, starting in 1840; 50 x 66 cm. 23 Roland Barthes, “The Reality Effect” (1969), trans. Richard Howard, in The Rustle of Language, ed. François Wahl (Berkeley, CA: University of California Press, 1989), 141–148. Mary Hunter discusses competing versions of realism circulating in art and medicine, mainly in the 1870s and 1880s, in The Face of Medicine: Visualizing Medical Masculinities in Late 19th-Century Paris(Manchester: Manchester University Press, 2016). For reasons of length, I bracket that question here. 24 Under the influence of Geoffroy Saint-Hilaire, Maclise held that organisms and species instantiate archetypal forms. 25 It’s unclear as to who performed the dissections: Quain, Maclise, and/or someone else. Richard Quain’s cousin—also named Richard Quain (and a physician, not a surgeon)—is said to have assisted in the organization and writing of the text. 26 Caroline Evans and Lorraine Gamman, “The Gaze Revisited, or Reviewing Queer Viewing”, in A Queer Romance: Lesbians, Gay Men and Popular Culture, ed. Paul Burston and Colin Richardson (London: Routledge, 1995), 13–56. 27 W.F. Bynum, “Quain, Richard (1800–1887), Anatomist and Surgeon”, Oxford Dictionary of National Biography(23 September 2004), https://doi-org/10.1093/ref:odnb/22940; Royal College of Surgeons of England, “Quain, Richard (1800–1887)”, Plarr’s Lives of the Fellows,https://livesonline.rcseng.ac.uk/client/en_GB/lives/search/detailnonmodal/ent:$002f$002fSD_ASSET$002f0$002fSD_ASSET:372381/one; [Obituary for Richard Quain], George David Pollock, Address of George David Pollock, F.R.C.S., President of the Royal Medical and Chirurgical Society of London at the Annual Meeting, March 1st, 1888(London, 1888), 20–22,https://wellcomecollection.org/works/yv6vfvs7; [Obituary], “Richard Quain”, British Medical Journal2, no. 1395 (24 September 1887): 694–695; and J.F. Clarke, Autobiographical Recollections of the Medical Profession(London: J. & A. Churchill, 1874), 145–146. 28 Richard Quain, On Some Defects in General Education: Being the Hunterian Oration of the Royal College of Surgeons for 1869(London: Macmillan, 1870). 29 Quain, Preface to The Anatomy of the Arteries of the Human Body, vi. 30 Adrian Desmond, The Politics of Evolution: Morphology, Medicine, and Reform in Radical London(Chicago, IL: University of Chicago Press, 1989), 360–364, 368–371, 376, and 425. “Transformism” was not only a term for what became “evolution”; it included adaptation, embryological and postnatal development, and the field of comparative anatomy. 31 Referring to James Elkins, The Object Stares Back. 32 For the trope of disembodied medical hands, see again, Stephens, “Touching Bodies”; and Hoffmann, “Sleeping Beauties in the Fairground”. 33 For extra grandness, the firm of Taylor & Walton puts the legend “lithographic printer to her Majesty” in small letters at the bottom of each plate. For performative aspects of grand atlas-making, see Carin Berkowitz, “Systems of Display: The Making of Anatomical Knowledge in Enlightenment Britain”, British Journal for the History of Science46, no. 3 (2013): 359–387, which faults Lorraine Daston and Peter Galison, Objectivity(New York: Zone Books, 2007), for decontextualizing the anatomical atlas and misunderstanding its place in the ensemble of materials mobilized in medical knowledge production and pedagogy. 34 Maclise, Preface to Surgical Anatomy, 1st ed. (1851). 35 In the mid-nineteenth century, the “lithotomy position” became the obstetrical term for a frequently used position for pelvic examination of women in pregnancy and childbirth, and is still used in obstetrics and gynecology. 36 John Bell, Principles of Surgery, 3 vols. (Edinburgh: Cadel & Davies & Longman, 1801–1808), Vol. 2, Fig. 1, facing page, 200. Charles Bell assisted his older brother and is said to have made all of the illustrations for the last two volumes of Principles of Surgery. Bell’s figure is perhaps based on an earlier illustrated work, François Tolet, A Treatise of Lithotomy: or, of the Extraction of the Stone Out of the Bladder … Written in French … Translated into English by A. Lovell(London: Printed by H.H. for William Cademan, 1683), which, in several plates, shows the tying of hands to feet. But the Bells were men of great surgical experience: their knowledge of the pose more likely derived from teachers or colleagues, and then their own considerable experience. Likewise, Maclise’s knowledge of the pose, and desire to show it, may have derived from his study of the Bell images, but also practical training and direct experience. Maclise may also have known other contemporary images of the operation, which were depicted in German and French surgical atlases. 37 John Bell, Engravings of the Bones, Muscles, and Joints, 2nd ed. (1804). Bell’s dedication to etching and sketchiness is an implicit rebuke to the polish of artful but inaccurate anatomical illustrations. But in the preface to Engravings of the Bones, Muscles, and Joints, he avows his admiration for the highly polished naturalist accuracy of Hunter’s and Haller’s anatomical engravings (and to a lesser extent those of Bidloo, who he says is true but disorganized). As for “taste”, he makes no critique of it; rather he aspires to a “higher taste”, and admires the dedication of artists who study anatomy for art’s sake, especially Leonardo and Michelangelo, who he says know more about anatomy than some anatomists. The hanging diaphragm on the wall in Bell’s etching is a visual quotation from Vesalius’s “muscle-man” series. 38 John Bell, Engravings of the Bones, Muscles, and Joints(1804), vi. See Carin Berkowitz, “Systems of Display”. 39 Quoted words are taken from Maclise, Surgical Anatomy, 2nd ed. (1856), Plate 26. 40 Daston and Galison, Objectivity.According to Carolyn Hannaway and Ann La Berge, “Paris Medicine: Perspectives Past and Present”, in Constructing Paris Medicine[ Clio Medica50], ed. Carolyn Hannaway and Ann La Berge (Amsterdam: Editions Rodopi, 1998), 23, in the mid-1860s, Charles Daremberg contrasted realism in pathological anatomical illustration againstbeautifying art. For Daremberg, pathological anatomy was the hardest case for “brutal exactitude” in medical illustration, as the field made its passage into the age of photography. But similar issues arose in gross and surgical anatomy. See C. Charles Daremberg, La médecine: Histoire et doctrines(Paris: Didier et cie, 1865), 290–304; Jacalyn Duffin, “Imaging Disease: The Illustration and Non-Illustration of Medical Texts, 1650–1850”, in “Muse and Reason: The Relation of Arts and Sciences, 1650–1850”, ed. B. Castel, J.A. Leith, and A.W. Riley, Queens Quarterly(1994): 79–108; Michael Sappol, “Anatomy’s Photography: Objectivity, Showmanship & the Reinvention of the Anatomical Image, 1860–1950”, REMEDIA, January 2017 (rev. 2018),https://www.academia.edu/31049714/Anatomy_s_photography_Objectivity_showmanship_and_the_reinvention_of_the_anatomical_image_1860_1950_Introduction. 41 Mechthild Fend, Fleshing Out Surfaces: Skin in French Art and Medicine 1650–1850(Manchester: University of Manchester Press, 2017), 250–251. 42 But Maclise never turns his anatomical subjects into hardened, stiff statues; on the contrary, they seem to live on the page (though sometimes posed as if resting or asleep). The issues at stake in Maclise’s lithographs of anatomized male nudes are similar to those in mid-nineteenth-century sculpture. See Michael Hatt, “Near and Far: Homoeroticism, Labour, and Hamo Thornycroft’s Mower”, Art History26, no. 1 (2003): 45–50. 43 Michael Hatt, “Space, Surface, Self: Homosexuality and the Aesthetic Interior”, Visual Culture in Britain8, no. 1 (2007): 105–128, quoted on 113. 44 Fend, Fleshing Out Surfaces, 5. 45 Quain, The Anatomy of the Arteries of the Human Body[text for the folio plates] (1844), 338. 46 Joseph Maclise, Comparative Osteology: Being Morphological Studies to Demonstrate the Archetype Skeleton of Vertebrated Animals(London: Taylor & Walton, 1847). For Geoffroy, see Toby A. Appel, The Cuvier-Geoffroy Debate: French Biology in the Decades before Darwin(New York: Oxford University Press, 1987); Adrian Desmond, The Politics of Evolution; and Valerie Racine, “Etienne Geoffroy Saint-Hilaire (1772–1844)”, Embryo Project Encyclopedia(5 August 2013),http://embryo.asu.edu/handle/10776/6045. Most of the literature on idealism (and Platonism) in Victorian Britain focuses on the late nineteenth century. The literature on the mid-century is thinner, but see Richard Thomas Eldridge, “Idealism in 19th-Century British and American Literature”, Aesthetics and Literature3 (2013): 121–144; and Rachel Kravetz, “The Art of Cognition: British Empiricism and Victorian Aesthetics”, PhD dissertation, City University of New York, 2017. 47 Philip F. Rehbock, “Transcendental Anatomy”, in Romanticism and the Sciences, ed. Andrew Cunningham and Nicholas Jardine (Cambridge: Cambridge University Press, 1990), 144–145. 48 Maclise, Preface to Surgical Anatomy(1856). 49 Quain is listed as the sole author of The Anatomy of the Arteries of the Human Body.Bourgery gave Jacob equal billing on the title pages of their collaborative atlas. Quain didn’t extend the same courtesy to Maclise, but did give him ample credit. (Some sources say that Quain was assisted in the writing of the text by his cousin, also named Richard Quain.) 50 Anon., “Review of New Books: The Anatomy of the Arteries of the Human Body”, 203. 51 According to Maclise, Comparative Osteology, iv: “While believing … that the subject of Anatomical Unity forms the goal of all comparative research, I sought also the opinions of … Cuvier, Goethe, Geoffroy, Oken, Spix, Carus and … Dr. Sharpey, Mr. Quain, and Mr. Owen … and from these I learned that the Law of a ‘unity of organization’ … admitted now-a-day of as little dispute as a mathematical axiom.” From which we can gather that, during the years of their collaboration, Maclise took Quain for a philosophical ally. One telling indicator of their divergence: variations on the keyword “symmetry” appear three times in Quain’s The Anatomy of the Arteries of the Human Body; but 37 times in Maclise’s Surgical Anatomy(1856). 52 Quotes are from obituary, “Richard Quain”, British Medical Journal2, no. 1395 (24 September 1887): 694–695; see also obituary, “Richard Quain”, The Lancet(1 October 1887): 687–688. Maclise received mention in Quain’s British Medical Journalobituary: “Without exaggeration, it may be said that Richard Quain’s life would have been well spent had he lived only to produce the Anatomy of the Arteries.” The obituary goes on to credit “the artistic powers of Mr. Joseph Maclise, F.R.C.S., author of a Surgical Anatomy, and an almost forgotten work on the homologies of the skeleton” [ Comparative Osteology]. 53 Liston was deceased; the other dedicatee, Samuel Cooper, was also one of Maclise’s deceased teachers. 54 Maclise, “Notice to the Second Edition”, Surgical Anatomy(1856). 55 According to Plarr’s Lives of the Fellows of the Royal College of Surgeons, following conventional wisdom, “The whole interest [of Joseph’s oeuvre] … is the relation of Joseph’s drawings to those of Daniel, as to how far the two brothers may be said to have viewed the human figure from the same point of view.” See Royal College of Surgeons of England, “Maclise, Joseph (1815–1880)”, Plarr’s Lives of the Fellows(London, 1912–1940),http://livesonline.rcseng.ac.uk/biogs/E002613b.htm. 56 Quain, The Anatomy of the Arteries of the Human Body, vii, cites Friedrich Tiedemann, Tabulae arteriarum corporis humani(Karlsruhe: C.F. Müller, 1822); the atlas also circulated in inferior British editions, Friedrich Tiedemann, Plates of the Arteries of the Human Body, ed. Thomas Wharton Jones and Robert Knox (Edinburgh: Printed for Maclachlan & Stewart, 1829–1836). Quain also cites Albrecht von Haller, Icones anatomicae quibus praecipuae aliquae partes corporis humani delineatae proponuntur et arteriarum potissimum historia continetur, 7 fascicles (Göttingen: Vandenhoeck, 1743–1756; many editions thereafter); and Antonio Scarpa, Sull’aneurisma riflessioni ed osservazioni anatomico-chirurgiche(Pavia, 1804)—all venerated works. Maclise, in the preface to Surgical Anatomy(London, 1851, 1856), namedrops Haller, Hunter (not specifying John or William), Astley Cooper, Robert Liston, Charles Bell, Scarpa, Cowper, Soemmerring, Cruveilhier, Magendie, Burdach, Carus, Meckel, and Camper, who we can take as members of Maclise’s pantheon of anatomical, physiological, and surgical greats. Maclise also praises anatomists who brilliantly illustrated their own works: Charles Bell, Petrus Camper, and Antonio Scarpa, a pantheon within a pantheon. 57 For the dialogics of touch (in relation to Victorian sculpture), see Hatt, “Near and Far”, 45–50; for touch in painting, see Fend, Fleshing Out Surfaces, 7, 74–75, and 77. 58 Fend, Fleshing Out Surfaces, 7. 59 The discussion here is heavily indebted to Fend, Fleshing Out Surfaces, 42, 65–66, 74–75, and 77, which discusses the caressing brushstrokes in Fragonard’s portraits de fantasie, and eighteenth- and nineteenth-century commentary on the expressive, sensual toucheof the painter’s hand. See also Rebecca Messbarger, The Lady Anatomist: The Work of Anna Morandi Manzolini(Chicago, IL: University of Chicago Press, 2010), 128–133, which discusses the evocation of touch in the work of anatomist/wax-modeler Anna Morandi Manzolini. 60 Maclise, “Commentary on Plate IX: The Surgical Dissection of the Wrist and Hand”, Surgical Anatomy, 1st ed. (1851); “Commentary on Plates XII & XIII”, Surgical Anatomy, 2nd ed. (1856). Maclise here is in dialogue with Charles Bell’s Bridgewater Treatise, The Hand, Its Mechanism and Vital Endowments as Evincing Design(London: William Pickering, 1833). 61 Anon., review of “ Surgical Anatomy. By Joseph Maclise, Surgeon. Fasciculus I. II. III. London, 1849. Twelve Lithograph Coloured Engravings”, Edinburgh Medical & Surgical Journal71, no. 180 (1849): 224–226. 62 Review of Joseph Maclise, Surgical Anatomy, fasc. 9 (1851) in British & Foreign Medico-Chirurgical Review, Or, Quarterly of Practical Medicine & Surgery8 (July–October 1851): 529–530. We unfortunately don’t know much about the production of the Philadelphia Surgical Anatomy(“T. Sinclair’s lith. 101 Chestnut St Phila”, 1851). 63 Fend, Fleshing Out Surfaces, 42 and 60, quoting and citing Louis Tocqué, “Reflexions sur la peinture et particulièrement sur le genre du portrait” [1750], in Le discours de Tocqué sur le genre du portrait, ed. Arnauld Doria (Paris: Jean Schemit, 1930), 22; see also Fend, Fleshing Out Surfaces, 193–195. 64 Anthea Callen, Looking at Men: Art, Anatomy and the Modern Male Body(New Haven, CT: Yale University Press, 2018), 16. 65 Richard Quain, Preface to The Anatomy of the Arteries of the Human Body(1844), v [the book of captions, printed separately from the lithographic folio], gives the number of bodies as 1,010. The preface published with the lithographic folio gives the figure of 930 bodies. 66 Maclise, preface to Surgical Anatomy, in all London editions (1851, 1856). 67 While we have some of Jacob’s preparatory drawings, no drawings by Maclise, preparatory or otherwise, have ever surfaced. 68 Edgar Allan Poe, “The Philosophy of Composition”, Graham’s American Monthly Magazine of Literature and Art(Philadelphia), April 1846. 69 The literature on the erotics of anatomy mostly focuses on the female body. For erotic elements and tropes in anatomical narrative, see Ludmilla Jordanova, Sexual Visions: Images of Gender in Science and Medicine from the 18th to the 20th Century(Madison, WI: University of Wisconsin Press, 1989); Jonathan Sawday, The Body Emblazon’d: Dissection and the Human Body in Renaissance Culture(London: Routledge, 1995); and Michael Sappol, A Traffic of Dead Bodies: Anatomy and Embodied Social Identity in 19th-Century America(Princeton, NJ: Princeton University Press, 2002). 70 Whitney Davis, Queer Beauty: Sexuality and Aesthetics from Winckelmann to Freud and Beyond(New York: Columbia University Press, 2010). 71 Abigail Solomon-Godeau, Male Trouble: A Crisis in Representation(London: Thames & Hudson, 1997), see especially, 177–185. 72 See Desmond, The Politics of Evolution, 360–363, 368–371, 376, 425, and 453. 73 The Lancet(London) 25 March 1852: 347; and 1 May 1852: 426–428. 74 Maclise, On Dislocations and Fractures, dedication: “I have founded my Thoracic Theory of Circulation [on] … Animal Mechanics. Modern Science is constrained to acknowledge that the doctrine of Harvey does not answer all important Anatomical facts and Physiological and Pathological phenomena.” The argument against Harvey is mounted in “Commentary on Plates 5, 6, 7 and 8, The Form and Mechanism of the Thorax as an Ingestive Apparatus—Fractures and Dislocations of the Ribs, Sternum, and Clavicle”: “In the beginning, when Harvey and originality (imitating that immortal fiat in Genesis which made Harvey ‘after its own image’) looked upon the Chaos of Animal Physiology, and saw that it was without form and void …” Maclise connected his critique of Harvey to a defense of “philosophical”, “transcendental”, and “comparative” anatomy and analysis of “homologies”. Critics in turn mocked him for “eccentric flights”; fancy philosophizing and overblown speechifying. The Lancet: 20 March 1858: 292; 5 May 1858: 470; 29 May 1858: 534; 4 September 1858: 258; 28 July 1860: 95–96; 4 August 1860: 120–122; 11 August 1860: 146; and 18 August 1860: 172. 75 Jessie Dobson, Conservators of the Hunterian Museum(London: Jackson, Ruston & Keeton, 1975), 25–26. 76 Joseph and Daniel may have also lived together at times in the 1850s at Russell Square, London. K.B. Roberts and J.D.W. Tomlinson, The Fabric of the Body: European Traditions of Anatomical Illustration(Oxford: Clarendon, 1992), 56–65; and John Turpin, “Daniel Maclise and Cork Society”, Journal of the Cork Historical and Archaeological Society85 (1985): 66–88. Daniel was said to have been deeply distressed by Isabella’s death. A third brother, William, was an army surgeon, whose health is said to have been destroyed in the Crimean War and who died upon his return to England or Ireland. So another sibling to mourn? There were two other siblings: Anna, who married, and Alexander, “a leather sales-man” ( London Gazette, 1836, Pt. 2, 2204), who is listed as residing at 14 Russell Place, Fitzroy Square, the same address as Daniel (and, in the 1850s, Joseph). 77 Maclise, Preface to Surgical Anatomy(1856). 78 Maclise, Surgical Anatomy(1856), from “Notice to the Second Edition”. The term anthropotomywas mostly associated with naturalist/anatomist Richard Owen. 79 Solomon-Godeau, Male Trouble, 93. 80 Dickens died a year after writing that explanation. W. Justin O’Donnell, A Memoir of Daniel Maclise, R.A.(London: Longmans, Green & Co., 1871), vi. Daniel Maclise illustrated a number of Dickens’s books and the two men were for many years close friends, although eventually they became somewhat estranged. Some of Dickens’s letters to Maclise survive; see John Turpin, “Daniel Maclise and Charles Dickens: A Study of their Friendship”, Studies: An Irish Quarterly Review73, no. 289 (Spring 1984): 47–66. 81 Obituary, “Richard Quain”, The Lancet, 1 October 1887, 687–688; and Obituary, “Richard Quain”, British Medical Journal2, no. 1395 (24 September 1887): 694–695: “Without exaggeration, it may be said that Richard Quain’s life would have been well spent had he lived only to produce the Anatomy of the Arteries.” The British Medical Journalobituary also credits “the artistic powers of Mr. Joseph Maclise, F.R.C.S, author of a Surgical Anatomy, and an almost forgotten work on the homologies of the skeleton” [ Comparative Osteology]. 82 O’Donnell, A Memoir of Daniel Maclise, 117–120. 83 Daston and Galison, Objectivity.The beginnings of the binary opposition of course go back much earlier. See Lyle Massey, “Against the ‘Statue Anatomized’: The ‘Art’ of 18th-Century Anatomy on Trial”, Art History40, no. 1 (2017): 68–103. 84 For the rhetorical opposition between truth and beauty in medical illustration and depiction of medical figures, see Mary Hunter, The Face of Medicine, 12, 16, 19–20, 24, 123, et passim. 85 Preface to Surgical Anatomy, 1st ed. (1851). 86 Bibliography Anon. “Jean-Baptiste Marc Bourgery”. Wikipedia,https://en.wikipedia.org/wiki/Jean-Baptiste_Marc_Bourgery. Anon. [Obituary] “Richard Quain”. British Medical Journal2, no. 1395 (24 September 1887): 694–695. Anon. [Obituary] “Richard Quain”. The Lancet, 1 October 1887: 687–688. Anon. “Review of New Books: The Anatomy of the Arteries of the Human Body, with its Applications to Pathology and Operative Surgery, in Lithographic Drawings, with Practical Commentaries. By Richard Quain, &c. &c. The Delineations byJoseph Maclise, Esq. Surgeon. Parts I and II. London: Taylor and Walton. 1840”. Provincial Medical & Surgical Journal1–1, no. 12 (19 December 1840): 203. doi:10.1136/bmj.s1-1.12.203. Anon. [Review of] Joseph Maclise, Surgical Anatomy, fasc. 9 (1851). British & Foreign Medico-Chirurgical Review, Or, Quarterly of Practical Medicine & Surgery8 (July–October 1851): 529–530. Anon. [Review of] “ Surgical Anatomy. By Joseph Maclise, Surgeon. Fasciculus I. II. III. London, 1849. Twelve Lithograph Coloured Engravings”. Edinburgh Medical & Surgical Journal71, no. 180 (1849): 224–226. Appel, Toby A. The Cuvier-Geoffroy Debate: French Biology in the Decades before Darwin. New York: Oxford University Press, 1987. Barthes, Roland. “The Reality Effect” (1969). Translated by Richard Howard. In The Rustle of Language. Edited by François Wahl, 141–148. Berkeley, CA: University of California Press, 1989. Bell, Charles. The Hand, Its Mechanism and Vital Endowments as Evincing Design. London: William Pickering, 1833. Bell, John. Engravings of the Bones, Muscles, and Joints, illustrating the first volume of the Anatomy of the Human Body, 2nd ed. London: Printed for Longman and Rees, 1804 (1794). Bell, John. Principles of Surgery, 3 vols. Edinburgh: Cadel & Davies & Longman, 1801–1808. Benjamin, Walter. “The Work of Art in the Age of Mechanical Reproduction”. In Illuminations.Translated by Harry Zohn. Edited by Hannah Arendt. New York: Schocken/Random House, 1969, 217–251.https://www.marxists.org/reference/subject/philosophy/works/ge/benjamin.htm. Berger, John. Ways of Seeing.London: Penguin, 1973. Berkowitz, Carin. “The Illustrious Anatomist: Authorship, Patronage and Illustrative Style in Anatomy Folios, 1700–1840”. Bulletin of the History of Medicine(2015): 171–208. Berkowitz, Carin “Systems of Display: The Making of Anatomical Knowledge in Enlightenment Britain”. British Journal for the History of Science46, no. 3 (2013): 359–387. Bersani, Leo. “Is the Rectum a Grave?” October43 (Winter 1987): 197–222. Bidloo, Govard. Anatomia humani corporis. 1685. Bourgery, Jean-Baptiste-Marc, and Nicolas Henri Jacob. Traité complet de l’anatomie de l’homme: Comprenant la médecine opératoire, avec planches lithographiées d’après nature, 8 vols .Paris: C.A. Delaunay, 1831–1854; 2nd ed., 1866–1871. Bynum, W.F. “Quain, Richard (1800–1887), Anatomist and Surgeon”. Oxford Dictionary of National Biography, 23 September 2004. doi:10.1093/ref:odnb/22940. Callen, Anthea. Looking at Men: Art, Anatomy and the Modern Male Body. New Haven, CT: Yale University Press, 2018. Clarke, J.F. Autobiographical Recollections of the Medical Profession. London: J. & A. Churchill, 1874. Daremberg, C. Charles. La médecine: Histoire et doctrines.Paris: Didier et cie, 1865. Daston, Lorraine, and Peter Galison. Objectivity. New York: Zone Books, 2007. Davis, Whitney. Queer Beauty: Sexuality and Aesthetics from Winckelmann to Freud and Beyond. New York: Columbia University Press, 2010. Desmond, Adrian. The Politics of Evolution: Morphology, Medicine, and Reform in Radical London. Chicago, IL: University of Chicago Press, 1989. Dobson, Jessie. Conservators of the Hunterian Museum. London: Jackson, Ruston & Keeton, 1975. Duffin, Jacalyn. “Imaging Disease: The Illustration and Non-Illustration of Medical Texts, 1650–1850”. In “Muse and Reason: The Relation of Arts and Sciences, 1650–1850”. Edited by B. Castel, J.A. Leith, and A.W. Riley. A Royal Society of Canada Symposium, Queens Quarterly(1994): 79–108. Edmondson, John. “Death and the Tourist: Dark Encounters in Mid-Nineteenth-Century London via the Paris Morgue”. In The Palgrave Handbook of Dark Tourism Studies. Edited by Philip R. Stone, Rudi Hartmann, Tony Seaton, Richard Sharpley, and Leanne White, 77–101. London: Palgrave, 2017. Eldridge, Richard Thomas. “Idealism in 19th-Century British and American Literature”. Aesthetics and Literature3 (2013): 121–144.https://works.swarthmore.edu/fac-philosophy/265/. Elkins, James. The Object Stares Back: On the Nature of Seeing. New York: Harcourt Brace, 1996. Evans, Caroline, and Lorraine Gamman. “The Gaze Revisited, or Reviewing Queer Viewing”. In A Queer Romance: Lesbians, Gay Men and Popular Culture. Edited by Paul Burston and Colin Richardson, 13–56. London: Routledge, 1995. Fend, Mechthild. Fleshing Out Surfaces: Skin in French Art and Medicine 1650–1850. Manchester: Manchester University Press, 2017. Foucault, Michel. The Birth of the Clinic: An Archaeology of Medical Perception. Translated by Alan M. Sheridan Smith (1973). New York: Routledge, 2003 (1963). Foucault, Michel. The History of Sexuality. Vol. 1, An Introduction. Translated by Robert Hurley. New York: Vintage, 1990. Gamelin, Jacques. Nouveau recueil d’ostéologie et de myologie, dessin‚ d’après nature … pour l’utilit‚ des sciences et des arts. Toulouse: J.F. Desclassan, 1779. Haller, Albrecht von. Icones anatomicae quibus praecipuae aliquae partes corporis humani delineatae proponuntur et arteriarum potissimum historia continetur, 7 fascicles. Göttingen: Vandenhoeck, 1743–1756. Hannaway, Carolyn, and Ann La Berge. “Paris Medicine: Perspectives Past and Present”. In Constructing Paris Medicine. Edited by Carolyn Hannaway and Ann La Berge. Amsterdam: Editions Rodopi, 1998, 1–69. Hatt, Michael. “Near and Far: Homoeroticism, Labour and Hamo Thornycroft’s Mower”. Art History26, no. 1 (February 2003): 26–55. doi:10.1111/1467-8365.d01-. Hatt, Michael. “Space, Surface, Self: Homosexuality and the Aesthetic Interior”. Visual Culture in Britain8, no. 1 (2007): 105–128. Hoffmann, Kathryn. “Sleeping Beauties in the Fairground: The Spitzner, Pedley, and Chemisé Exhibits”. Early Popular Visual Culture4, no. 2 (July 2006): 139–159. Hunter, Mary. The Face of Medicine: Visualizing Medical Masculinities in Late 19th-Century Paris.Manchester: Manchester University Press, 2016. Hunter, William. The Anatomy of the Human Gravid Uterus.Birmingham: John Baskerville, 1774. Johnson, Dorothy. Jacques-Louis David: The Art of Metamorphosis. Princeton, NJ: Princeton University Press, 1993. Jordanova, Ludmilla. Sexual Visions: Images of Gender in Science and Medicine from the 18th to the 20th Century. Madison, WI: University of Wisconsin Press, 1989. Kosofsky Sedgwick, Eve. Between Men: English Literature and Male Homosocial Desire.New York: Columbia University Press, 1985. Kosofsky Sedgwick, Eve. Epistemology of the Closet. Berkeley, CA: University of California Press, 2008 (1990). Kravetz, Rachel. “The Art of Cognition: British Empiricism and Victorian Aesthetics”. PhD dissertation, City University of New York, 2017. Lo, Melissa. “The Wounded Ideal Goes Anatomical”. In Hidden Treasure. Edited by Michael Sappol, 44–49. New York: Blast Books, 2011. Maclise, Joseph. Comparative Osteology: Being Morphological Studies to Demonstrate the Archetype Skeleton of Vertebrated Animals. London: Taylor & Walton, 1847. Maclise, Joseph. On Dislocations and Fractures. London: John Churchill, 1859. Maclise, Joseph. Surgical Anatomy, 1st ed. London: John Churchill, 1851. Internet Archive:https://archive.org/details/b32723684/page/n3/mode/2up. Maclise, Joseph. Surgical Anatomy, 2nd ed. London: John Churchill, 1856. Internet Archive:https://archive.org/details/surgicalanatomy00macl_0. McCormack, Helen. William Hunter and his Eighteenth-Century Cultural Worlds: The Anatomist and the Fine Arts. London: Routledge, 2017. Mascagni, Paolo. Anatomia universaor Grande anatomia del corpo umano. Pisa: Presso Niccolò Capurro, 1823–1830. Massey, Lyle. “Against the ‘Statue Anatomized’: The ‘Art’ of 18th-Century Anatomy on Trial”. Art History40, no. 1 (2017): 68–103. Messbarger, Rebecca. The Lady Anatomist: The Work of Anna Morandi Manzolini.Chicago, IL: University of Chicago Press, 2010. O’Donnell, W. Justin. A Memoir of Daniel Maclise, R.A. London: Longmans, Green & Co., 1871. Poe, Edgar Allan. “The Philosophy of Composition”. Graham’s American Monthly Magazine of Literature and Art(Philadelphia), April 1846. Pollock, George David. [Obituary of Richard Quain], Address of George David Pollock, F.R.C.S., President of the Royal Medical and Chirurgical Society of London at the Annual Meeting, March 1st, 1888. London, 1888, 20–22.https://wellcomecollection.org/works/yv6vfvs7. Quain, Richard. The Anatomy of the Arteries of the Human Body and its Application to Pathology and Operative Surgery. London: Taylor & Walton, 1844. Quain, Richard. On Some Defects in General Education: Being the Hunterian Oration of the Royal College of Surgeons for 1869. London: Macmillan, 1870. Racine, Valerie. “Etienne Geoffroy Saint-Hilaire (1772–1844)”. Embryo Project Encyclopedia(5 August 2013).http://embryo.asu.edu/handle/10776/6045. Rehbock, Philip F. “Transcendental Anatomy”. In Romanticism and the Sciences. Edited by Andrew Cunningham and Nicholas Jardine. Cambridge: Cambridge University Press, 1990, 144–160. Roberts, K.B., and J.D.W. Tomlinson. The Fabric of the Body: European Traditions of Anatomical Illustration.Oxford: Clarendon, 1992. Royal College of Surgeons of England. “Maclise, Joseph (1815–1880)”. Plarr’s Lives of the Fellows(London, 1912–1940).http://livesonline.rcseng.ac.uk/biogs/E002613b.htm. Royal College of Surgeons of England. “Quain, Richard (1800–1887)”. Plarr’s Lives of the Fellows.https://livesonline.rcseng.ac.uk/client/en_GB/lives/search/detailnonmodal/ent:$002f$002fSD_ASSET$002f0$002fSD_ASSET:372381/one. Sappol, Michael. “Anatomy’s Photography: Objectivity, Showmanship & the Reinvention of the Anatomical Image, 1860–1950”. REMEDIA, January 2017 (rev. 2018).https://www.academia.edu/31049714/Anatomy_s_photography_Objectivity_showmanship_and_the_reinvention_of_the_anatomical_image_1860_1950_Introduction. Sappol, Michael. Dream Anatomy. Washington, DC: GPO, 2003. Sappol, Michael. A Traffic of Dead Bodies: Anatomy and Embodied Social Identity in 19th-Century America.Princeton, NJ: Princeton University Press, 2002. Sawday, Jonathan. The Body Emblazon’d: Dissection and the Human Body in Renaissance Culture. London: Routledge, 1995. Scarpa, Antonio. Sull’aneurisma riflessioni ed osservazioni anatomico-chirurgiche. Pavia, 1804. Schwartz, Vanessa R. Spectacular Realities: Early Mass Culture in Fin-de-Siècle Paris. Berkeley, CA: University of California Press, 1999. Slipp, Naomi. “International Anatomies: Teaching Visual Literacy in the Harvard Lecture Hall”. In Bodies Beyond Borders: Moving Anatomies, 1750–1950. Edited by Kaat Wils, Raf de Bont, and Sokhieng Au, 197–230. Leuven: Leuven University Press, 2017. Solomon-Godeau, Abigail. Male Trouble: A Crisis in Representation. London: Thames & Hudson, 1997. Stephens, Elizabeth. “Touching Bodies: Tact/ility in 19th-Century Medical Photographs and Models”. In Bodies, Sex and Desire from the Renaissance to the Present. Edited by Kate Fisher and Sarah Toulalan, 87–101. London: Palgrave Macmillan, 2011. Tiedemann, Friedrich. Plates of the Arteries of the Human Body. Edited by Thomas Wharton Jones and Robert Knox. Edinburgh: Printed for Maclachlan & Stewart, 1829–1836. Tiedemann, Friedrich. Tabulae arteriarum corporis humani. Karlsruhe: C.F. Müller, 1822. Tocqué, Louis. “Reflexions sur la peinture et particulièrement sur le genre du portrait” (1750). In Le discours de Tocqué sur le genre du portrait. Edited by Arnauld Doria, 1–40. Paris: Jean Schemit, 1930. Tolet, François. A Treatise of Lithotomy: or, of the Extraction of the Stone Out of the Bladder … Written in French … Translated into English by A. Lovell. London: Printed by H.H. for William Cademan, 1683. Turpin, John. “Daniel Maclise and Charles Dickens: A Study of their Friendship”. Studies: An Irish Quarterly Review73, no. 289 (Spring 1984): 47–66. Turpin, John. “Daniel Maclise and Cork Society”. Journal of the Cork Historical and Archaeological Society85 (1985): 66–88. Zitin, Abigail. “Wantonness: Milton, Hogarth, and The Analysis of Beauty”. Differences27, no. 1 (2016): 25–46. Imprint Author Michael Sappol Date 18 July 2021 Category One Object Review status Peer Reviewed (Double Blind) Licence CC BY-NC International 4.0 Downloads PDF format Article DOI https://doi.org/10.17658/issn.2058-5462/issue-20/msappol Cite as Michael Sappol, "Mr Joseph Maclise and the Epistemology of the Anatomical Closet", British Art Studies, Issue 20, https://doi.org/10.17658/issn.2058-5462/issue-20/msappol NextJoseph Maclise, Taylor & Walton, and Publishing on Gower Street in the 1840s

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LY v. STATE | Civil Action No. 07-3259-CM. | 20091203933 | Leagle.com MEMORANDUM AND ORDER CARLOS MURGUIA District Judge. Petitioner Binh Ly a prisoner at the Lansing Correctional Facility in Lansing...20091203933 LY v. STATE Civil Action No. 07-3259-CM. View Case Cited Cases BINH LY, Petitioner, v. STATE OF KANSAS, et al., Respondents. United States District Court, D. Kansas. https://leagle.com/images/logo.png December 2, 2009. December 2, 2009. United States District Court, D. Kansas. MEMORANDUM AND ORDER CARLOS MURGUIA, District Judge. Petitioner Binh Ly, a prisoner at the Lansing Correctional Facility in Lansing, Kansas, files this Petition Under 28 U.S.C. § 2254 for Writ of Habeas Corpus By A Person in State Custody (Doc. 1). Petitioner was convicted by a jury of felony murder and aggravated burglary, and is serving a controlling sentence of life imprisonment. His convictions were affirmed on appeal by the Kansas Supreme Court in State v. Ly,85 P.3d 1200(Kan. 2004), cert. denied, 541 U.S. 1090 (2004). His petition pursuant to Kan. Stat. Ann. § 60-1507 was denied without a hearing, and the denial was affirmed by the Kansas Court of Appeals. See Ly v. State, No. 94,732, 2006 WL 3231373 (Kan. Ct. App. Nov. 3, 2006), rev. denied(Kan. Feb 14, 2007). This habeas petition followed. Petitioner requests federal relief on a number of grounds. For the following reasons, the petition is denied. I. Procedural Background 1 On May 31, 2001, petitioner was charged in the District Court of Sedgwick County, Kansas with one count of first-degree murder in violation of Kan. Stat. Ann. § 21-3401. The State later added an alternative charge of aggravated burglary in violation of Kan. Stat. Ann. § 21-3716 and felony murder in violation of Kan. Stat. Ann. § 21-3401(b). On September 13, 2001, petitioner was convicted by a jury of aggravated burglary and felony murder. He was sentenced to a controlling term of life imprisonment plus forty-nine (49) months in the Kansas Department of Corrections. On direct appeal, petitioner's convictions were affirmed by the Kansas Supreme Court. 2 State v. Ly,85 P.3d 1200(2004), cert. denied541 U.S. 1090 (2004). On November 5, 2004, petitioner filed for state post-conviction relief pursuant to Kan. Stat. Ann. § 60-1507. 3The Sedgwick County District Court denied his petition without a hearing, and the denial was affirmed by the Kansas Court of Appeals. 4 See Ly v. State, No. 94,732, 2006 WL 3231373 (Kan. Ct. App. Nov. 3, 2006), rev. deniedFeb 14, 2007. In this federal habeas petition, petitioner raises the following claims, all of which were presented to the state courts either on direct appeal or through petitioner's motion for post-conviction relief: (1) The trial court committed reversible error in denying petitioner's pretrial motion for continuance; (2) Prosecutorial misconduct in closing argument requires reversal of petitioner's conviction; (3) The evidence was insufficient to support petitioner's conviction of felony murder and aggravated burglary; (4) The district court erred in ruling on petitioner's newly discovered evidence claim; (5) The aiding and abetting instruction was fatally defective; (6) Trial counsel was ineffective for failing to conduct independent firearm testing or otherwise challenge the State's firearm expert; (7) Trial counsel was ineffective for allowing a friend of the prosecutor's family to remain on the jury; (8) Trial counsel was ineffective because he "tricked and c[a]joled" petitioner into making statements to law enforcement; (9) Trial and appellate counsel were ineffective in failing to object to or challenge the trial court's response to the jury's question during deliberations; (10) Trial counsel was ineffective in his closing argument to the jury by conceding petitioner's guilt; and (11) The trial court erred in finding petitioner's trial and/or appellate counsel effective, and in failing to conduct an evidentiary hearing related to issues number (6), (7), (8), and (9). II. Factual Background In a federal habeas proceeding, the state court's factual findings are presumed correct and petitioner bears the burden of rebutting that presumption by clear and convincing evidence. 28 U.S.C. § 2254(e)(1). Petitioner does not challenge the factual background set out by the state court. 5Accordingly, the court incorporates the Kansas Supreme Court's version of the facts: On May 24, 2001, 20-year-old Chanh Chantivong and his younger twin brothers, Chin and Done, were at Woody Sim's house for haircuts. Several other young men, who were suspected to be members of the East Side Crips gang, were also at Sim's house. Because Sim's house had formerly been a duplex, his bedroom had an exterior door leading onto the front porch. The young men were hanging out in Sim's bedroom and on the front porch. During the evening, the young men had a conflict with a rival gang member who had fired shots at Sim's house. Afterwards, Chanh sent Done home to get Chanh's gun. When Done returned, the group of young men at Sim's house sat on the deck and discussed the earlier shooting incident. They kept all of the lights off because the police were patrolling the area and they did not want to attract attention. At about midnight, Done saw two cars following each other down the street. The two cars parked near Sim's house. A group of people ran from the cars, kicked in the door to Sim's bedroom, and turned on the lights. Some of the young men ran from the deck to the backyard, but Chanh ran into the house with his gun. A round of gunshots followed, and the group ran back to their cars and drove away. Witnesses estimated that the shooting lasted about 1 minute, but when it was over, Chanh lay dead on the floor. He had been shot 16 times and stabbed once. Police found 26 cartridge casings in Sim's bedroom from four different caliber weapons. The ballistics report from the State's expert indicated that Chanh's attackers used five different guns, but the only gun police recovered from the scene was Chanh's. Ly became a suspect in Chanh's murder when Salina police responded to a statewide alert for anyone reporting to a hospital with gunshot wounds. Ly sought treatment at the Salina Hospital for two gunshot wounds. A hospital employee notified the Salina police, who questioned Ly and his companions about Ly's injuries. Both Ly and his companions told the Salina police that Ly had been shot while he was urinating near the highway in Salina. However, a firearms expert analyzed the bullet removed from one of Ly's wounds and determined that it had been fired from Chanh's gun. After his arrest, Ly admitted to being present at the shooting but told police that he did not have a weapon. Ly gave the police the names of three individuals who were armed. The State charged Ly with one count of aggravated burglary and one count of first-degree murder under alternative theories of premeditated murder and felony murder. The case was originally set for trial on August 13, 2001, but the State received a continuance, over Ly's objection, until September 10, 2001, to get the ballistics report completed. On September 5, 2001, the State orally advised Ly's counsel of the results from the ballistics report. The next day, Ly's counsel orally moved for a continuance to get a ballistics report from an independent firearms expert. The court denied Ly's request. At trial, Ly claimed ignorance as a defense. He told police that he thought he was going to a party and that no one had talked about any shooting. Disbelieving Ly's claim of ignorance, the jury found him guilty of felony murder and aggravated burglary. Ly, 85 P.3d at 1203. Petitioner now seeks federal habeas corpus relief pursuant to 28 U.S.C. § 2254. III. Standard of Review Because petitioner has satisfied the exhaustion requirement the Antiterrorism and Effective Death Penalty Act of 1996, 28 U.S.C. § 2254(b)(1)(A), (c), the court reviews petitioner's claims pursuant to the provisions of the Act. Wallace v. Ward,191 F.3d 1235, 1240 (10th Cir. 1999). The Act permits a court to grant a writ of habeas corpus only if one of two circumstances is present: (1) the state court's decision "was contrary to, or involved an unreasonable application of, clearly established Federal law, as determined by the Supreme Court of the United States," 28 U.S.C. § 2254(d)(1); or (2) the state court's decision "was based on an unreasonable determination of the facts in light of the evidence presented in the State court proceeding," id.§ 2254(d)(2). Absent clear and convincing evidence to the contrary, the court presumes that state court factual findings are correct. Id.§ 2254(e)(1). Under the first alternative, the court will find that a state court decision is contrary to clearly established law "if the state court arrives at a conclusion opposite to that reached by [the Supreme Court] on a question of law or if the state court decides a case differently than [the Supreme Court] has on a set of materially indistinguishable facts." Williams v. Taylor,529 U.S. 362, 413 (2000). Under the second alternative, the court will find that a state court decision is an unreasonable application of clearly established federal law "if the state court identifies the correct governing legal principle from [the Supreme Court's] decisions but unreasonably applies that principle to the facts of the prisoner's case." Id.The key inquiry is whether the state court's application of the law was objectively unreasonable. Id.at 409; see also Lockyer v. Andrade,538 U.S. 63, 75-76 (2003) (observing that the "objectively unreasonable" standard of review is more deferential than the "clear error" standard). But the petitioner need not show that "all reasonable jurists" would disagree with the decision of the state court. Williams, 529 U.S. at 409-10. IV. Discussion Under the standard set out above, the scope of this court's review is to determine whether, as to each of petitioner's claims, the Kansas courts either (1) arrived at a conclusion contrary to clearly established federal law; or (2) applied the law in an objectively unreasonable manner. In doing so, this court presumes the correctness of the state court's factual findings, absent clear and convincing evidence to the contrary. 28 U.S.C. § 2254(e)(1). However, the court does not correct errors of state law, and is bound by a state court's interpretation of its own law. See Bradshaw v. Richey,546 U.S. 74, 76 (2005). Trial Court's Denial of Motion for Continuance Noting that the trial court has discretion to grant or deny continuances pursuant to Kan. Stat. Ann. § 22-3401, the Kansas Supreme Court looked at "the opportunities Ly had to dispute the evidence both before and after trial" and petitioner's failure to demonstrate any prejudice because the State's case was not based purely on the theory that petitioner shot the victim. Ly, 85 P.3d at 1204 (citing State v. Snodgrass,843 P.2d 720, 728-29 (Kan. 1992);) see also United States v. West,828 F.2d 1468, 1469-70 (10th Cir. 1987). The state court found no abuse of discretion in the denial of petitioner's request for a continuance to allow him time to locate a firearms expert. Ly, 85 P.3d at 1204. This conclusion was based on state law, and was consistent with precedent. Additionally, the state court's standard is consistent with that applied in the federal courts. See United States v. Riviera,900 F.2d 1462, 1475 (10th Cir. 1990). The state court did not address whether petitioner's constitutional rights were violated by the trial court's denial of the motion for continuance and the motion to dismiss. To warrant federal habeas relief, the denial of a continuance must be so arbitrary and fundamentally unfair as to constitute a violation of due process. Case v. Mondragon,887 F.2d 1388, 1396-97 (10th Cir. 1989), cert. denied, 494 U.S. 1035 (1990); Scott v. Roberts,777 F.Supp. 897, 899 (D. Kan. 1991). There are no mechanical tests to be applied in determining whether the denial of a continuance is so arbitrary as to violate due process. Ungar v. Sarafite,376 U.S. 575, 589 (1964). In making such a fundamental fairness evaluation, the court must focus on the need for the continuance and actual prejudice resulting from its denial. Case, 887 F.2d at 1397. Construing the petition liberally, the court discerns allegations that the failure to grant a continuance violated petitioner's right to assistance of counsel, due process, and a fair trial. However, petitioner fails to establish how he was prejudiced by not having an expert to rebut the report of the state's expert. On the record presented, petitioner fails to establish actual prejudice. Prosecutorial Misconduct in Closing In reviewing a prosecutorial misconduct claim, the Kansas courts apply a two-step inquiry: First, the court considers whether the challenged remarks were outside the considerable latitude allowed to the prosecutor in discussing the evidence. If so, then the court determines whether the remarks constituted plain error, that is, whether "the prosecutor's remarks are so gross and flagrant as to prejudice the jury against the defendant and deny the defendant a fair trial, requiring reversal." Id., at 1206 (citing State v. McCorkendale,979 P.2d 1239, 1250 (Kan. 1999), overruled on other grounds by State v. King, 204 P.3d 585 (Kan. 2009)). As for petitioner's challenge to the prosecutor's statement that five people entered Sim's room, the court found that the statement was supported by the evidence and that petitioner's failure to object to it rendered review unnecessary. However, the court found that the prosecutor misstated the evidence in closing by stating that Sovong Vann, a material witness in the case, "`saw one of the people, and they had on a yellow shirt and that that person with the yellow shirt had a gun. [Vann] says all of them had guns.'" Id. at 1206. Vann had not made any statements to indicate that all of the intruders had guns. Petitioner claimed that the error required reversal because without it, the prosecutor could not establish that petitioner had a weapon. Applying the plain error standard, however, the court found that (1) the prosecutor's statement had no significance to the State's case because Vann had testified that the yellow-shirted intruder had a gun, and petitioner admitted to having worn a yellow shirt; and (2) the statement was unnecessary under the aiding and abetting theory. Therefore, the error, when viewed in light of the whole record, would not have changed the verdict or the result of the trial. This court concludes that petitioner is not entitled to relief on his argument that prosecutorial misconduct requires reversal of his conviction. See Cummings v. Evans,161 F.3d 610, 618 (10th Cir. 1998) (employing identical standard for review of prosecutorial misconduct claims, requiring reversal only if misconduct is so egregious that it renders the entire trial fundamentally unfair.). The Kansas Supreme Court's conclusion is not contrary to clearly established federal law, nor was it applied in an objectively unreasonable manner. Insufficient Evidence In evaluating petitioner's insufficient evidence argument on direct appeal, the Kansas Supreme Court looked to whether, after review of all the evidence, viewed in a light most favorable to the prosecution, a rational factfinder could have found petitioner guilty beyond a reasonable doubt. Ly, 85 P.3d at 1207 (citing State v. Beach,67 P.3d 121, 127-28 (Kan. 2003)). The court first rejected petitioner's argument that his mere presence at the scene was insufficient to establish his guilt on an aiding and abetting theory. The evidence, viewed in the light most favorable to the prosecution, established that petitioner was not merely along for the ride. Petitioner was shot twice by the victim, and the wounds indicated petitioner was facing the victim when petitioner was shot. Second, a witness described a man in a yellow shirt with a gun. Petitioner admitted that he was wearing a yellow shirt at the time of the shooting, making it reasonable to infer that he was armed. Third, evidence of the circumstances surrounding the timing of the shooting, the conditions of the premises, and involvement of various parties tended to discredit petitioner's version of events. From this evidence, the state court found that the jury could have inferred that the group of shooters coordinated their events prior to arriving at Sim's house, and that petitioner encouraged and participated in the incident. Other than petitioner's own self-serving statements, there was no evidence tending to support his version of the events. Considering this under the applicable standard, the court concluded that there was sufficient evidence to support the verdict. This conclusion is not contrary to clearly established federal law, nor was it applied in an objectively unreasonable manner. Newly Discovered Evidence In petitioner's application for post-conviction relief, he claims that, after his trial, he came in contact with a codefendant, Sashada Makthepharak, and obtained an affidavit from him stating that petitioner did not have anything to do with the murder. Specifically, Makthepharak's affidavit states that Makthepharak told the prosecutor that "Binh Ly had nothing to do with the crime [and] did not know what was going on"; that the prosecutor "chose to ignore" the information and stated that was "too bad for [petitioner.]" (Doc. 1, at 15.) The Sedgwick County District Court denied relief and the Kansas Court of Appeals affirmed. The appellate court noted that new trials on the ground of newly discovered evidence are "not favored and are viewed with great caution. The defendant bears the burden of establishing that the newly proffered evidence is indeed `new'; that is, it could not, with reasonable diligence, have been produced at trial; and then the evidence must be of such a materiality that there is a reasonable probability that it would have produced a different result at trial." Ly, 2006 WL 3231373, at *1 (citing State v. Moncla,4 P.3d 618(Kan. 2000)). The court determined that the record "clearly show[ed] that [petitioner] knew [Makthepharak] was a codefendant," and that petitioner "could have discovered the information prior to trial." Id. at *2. Before this court, petitioner suggests that, "[i]f [Makthepharak] specifically told [the prosecutor] that petitioner was not involved in the incident and that he had no prior knowledge of what was going on and the prosecutor failed to share that information with petitioner," the petitioner is entitled to relief under Brady v. Maryland,373 U.S. 83, 87 (1963). The state court did not address petitioner's Bradyargument, although it was briefed. In a Bradyclaim, the petitioner bears the burden of showing that the prosecution suppressed material evidence favorable to the petitioner. Smith v. Workman,550 F.3d 1258, 1273 (10th Cir. 2008); Moore v. Gibson,195 F.3d 1152, 1164 (10th Cir. 1999). In order for the evidence to be material, there must be a "reasonable probability that, had the evidence been disclosed to the defense, the result of the proceeding would have been different. A `reasonable probability' is a probability sufficient to undermine confidence in the outcome." Id. at 1165 (quoting Pennsylvania v. Ritchie,480 U.S. 39, 57 (1987)) (internal quotation marks omitted). After careful review of the record and briefs, this court finds that petitioner fails to meet his burden. The jury rejected petitioner's claim of innocence based on ignorance. Petitioner fails to show that there is a reasonable probability that, had the prosecutor informed petitioner that Makthepharak would also attest to his ignorance, Makthepharak would have so testified, and the jury would have reached a different outcome. Indeed, the petitioner does not allege that Makthepharak in fact made this statement to the prosecutor prior to petitioner's trial. He argues that, " [i]f[Makthepharak] specifically told [the prosecutor] that petitioner was not involved . . . and the prosecutor failed to share that information with petitioner," then there would be a Bradyproblem. (Doc. 9, at 12) (emphasis added). Moreover, it appears that petitioner would be in the best position to know what his codefendant might say concerning his guilt or innocence, and would be entirely capable of investigating that. Such a burden should not fall to the prosecution. Petitioner's arguments are insufficient to entitle him to relief. Aiding and Abetting Instruction The state appellate courts did not evaluate petitioner's claim that the aiding and abetting instruction was erroneous. However, this claim was raised in petitioner's state post-conviction petition and was addressed by the Sedgwick County District Court. That court concluded the claim was barred because, as a mere trial error, it could have been raised in his direct appeal. (Sedgwick County District Court, Order Denying Relief Pursuant to K.S.A. 60-1507, filed stamped June 29, 2005, at 2-3) (citing Kan. Sup. Ct. R. 183(c)(3)). Regardless, the court held the instruction mirrored Kansas Pattern Instruction P.I.K. Crim. 3d § 56.02. Id. (citing State v. Butler, 897 P.2d 1007 (Kan. 1995)). The court therefore finds that the claim is procedurally defaulted under the independent and adequate state ground doctrine. Petitioner fails to demonstrate cause for the default and actual prejudice, or that a fundamental miscarriage of justice will occur if the court does not consider the claim. Coleman v. Thompson,501 U.S. 722, 750 (1991). Ineffective Assistance of Counsel Petitioner raised a number of claims regarding the effectiveness of his trial and appellate counsel. Specifically, he argued that his counsel was ineffective for (1) failing to conduct independent firearm testing or challenge the State's firearm expert; (2) allowing a juror who had a special relationship with the prosecutor to remain on the jury; (3) tricking him into giving up his right to remain silent; (4) failing to object to or challenge the trial court's response to a question from the jury during its deliberations; and (5) conceding petitioner's guilt in closing. Respondent argues that these claims are not supported by the record, do not entitle petitioner to relief, or are not properly before the court. In evaluating petitioner's claims that his counsel provided ineffective assistance, the appropriate standard is that identified in Strickland v. Washington,466 U.S. 668(1984). See Romano v. Gibson,278 F.3d 1145, 1151 (10th Cir. 2002) (applying Strickland) .Under Strickland,a petitioner bears the burden of satisfying a two-pronged test. First, he must show that his attorney's "performance was deficient" and "fell below an objective standard of reasonableness." Strickland,466 U.S. at 687-88. The court affords considerable deference to an attorney's strategic decisions and "recognize[s] that counsel is strongly presumed to have rendered adequate assistance and made all significant decisions in the exercise of reasonable professional judgment." Id.at 690. Second, a habeas petitioner must demonstrate prejudice, which requires a showing that there is "a reasonable probability that, but for counsel's unprofessional errors, the result of the proceeding would have been different." Id.at 694. "A reasonable probability is a probability sufficient to undermine confidence in the outcome." Id. There is no reason for a court deciding an ineffective assistance claim to address both components of the inquiry if the petitioner makes an insufficient showing on one. If it is easier to dispose of an ineffectiveness claim on the ground of lack of sufficient prejudice, that course should be followed. Strickland,466 U.S. at 697. To the extent the Kansas Court of Appeals addressed the merits of petitioner's arguments in its November 3, 2006 Memorandum Opinion, this court confines its review to whether the appellate court unreasonably applied Stricklandor whether its factual determination was unreasonable in light of the evidence presented. In its opinion, the Kansas Court of Appeals set forth the Stricklandstandard cited above, identified petitioner's allegations of ineffective assistance of counsel, and rejected petitioner's claims. Independent Firearm Testing or Challenging State's Expert As to his claim that counsel was ineffective for failing to conduct independent firearm testing or challenge the State's firearm expert, the court noted that counsel had sought a continuance—it was the trial court that denied the motion. The appellate court's review of the record led it to agree with the district court that no amount of expert testimony from a defense ballistics expert would refute the State's aiding and abetting theory of the case. The court found that petitioner's theory of defense was not harmed by the State's expert's testimony, because the State never sought to put the murder weapon in petitioner's hand. Ly, 2006 WL 3231373 at *3-4. The court's conclusion is not contrary to clearly established federal law, nor was it applied in an objectively unreasonable manner. Juror's Relationship to Prosecutor The court also considered petitioner's claim that his counsel was ineffective for allowing a juror to remain on the jury after the juror indicated that he knew the prosecutor's brother, although not the prosecutor. Petitioner faulted his counsel for failing to ask more questions on voir dire regarding impartiality, and argued that an evidentiary hearing was necessary. The court rejected these arguments, stating, "[w]e do not know what else trial counsel could have asked, other than to repeat the same question" the prosecutor asked. Id. at *3. The Court of Appeals noted that, "[w]here the prosecutor asked the necessary questions regarding potential bias, defense counsel's failure to repeat them does not constitute ineffective assistance." Id. at *3. The court's conclusion is not contrary to clearly established federal law, nor was it applied in an objectively unreasonable manner. Statements to Law Enforcement As to his claim that counsel tricked him into making statements to law enforcement, that his statements were not voluntary, and that an evidentiary hearing was required in order for the district court to deny relief, the court determined that none of the information provided by petitioner to police was damaging. Moreover, the court noted that all of the information came into evidence through the testimony of other witnesses. And finally, petitioner's counsel had attempted to use petitioner's cooperation with authorities to cast him in a positive light to the jury. See id. at *2. The court's conclusion is not contrary to clearly established federal law, nor was it applied in an objectively unreasonable manner. Response to Jury Questions The court also addressed petitioner's claim that his counsel was ineffective for failing to object or challenge the trial court's response to a question from the jury during its deliberations. According to the Court of Appeals, the jury sent several questions to the trial court during its deliberations. The first asked for a definition of "intentionally," which the court's response provided. The second question, after clarification, asked "[d]o we have to select felony murder if we select [second]-degree murder and aggravated burglary?" The trial court responded by asking the jurors to carefully read the instructions. Petitioner's counsel agreed that the instructions held the answer to the jury's question. The Court of Appeals rejected petitioner's claim that this constituted ineffective assistance, or that his appellate counsel was ineffective for raising it on direct appeal. The court found that the trial court's responses were a correct recitation of the law, and that petitioner was unable to establish prejudice. The instructions told the jury that it must decide each charge independently of the others, and that petitioner may be convicted or acquitted on any or all of the charges. The court's conclusion is not contrary to clearly established federal law, nor was it applied in an objectively unreasonable manner. Conceding Petitioner's Guilt in Closing Argument The state appellate courts did not evaluate petitioner's claim that his counsel was ineffective for conceding petitioner's guilt in closing. However, this claim was raised in petitioner's state post-conviction petition and was addressed by the Sedgwick County District Court. That court rejected the claim, finding that, "when the closing argument is viewed in its entirety, it is evident trial counsel did not ask the jury to convict." (Sedgwick County District Court, Order Denying Relief Pursuant to K.S.A. 60-1507, filed stamped June 29, 2005, at 5.) The denial of petitioner's ineffective assistance argument was proper because petitioner failed to show that counsel's performance was deficient. Petitioner is entitled to a fair trial, not a perfect one. See Ross v. Oklahoma,487 U.S. 81, 91 (1988); State v. Crawford,872 P.2d 293, 298 (Kan. 1994). And this court's review in the context of a habeas petition is limited. See Turrentine v. Mullin,390 F.3d 1181, 1201 (10th Cir. 2004). Based on this court's review of the record, which includes briefs in this court as well as in the state appellate courts; the transcripts of the trial and motions hearings; and the orders and opinions of the Kansas courts, petitioner received a fair trial. For all the reasons set out above, petitioner is not entitled to habeas relief. IT IS THEREFORE ORDERED that the Petition Under 28 U.S.C. § 2254 for Writ of Habeas Corpus By A Person in State Custody (Doc. 1) is denied. FootNotes 1. This procedural background is drawn largely from the State of Kansas's Answer and Return, with which petitioner agrees. 2. On direct appeal, petitioner raised the following three claims: (1) the trial court committed reversible error when it denied petitioner's motion for continuance; (2) prosecutorial misconduct in closing argument required reversal of the convictions; and (3) evidence was insufficient to support the convictions of felony murder and aggravated burglary. 3. Ly v. State , Sedgwick County District Court Case No. 04 CV 4450. In his motion pursuant to Kan. Stat. Ann. § 60-1507, petitioner raised the following grounds for relief: (1) The trial court's instructions to the jury were fatally and fundamentally defective on aiding and abetting; (2) trial and appellate counsel were ineffective for their failure to raise the constructive amendment of the fatally defective jury instructions; (3) newly discovered evidence tends to completely exonerate petitioner of all charges; (4) the conviction for felony murder cannot stand because of the merger doctrine; (5) counsel was ineffective in his closing arguments to the jury by conceding the guilt of his client; (6) trial counsel was ineffective because he "tricked and [cajoled]" petitioner into giving up his right to remain silent; (7) trial counsel was ineffective for failing to investigate or seek a continuance prior to trial on scientific evidence (regarding firearms); (8) trial counsel was ineffective for allowing a close friend and associate of the prosecutor's family to remain seated on the jury; (9) trial counsel was ineffective in failing to object to the judge's answer to a question asked by the jury; (10) trial counsel was ineffective for the failure to object to Instruction No. 14 (aiding and abetting); and (11) counsel was ineffective for failure to object to the trial court's constructively amending the information by instructing the jury on an aggravated burglary charge not set out in the information. 4. On post-conviction appeal, petitioner raised the following grounds: (1) the district court erred regarding petitioner's claim of new evidence by making incorrect legal and factual findings; and (2) the district court erred in finding trial counsel effective based on erroneous findings of fact and conclusions of law. 5. Petitioner's statement of factual history merely adopts "the facts as layed [sic] out in his direct appeal brief." That brief is included in the record and, although it is incomplete, contains a recitation of evidence that is not inconsistent with the facts set out in the Kansas Supreme Court's opinion.

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1107 0.3376 1197 2.9207 17/359-04:32:49.063 ON ON ON ON ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 1154 0.3520 1187 2.8963 17/359-04:34:19.066 ON ON ON ON ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 1155 0.3523 1179 2.8768 17/359-04:35:49.070 ON ON ON ON ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 1156 0.3526 1172 2.8597 17/359-04:37:19.074 ON ON ON ON ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 1157 0.3529 1168 2.8499 17/359-04:38:49.078 ON ON ON ON ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 1158 0.3532 1164 2.8402 17/359-04:40:19.082 ON ON ON ON ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 1158 0.3532 1161 2.8328 17/359-04:41:49.086 ON ON ON ON ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 1159 0.3535 1158 2.8255 17/359-04:43:19.090 ON ON ON ON ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 1160 0.3538 1156 2.8206 17/359-04:43:19.094 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1160 0.3538 1156 2.8206 17/359-04:44:49.094 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1160 0.3538 1155 2.8182 17/359-04:46:08.235 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1160 0.3538 1154 2.8158 17/359-04:47:49.098 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1161 0.3541 1152 2.8109 17/359-04:50:49.106 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1162 0.3544 1151 2.8084 17/359-04:52:19.110 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1162 0.3544 1150 2.8060 17/359-04:55:19.118 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1162 0.3544 1149 2.8036 17/359-04:58:19.130 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1162 0.3544 1149 2.8036 17/359-04:59:49.130 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1163 0.3547 1149 2.8036 17/359-05:01:19.134 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1162 0.3544 1149 2.8036 17/359-05:02:49.138 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1163 0.3547 1149 2.8036 17/359-05:08:49.154 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1163 0.3547 1148 2.8011 17/359-05:13:19.166 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF 1163 0.3547 1147 2.7987 17/359-05:13:19.170 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1163 0.3547 1147 2.7987 17/359-05:14:49.166 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1163 0.3547 1148 2.8011 17/359-05:22:19.186 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1163 0.3547 1147 2.7987 17/359-05:26:08.358 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1164 0.3550 1147 2.7987 17/359-05:26:49.198 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1163 0.3547 1148 2.8011 17/359-05:28:19.206 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1163 0.3547 1148 2.8011 17/359-05:29:49.206 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1163 0.3547 1149 2.8036 17/359-05:42:08.398 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1163 0.3547 1150 2.8060 17/359-05:43:19.246 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1163 0.3547 1150 2.8060 17/359-05:49:19.254 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1163 0.3547 1151 2.8084 17/359-05:52:19.262 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1163 0.3547 1150 2.8060 17/359-05:56:49.274 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1163 0.3547 1151 2.8084 17/359-05:58:08.415 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1163 0.3547 1150 2.8060 17/359-05:58:19.282 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1163 0.3547 1150 2.8060 17/359-06:01:19.286 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1164 0.3550 1150 2.8060 17/359-06:02:49.286 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1163 0.3547 1149 2.8036 17/359-06:04:19.294 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1164 0.3550 1149 2.8036 17/359-06:05:49.294 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1163 0.3547 1149 2.8036 17/359-06:06:08.435 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1164 0.3550 1149 2.8036 17/359-06:13:19.314 ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1164 0.3550 1148 2.8011 17/359-06:13:19.318 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1164 0.3550 1148 2.8011 17/359-06:14:49.318 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1164 0.3550 1149 2.8036 17/359-06:19:19.330 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1165 0.3553 1148 2.8011 17/359-06:28:19.358 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1165 0.3553 1148 2.8011 17/359-06:29:49.354 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1165 0.3553 1149 2.8036 17/359-06:34:19.366 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1166 0.3556 1148 2.8011 17/359-06:43:19.394 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1166 0.3556 1148 2.8011 17/359-06:44:49.394 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1166 0.3556 1149 2.8036 17/359-06:49:19.406 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1166 0.3556 1150 2.8060 17/359-06:50:49.406 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1167 0.3559 1150 2.8060 17/359-06:52:19.410 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1166 0.3556 1150 2.8060 17/359-06:55:19.418 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1167 0.3559 1149 2.8036 17/359-06:56:49.422 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1166 0.3556 1150 2.8060 17/359-06:58:19.426 ON ON ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF OFF OFF OFF OFF 1167 0.3559 1150 2.8060 17/359-06:58:19.430 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1167 0.3559 1150 2.8060 17/359-07:07:19.450 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1167 0.3559 1149 2.8036 17/359-07:08:49.450 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1123 0.3425 1148 2.8011 17/359-07:10:08.594 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1043 0.3181 1149 2.8036 17/359-07:10:19.458 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 1033 0.3151 1149 2.8036 17/359-07:11:49.458 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 918 0.2800 1151 2.8084 17/359-07:13:19.462 ON ON ON OFF ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 785 0.2394 1155 2.8182 17/359-07:13:19.466 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 785 0.2394 1155 2.8182 17/359-07:14:49.466 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 641 0.1955 1160 2.8304 17/359-07:16:19.470 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 498 0.1519 1165 2.8426 17/359-07:17:49.474 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 467 0.1424 1177 2.8719 17/359-07:18:08.614 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 467 0.1424 1179 2.8768 17/359-07:19:19.478 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 467 0.1424 1186 2.8938 17/359-07:20:49.482 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 467 0.1424 1193 2.9109 17/359-07:22:19.486 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 466 0.1421 1199 2.9256 17/359-07:23:49.490 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 466 0.1421 1204 2.9378 17/359-07:25:19.494 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 466 0.1421 1209 2.9500 17/359-07:26:08.634 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 465 0.1418 1211 2.9548 17/359-07:26:49.498 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 466 0.1421 1213 2.9597 17/359-07:28:19.502 ON ON ON ON ON ON ON OFF ON OFF ON OFF ON OFF ON OFF ON OFF OFF OFF 465 0.1418 1216 2.9670 17/359-07:28:19.506 ON ON ON OFF ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 465 0.1418 1216 2.9670 17/359-07:29:49.502 ON ON ON OFF ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 465 0.1418 1219 2.9744 17/359-07:31:19.510 ON ON ON OFF ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 465 0.1418 1222 2.9817 17/359-07:32:49.510 ON ON ON OFF ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 465 0.1418 1224 2.9866 17/359-07:34:08.654 ON ON ON OFF ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 465 0.1418 1226 2.9914 17/359-07:34:19.514 ON ON ON OFF ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 465 0.1418 1227 2.9939 17/359-07:35:49.518 ON ON ON OFF ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 465 0.1418 1229 2.9988 17/359-07:37:19.522 ON ON ON OFF ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 465 0.1418 1231 3.0036 17/359-07:38:49.526 ON ON ON OFF ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 465 0.1418 1232 3.0061 17/359-07:40:19.530 ON ON ON OFF ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 465 0.1418 1234 3.0110 17/359-07:41:49.534 ON ON ON OFF ON ON ON OFF ON ON ON ON ON OFF ON OFF ON OFF ON OFF 465 0.1418 1235

https://lasp.colorado.edu/maven/sdc/public/data/sites/site-20190821T223129/anc/eng/sasm1/sci_anc_sasm117_357_361.drf

United States v. Leeper | Eighth Circuit | 05-13-1992 | www.anylaw.com Research the case of United States v. Leeper, from the Eighth Circuit, 05-13-1992. AnyLaw is the FREE and Friendly legal research service that gives you unlimited access to massive amounts of valuable legal data. This case cites: United States v. Leeper 964 F.2d 751 (1992) | Cited 8 times | Eighth Circuit | May 13, 1992 HEANEY, Senior Circuit Judge. Lanny Leeper appeals his conviction for being a felon in possession of a firearm and his career offender sentence. We affirm. BACKGROUND On March 4, 1990, residents of Des Moines, Iowa alerted the police that an armed man had approached their home. By the time the police arrived at the residence, the man had fired three shots into the home and bled in a blue tow truck. In response, the officers radioed their colleagues to apprehend the driver of the vehicle. Soon thereafter, other officers spotted the truck and observed the driver of the truck sticking his arm out of the driver's side window and lifting it in an upward motion over the cab. The police pulled the truck over and arrested the driver, who identified himself as Lanny Leeper. In the meantime, a private citizen had noticed a firearm laying in the street, honked his horn to gain the officers' attention, and directed them to the firearm. One of the officers then recovered the firearm, a Colt .45, and noticed that it had scrape marks on it. The officers next proceeded to the residence where the investigation began. Once at the home, the officers observed three bullet holes in a rear window and numerous bullet holes in the home's interior. One of the officers also retrieved three empty shell casings from the back yard. A gunshot residue test of Leeper's hand revealed concentrations of antimony, barium, and lead, suggesting that Leeper had physical contact with a recently fired gun. Moreover, prints of Leeper's palm print matched those found on the Colt .45's magazine. Finally, the retrieved shell casings were compared with a test-fired cartridge from the Colt .45. Based on the casings' firing-pin impressions, a criminalist from the Iowa Division of Criminal Investigation concluded that the spent shell casings had been fired from the Colt .45. Based on this evidence (and on his prior record), a jury found Leeper guilty of being a felon in possession of a firearm in violation of 18 U.S.C. § 922(g). Pursuant to 18 U.S.C. § 924(e) and U.S.S.G. § 4B1.1, the district court sentenced Leeper to 30 years imprisonment. Leeper challenges his conviction and sentence. Discussion I. Mistrial Leeper seeks a new trial on two grounds. First, he contends that the trial court prejudiced the jury by orally instructing it that Leeper had been previously convicted of manslaughter. This disclosure violated the parties' stipulation that the jury would be informed only that Leeper had been "convicted of a crime punishable by imprisonment for a term exceeding one year." The trial court offered to orally correct the instruction, but Leeper declined. The court, however, conformed the written instructions submitted to the jury to the parties' stipulation, and throughout the entire trial the government honored the stipulation by never mentioning the nature of the predicate felony. Given these facts and the overwhelming nature of the evidence recited above, we are convinced that the inadvertent disclosure was harmless error. See Fed. R. Crim. Pro. 52(a). Leeper also argues that the trial court erred in refusing to let him use the magazine with his palm print on it for demonstration purposes. Leeper intended to demonstrate that the officers mishandled the Colt .45 and the magazine so as to taint the palm print evidence. The prosecution countered that using the magazine in this manner would destroy the prints already on it, thereby rendering this evidence useless if the case were retried. The parties then stipulated that the demonstration could be accomplished with the original gun and a substitute magazine. The court informed the jury of this arrangement, and Leeper performed the demonstration. Leeper now contends that the district court effectively denied him his right to confront the evidence against him. We find no merit in this argument. II. Sentence Leeper next attacks his career offender sentence. Finding that Leeper had been convicted of three violent felonies (manslaughter in 1977, terrorism in 1982, and robbery in 1983), the district court sentenced Leeper to 360 months as a career offender pursuant to 18 U.S.C. § 924(e) 1and U.S.S.G. § 4B1.1(A). 2Leeper contends that he should not have been sentenced as a career offender because neither his 1977 manslaughter nor his 1983 robbery convictions should be considered for enhancement purposes, and because his present conviction for being a felon in possession of 'a firearm is not a "crime of violence." 3 Leeper argues that his manslaughter conviction cannot be a predicate felony under 18 U.S.C. § 924(e) and U.S.S.G. § 4B1.1, because this crime did not require an element of intent. Neither of these provisions limit "crimes of violence" to intentional acts. 18 U.S.C. §§ 924(e)(2)(B)(i)-(ii) define "violent felony" as any crime punishable by imprisonment for a term exceeding one year that either "has as an element the use, attempted use, or threatened use of physical force against the person of another" or "involves conduct that presents a serious risk of physical injury to another." The guidelines deploy this same definition. See U.S.S.G. §§ 4B1.2(1)(i)-(ii). By definition, manslaughter means that someone has been killed. See Iowa Code §§ 707(4)-(5) (1977). Leeper was convicted because he shot someone through the head at close range and the victim eventually died. Thus, whether we focus on the elements of the crime or the offense's underlying facts, manslaughter qualifies as a crime of violence. See United States v. Wright, 1992 WL 26167 (8th Cir. 1992); see also U.S.S.G. § 4B1.2 n.2 (identifying manslaughter as a crime of violence). As for the robbery conviction, this court has already ruled that "robbery is a crime of violence under U.S.S.G. § 4B1.2(1)(i)." United States v. Wright, 957 F.2d 520 (8th Cir. 1992). This court concluded "because robbery cannot be committed without violence within the meaning of section 4B1.1, courts cannot examine the facts underlying each robbery." Id. (citations omitted). This precedent binds us. Thus, Leeper has the three prior felony convictions necessary to trigger 18 U.S.C. § 924(e). 4 Accordingly, we affirm Leeper's conviction and sentence. Disposition Affirmed. 1. Subsection (1) of this statute reads: In the case of a person who violates section 922(g) of this title and has three previous convictions . . . for a violent felony or a serious drug offense or both . . . [such person] shall be . . . imprisoned not less than fifteen years . . . . 2. Section 4B1.1 of the guidelines provides: A defendant is a career offender if (1) the defendant was at least eighteen years old at the time of instant offense, (2) the instant offense of conviction is a felony that is either a crime of violence or a controlled substance offense, and (3) the defendant has at least two prior felony convictions of either a crime of violence or a controlled substance offense. 3. Leeper also urges that the district court erred in refusing to depart downward from the minimum guidelines sentence of 360 months. We find this contention to be without merit.

https://www.anylaw.com/case/united-states-v-leeper/eighth-circuit/05-13-1992/gIeiP2YBTlTomsSB8FVh

Gold's Relative Strength and What it Means Technician Jordan Roy-Byrne of The Daily Gold charted gold's performance against foreign currencies, equities, oil, industrial metals, and bonds and shares his analysis and conclusions. COMMODITIES Gold's Relative Strength and What it Means Technician Jordan Roy-Byrne of The Daily Gold charted gold’s performance against foreign currencies, equities, oil, industrial metals, and bonds and shares his analysis and conclusions. Longtime readers of our editorials know that we are big fans ofintermarket analysisas well as ardent believers in the real price of gold (real POG) or relative gold. No, this has nothing to do with the paper market versus the physical market. The real POG is essentially the POG relative to other assets and markets. There are two reasons why we track this. First, gold priced against the other currencies (specifically the euro) has been a leading indicator for gold in US$ terms. Second, gold’s performance against commodities (in general) is a leading indicator for margins of the miners. Over the weekend, I reviewed gold’s performance in relative terms and it seems to have reasserted its uptrend. In the chart below we graph gold against six other markets (foreign currencies, equities, oil, industrial metals, and bonds). Note that gold is in a steady uptrend against each market with the exception of bonds. Gold priced in the inverse of the US$ basket is only 4% off its all-time high while relative to oil and industrial metals, it just reached a one-year high. Gold has turned down relative to bonds as they are close to testing their all-time high. Meanwhile, the yellow metal just matched a six-month high relative to the S&P 500. So why should we care about all of this? First, it tells us that gold is in a healthy bullish position because it’s trending higher against all major markets with the exception of bonds. In other words, gold is showing broad strength and is only being held back by the strength in bonds, which happens to be the largest market by a mile. Thus, when we see bonds soften, gold should have a shot to retest its recent high. |pagebreak| The strength in the real POG usually reflects economic contraction or deceleration. After all, if things were going well we’d expect equities and economically sensitive commodities to outperform gold. A rise in the real POG is a negative signal for the economy and asset markets. That in itself is a catalyst for central bank action, which gives liftoff to precious metals and also explains why the real POG is a trusty leading indicator. The current interpretation of the real POG bodes well for the underlying cyclical or intermediate term trend, which turned bullish in May. While we are here, here are a few quick thoughts on the gold and silver shares. In the chart below we plot the two support lines forGDX,GDXJ, andSIL. It appears these markets will test the lower support line. Short-term breadth indicators (courtesy of sentimentrader.com) show only 15% of gold stocks trading above their 10-day moving average and only 23% trading above their 50-day moving average. The last time both these ratios were beneath 20% was July, in which a tremendous rebound began. CHART3 To conclude, the real price of gold is trending bullish, which implies good times ahead for precious metals in the coming months. The poor outlook for oil and industrial prices is a good thing for gold and silver producers as their margins could expand even further in the quarters ahead.The correction that began at the end of September is likely within days of ending. Now, with mining equities trading off their highs, is the time to do your research and find the companies that will lead the next leg higher and outperform the gold stock sector. Jordan Roy-Byrne can be found at The Daily Gold .

https://www.moneyshow.com/articles/DAYTRADERS-29743/

Determinism - Wordsmith.org Re: Determinism jenny jenny # 209354 02/01/13 09:11 PM so write it offline in an editor (e.g., Notepad) and paste it in your little post box, viz.: Determinism From Wikipedia, the free encyclopedia This article is about the general notion of determinism in philosophy. For other uses, see Determinism (disambiguation). Not to be confused with Fatalism, Predeterminism, or Predictability. Determinism is a metaphysical philosophical position stating that for everything that happens there are conditions such that, given those conditions, nothing else could happen. "There are many determinisms, depending upon what pre-conditions are considered to be determinative of an event."[1] Determinism throughout the history of philosophy has sprung from diverse considerations, some of which overlap. Some forms of determinism can be tested empirically with ideas stemming from physics and the philosophy of physics. The opposite of determinism is some kind of indeterminism (otherwise called nondeterminism). Determinism is often contrasted with free will.[2] Determinism often is taken to mean simply causal determinism, that is, basing determinism upon the idea of cause-and-effect. It is the concept that events within a given paradigm are bound by causality in such a way that any state (of an object or event) is completely determined by prior states. This meaning can be distinguished from other varieties of determinism mentioned below. The introduction of "cause-and-effect" introduces unnecessary complications related to what is meant by a 'cause' and how the presence of a 'cause' might be established, the interpretation of which varies from one physical theory to another. These complications are avoided by a more general formulation based upon connections between 'events' supplied by a theory: "a theory is deterministic if, and only if, given its state variables for some initial period, the theory logically determines a unique set of values for those variables for any other period."[3] �Ernest Nagel, Alternative descriptions of physical state p. 292 This quote replaces the idea of 'cause-and-effect' with that of 'logical implication' according to one or another theory that connects events. In addition, an 'event' is related by the theory itself to formalized states described using the parameters defined by that theory. Thus, the details of interpretation are placed where they belong, fitted to the context in which the chosen theory applies. Other debates often concern the scope of determined systems, with some maintaining that the entire universe (or multiverse) is a single determinate system and others identifying other more limited determinate systems. For example, using the definition of physical determinism above, the limitations of a theory to some particular domain of experience also limits the associated definition of 'determinism' to that same domain. There are numerous historical debates involving many philosophical positions and varieties of determinism. They include debates concerning determinism and free will, technically denoted as compatibilistic (allowing the two to coexist) and incompatibilistic (denying their coexistence is a possibility). Determinism should not be confused with self-determination of human actions by reasons, motives, and desires. Determinism rarely requires that perfect prediction be practically possible � merely predictable in theory. Many philosophical theories of determinism frame themselves with the idea that reality follows a sort of predetermined path Causal determinism is "the idea that every event is necessitated by antecedent events and conditions together with the laws of nature".[4] However, causal determinism is a broad enough term to consider that "one's deliberations, choices, and actions will often be necessary links in the causal chain that brings something about. In other words, even though our deliberations, choices, and actions are themselves determined like everything else, it is still the case, according to causal determinism, that the occurrence or existence of yet other things depends upon our deliberating, choosing and acting in a certain way".[5] Causal determinism proposes that there is an unbroken chain of prior occurrences stretching back to the origin of the universe. The relation between events may not be specified, nor the origin of that universe. Causal determinists believe that there is nothing uncaused or self-caused. Historical determinism (a sort of path dependence) can also be synonymous with causal determinism. Nomological determinism (sometimes called 'scientific' determinism, although that is a misnomer) is the most common form of causal determinism. It is the notion that the past and the present dictate the future entirely and necessarily by rigid natural laws, that every occurrence results inevitably from prior events. Quantum mechanics and various interpretations thereof pose a serious challenge to this view. Nomological determinism is sometimes illustrated by the thought experiment of Laplace's demon.[6] Physical determinism holds holds that all physical events occur as described by physical laws.[7][8] Depending upon definitions, there is some room here for the view that not everything in the universe must be tied to some physical state, but that view is not usually emphasized by adherents of physical determinism because of the widely accepted scientific view that the operation of all physical systems (often unnecessarily taken to mean everything) can be explained entirely in physical terms, the assumed causal closure of physics. Necessitarianism is very related to the causal determinism described above. It is a metaphysical principle that denies all mere possibility; there is exactly one way for the world to be. Leucippus claimed there were no uncaused events, and that everything occurs for a reason and by necessity.[9] Predeterminism is the idea that all events are determined in advance.[10][11] The concept of predeterminism is often argued by invoking causal determinism, implying that there is an unbroken chain of prior occurrences stretching back to the origin of the universe. In the case of predeterminism, this chain of events has been pre-established, and human actions cannot interfere with the outcomes of this pre-established chain. Predeterminism can be used to mean such pre-established causal determinism, in which case it is categorised as a specific type of determinism.[10][12] It can also be used interchangeably with causal determinism - in the context of its capacity to determine future events.[10][13] Despite this, predeterminism is often considered as independent of causal determinism.[14][15] The term predeterminism is also frequently used in the context of biology and hereditary, in which case it represents a form of biological determinism.[16] Fatalism is normally distinguished from "determinism".[17] Fatalism is the idea that everything is fated to happen, so that humans have no control over their future. Fate has arbitrary power, and need not follow any causal or otherwise deterministic laws.[18] Types of Fatalism include hard theological determinism and the idea of predestination, where there is a God who determines all that humans will do. This may be accomplished either by knowing their actions in advance, via some form of omniscience[19] or by decreeing their actions in advance.[20] Theological determinism is a form of determinism which states that all events that happen are pre-ordained, or predestined to happen, by a monotheistic deity, or that they are destined to occur given its omniscience. Two forms of theological determinism exist, here referenced as strong and weak theological determinism.[21] The first one, strong theological determinism, is based on the concept of a creator deity dictating all events in history: "everything that happens has been predestined to happen by an omniscient, omnipotent divinity".[22] The second form, weak theological determinism, is based on the concept of divine foreknowledge - "because God's omniscience is perfect, what God knows about the future will inevitably happen, which means, consequently, that the future is already fixed".[23] There exist slight variations on the above categorisation. Some claim that theological determinism requires predestination of all events and outcomes by the divinity (i.e. they do not classify the weaker version as 'theological determinism' unless libertarian free will is assumed to be denied as a consequence), or that the weaker version does not constitute 'theological determinism' at all.[24] With respect to free will, "theological determinism is the thesis that God exists and has infallible knowledge of all true propositions including propositions about our future actions", more minimal criteria designed to encapsulate all forms of theological determinism.[25] Theological determinism can also be seen as a form of causal determinism, in which the antecedent conditions are the nature and will of God.[5] Logical determinism or Determinateness is the notion that all propositions, whether about the past, present, or future, are either true or false. Note that one can support Causal Determinism without necessarily supporting Logical Determinism and vice versa (depending on one's views on the nature of time, but also randomness). The problem of free will is especially salient now with Logical Determinism: how can choices be free, given that propositions about the future already have a truth value in the present (i.e. it is already determined as either true or false)? This is referred to as the problem of future contingents. Adequate determinism focuses on the fact that, even without a full understanding of microscopic physics, we can predict the distribution of 1000 coin tosses Often synonymous with Logical Determinism are the ideas behind Spatio-temporal Determinism or Eternalism: the view of special relativity. J. J. C. Smart, a proponent of this view, uses the term "tenselessness" to describe the simultaneous existence of past, present, and future. In physics, the "block universe" of Hermann Minkowski and Albert Einstein assumes that time is a fourth dimension (like the three spatial dimensions). In other words, all the other parts of time are real, like the city blocks up and down a street, although the order in which they appear depends on the driver (see Rietdijk�Putnam argument). Adequate determinism is the idea that quantum indeterminacy can be ignored for most macroscopic events. This is because of quantum decoherence. Random quantum events "average out" in the limit of large numbers of particles (where the laws of quantum mechanics asymptotically approach the laws of classical mechanics).[26] Stephen Hawking explains a similar idea: he says that the microscopic world of quantum mechanics is one of determined probabilities. That is, quantum effects rarely alter the predictions of classical mechanics, which are quite accurate (albeit still not perfectly certain) at larger scales.[27] Something as large as an animal cell, then, would be "adequately determined" (even in light of quantum indeterminacy). Nature and nurture interact in humans. A scientist looking at a sculpture after some time does not ask whether we are seeing the effects of the starting materials OR environmental influences. Although some of the above forms of determinism concern human behaviors and cognition, others frame themselves as an answer to the Nature or Nurture debate. They will suggest that one factor will entirely determine behavior. As scientific understanding has grown, however, the strongest versions of these theories have been widely rejected as a single cause fallacy.[28] In other words, the modern deterministic theories attempt to explain how the interaction of both nature and nurture is entirely predictable. The concept of heritability has been helpful to make this distinction. Biological determinism, sometimes called Genetic determinism, is the idea that each of our behaviors, beliefs, and desires are fixed by our genetic nature. Behaviorism is the idea that all behavior can be traced to specific causes�either environmental or reflexive. This Nurture-focused determinism was developed by John B. Watson and B. F. Skinner. Cultural determinism or social determinism is the nurture-focused theory that it is the culture in which we are raised that determines who we are. Environmental determinism is also known as climatic or geographical determinism. It holds the view that the physical environment, rather than social conditions, determines culture. Supporters often also support Behavioral determinism. Key proponents of this notion have included Ellen Churchill Semple, Ellsworth Huntington, Thomas Griffith Taylor and possibly Jared Diamond, although his status as an environmental determinist is debated.[29] A technological determinist might suggest that technology like the mobile phone is the greatest factor shaping human civilization. Other 'deterministic' theories actually seek only to highlight the importance of a particular factor in predicting the future. These theories often use the factor as a sort of guide or constraint on the future. They need not suppose that complete knowledge of that one factor would allow us to make perfect predictions. Psychological determinism can mean that humans must act according to reason, but it can also be synonymous with some sort of Psychological egoism. The latter is the view that humans will always act according to their perceived best interest. Linguistic determinism claims that our language determines (at least limits) the things we can think and say and thus know. The Sapir�Whorf hypothesis argues that individuals experience the world based on the grammatical structures they habitually use. Economic determinism is the theory which attributes primacy to the economic structure over politics in the development of human history. It is associated with the dialectical materialism of Karl Marx. Technological determinism is a reductionist theory that presumes that a society's technology drives the development of its social structure and cultural values. Media determinism, a subset of technological determinism, is a philosophical and sociological position which posits the power of the media to impact society. Two leading media determinists are the Canadian scholars Harold Innis and Marshall McLuhan. [snip]

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Insomnia Global Clinical Trials Review, H1, 2020 Insomnia Global Clinical Trials Review, H1, 2020 Insomnia Global Clinical Trials Review, H1, 2020 Summary GlobalData's clinical trial report, “Insomnia Global Clinical Trials Review, H1, 2020 provides - Market research report and industry analysis - 13025270 Insomnia Global Clinical Trials Review, H1, 2020 Best Price Guarantee Price from $2,500 Length 524 Pages Publisher GlobalData Published Date January, 2020 SKU GBDT16163618 Table of Contents Description Insomnia Global Clinical Trials Review, H1, 2020 SummaryGlobalData's clinical trial report, “Insomnia Global Clinical Trials Review, H1, 2020 provides an overview of Insomnia Clinical trials scenario. This report provides top line data relating to the clinical trials on Insomnia. Report includes an overview of trial numbers and their average enrollment in top countries conducted across the globe. The report offers coverage of disease clinical trials by region, country (G7 & E7), phase, trial status, end points status and sponsor type. Report also provides prominent drugs for in-progress trials (based on number of ongoing trials). GlobalData Clinical Trial Reports are generated using GlobalData’s proprietary database - Pharma eTrack Clinical trials database. Clinical trials are collated from 80+ different clinical trial registries, conferences, journals, news etc across the globe. Clinical trials database undergoes periodic update by dynamic process.The report enhances the decision making capabilities and helps to create an effective counter strategies to gain competitive advantage. Note:Certain sections in the report may be removed or altered based on the availability and relevance of data for the indicated disease. Scope The report provides a snapshot of the global clinical trials landscape Report provides top level data related to the clinical trials by Region, Country (G7 & E7), Trial Status, Trial Phase, Sponsor Type and End point status The report reviews top companies involved and enlists all trials (Trial title, Phase, and Status) pertaining to the company The report provides all the unaccomplished trials (Terminated, Suspended and Withdrawn) with reason for unaccomplishment The Report provides enrollment trends for the past five years Report provides latest news for the past three months Note: Certain sections in the report may be removed or altered based on the availability and relevance of data for the indicated disease. Reasons to buy Assists in formulating key business strategies with regards to investment Helps in identifying prominent locations for conducting clinical trials which saves time and cost Supports understanding of trials count and enrollment trends by country in global therapeutics market Aids in interpreting the success rates of clinical trials by providing a comparative scenario of completed and uncompleted (terminated, suspended or withdrawn) trials Facilitates clinical trial assessment of the indication on a global, regional and country level Note: Insomnia Therapeutics Clinical Trials, Europe, Top Five Countries, 2020* Insomnia Therapeutics Clinical Trials, North America, Top Countries, 2020* Insomnia Therapeutics Clinical Trials, Middle East and Africa, Top Countries, 2020* Insomnia Therapeutics Clinical Trials, Central and South America, Top Five Countries, 2020* Proportion of Insomnia to Central Nervous System Clinical Trials, G7 Countries (%), 2020* Insomnia Therapeutics, G7 Countries, Clinical Trials by Phase, 2020* Insomnia Therapeutics, G7 Countries, Clinical Trials by Trial Status, 2020* Proportion of Insomnia to Central Nervous System Clinical Trials, E7 Countries (%), 2020* Insomnia Therapeutics, E7 Countries, Clinical Trials by Phase, 2020* Insomnia Therapeutics, E7 Countries, Clinical Trials by Trial Status, 2020* Insomnia Therapeutics, Global, Clinical Trials by Phase, 2020* Insomnia Therapeutics, Global, Clinical Trials In Progress by Phase 2020* Insomnia Therapeutics, Global, Clinical Trials by Trial Status, 2020* Insomnia Therapeutics Clinical Trials, Global, by End Point Status, 2020* Insomnia Therapeutics Clinical Trials, Global, Average Enrollment Target Trends, 2014-2018 Insomnia Therapeutics Market, Global, Clinical Trials by Sponsor Type, 2020* Insomnia Therapeutics Clinical Trials, Global, Key Sponsors, 2020* Insomnia Therapeutics Clinical Trials, Global, Top Companies by Phase, 2020* Insomnia Therapeutics Clinical Trials, Global, Ongoing Clinical Trials by Prominent Drugs, 2020* List of Figures Insomnia Therapeutics, Global, Clinical Trials by Region, 2020* Insomnia Therapeutics, Global, Clinical Trials and Average Enrollment by Top Countries, 2020* Insomnia Therapeutics Clinical Trials, Asia-Pacific, Top Five Countries, 2020* Insomnia Therapeutics Clinical Trials, Europe, Top Five Countries, 2020* Insomnia Therapeutics Clinical Trials, North America, Top Countries, 2020* Insomnia Therapeutics Clinical Trials, Middle East and Africa, Top Countries, 2020* Insomnia Therapeutics Clinical Trials, Central and South America, Top Five Countries, 2020* Proportion of Insomnia to Central Nervous System Clinical Trials, G7 Countries (%), 2020* Insomnia Therapeutics, G7 Countries, Clinical Trials by Phase, 2020* Insomnia Therapeutics, G7 Countries, Clinical Trials by Trial Status, 2020* Proportion of Insomnia to Central Nervous System Clinical Trials, E7 Countries (%), 2020* Insomnia Therapeutics, E7 Countries, Clinical Trials by Phase, 2020* Insomnia Therapeutics, E7 Countries, Clinical Trials by Trial Status, 2020* Insomnia Therapeutics, Global, Clinical Trials by Phase, 2020* Insomnia Therapeutics, Global, Clinical Trials In Progress by Phase 2020* Insomnia Therapeutics, Global, Clinical Trials by Trial Status, 2020* Insomnia Therapeutics Clinical Trials, Global, by End Point Status, 2020* Insomnia Therapeutics Clinical Trials, Global, Average Enrollment Target Trends, 2014-2018 Insomnia Therapeutics Market, Global, Clinical Trials by Sponsor Type, 2020* Insomnia Therapeutics Clinical Trials, Global, Key Sponsors, 2020* Insomnia Therapeutics Clinical Trials, Global, Top Companies by Phase, 2020* Insomnia Therapeutics Clinical Trials, Global, Ongoing Clinical Trials by Prominent Drugs, 2020* Delirium Disease - Global Clinical Trials Review, H2, 2020 Diphtheria Disease - Global Clinical Trials Review, H2, 2020 Bronchitis Disease - Global Clinical Trials Review, H2, 2020 Aspergillosis Disease - Global Clinical Trials Review, H2, 2020 Hypertriglyceridemia Disease - Global Clinical Trials Review H2, 2020 13025271 Single User License: $2,500 Site License: $5,000 Global Site License: $7,500 Download our eBook: How to Succeed Using Market Research Learn how to effectively navigate the market research process to help guide your organization on the journey to success. 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Publications - Global Disability Innovation Hub The GDI Hub brings together academic excellence, innovative practice and co-creation; harnessing the power of technology for good. Type Journal Paper Themes Assistive & Accessible Technology Research Group Local Productions A review of innovation strategies and processes to improve access to AT: Looking ahead to open innovation ecosystems Catherine Holloway, Dafne Zuleima Morgado Ramirez,Tigmanshu Bhatnagar,Ben Oldfrey, Priya Morjaria, Soikat Ghosh Moulic, Ikenna D. Ebuenyi,Giulia Barbareschi,Fiona Meeks, Jessica Massie, Felipe Ramos-Barajas, Joanne McVeigh, Kyle Keane, George Torrens, P. V.M. Rao, Malcolm MacLachlan,Victoria Austin, Rainer Kattel, Cheryl D Metcalf & Srinivasan Sujatha It is essential to understand the strategies and processes which are deployed currently across the Assistive Technology (AT) space toward measuring innovation. The main aim of this paper is to identify functional innovation strategies and processes which are being or can be deployed in the AT space to increase access to AT globally. Assistive Technology The Official Journal of RESNA Abstract A review of innovation strategies and processes to improve access to AT: Looking ahead to open innovation ecosystems It is essential to understand the strategies and processes which are deployed currently across the Assistive Technology (AT) space toward measuring innovation. The main aim of this paper is to identify functional innovation strategies and processes which are being or can be deployed in the AT space to increase access to AT globally. We conducted a scoping review of innovation strategies and processes in peer-reviewed literature databases and complemented this by identifying case studies demonstrating innovation strategies. The review includes WHO world region, publication year, AT type and a sector analysis against the Systems-Market for Assistive and Related Technologies Framework. We analyzed the case studies and interviews using thematic analysis. We included 91 papers out of 3,127 after review along with 72 case studies. Our results showed that product innovations were more prevalent than provision or supply innovations across papers and case studies. Case studies yielded two themes: open innovation (OI); radical and disruptive innovation. Financial instruments which encourage OI are needed and we recommend pursuing OI for AT innovation. Embedding AT within larger societal missions will be key to success governments and investors need to understand what AT is and their translational socioeconomic value. Cite A review of innovation strategies and processes to improve access to AT: Looking ahead to open innovation ecosystems Catherine Holloway, Dafne Zuleima Morgado Ramirez, Tigmanshu Bhatnagar, Ben Oldfrey, Priya Morjaria, Soikat Ghosh Moulic, Ikenna D. Ebuenyi, Giulia Barbareschi, Fiona Meeks, Jessica Massie, Felipe Ramos-Barajas, Joanne McVeigh, Kyle Keane, George Torrens, P. V.M. Rao, Malcolm MacLachlan, Victoria Austin, Rainer Kattel, Cheryl D Metcalf & Srinivasan Sujatha (2021) A review of innovation strategies and processes to improve access to AT: Looking ahead to open innovation ecosystems, Assistive Technology, 33:sup1, 68-86, DOI:10.1080/10400435.2021.1970653 A review of innovation strategies and processes to improve access to AT: Looking ahead to open innovation ecosystems Type Journal Paper Themes Assistive & Accessible Technology Research Group Local Productions Additive manufacturing techniques for smart prosthetic liners B Oldfrey, A Tchorzewska, R Jackson, M Croysdale, R Loureiro,C Holloway, M Miodownik Elastomeric liners are commonly worn between socket and limb by prosthetic wearers. This is due to their superior skin adhesion, load distribution and their ability to form a seal. Laboratory tests suggest that elastomeric liners allow reduced shear stress on the skin and give a higher cushioning effect on bony prominences, since they are soft in compression, and similar to biological tissues [1]. However, they also increase perspiration reducing hygiene and increasing skin irritations. Prosthetic users in general face a myriad of dermatological problems associated with lower limb prosthesis such as ulcers, cysts, and contact dermatitis, which are exacerbated by the closed environment of a fitted socket where perspiration is trapped and bacteria can proliferate [2]. Medical Engineering & Physics; 2021 Abstract Additive manufacturing techniques for smart prosthetic liners Elastomeric liners are commonly worn between the prosthetic socket and the limb. A number of improvements to the state of the art of liner technology are required to address outstanding problems. A liner that conforms to the residuum more accurately, may improve the skin health at the stump-socket interface. Previous work has shown that for effective thermal management of the socket environment, an active heat removal system is required, yet this is not available. Volume tracking of the stump could be used as a diagnostic tool for looking at the changes that occur across the day for all users, which depend on activity level, position, and the interaction forces of the prosthetic socket with the limb. We believe that it would be advantageous to embed these devices into a smart liner, which could be replaced and repaired more easily than the highly costly and labour-intensive custom-made socket. This paper presents the work to develop these capabilities in soft material technology, with: the development of a printable nanocomposite stretch sensor system; a low-cost digital method for casting bespoke prosthetic liners; a liner with an embedded stretch sensor for growth / volume tracking; a model liner with an embedded active cooling system. Cite Additive manufacturing techniques for smart prosthetic liners , Additive manufacturing techniques for smart prosthetic liners, Medical Engineering & Physics, Volume 87, 2021, Pages 45-55, ISSN 1350-4533,https://doi.org/10.1016/j.mede.... Additive manufacturing techniques for smart prosthetic liners Type Journal Paper Themes Assistive & Accessible Technology Applying market shaping approaches to increase access to assistive technology in low- and middle-income countries Margaret Savage, Sarah Albala, Frederic Seghers, Rainer Katte, Cynthia Liao, Mathilde Chaudron, Novia Afdhila Development outcomes are inextricably linked to the health of the marketplace that delivers products and services to people in low- and middle- income countries (LMIC). Shortcomings in the market for assistive technology (AT) contribute to low access in LMIC. Market shaping is aimed at improving a market’s specific outcomes, such as access to high quality, affordable AT, by targeting the root causes of these shortcomings. The paper summarizes the findings of a market and sector analysis that was conducted under the UK aid funded AT2030 programme and aims to discuss how market shaping can help more people gain access to the AT that they need and what are the best mechanisms to unlock markets and commercial opportunity in LMICs. Assistive Technology The Official Journal of RESNA; 2021 Abstract Applying market shaping approaches to increase access to assistive technology in low- and middle-income countries Development outcomes are inextricably linked to the health of the marketplace that delivers products and services to people in low- and middle-income countries (LMIC). Shortcomings in the market for assistive technology (AT) contribute to low access in LMIC. Market shaping is aimed at improving a market’s specific outcomes, such as access to high quality, affordable AT, by targeting the root causes of these shortcomings. The paper summarizes the findings of market analyses conducted under the UK aid funded AT2030 programme in support of ATscale and aims to discuss how market shaping can help more people gain access to the AT that they need and what are the best mechanisms to unlock markets and commercial opportunity in LMICs. The paper also explores how market shaping for AT markets could be part of a mission-oriented approach AT policy. A mission-oriented approach can help accelerate progress toward a common objective among stakeholders, at country or global level. While market-shaping activities direct the outcomes of the market toward a specific end goal, such as access to quality, affordable products and services, missions are more comprehensive and include other policy interventions and stakeholder collaborations in order to create a robust and sustainable structure. Cite Applying market shaping approaches to increase access to assistive technology in low- and middle-income countries Margaret Savage, Sarah Albala, Frederic Seghers, Rainer Kattel, Cynthia Liao, Mathilde Chaudron & Novia Afdhila (2021) Applying market shaping approaches to increase access to assistive technology in low- and middle-income countries, Assistive Technology, 33:sup1, 124-135, DOI: 10.1080/10400435.2021.1991050 Applying market shaping approaches to increase access to assistive technology in low- and middle-income countries Type Journal Paper Themes Assistive & Accessible Technology Assistive technology access in longitudinal datasets: a global review Jamie Danemayer, Sophie Mitra,Cathy Holloway, Shereen Hussein A person’s access to assistive products such as hearing aids, wheelchairs, and glasses, is an essential part of their ability to age in a healthy way. But, according to the World Health Organization, a staggering 90% of people who need assistive products worldwide, do not have access to them. In many instances access is limited or simply non-existent. This is often due to assistive products being too expensive, demand outweighing supply, not always being suitable to use in different environments, or even the lack of availability of trained providers. In such circumstances, people are more likely to age ‘unhealthily’ if they do not have access to assistive products that are designed to support their day to day functioning and independence. International Journal of Population Data Science; 2023 Abstract Assistive technology access in longitudinal datasets: a global review Functional limitations become more prevalent as populations age, emphasising an increasingly urgent need for assistive technology (AT). Critical to meeting this need trajectory is understanding AT access in older ages. Yet few publications examine this from a longitudinal perspective. This review aims to identify and collate what data exist globally, seeking all population-based cohorts and repeated cross-sectional surveys through the Maelstrom Research Catalogue (searched May 10, 2022) and the Disability Data Report (published 2022), respectively. Datasets incorporating functional limitations modules and question(s) dedicated to AT, with a wave of data collection since 2009, were included. Of 81 cohorts and 202 surveys identified, 47 and 62 meet inclusion criteria, respectively. Over 40% of cohorts were drawn from high-income countries which have already experienced significant population ageing. Cohorts often exclude participants based on pre-existing support needs. For surveys, Africa is the most represented region (40%). Globally, 73% of waves were conducted since 2016. 'Use' is the most collected AT access indicator (69% of cohorts and 85% of surveys). Glasses (78%) and hearing aids (77%) are the most represented AT. While gaps in data coverage and representation are significant, collating existing datasets highlights current opportunities for analyses and methods for improving data collection across the sector. Assistive technology access in longitudinal datasets: a global review Danemayer, J., Mitra, S., Holloway, C. and Hussein, S. (2023) “Assistive technology access in longitudinal datasets: a global review”, International Journal of Population Data Science, 8(1). doi: 10.23889/ijpds.v8i1.1901. Assistive technology access in longitudinal datasets: a global review Type Report Themes Assistive & Accessible Technology Assistive Technology Changes Lives: an assessment of AT need and capacity in England Vicki Austin,Dilisha Patel,Jamie Danemayer,Kate Mattick,Anna Landre,Marketa Smitova,Maryam Bandukda, Aoife Healy, Nachiappan Chockalingam,Diane Bell,Cathy Holloway This report was prepared by Global Disability Innovation (GDI) Hub for the Disability Unit in the Cabinet Office His Majesty’s Government (HMG). The report presents findings from a Country Capacity Assessment (CCA) of AT access in England. Findings illustrate a complex state of AT in England. While delivery systems tend to provide quality products that have a strong, positive impact on people’s lives, processes are often slow and stressful for users and providers alike. Startlingly, there is also an AT access gap of 31% of disabled people not having the assistive products they need to flourish, thrive, or even participate in daily life. The Cabinet Office; 2023 Abstract Assistive Technology Changes Lives: an assessment of AT need and capacity in England The aim of this research was to undertake a Country Capacity Assessment (CCA) to inform a more integrated approach to Assistive Technology (AT) provision in England. The results aim to support policymakers in identifying actions to strengthen service delivery to better meet disabled people’s needs, improving outcomes for AT users and reducing inefficiencies in the current approach. The research was undertaken from November 2022 to March 2023 and led by the Global Disability Innovation (GDI) Hub, which is the World Health Organization (WHO) Global Collaborating Centre on AT access, using WHO tools in the Assistive Technology Assessment (ATA) suite. Cite Assistive Technology Changes Lives: an assessment of AT need and capacity in England Suggested Citation: Austin, V, Patel, D, Danemayer, J, Mattick, K, Landre, A, Smitova, M, Bandukda, M, Healy, A, Chockalingam, N, Bell D, and Holloway, C; Assistive Technology Changes Lives: an assessment of AT need and capacity in England; Cabinet Office, HMG; 2023 Assistive Technology Changes Lives: an assessment of AT need and capacity in England Type Journal Paper Themes Assistive & Accessible Technology Assistive Technology Use and Provision During COVID19: Results From a Rapid Global Survey Emma M Smith, Maria Luisa Toro Hernandez, Ikenna D Ebuenyi, Elena V Syurina,Giulia Barbareschi, Krista L Best,Jamie Danemayer,Ben Oldfrey, Nuha Ibrahim,Catherine Holloway, Malcolm MacLachlan The coronavirus disease 2019 (COVID-19) pandemic has impacted all segments of society, but it has posed particular challenges for the inclusion of persons with disabilities, those with chronic illness and older people regarding their participation in daily life. These groups often benefit from assistive technology (AT) and so it is important to understand how use of AT may be affected by or may help to mitigate the impacts of COVID-19.Objective:The objectives of this study were to explore the how AT use and provision have been affected during the initial stages of the COVID-19 pandemic, and how AT policies and systems may be made more resilient based on lessons learned during this global crisis. International Journal of Health Policy and Management; 2020 Abstract Assistive Technology Use and Provision During COVID19: Results From a Rapid Global Survey Abstract Background:The coronavirus disease 2019 (COVID-19) pandemic has impacted all segments of society, but it has posed particular challenges for the inclusion of persons with disabilities, those with chronic illness and older people regarding their participation in daily life. These groups often benefit from assistive technology (AT) and so it is important to understand how use of AT may be affected by or may help to mitigate the impacts of COVID-19.Objective:The objectives of this study were to explore the how AT use and provision have been affected during the initial stages of the COVID-19 pandemic, and how AT policies and systems may be made more resilient based on lessons learned during this global crisis. Methods:This study was a rapid, international online qualitative survey in the 6 United Nations (UN) languages (English, French, Spanish, Russian, Arabic, Mandarin Chinese) facilitated by extant World Health Organization (WHO) and International Disability Alliance networks. Themes and subthemes of the qualitative responses were identified using Braun and Clarke's 6-phase analysis. Results:Four primary themes were identified in in the data: Disruption of Services, Insufficient Emergency Preparedness, Limitations in Existing Technology, and Inadequate Policies and Systems. Subthemes were identified within each theme, including subthemes related to developing resilience in AT systems, based on learning from the pandemic. Conclusion:COVID-19 has disrupted the delivery of AT services, primarily due to infection control measures resulting in lack of provider availability and diminished one-to-one services. This study identified a need for stronger user-centred development of funding policies and infrastructures that are more sustainable and resilient, best practices for remote service delivery, robust and accessible tools and systems, and increased capacity of clients, caregivers, and clinicians to respond to pandemic and other crisis situations. Cite Assistive Technology Use and Provision During COVID19: Results From a Rapid Global Survey Smith EM, Toro Hernandez ML, Ebuenyi ID, Syurina EV, Barbareschi G, Best KL, Danemayer J, Oldfrey B, Ibrahim N, Holloway C, MacLachlan M. Assistive Technology Use and Provision During COVID-19: Results From a Rapid Global Survey. Int J Health Policy Manag. 2022 Jun 1;11(6):747-756. doi: 10.34172/ijhpm.2020.210. PMID: 33201656; Assistive Technology Use and Provision During COVID19: Results From a Rapid Global Survey

https://www.disabilityinnovation.com/publications?order=title&type=workshop+journal-paper+report+phd+book

The Immunology of Posttransplant CMV Infection | Request PDF Request PDF | The Immunology of Posttransplant CMV Infection | The immune response to cytomegalovirus (CMV) infection is highly complex, including humoral, cellular, innate, and adaptive immune responses.... | Find, read and cite all the research you need on ResearchGate March 2016 Transplantation 100(Suppl 3):S11-S18 DOI: 10.1097/TP.0000000000001095 Authors: Javier Carbone Hospital Universitario Puerta de Hierro-Majadahonda Request full-text Download citation Copy link Link copied Request full-text Download citation Copy link Link copied To read the full-text of this research, you can request a copy directly from the author. Citations (47) References (77) Abstract The immune response to cytomegalovirus (CMV) infection is highly complex, including humoral, cellular, innate, and adaptive immune responses. Detection of CMV by the innate immune system triggers production of type I IFNs and inflammatory cytokines which initiate cellular and humoral responses that are critical during the early viremic phase of CMV infection. Sustained control of CMV infection is largely accounted for by cellular immunity, involving various T-cell and B-cell subsets. In solid organ transplant patients, global suppression of innate and adaptive immunities by immunosuppressive agents limits immunological defense, including inhibition of natural killer cell activity with ongoing lowering of Ig levels and CMV-specific antibody titers. This is coupled with a short-term suppression of CMV-specific T cells, the extent and duration of which can predict risk of progression to CMV viremia. CMV immunoglobulin (CMVIG) preparations have the potential to exert immunomodulatory effects as well as providing passive immunization. Specific CMVIG antibodies and virus neutralization might be enhanced by modulation of dendritic cell activity and by a decrease in T-cell activation, effects which are of importance during the initial phase of infection. In summary, the role of CMVIG in reconstituting specific anti-CMV antibodies may be enhanced by some degree of modulation of the innate and adaptive immune responses, which could help to control some of the direct and indirect effects of CMV infection. Discover the world's research 25+ million members 160+ million publication pages 2.3+ billion citations Join for free No full-text available To read the full-text of this research, you can request a copy directly from the author. Request full-text PDF Citations (47) References (77) ... After entering the body, CMV is initially detected by the innate immune system via Toll-like receptors and Natural killer (NK) cells initiating a humoral response [3]. However, long-term control of CMV is predominantly through cell-mediated immunity [4, 5] . CMV remains latent in peripheral blood leukocytes and bone marrow cells of hosts. ... ... CMV retinitis is an ocular opportunistic infection and a leading cause of ocular morbidity associated with AIDS. It is categorized as an AIDS-defining illness and is usually associated with CD 4 counts < 50 cells/ mm 3 in patients with HIV [3, 5] . However, apart from AIDS, even malignancies and chemotherapies that target T cells are known to increase the risk of CMV disease [3][4][5]. ... ... It is categorized as an AIDS-defining illness and is usually associated with CD 4 counts < 50 cells/ mm 3 in patients with HIV [3,5]. However, apart from AIDS, even malignancies and chemotherapies that target T cells are known to increase the risk of CMV disease [3][4] [5] . ... Cytomegalovirus retinitis in patients of non-Hodgkin’s lymphoma: clinical presentations and outcomes Article Full-text available Oct 2021 Shashwat Behera Subhakar Reddy Mudit Tyagi Hitesh Agrawal Background Cytomegalovirus (CMV) retinitis in patients with Non-Hodgkin’s Lymphoma (NHL) can occur even in the presence of high CD 4 counts and can behave differently when compared to CMV retinitis in human immunodeficiency (HIV) patients. It, therefore, becomes important to understand its varied presentations and the challenges in management of these cases. The aim of this study was to analyse the various patterns of presentations and outcomes of CMV Retinitis in patients with NHL. Study design A retrospective chart review of seven eyes of four patients of NHL presenting with CMV retinitis between June 2017 and May 2020 was done. Methods Clinical patterns of CMV Retinitis, CD4 counts at the time of presentation and the duration of treatment along with recurrences and time for recurrence of retinitis were assessed. Results Granular or indolent retinitis (6 out of 7 eyes) was the commonest form of CMV retinitis in patients of NHL. Three patients had a presenting CD4 count above 150 cells/mm³ and none of them were below 50 cells/mm³. Floaters were the commonest presenting complaint. All patients had vitritis and majority of the patients (3 out of 4) had anterior chamber (AC) inflammation. Two out of the 4 patients had a recurrence (mean time 33.8 days) after stopping the maintenance phase of ganciclovir and one patient had significant myelosuppression related to oral valganciclovir which required discontinuation of the drug. Conclusion CMV retinitis in NHL patients is usually of an indolent or granular type and can occur even in the presence of high CD4 counts as compared to patients with HIV. These patients may require a long term maintenance in view of frequent recurrences after discontinuation of treatment. View Show abstract ... After entering the body, CMV is initially detected by the innate immune system via Toll-like receptors and Natural killer (NK) cells initiating a humoral response [3]. However, long-term control of CMV is predominantly through cell-mediated immunity [4] [5] . CMV remains latent in peripheral blood leukocytes and bone marrow cells of hosts. ... ... CMV retinitis is an ocular opportunistic infection and a leading cause of ocular morbidity associated with AIDS. It is categorized as an AIDS-de ning illness and is usually associated with CD 4 counts < 50 cells/ mm 3 in patients with HIV [3, 5] . However, apart from AIDS, even malignancies and chemotherapies that target T cells are known to increase the risk of CMV disease [3][4][5]. ... ... It is categorized as an AIDS-de ning illness and is usually associated with CD 4 counts < 50 cells/ mm 3 in patients with HIV [3,5]. However, apart from AIDS, even malignancies and chemotherapies that target T cells are known to increase the risk of CMV disease [3][4] [5] . ... Cytomegalovirus Retinitis in Patients of Non-Hodgkin’s Lymphoma: Clinical Presentations and Outcomes Preprint Full-text available Jun 2021 Subhakar Reddy Mudit Tyagi Shaswat Behera Hitesh Agrawal Background Cytomegalovirus (CMV) retinitis in patients with Non-Hodgkin’s Lymphoma (NHL) can occur even in presence of high CD 4 counts and can behave differently when compared to CMV retinitis in human immunodeficiency (HIV) patients. It, therefore, becomes important to understand its varied presentations and challenges in the management of these cases. The aim of this study was to analyse the various patterns of presentations and outcomes of CMV Retinitis in patients with NHLStudy designA retrospective chart review of seven eyes of four patients of NHL presenting with CMV retinitis between June 2017 and May 2020 was done. Methods Clinical patterns of CMV Retinitis, the CD4 counts at the time of presentation and the duration of treatment along with recurrences and time for recurrence of retinitis were assessed.ResultsGranular or indolent retinitis (6 out of 7 eyes) was the commonest form of CMV retinitis in patients of NHL. Three patients had a presenting CD4 count above 150 cells/mm ³ and none of them were below 50 cells/mm ³ . Floaters were the commonest presenting complaint. All patients had vitritis and majority of the patients (3 out of 4) had anterior chamber (AC) inflammation. Two out of the 4 patients had a recurrence (mean time 33.8 days) after stopping the maintenance phase of ganciclovir and one patient had significant myelosuppression related to oral valganciclovir which required discontinuation of the drug. ConclusionCMV retinitis in NHL patients is usually of an indolent or granular type and can occur even in the presence of high CD4 counts as compared to patients with HIV. These patients may require a long term maintenance in view of frequent recurrences after discontinuation of treatment View Show abstract ... Although tolerability and antiviral efficacies of CMVIg have clearly been confirmed in the past [13], its use is currently not a recommended standard for antiviral prophylaxis or treatment in the LT setting [14]. In recent years, there seems to be growing suggestive evidence that, apart from antiviral activities, CMVIg may provide anti-inflammatory and immunoregulatory properties that could be valuable to counteract CMV-related immune reactions [12,[15][16] [17] . We hypothesized that critically ill liver recipients, who are particularly threatened by early inflammatory events, might benefit from these positive immunomodulatory capabilities. ... ... MELD score at LT). Diagnosis and grading (grades 1-3) of ACLF and related MOF were based on the European Association for the Study of the Liver-Chronic Liver Failure (EASL-CLIF) Consortium consensus criteria. The respective CLIF-C organ failure (OF) score (range: [11][12][13][14][15][16] [17] [18] at LT was calculated for each patient [18]. Preoperative serum lactate levels (normal range: <1.8 mmol/L) were used for describing severity of critical illness and tissue hypoxia [19,20], whereas values of C-reactive protein (CRP; normal range: <0.5 mg/dL), interleukin-6 (IL-6; normal range: <7 pg/mL) and procalcitonin (PCT; normal range: <0.1 ng/mL) reflected extent of systemic inflammatory response reaction prior to LT [21]. ... ... Treatment was well-tolerated in all cases, without need of dose reduction. Median total number of CMVIg doses/patient was 7 (range: [7][8][9][10][11][12][13][14][15][16] [17] [18][19][20]. The incidence of intra-abdominal infections was significantly higher in the non-CMVIg cohort, while rates of systemic bacterial infections were comparable (Table 2). ... Prophylactic Anti-Cytomegalovirus Hyperimmunoglobulin in Critically Ill Liver Transplant Patients: Impact on Early Immunology and Survival Article Full-text available Feb 2020 Arno Kornberg Ulrike Witt Jennifer Kornberg Katharina Thrum Background: Anti-cytomegalovirus hyperimmunoglobulin (CMVIg) was shown to provide beneficial immunodulatory properties beyond antiviral efficacies. The aim of this retrospective study was to assess the impact of prophylactic CMVIg treatment on early outcome following liver transplantation (LT) in critically ill patients. Methods: Forty-three cirrhotic patients requiring pre-LT intensive care due to multiorgan failure were analyzed. Twenty-eight patients with enhanced CMV risk (D+/R+; D+/R-; D-/R+) received prophylactic CMVIg for a minimum of 7 days, while 15 patients (D-/R-) did not. Results: Post-transplantation rates of intra-abdominal infections (28% vs. 61.1%; p = 0.03), Epstein-Barr virus infections (0% vs. 33.3%; p = 0.034), allograft rejections (0% vs. 22.2%; p = 0.013) and sepsis-related mortality (4% vs. 27.8%; p = 0.026) were significantly lower, whereas incidence of CMV infections (4% vs. 22.2%; p = 0.066) tended to be lower in the CMVIg subset. In multivariate analysis, only pretransplant elevated serum lactate level (hazard ratio = 34.63; p = 0.009) and absence of CMVIg therapy (hazard ratio = 21.76; p = 0.023) were identified as independent promoters of 3-month mortality. Conclusion: Prophylactic treatment with CMVIg reduces predisposition for severe immunological and septic events and, thereby, early mortality in critically ill liver recipients. View Show abstract ... Several studies have focused on the efficacy of hyperimmunoglobulin (HIG) to prevent maternal-fetal transmission with disparate results [9][10][11][12][13][14][15][16]. Despite the interest of hyperimmune immunoglobulins in the preventive treatment of transplanted patients [17, 18] and the efficacy of the molecule in the prevention of congenital CMV infection in published retrospective studies, anti-CMV antibodies (Cytogam ® and Cytotect ® ) have not proven their efficacy in the only two randomized trials published to date on the subject [9,13]. Differences in the recruitment of subjects based on the time of seroconversion, HIG concentration, and frequency of treatment administration determine the efficacy [10,19]. ... ... Thus, understanding the HIG mechanism of action remains an important goal. Innate and adaptive immune responses could be mediated by the immunomodulatory effects of anti-CMV immunoglobulin (CMVIG) preparations that would also provide passive immunization [17] and could help to control some of the direct and indirect effects of CMV infection. However, these preparations are not necessarily identical in terms of avidity, neutralizing potential, specific anti-CMV IgG content, or immunomodulation. ... Potential of Anti-CMV Immunoglobulin Cytotect CP® In Vitro and Ex Vivo in a First-Trimester Placenta Model Article Full-text available Mar 2022 Perrine Coste Mazeau Chloé Jacquet Clotilde Muller Sophie Alain Background: Congenital CMV infection is the leading cause of neonatal neurological deficit. We herein studied in vitro and ex vivo the potential of the hyperimmune globulin Cytotect CP® (Biotest, Germany) for congenital infection prevention and treatment. Methods: In vitro neutralization assays were conducted in fibroblasts and retinal epithelial cells on the CMV strains TB40/E and VHL/E to determine the 50% and 90% neutralizing doses (ND50 and ND90). The toxicity was assessed by measuring LDH release. Ex vivo assays were conducted in first-trimester villi explants with the TB40/E strain, namely, neutralization assays, the prevention of villi infection, and the inhibition of viral replication in infected villi. Viability was assessed by β-HCG quantification in supernatants. Results: The in vitro neutralization tests showed that Cytotect CP®® inhibits the development of infection foci (DN50: 0.011-0.014 U/mL for VHL/E and 0.032-0.033 U/mL for TB40E) without any toxicity. In the ex vivo neutralization assays, the DN50 were 0.011 U/mL on day 7 and 0.093 U/mL on day 14. For the prevention of villi infection, the EC50 was 0.024 U/mL on day 7. Cytotect-CP® did not inhibit viral growth in infected villi. No impact on villi viability was observed. Conclusions: These results sustained that Cytotect CP® has the potential to prevent CMV congenital infection. View Show abstract ... The primary function of CMVIG is to provide passive immunity by neutralizing circulating CMV particles and facilitating their elimination [17]. CMVIG also exerts enhancing and suppressive immunomodulatory functions that might contribute to the control of the direct and indirect effects of post-transplant CMV infection [16] [17][18]61]. Immunocompromised patients do not produce the cellular and humoral immune responses that would typically defend against CMV infection or reactivation. ... ... Immunocompromised patients do not produce the cellular and humoral immune responses that would typically defend against CMV infection or reactivation. Immunomodulatory actions proposed for human CMVIG include the suppression of functional dendritic cell maturation and the inhibition of T-cell proliferation, potentially leading to lower rates of transplant rejection [16] . Some reports have suggested that CMVIG may not only be useful as prophylaxis but also as rescue therapy upon the detection of CMV infection, but the current analysis focused on studies evaluating the labeled use in prophylaxis, in which CMVIG prophylaxis was started around the day of transplantation. ... Effectiveness of Prophylactic Human Cytomegalovirus Hyperimmunoglobulin in Preventing Cytomegalovirus Infection following Transplantation: A Systematic Review and Meta-Analysis Article Full-text available Mar 2022 Markus J Barten Fausto Baldanti Alexander Staus Andreas Zuckermann Cytomegalovirus (CMV) is a common infection occurring in patients undergoing solid organ transplantation (SOT) or hematopoietic stem cell transplantation (HSCT). CMV-specific hyperimmunoglobulin (CMVIG) has been used for the past four decades and is typically administered either prophylactically or pre-emptively. The present meta-analysis evaluated CMV infection rates in SOT patients who received prophylactic CMVIG. PubMed and the Cochrane Library were searched for studies published up to October 2021. The primary endpoint was CMV infection rate. Thirty-two SOT studies were identified (n = 1521 CMVIG-treated and n = 1196 controls). Prophylactic CMVIG treatment was often associated with a lower risk of CMV infection in transplant recipients. The average CMV infection rate was 35.8% (95% confidence interval [CI]: 33.4–38.2%) in patients treated prophylactically with CMVIG and 41.4% (95% CI: 38.6–44.2%) in the control group not receiving CMVIG (p = 0.003). Similar results were observed in analyses limited to publications evaluating currently available CMVIG products (Cytotect CP and Cytogam; p < 0.001). In combination with the established safety profile for CMVIG, these results suggest that prophylactic CMVIG treatment in patients undergoing solid organ transplantation may be beneficial, particularly in those at high risk of CMV infection or disease. View Show abstract ... 18 In addition, CMVIG exerts enhancing and suppressive immunomodulatory functions that might help control some of the direct and indirect effects of posttransplant CMV infection. 18,[44][45][46][47][48][49] [50] [51] However, to date, the immunomodulatory properties of CMVIG remain poorly characterized. ... ... Second, this study aimed at investigating early events following treatment with CMVIG and thus did not consider the late antiinflammatory effects associated with CMVIG that contribute to reduction of organ rejection after transplantation. 18,[48][49] [50] [51]73 Finally, the characterization of the stimulatory effect of CMVIG on innate immunity conducted in CMV-seropositive individuals by intracellular staining and flow cytometry could not be compared with that in CMV-seronegative subjects. ... Stimulatory Effect of CMV Immunoglobulin on Innate Immunity and on the Immunogenicity of CMV Antigens Article Full-text available Oct 2021 Ludwig Deml Christian M. Hüber Sascha Barabas Paolo Grossi Background: Cytomegalovirus (CMV) immunoglobulin (CMVIG) is used for the prophylaxis of CMV infection after transplantation. Beyond providing passive CMV-specific immunity, CMVIG exerts enhancing and suppressive immunomodulatory functions. Although the anti-inflammatory activities of CMVIG have been extensively documented, its immunostimulatory activities remain poorly characterized. Methods: This exploratory study analyzed the capacity of CMVIG to modulate cell-mediated innate and adaptive immunities in vitro on freshly isolated peripheral blood mononuclear cells (PBMCs) of CMV-seropositive and -seronegative healthy individuals, using interferon-γ (IFN-γ) enzyme-linked immunospot and intracellular cytokine staining assays. Results: We showed that CMVIG treatment increases the number of IFN-γ-secreting PBMCs of both CMV-seronegative and -seropositive individuals, indicating a global stimulatory effect on innate immune cells. Indeed, CMVIG significantly increased the frequency of natural killer cells producing the T helper cell 1-type cytokines tumor necrosis factor and IFN-γ. This was associated with the induction of interleukin-12-expressing monocytes and the activation of cluster of differentiation (CD) 4+ and CD8+ T cells, as measured by the expression of tumor necrosis factor and IFN-γ. Interestingly, stimulation of PBMCs from CMV-seropositive subjects with CMVIG-opsonized CMV antigens (phosphoprotein 65, CMV lysate) enhanced CD4+ and CD8+ T-cell activation, suggesting that CMVIG promotes the immunogenicity of CMV antigens. Conclusions: Our data demonstrate that CMVIG can stimulate effector cells of both innate and adaptive immunities and promote the immunogenicity of CMV antigens. These immunostimulatory properties might contribute to the protective effect against CMV infection mediated by CMVIG. View Show abstract ... Los medicamentos alternos ante resistencia son cidofovir y foscarnet. 201,202,203,204,205,206, 207 Dada la complejidad de la respuesta inmune del huésped contra el CMV, en la que participa activamente la respuesta inmune celular y humoral, innata y adaptativa, se han realizado estudios con inmunoglobulina humana específica para CMV que han mostrado que esta inmunoglobulina puede tener mejor efecto inmunomodulador. 201,202,203,204,207 También se ha informado su uso en resistencia del CMV a ganciclovir. ... ... 201,202,203,204,205,206,207 Dada la complejidad de la respuesta inmune del huésped contra el CMV, en la que participa activamente la respuesta inmune celular y humoral, innata y adaptativa, se han realizado estudios con inmunoglobulina humana específica para CMV que han mostrado que esta inmunoglobulina puede tener mejor efecto inmunomodulador. 201,202,203,204, 207 También se ha informado su uso en resistencia del CMV a ganciclovir. 208 En la actualidad se acepta que la hipogammaglobulinemia es un factor de riesgo para CMV después de TOS, especialmente en trasplante de corazón. ... Guía colombiana de práctica clínica para el uso de inmonuglobulinas en el tratamiento de reemplazo e inmunomodulación Article Full-text available Sep 2017 Rev Alerg Mex Margarita Olivares Carlos Eduardo Olmos-Olmos Martha I. Álvarez Sergio Francisco Ramírez Las inmunoglobulinas son proteínas heterodiméricas compuestas de 2 cadenas pesadas y 2 cadenas ligeras. La inmunoglobulina G humana es un derivado del plasma y contiene más de 95 % de IgG. La composición de las subclases de IgG es similar a la del plasma humano normal. El tratamiento con inmunoglobulina comenzó hace más de 50 años y su uso se ha descrito en numerosas enfermedades. En Colombia, la importancia de este recurso inmunomodulador condujo a la necesidad de contar con una guía de práctica clínica para su uso, para lo cual se reunió un grupo multidisciplinario de expertos, quienes se distribuyeron en mesas de trabajo, por especialidad, para redactar un texto base. Se llevaron a cabo búsquedas bibliográficas sistemáticas; las evidencias identificadas se valoraron y clasificaron para sustentar un texto preliminar que fue discutido, analizado y corregido. Se emitieron recomendaciones de uso de la inmunoglobulina intravenosa en patologías que abarcan inmunodeficiencias primarias y secundarias, enfermedades autoinmunes, alteraciones neurológicas, infecciones, trasplantes y enfermedades misceláneas; se asignaron calificaciones según el sistema GRADE para cada una. El resultado final se tradujo en las recomendaciones que se presentan con la finalidad de informar, orientar y apoyar en el uso óptimo de dicho recurso inmunomodulador. View Show abstract ... Also, all patients received HCMV immunoglobulin after transplantation to improve the virologic outcome. 10 This could have affected the Ab functions immediately post-transplantation, but at later time-points no effect of immunoglobulin would be expected due to its limited biologic half-life. 10 In conclusion, our data show that, in HCMV-seropositive LTRs, Ab functions are low early after transplantation and recover thereafter. ... ... 10 This could have affected the Ab functions immediately post-transplantation, but at later time-points no effect of immunoglobulin would be expected due to its limited biologic half-life. 10 In conclusion, our data show that, in HCMV-seropositive LTRs, Ab functions are low early after transplantation and recover thereafter. Neutralizing Ab levels at the end of the (val-)ganciclovir prophylaxis seem to predict to a certain degree freedom from viremia. ... Association Between Antibody Functions And HCMV Replication After Lung Transplantation In HCMV Seropositive Patients Article Jul 2017 Hannes Vietzen Irene Görzer Claudia Honsig Elisabeth Puchhammer-Stöckl View ... The most extensive experience is with CMV immunoglobulin (CMVIG), a high-titer preparation derived from donors with elevated CMV antibody titers. CMVIG is thought to act by inhibiting both the direct and indirect effects of CMV infection, via modulation of the innate and adaptive immune responses [5] . It neutralizes the CMV virus with high avidity antibodies and possibly reduces cytokine-mediated cellular immune responses [5]. ... ... CMVIG is thought to act by inhibiting both the direct and indirect effects of CMV infection, via modulation of the innate and adaptive immune responses [5]. It neutralizes the CMV virus with high avidity antibodies and possibly reduces cytokine-mediated cellular immune responses [5] . CMVIG administration in pregnant women has been shown to induce a significant increase in the concentration and avidity of CMV-specific IgG [6]. ... Successful treatment of intrauterine cytomegalovirus infection with an intraventricular cyst in a dichorionic diamniotic twin gestation using cytomegalovirus immunoglobulin Article Jun 2017 Maria De la Calle Fernando Baquero Roberto Rodríguez José Luis Bartha Congenital cytomegalovirus (CMV) infection is the leading cause of severe congenital abnormalities. CMV immunoglobulin (CMVIG) may lower risk for symptomatic disease in congenital CMV infection. In a twin pregnancy, only one fetus showing CMV infection, raising a dilemma about intervention since the uninfected fetus would be exposed to treatment unnecessarily. CMVIG (2 × 200 U/kg) was given due to high viral load and development of an intraventricular cyst. The cyst growth plateaued, no other brain damage developed, and at 8 months the infant was symptom-free. CMVIG appears appropriate to treat intrauterine CMV infection in this setting. View Show abstract ... After primary infection, glycoproteins trigger the innate immune system response leading to the production of cytokines that activate B cells and result in the production of CMV antibodies [5]. However, as a result of strain variability, such antibodies may not confer organ recipients adequate protection against donor-derived CMV [6] . Furthermore, CMV-antibodies cross-react with recipients' epithelial cells leading to sustained inflammation, thrombosis, atherosclerosis, and eventually chronic graft dysfunction [7]. ... ... Furthermore, calcineurin inhibitors' dosedependent inhibitory effect on CMV-specific T cell responses is especially profound during the early posttransplant period [10,11]. On the other hand, mycophenolate mofetil suppresses both B cell and T cell functions, further blunting the immune response to CMV infection in organ transplant recipients [6] . ... Cytomegalovirus Infection in Lung Transplant Recipients Article Apr 2017 Expet Rev Respir Med Reem S. Almaghrabi Ali S Omrani Ziad Memish Introduction: Cytomegalovirus (CMV) infection is a major cause of morbidity and mortality in solid organ transplant (SOT) patients. Lung transplant recipients are particularly at risk given the intense immunosuppression required. Areas covered: The Detailed review of the literature related to CMV infection, its direct and indirect effect on lung allograft function, as well as diagnosis, immune monitoring, treatment options and prevention strategies. Expert commentary: In lung transplant recipients, CMV infection is associated with pro-inflammatory and immune inhibitory effects that increase the risk of graft dysfunction and loss. Diagnosis of CMV infection remains challenging. Treatment options remain relatively limited. View Show abstract ... Antibody. Antiviral antibody functions as one of the early defense mechanisms against viral infection in sero(+) individuals through neutralizing viruses and eliminating virus-infected cells [69] [70][71]. It has been reported that low anti-CMV titer before transplant or at 1 month after transplant was associated with a higher risk of CMV disease in heart transplant recipients [72]. ... ... Elevated risk for CMV infection during the 1st year posttransplant was also reported in solid organ transplant recipients with severe hypogammaglobulinemia [73]. In addition, we and others have reported on the benefit of CMV immunoglobulin or IVIg use in the prevention and treatment of viral complications of transplantation including CMV [40, 69, 74], EBV/PTLD [75], parvovirus B19 [76], and BKV infections [21,77]. These study results demonstrate an important role of antiviral antibody in antiviral immunity in transplant recipients although neutralizing antibodies may not prevent subsequent rounds of infection and the cellular immune response eventually evolves to eradicate the infection [78]. ... Impact of Desensitization on Antiviral Immunity in HLA-Sensitized Kidney Transplant Recipients Article Full-text available Feb 2017 Mieko Toyoda Bong-Ha Shin Shili Ge Stanley C. Jordan Viral infections represent significant morbidity and mortality factors in kidney transplant recipients, with CMV, EBV, and BKV infections being most common. Desensitization (DES) with IVIg and rituximab with/without plasma exchange followed by kidney transplantation with alemtuzumab induction increased successful transplant rates in HLA-sensitized patients but may represent an increased risk for viral infections due to severe lymphocyte depletion. Here, we report on the posttransplant viral infection status in 372 DES versus 538 non-DES patients. CMV and EBV viremia were significantly lower in DES patients, while BKV viremia was similar. This trend was observed primarily in CMV sero(−), EBV sero(+), and sero(−) patients. No patient developed PTLD. The incidence of BKAN, allograft, and patient survival was similar in both groups. These viral infections were not associated with subsequent allograft rejection which occurred within 6 months after the infection. Conclusions. The IVIg + rituximab desensitization combined with alemtuzumab induction with triple immunosuppression maintenance does not increase the risk for CMV, EBV, and BKV infections. Possible factors include, in addition to posttransplant antiviral prophylaxis and PCR monitoring, presence of memory T cells and antibodies specific to CMV and likely EBV, NK cell-mediated ADCC despite lymphocyte depletion, elimination of EBV and CMV reservoirs by rituximab and alemtuzumab, and use of IVIg with antiviral properties. View Show abstract ... In transplant recipients with immunosuppressants, we have a brake and a limited attenuation of the primary responses, both innate and adaptive, slowing down the response of NKs cells and causing a continuous decrease in immunoglobulin levels and the levels of specific antibodies against CMV [13] . BCMA levels may, in this case, be essential to modulating this production of CMV-specific antibodies. ... Early Cytomegalovirus Reactivation in Renal Recipients Is Associated with High Levels of B Cell Maturation Antigen Transcript Expression Prior to Transplantation Article Full-text available Jun 2023 INT J MOL SCI Rafael Alfaro Luis Rodríguez-Aguilar Santiago Llorente Manuel Muro Cytomegalovirus (CMV) infection is the most frequent infection episode in kidney transplant (KT) recipients. Reactivation usually occurs in the first three months after transplantation and is associated with higher cellular and/or antibody-mediated rejection rates and poorer graft performance. CMV induces the expression of BAFF (B-cell-activating factor, a cytokine involved in the homeostasis of B cells), which communicates signals for survival and growth to B cells and virus-specific plasma cells via the R-BAFF (BAFF receptor), TACI (the calcium modulator, the cyclophilin ligand interactor), and BCMA (B cell maturation antigen) receptors. These molecules of the BAFF system have also been suggested as biomarkers for the development of alloantibodies and graft dysfunction. This prospective study included 30 CMV-IgG seropositive KT recipients. The expression levels of the genes BAFF-R, transmembrane activator and CAML interactor (TACI), and B cell maturation antigen (BCMA) in peripheral blood leukocytes (PBL) pre-KT were determined using qPCR. qPCR was also used to monitor CMV reactivation in the first three months following KT. The remainder of the KT recipients were classified as CMV− reactivation, and those with more than 500 copies/mL in at least one sample were classified as CMV+ reactivation. There were no discernible variations in the BAFF-R and TACI transcript expression levels. In the CMV+ group, we examined the relationship between the transcript levels and peak viremia. Peak viremia levels and BCMA transcript levels showed a strong correlation. BAFF-R and TACI expressions showed no measurable differences. In patients with early CMV reactivation, high BCMA receptor expression was associated with increased plasmablast, lymphocyte B cell class-switched levels (LBCS), and viral load. Our findings demonstrate that pre-KT BCMA transcript levels increased in KT recipients with early CMV reactivation. These transcript levels positively correlate with peak viremia and weakly with plasmablast and LBCS levels in PBLs. View Show abstract ... Decreased antibody (Ig) production significantly impacts human health by weakening the ability to maintain immunocompetence and survive infectious diseases (1)(2) (3) (4). Conversely, antibodies against selfproteins play a major role in autoimmune diseases (5)(6)(7). The immunoglobulin heavy chain (IgH) gene is responsible for the expression of all Ig classes/isotypes, i.e. ... Evolutive emergence and divergence of an Ig regulatory node: An environmental sensor getting cues from the aryl hydrocarbon receptor? Article Full-text available Feb 2023 Pietro D'Addabbo Domenico Frezza Courtney E W Sulentic One gene, the immunoglobulin heavy chain (IgH) gene, is responsible for the expression of all the different antibody isotypes. Transcriptional regulation of the IgH gene is complex and involves several regulatory elements including a large element at the 3’ end of the IgH gene locus (3’RR). Animal models have demonstrated an essential role of the 3’RR in the ability of B cells to express high affinity antibodies and to express different antibody classes. Additionally, environmental chemicals such as aryl hydrocarbon receptor (AhR) ligands modulate mouse 3’RR activity that mirrors the effects of these chemicals on antibody production and immunocompetence in mouse models. Although first discovered as a mediator of the toxicity induced by the high affinity ligand 2,3,7,8-tetracholordibenzo-p-dioxin (dioxin), understanding of the AhR has expanded to a physiological role in preserving homeostasis and maintaining immunocompetence. We posit that the AhR also plays a role in human antibody production and that the 3’RR is not only an IgH regulatory node but also an environmental sensor receiving signals through intrinsic and extrinsic pathways, including the AhR. This review will 1) highlight the emerging role of the AhR as a key transducer between environmental signals and altered immune function; 2) examine the current state of knowledge regarding IgH gene regulation and the role of the AhR in modulation of Ig production; 3) describe the evolution of the IgH gene that resulted in species and population differences; and 4) explore the evidence supporting the environmental sensing capacity of the 3’RR and the AhR as a transducer of these cues. This review will also underscore the need for studies focused on human models due to the premise that understanding genetic differences in the human population and the signaling pathways that converge at the 3’RR will provide valuable insight into individual sensitivities to environmental factors and antibody-mediated disease conditions, including emerging infections such as SARS-CoV-2. View Show abstract ... Sustained control of CMV infection is largely due to cellular immunity with a diverse T-cell response occurring that is targeted toward CMV-specific CD4+ and CD8+ cells. 51 In the first days after SOT, there is a marked decrease in CD8+ T cells and immunoglobulin G (IgG) levels so that immunity against CMV is reduced. However, the immune response varies depending on the organ being transplanted and the CMV status of the donor and recipient. ... Cytomegalovirus in the transplant setting: Where are we now and what happens next? A report from the International CMV Symposium 2021 Article Full-text available Oct 2022 Camille Kotton Julián Torre-Cisneros The CMV Symposium in September 2021 was an international conference dedicated to cytomegalovirus (CMV) infection after solid organ or haematopoietic stem cell transplantation. This review provides an overview of the presentations given by the expert faculty, supplemented with educational clinical cases. Topics discussed include CMV epidemiology and diagnosis, the burden of CMV infection and disease, CMV-specific immunity and management of CMV in transplant settings. Major advances in the prevention and treatment of CMV in the past decade and increased understanding of CMV immunity has led to improved patient outcomes. In the future, management algorithms may be individualised based on the transplant recipient's immune profile which will mark the start of a new era for patients with CMV. This article is protected by copyright. All rights reserved. View Show abstract ... Lack of either global innate or CMV-specific adaptive immunity has been described as poor prognostic factors for CMV reactivation after SOT (14) . The restoration of viral-specific cell-mediated immunity is associated with viral clearance and, conversely, the failure of this immunity is associated with uncontrolled infection by viruses such as adenovirus, BK polyomavirus, and CMV (8,15,16). ... Predictors of CMV Infection in CMV-Seropositive Kidney Transplant Recipients: Impact of Pretransplant CMV-Specific Humoral Immunity Article Full-text available Apr 2021 Similan Kirisri Apirom Vongsakulyanon Surasak Kantachuvesiri Jackrapong Bruminhent Introduction Although cytomegalovirus (CMV)-seropositive solid organ transplant recipients have a relatively lower risk of CMV infection than CMV-seronegative recipients who receive allograft from CMV-seropositive donors, some patients remain at risk of CMV infection after transplant. We investigated the pre-transplant CMV-specific humoral immunity (CHI) and other CMV infection predictors in CMV-seropositive kidney transplant (KT) recipients. Methods This retrospective study was conducted on adult CMV-seropositive KT recipients during 2017 and 2018. The cumulative incidence of CMV infection was estimated with Kaplan–Meier methodology. CHI, measured with an enzyme-linked fluorescent immunoassay and other predictors for CMV infection, was analyzed using Cox proportional hazards models. Results Of the 340 CMV-seropositive KT recipients (37% female; age [mean ± SD]: 43±11 years), 69% received deceased-donor allograft and 64% received induction therapy. During a mean follow-up of 14 months, the cumulative incidence of CMV infection was 14.8%. In multivariate analysis, low pre-transplant CHI (defined as anti-CMV IgG titer <20 AU/ml) was significantly associated with CMV infection (HR, 2.98; 95% CI, 1.31–6.77, [p=0.009]). Other significant predictors of CMV infection included older donor age (HR, 1.03; 95% CI, 1.01–1.06, [p=0.005]), anti-thymocyte induction therapy (HR, 2.90; 95% CI 1.09–7.74, [p=0.033]), and prolonged cold ischemic time (HR, 1.06; 95% CI, 1.02–1.10, [p=0.002]). Conclusion A low pre-transplant CHI is independently associated with post-transplant CMV infection in CMV-seropositive KT recipients. A quantitative anti-CMV IgG assay could potentially stratify CMV-seropositive patients at risk of CMV infection after KT. View Show abstract ... 6 Immunological reactions to CMV infection are highly complex and obtain humoral, cellular, innate, and adaptive immune responses. 7 Lung transplant patients are challenged to control CMV infection and gain immunological defense while deliberately impairing immune recognition of their allograft. Many of the same mechanisms that are required to control latent or acute infection are disabled by the immunosuppression that prevents allograft rejection. ... Association of CMV‐specific‐T‐cell immunity and risk of CMV infection in lung transplant recipients Article Full-text available Mar 2021 Tobias Veit Ming Pan Dieter Munker Teresa Kauke Background Protecting against CMV infection and maintaining CMV in latent state are largely provided by CMV‐specific T cells in lung transplant recipients. The aim of the study was to assess whether a specific T cell response is associated with the risk for CMV infection in seronegative patients who are at high risk for delayed CMV infection. Methods All CMV seronegative recipients (R‐) from CMV seropositive donors (D+) between January 2018 and April 2019 were included and retrospectively screened for CMV‐infection before and after assessment of CMV‐specific cell‐mediated immunity. Results Thirty‐one of the 50 patients (62%) developed early onset CMV infection. Lower absolute neutrophil counts were significantly associated with early onset CMV infection. Antiviral prophylaxis was ceased after 137.2 ± 42.8 days. CMV‐CMI were measured at a median of 5.5 months after LTx. 19 patients experienced early and late onset CMV infection after prophylaxis withdrawal within 15 months post transplantation. Positive CMV‐CMI was significantly associated with lower risk of late onset CMV infection after transplantation in logistic and cox‐regression analysis (OR=0.05, p=0.01; OR=2,369, p=0.026). Conclusion D+/R‐ lung transplant recipients are at high risk of developing early and late onset CMV infection. Measurement of CMV‐CMI soon after transplantation might further define the CMV infection prediction risk in LTx recipients being at high risk for CMV viremia. View Show abstract ... Passive immunity utilizing intravenous immunoglobulin has been a longstanding component of the treatment of CMV infection. 76 CMV hyperimmune globulin (CMVIg) has been the most extensively studied in this role but due to cost and the prevalence of CMV in the general population, standard IVIG is frequently used interchangeably, as no literature exists supporting clinical superiority of either product. However, given the lack of rigorously designed studies, it is difficult to tease apart the direct effects of IVIG on CMV treatment, thus recent consensus guidelines do not give a strong recommendation for or against its use beyond stating it may have a role in some clinical scenarios. ... Cytomegalovirus Antiviral Stewardship in the COVID‐19 Era: Increasing Complexity of Prophylaxis and Treatment and Potential Mitigation Strategies Article Feb 2021 Cynthia Wong Chris Saddler Margaret R. Jorgenson Jillian L. Descourouez Cytomegalovirus (CMV) infection is one of the most common and significant complications after solid organ transplant (SOT). Severe acute respiratory coronavirus 2 (SARS‐CoV‐2), which causes the novel betacoronavirus 2019 disease (COVID‐19), has become the first global pandemic in 100 years. The world’s attention has turned to address this unanticipated development; however, the viral infection that has long plagued outcomes after solid organ transplantation still requires vigilance. With physical distancing as the key intervention to reduce the healthcare burden, and the unease related to healthcare contact within the transplant population given the associated morbidity and mortality of COVID‐19 in transplant recipients, providers have struggled to evaluate and streamline essential in‐person healthcare contact, including laboratory visits. Due to this, the COVID‐19 pandemic has placed a significant strain on the delivery of CMV prophylaxis and treatment after solid organ transplantation. In this piece, we will describe issues our CMV antiviral stewardship service has encountered in the care of the transplant recipient with CMV during the this unprecedented time and share our expert opinion to approaches to providing optimal, evidenced based care during a pandemic associated with a seemingly unrelated viral infection. View Show abstract ... In transplant patients, the use of IVIG (400 mg/kg on days 1, 2, and 7 and 200 mg/kg on day 14) combined with antiviral drugs such as acyclovir and gancyclovir, seems to prevent CMV related complications, such as pneumonitis, whereas either treatment alone does not [52][53][54]. In the meanwhile, the same combination does not seem to give benefit in case of CMV gastrointestinal involvement [55, 56]. IVIG were also used as prophylaxis for VZV infection in newborns exposed to the virus after birth and were effective for the treatment of a disseminated VZV infection [57,58]. ... Intravenous Immunoglobulins at the Crossroad of Autoimmunity and Viral Infections Article Full-text available Jan 2021 Carlo Perricone Paola Triggianese Roberto Bursi Roberto Perricone Intravenous immunoglobulins (IVIG) are blood preparations pooled from the plasma of donors that have been first employed as replacement therapy in immunodeficiency. IVIG interact at multiple levels with the different components of the immune system and exert their activity against infections. Passive immunotherapy includes convalescent plasma from subjects who have recovered from infection, hyperimmune globulin formulations with a high titer of neutralizing antibodies, and monoclonal antibodies (mAbs). IVIG are used for the prevention and treatment of several infections, especially in immunocompromised patients, or in case of a poorly responsive immune system. The evolution of IVIG from a source of passive immunity to a powerful immunomodulatory/anti-inflammatory agent results in extensive applications in autoimmune diseases. IVIG composition depends on the antibodies of the donor population and the alterations of protein structure due to the processing of plasma. The anti-viral and anti-inflammatory activity of IVIG has led us to think that they may represent a useful therapeutic tool even in COVID-19. The human origin of IVIG carries specific criticalities including risks of blood products, supply, and elevated costs. IVIG can be useful in critically ill patients, as well as early empirical treatment. To date, the need for further well-designed studies stating protocols and the efficacy/tolerability profile of IVIG and convalescent plasma in selected situations are awaited. View Show abstract ... 15,16 On the other hand, the monthly administration of high-titre CMV IgG provided a passive specific immunity, playing an important role in the immunomodulation of a specific protective response. 17 Aim of the study In a cohort of lung transplanted patients who received a combined CMV prophylaxis scheme we aimed to: (a) evaluate the potential use of a CMV ELISPOT assay to identify patients at risk of CMV infections after lung transplantation; (b) measure the concordance between CMV ELISPOT response and CMV serology; (c) describe concordance between CMV ELISPOT response and CMV serology during CMV infections; (d) describe changes in CMV ELISPOT response, CMV serology and titre during the observation period after lung transplantation. ... Cellular and humoral cytomegalovirus immunity changes in one-year combined prophylaxis after lung transplantation: suggestions from and for clinical practice Article Full-text available Dec 2020 Ther Adv Respir Dis Massimo Boffini Carlo Albera Paolo Solidoro Filippo Patrucco Background Immune responses, both cellular and humoral, against cytomegalovirus (CMV) are used to predict CMV manifestations in solid organ recipients. The aim of this study is to evaluate CMV enzyme-linked immunospot (ELISPOT) assay and serology during CMV infections, their concordance and variations after lung transplantation (LTx). Methods We retrospectively analysed in one year the follow-up data of 43 patients receiving combined CMV prophylaxis with antiviral agents and CMV-specific immunoglobulin G (IgG). CMV infections were investigated by using molecular analyses on both 167 bronchoalveolar lavage and biopsy specimens and 1134 blood samples. Cellular CMV immunity was assessed with specific ELISPOT whereas the humoral one was assessed by quantifying specific immunoglobulins. Results At the first month after LTx the majority of patients were ELISPOT responders (52.3%) and 30.9% were non-responders. ELISPOT responders had a lower incidence of CMV viremia ( p = 0.047), whereas neither effects on CMV pulmonary asymptomatic infection nor on acute rejection were observed. Responders had a higher CMV IgG titre ( p < 0.0001) in particular at the first month after LTx ( p = 0.0001). Concordance among CMV ELISPOT assay and IgG levels was moderate (Cohen’s K 0.524), with an agreement of 89.8%. All ELISPOT responders maintained their status and almost all non-responders became responders during follow-up (92.3%); the percentage of IgG seropositive subjects increased from 74.4% at the first month of follow-up to 97.4% after 1 year. Conclusions Despite a moderate concordance with serology, ELISPOT response predicted a lower incidence of CMV viremia in LTx patients; no effects were reported on pulmonary clinical manifestations nor on acute rejection. The ELISPOT response as well as serology changed during the follow-up, not only after first CMV contact. The reviews of this paper are available via the supplemental material section. View Show abstract ... All rights reserved antibodies are being used for the treatment and prevention of other diseases (106). This is similar to the concept of using hyper-immune globulin for various indications, including viral diseases in immunocompetent and immunocompromised hosts (107)(108) (109) . A "cocktail antibody approach" for SARS-CoV-2 was proposed based on the studies suggesting that the combination of antibodies from diverse donors may exert a synergistic neutralization effect (36) . ... Antibody response to SARS‐Co‐V‐2, diagnostic and therapeutic implications Article Full-text available Aug 2020 Yuval Ishay Asa Kessler Asaf Schwarts Yaron Ilan The immune response against SARS‐CoV‐2 is comprised of both cellular and humoral arms. While current diagnostic methods are mainly based on PCR, they suffer from insensitivity. Therefore, antibody‐based serological tests are being developed to achieve higher sensitivity and specificity. Current efforts in treating SARS‐CoV‐2 infection include blocking of viral entry into the host cells, prohibiting viral replication and survival in the host cells, or reducing the exaggerated host immune response. Administration of convalescent plasma containing anti‐viral antibodies was proposed to improve the outcome in severe cases. In this paper, we review some of the aspects associated with the development of antibodies against SARS‐CoV‐2 and their potential use for improved diagnosis and therapy. View Show abstract ... In fact, this new therapeutic modality offers the advantage of targeting the very early stage of viral infection, in contrast to the currently present antivirals that target DNA polymerase during the initiation of the infection or reactivation, which could prevent the immunopathological effect of some expressed IE genes, like in HCMV infection. 220 Moreover, as those "epidrugs" target the host rather than viral components, the emergence of strains that can develop drug resistance or hijack and manipulate the host machinery to escape immune surveillance is minimized. Besides, as those agents are disclosing new horizons in various viral infection, their use in coinfection scenario (e.g. ... Targeting histone epigenetics to control viral infections Chapter Full-text available Jan 2020 Zeina Nehme Sébastien Pasquereau Georges Herbein During the past decades, many studies have significantly broadened our understanding of complex virus-host interactions to control chromatin structure and dynamics.1, 2 However, the role and impact of such modifications during viral infections is not fully revealed. Indeed, this type of regulation is bidirectional between the virus and the host. While viral replication and gene expression are significantly impacted by histone modifications on the viral chromatin,³ studies have shown that some viral pathogens dynamically manipulate cellular epigenetic factors to enhance their own survival and pathogenesis, as well as escape the immune system defense lines.⁴ In this dynamic, histone posttranslational modifications (PTMs) appear to play fundamental roles in the regulation of chromatin structure and recruitment of other factors.⁵ Genuinely, those PTMs play a vital role in lytic infection, latency reinforcement, or, conversely, viral reactivation.⁶ In this chapter, we will examine and review the involvement of histone modifications as well as their potential manipulation to control infections during various viral life cycle stages, highlighting their prospective implications in the clinical management of human immunodeficiency virus (HIV), herpes simplex virus (HSV), human cytomegalovirus (HCMV), hepatitis B and C viruses (HBV and HCV, respectively), Epstein–Barr virus (EBV), and other viral diseases. Targeting histone modifications is critical in setting the treatment of chronic viral infections with both lytic and latent stages (HIV, HCMV, HSV, RSV), virus-induced cancers (HBV, HCV, EBV, KSHV, HPV), and epidemic/emerging viruses (e.g. influenza virus, arboviruses). View Show abstract ... The immune response to CMV infection is highly complex and includes innate and adaptive immune responses (24) . Accordingly, CMV infection is first detected by the innate immune system, which seems to be crucial during the early phase of an CMV infection (25). ... The Aquaporin 5 −1364A/C Promoter Polymorphism Is Associated With Cytomegalovirus Infection Risk in Kidney Transplant Recipients Article Full-text available Dec 2019 Tim Rahmel Hartmuth Nowak Katharina Rump Lars Bergmann Background: The aquaporin 5 ( AQP5) −1364A/C promoter single nucleotide polymorphism affects key mechanisms of inflammation and immune cell migration. Thus, it could be involved in the pathogenesis of cytomegalovirus infection. Accordingly, we tested the hypothesis that the AQP5 promoter −1364A/C polymorphism is associated with the risk of cytomegalovirus infection in kidney transplantation recipients. Methods: We included 259 adult patients who received a kidney transplant from 2007 and 2014 in this observational study. Patients were genotyped for the AQP5 promoter −1364A/C single nucleotide polymorphism and followed up for 12 months after transplantation. Kaplan–Meier plots and multivariable proportional hazard analyses were used to evaluate the relationship between genotypes and the incidence of cytomegalovirus infection. Results: The incidences of cytomegalovirus infection within 12 months after kidney transplantation were 22.9% for the AA genotypes (43/188) and 42.3% for the AC/CC genotypes (30/71; p = 0.002). Furthermore, multivariable COX regression revealed the C-allele of the AQP5 −1364A/C polymorphism to be a strong and independent risk factor for cytomegalovirus infection. In this analysis, AC/CC subjects demonstrated a more than 2-fold increased risk for cytomegalovirus infection within the first year after kidney transplantation (hazard ratio: 2.28; 95% CI: 1.40–3.73; p = 0.001) compared to that in individuals with homozygous AA genotypes. Conclusions: With respect to opportunistic cytomegalovirus infections (attributable to immunosuppression after kidney transplantation), the C-allele of the AQP5 −1364A/C promoter polymorphism is independently associated with an increased 12-months infection risk. These findings emphasize the importance of genetic variations as additional risk factors of cytomegalovirus infection after solid organ transplantations and might also facilitate the discovery of novel therapeutic targets. View Show abstract ... In immunocompetent individuals, both primary infection and reactivation events are well controlled by a broad and robust immune response (9). However, HCMV reactivation is a major cause of morbidity and mortality in immunocompromised patients, including stem cell and organ transplant recipients (10, 11) . ... Interferon-Responsive Genes Are Targeted during the Establishment of Human Cytomegalovirus Latency Article Full-text available Dec 2019 Elizabeth G. Elder Ben Krishna James C Williamson John Sinclair Human cytomegalovirus (HCMV) latency is an active process which remodels the latently infected cell to optimize latent carriage and reactivation. This is achieved, in part, through the expression of viral genes, including the G-protein-coupled receptor US28. Here, we use an unbiased proteomic screen to assess changes in host proteins induced by US28, revealing that interferon-inducible genes are downregulated by US28. We validate that major histocompatibility complex (MHC) class II and two pyrin and HIN domain (PYHIN) proteins, myeloid cell nuclear differentiation antigen (MNDA) and IFI16, are downregulated during experimental latency in primary human CD14 ⁺ monocytes. We find that IFI16 is targeted rapidly during the establishment of latency in a US28-dependent manner but only in undifferentiated myeloid cells, a natural site of latent carriage. Finally, by overexpressing IFI16, we show that IFI16 can activate the viral major immediate early promoter and immediate early gene expression during latency via NF-κB, a function which explains why downregulation of IFI16 during latency is advantageous for the virus. IMPORTANCE Human cytomegalovirus (HCMV) is a ubiquitous herpesvirus which infects 50 to 100% of humans worldwide. HCMV causes a lifelong subclinical infection in immunocompetent individuals but is a serious cause of mortality and morbidity in the immunocompromised and neonates. In particular, reactivation of HCMV in the transplant setting is a major cause of transplant failure and related disease. Therefore, a molecular understanding of HCMV latency and reactivation could provide insights into potential ways to target the latent viral reservoir in at-risk patient populations. View Show abstract ... Hence, we inferred that elevated serum LDH levels were associated with pneumonia and not attributed to renal, cardiac and hepatic dysfunction. With comprehensive suppression of the innate and adaptive immune responses by immunosuppressive agents, it is relatively difficult for renal transplant recipients to mount an effective immune response (31) . This may partially explain the higher mortality of these patients with severe CAP. ... Lactate dehydrogenase as a prognostic marker of renal transplant recipients with severe community-acquired pneumonia: a 10-year retrospective study Article Full-text available Nov 2019 Ying Su Min-Jie Ju Jie-fei Ma Zhe Luo Background: Lactate dehydrogenase (LDH) is an easily accessible biological marker that has been associated with several pulmonary disorders. The aim of this study was to investigate the prognostic value of serum LDH in renal transplant recipients with severe community-acquired pneumonia (CAP). Methods: A total of 77 renal transplant recipients with severe CAP admitted to the intensive care unit (ICU) were screened for eligibility in this retrospective study. Patient characteristics and laboratory tests, such as LDH on day 1 (LDHday 1) and day 3 (LDHday 3) were recorded. Cox regression models were used to assess the performance of LDH to predict 90-day mortality. Results: Median LDH level was higher on day 1 in 90-day nonsurvivors (440 U/L, IQR, 362-1,055 U/L) than in survivors (334 U/L, IQR, 265-432 U/L; P<0.001); median LDH level on day 3 in nonsurvivors was 522.5 U/L (IQR, 457.5-1,058.5 U/L) and in survivors 290 U/L (IQR, 223-387.5 U/L; P<0.001). Analysis of LDH kinetics from day 1 to day 3 showed an increase in nonsurvivors and a decrease in survivors. Moreover, Multivariate Cox analysis showed that LDHday 1 (increase per 100 U/L), LDHday 3 (increase per 100 U/L) and LDH kinetics (increase per 10%) were independently associated with 90-day mortality. Conclusions: Serum LDH levels and LDH kinetics early were independently associated with 90-day mortality in renal transplant recipients with severe CAP. In future, the prognostic role of LDH needs to be warranted. View Show abstract ... Cellular immune response is the main mechanism for the body to control CMV replication [11][12][13][14][15]. Therefore, if the strength of one's CMV antigen-specific T cell immunity is attainable, it is possible to predict the host's CMV reactivation risk [16][17][18][19]. Existing research mainly focuses on transplant recipients [20][21] [22] [23][24]. However, CMV-related immunological tests commonly used in clinical practice lack direct indicators of specific cellular immunity. ... The establishment of CMV antigen-specific enzyme-linked immunospot assay among Chinese: A pilot study Article Oct 2019 CLIN CHIM ACTA Yutong Liu Susu Ye Baotong Zhou Xiaoqing Liu Background: The prevalence of cytomegalovirus in China is high and the infection could result in disastrous consequences. Cellular immunity is the main mechanism for virus controlling. We explored the performance of cytomegalovirus antigen-specific enzyme-linked immunospot assay in patients with different infection state in endemic area. Methods: Twenty-eight patients with reliable results were included in statistical analysis. Peripheral blood mononuclear cells were extracted and were stimulated by cytomegalovirus phosphoprotein 65 or immediate early protein-1 antigen respectively. Spot forming cells (SFCs) were counted to evaluate the cellular immune response elicited by antigens. Results: Spots could be clearly displayed, and evenly dispersed with a clean background. The numbers of SFCs were 0 [0-0], 426 [210-601] and 230 [57-513] for uninfected individuals (n=7), latently infected patients (n=11) and actively infected patients (n=10) respectively, which were statistically different. The number of SFCs stimulated by phosphoprotein 65 was significantly higher than that by immediate early protein-1. Conclusions: The number of SFCs was significantly different among patients with different infection state. The stimulatory effect of phosphoprotein 65 was better than that of immediate early protein-1. View Show abstract ... In immunocompetent individuals, both primary infection and reactivation events are well controlled by a broad and robust immune response (9). However, HCMV reactivation is a major cause of morbidity and mortality in immunocompromised patients, including stem cell and organ transplant recipients (10, 11) . ... Interferon-responsive genes are targeted during the establishment of human cytomegalovirus latency Preprint Oct 2019 Elizabeth G. Elder Ben Krishna James C Williamson John Sinclair Human cytomegalovirus (HCMV) latency is an active process which remodels the latently infected cell to optimise latent carriage and reactivation. This is achieved, in part, through the expression of viral genes, including the G-protein coupled receptor US28. Here, we use an unbiased proteomic screen to assess changes in host proteins induced by US28, revealing that interferon-inducible genes are downregulated by US28. We validate that MHC Class II and two PYHIN proteins, MNDA and IFI16, are downregulated during experimental latency in primary human CD14 ⁺ monocytes. We find that IFI16 is targeted rapidly during the establishment of latency in a US28-dependent manner, but only in undifferentiated myeloid cells, a natural site of latent carriage. Finally, by overexpressing IFI16, we show that IFI16 can activate the viral major immediate early promoter and immediate early gene expression during latency via NF-κB, a function which explains why downregulation of IFI16 during latency is advantageous for the virus. Importance Human cytomegalovirus (HCMV) is a ubiquitous herpesvirus which infects 50-100% of humans worldwide. HCMV causes a lifelong subclinical infection in immunocompetent individuals, but is a serious cause of mortality and morbidity in the immunocompromised and in neonates. In particular, reactivation of HCMV in the transplant setting is a major cause of transplant failure and related disease. Therefore, a molecular understanding of HCMV latency and reactivation could provide insights into potential ways to target the latent viral reservoir in at-risk patient populations. View Show abstract ... High-titer CMV IgG provided a passive CMVspecific immunity and seemed to play an important role in immunomodulation of specific responses, showing an antiviral effect similar to that of antivirals. 45 Although the use of these preparations seems to be effective in reducing CMV pneumonia and to have some effect on acute rejection, the high variability in dosages and administration schedules has undoubtedly contributed to downplaying their evidence-based effectiveness despite the existence of many single-center or anecdotal studies. 36,37 The main point of discussion of our results are the following ... Tailored combined cytomegalovirus management in lung transplantation: a retrospective analysis Article Full-text available Sep 2019 Ther Adv Respir Dis Daniela Libertucci Cristina Costa Paolo Solidoro Filippo Patrucco Background There is no univocal prophylactic regimen to prevent cytomegalovirus (CMV) infection/disease in lung transplantation (LT) recipients. The aim of this study is to evaluate short-term clinical outcomes of a tailored combined CMV management approach. Methods After 1-year follow up, 43 LT patients receiving combined CMV prophylaxis with antiviral agents and CMV-specific IgG were evaluated in a retrospective observational study. Systemic and lung viral infections were investigated by molecular methods on a total of 1134 whole blood and 167 bronchoalveolar lavage (BAL) and biopsy specimens. CMV immunity was assessed by ELISPOT assay. Clinical and therapeutic data were also evaluated. Results We found 2/167 cases of CMV pneumonia (1.2%), both in the donor-positive/recipient-positive (D ⁺ /R ⁺ ) population, and 51/167 cases of CMV pulmonary infection (BAL positivity 30.5%). However, only 32/167 patients (19.1%) were treated due to their weak immunological response at CMV ELISPOT assay. Viremia ⩾100,000 copies/mL occurred in 33/1134 specimens (2.9%). Regarding CMV-serological matching (D/R), the D ⁺ /R – population had more CMV viremia episodes ( p < 0.05) and fewer viremia-free days ( p < 0.001). Conclusions Compared to previous findings, our study shows a lower incidence of CMV pneumonia and viremia despite the presence of a substantial CMV load. In addition, our findings further confirm the D ⁺ /R – group to be a high-risk population for CMV viremia. Overall, a good immunological response seems to protect patients from CMV viremia and pneumonia but not from CMV alveolar replication. The reviews of this paper are available via the supplemental material section. View Show abstract ... Nevertheless, these drugs have considerable side effects (including immunological effects) in certain patient populations and do not protect patients against later onset HCMV infections. Passive immunization by administration of HCMV immunoglobulin (CMVIG) preparations aims to restore normal concentrations of HCMV-specific immunoglobulin in the immunocompromised patient but data as monotherapy is still limited and its mode of action in the immune system is not yet fully understood (60, 61) . Several HCMV-specific monoclonal antibodies are in the research pipeline (62,63) or entering clinical trials for assessing efficacy, pharmacokinetic and anti-drug antibody evaluations (64). ... Signatures of T and B Cell Development, Functional Responses and PD-1 Upregulation After HCMV Latent Infections and Reactivations in Nod.Rag.Gamma Mice Humanized With Cord Blood CD34+ Cells Article Full-text available Nov 2018 Sebastian J. Theobald Sahamoddin Khailaie Michael E Meyer-Hermann Renata Stripecke Human cytomegalovirus (HCMV) latency is typically harmless but reactivation can be largely detrimental to immune compromised hosts. We modeled latency and reactivation using a traceable HCMV laboratory strain expressing the Gaussia luciferase reporter gene (HCMV/GLuc) in order to interrogate the viral modulatory effects on the human adaptive immunity. Humanized mice with long-term (more than 17 weeks) steady human T and B cell immune reconstitutions were infected with HCMV/GLuc and 7 weeks later were further treated with granulocyte-colony stimulating factor (G-CSF) to induce viral reactivations. Whole body bio-luminescence imaging analyses clearly differentiated mice with latent viral infections vs. reactivations. Foci of vigorous viral reactivations were detectable in liver, lymph nodes and salivary glands. The number of viral genome copies in various tissues increased upon reactivations and were detectable in sorted human CD14+, CD169+, and CD34+ cells. Compared with non-infected controls, mice after infections and reactivations showed higher thymopoiesis, systemic expansion of Th, CTL, Treg, and Tfh cells and functional antiviral T cell responses. Latent infections promoted vast development of memory CD4+ T cells while reactivations triggered a shift toward effector T cells expressing PD-1. Further, reactivations prompted a marked development of B cells, maturation of IgG+ plasma cells, and HCMV-specific antibody responses. Multivariate statistical methods were employed using T and B cell immune phenotypic profiles obtained with cells from several tissues of individual mice. The data was used to identify combinations of markers that could predict an HCMV infection vs. reactivation status. In spleen, but not in lymph nodes, higher frequencies of effector CD4+ T cells expressing PD-1 were among the factors most suited to distinguish HCMV reactivations from infections. These results suggest a shift from a T cell dominated immune response during latent infections toward an exhausted T cell phenotype and active humoral immune response upon reactivations. In sum, this novel in vivo humanized model combined with advanced analyses highlights a dynamic system clearly specifying the immunological spatial signatures of HCMV latency and reactivations. These signatures can be merged as predictive biomarker clusters that can be applied in the clinical translation of new therapies for the control of HCMV reactivation. View Show abstract ... In fact, some in vitro studies suggest that the binding of Megalotect to the viral antigens may prevent the CMV binding to target cells, thus modulating CMV infection and disease, until anti-CMV CD8+ T-cells are present. 30 It should be noticed that the dose, the schedule, and the number of administrations of Megalotect in the prophylaxis setting is widely variable in this series. This heterogeneity is due to the lack of published data and reflects the different Centers' policy and internal guidelines for CMV management. ... CMV Management with Specific Immunoglobulins: A Multicentric Retrospective Analysis on 92 Allotransplanted Patients Article Full-text available Aug 2019 Michele Malagola Raffaella Greco Stella Santarone Jacopo Peccatori CMV represents one of the most serious life-threatening complications of allogeneic stem cell transplantaion (allo-SCT). Pre-emptive treatment is highly effective, but toxicity and repetitive reactivation of CMV represent a major challenge in the clinical practice. The use of anti-CMV specific immunoglobulins (Megalotect) is controversial. We retrospectively collected data on 92 patients submitted to allo-SCT for hematological malignancies, in whom Megalotect was used either for prophylaxis (n=14) or with pre-emptive therapy (n=78). All the patients were considered at high-risk of developing CMV reactivation and CMV disease. The treatment was well tolerated, with no reported infusion reactions, nor other adverse events. None of the 14 cases treated with Megalotect as prophylaxis developed CMV reactivation. 51/78 (65%) patients who received Megalotect during pre-emptive treatment achieved complete clearance of CMV viremia, and 14/51 patients (29%) developed a breakthroug CMV infection. 7/78 patients (9%) developed CMV disease. The projected 1-year OS, 1-year TRM and 1-year RR is 74%, 15% and 19%, respectively. No differences were observed in terms of OS, TRM and RR by comparing patients who achieved a complete response after treatment versus those who did not.. These retrospective data suggest that Megalotect is safe and well tolerated. When used as prophylaxis, no CMV reactivation was recorded. We have no conclusive data regarding its efficacy in reducing the cumulative dose of anti-CMV specific drugs in the pre-emptive setting. Further prospective trials are warrented to identify the best setting of patients who can benefit from Megalotect alone or in addition to anti-CMV specific drugs. View Show abstract ... It is worth to mention that the current antivirals used in HCMV management are ganciclovir, and its oral prodrug valganciclovir, cidofovir, and foscavir that target the viral DNA polymerase, in addition to fomivirsen, an antisense antiviral drug used in the treatment of CMV retinis [68], and the recently FDA-approved letermovir used to prevent viral infection following allogenic hematopoietic stem cell transplant [69]. Since those antivirals target the viral DNA replication step, the function and expression of the immediate early (IE) and early (E) HCMV genes during the early stages of infection are not blocked, paving the road to immunopathology and raise the risk of graft rejection [70] . Thus, this mandates and sheds the light on the urgent necessity of developing new antiviral drugs with novel mechanisms of action based on new potential viral or cellular targets. ... Control of viral infections by epigenetic-targeted therapy Article Full-text available Mar 2019 Zeina Nehme Sébastien Pasquereau Georges Herbein Epigenetics is defined as the science that studies the modifications of gene expression that are not owed to mutations or changes in the genetic sequence. Recently, strong evidences are pinpointing toward a solid interplay between such epigenetic alterations and the outcome of human cytomegalovirus (HCMV) infection. Guided by the previous possibly promising experimental trials of human immunodeficiency virus (HIV) epigenetic reprogramming, the latter is paving the road toward two major approaches to control viral gene expression or latency. Reactivating HCMV from the latent phase (“shock and kill” paradigm) or alternatively repressing the virus lytic and reactivation phases (“block and lock” paradigm) by epigenetic-targeted therapy represent encouraging options to overcome latency and viral shedding or otherwise replication and infectivity, which could lead eventually to control the infection and its complications. Not limited to HIV and HCMV, this concept is similarly studied in the context of hepatitis B and C virus, herpes simplex virus, and Epstein-Barr virus. Therefore, epigenetic manipulations stand as a pioneering research area in modern biology and could constitute a curative methodology by potentially consenting the development of broad-spectrum antivirals to control viral infections in vivo. View Show abstract ... Nevertheless, these drugs have considerable side effects (including immunological effects) in certain patient populations and do not protect patients against later onset HCMV infections. Passive immunization by administration of HCMV immunoglobulin (CMVIG) preparations aims to restore normal concentrations of HCMV-specific immunoglobulin in the immunocompromised patient but data as monotherapy is still limited and its mode of action in the immune system is not yet fully understood (60, 61) . Several HCMV-specific monoclonal antibodies are in the research pipeline (62,63) or entering clinical trials for assessing efficacy, pharmacokinetic and anti-drug antibody evaluations (64). ... Multidimensional signatures of T cell maturation, PD-1 upregulation and B cell class-switch after HCMV latent infections and reactivations in humanized mice Article Full-text available Nov 2018 Valery Volk Simon Danisch Human cytomegalovirus (HCMV) latency is typically harmless but reactivation can be largely detrimental to immune compromised hosts. We modeled latency and reactivation using a traceable HCMV laboratory strain expressing the Gaussia luciferase reporter gene (HCMV/GLuc) in order to interrogate the viral modulatory effects on the human adaptive immunity. Humanized mice with long-term (more than 17 weeks) steady human T and B cell immune reconstitutions were infected with HCMV/GLuc and 7 weeks later were further treated with granulocyte-colony stimulating factor (G-CSF) to induce viral reactivations. Whole body bio-luminescence imaging analyses clearly differentiated mice with latent viral infections vs. reactivations. Foci of vigorous viral reactivations were detectable in liver, lymph nodes and salivary glands. The number of viral genome copies in various tissues increased upon reactivations and were detectable in sorted human CD14+, CD169+, and CD34+ cells. Compared with non-infected controls, mice after infections and reactivations showed higher thymopoiesis, systemic expansion of Th, CTL, Treg, and Tfh cells and functional antiviral T cell responses. Latent infections promoted vast development of memory CD4+ T cells while reactivations triggered a shift toward effector T cells expressing PD-1. Further, reactivations prompted a marked development of B cells, maturation of IgG+ plasma cells, and HCMV-specific antibody responses. Multivariate statistical methods were employed using T and B cell immune phenotypic profiles obtained with cells from several tissues of individual mice. The data was used to identify combinations of markers that could predict an HCMV infection vs. reactivation status. In spleen, but not in lymph nodes, higher frequencies of effector CD4+ T cells expressing PD-1 were among the factors most suited to distinguish HCMV reactivations from infections. These results suggest a shift from a T cell dominated immune response during latent infections toward an exhausted T cell phenotype and active humoral immune response upon reactivations. In sum, this novel in vivo humanized model combined with advanced analyses highlights a dynamic system clearly specifying the immunological spatial signatures of HCMV latency and reactivations. These signatures can be merged as predictive biomarker clusters that can be applied in the clinical translation of new therapies for the control of HCMV reactivation. View Show abstract ... These responses are essential in controlling viral replication and dissemination as shown by primary infection in either the immune-naive or immunosuppressed. Here, uncontrolled virus replication leads to end organ disease and morbidity and if left uncontrolled, mortality (Carbone 2016; Chan and Logan 2017;Kagan and Hamprecht 2017). Primary HCMV infection has a profound effect on the human immune system, leaving a permanent signature in the form of phenotypically distinct T and NK cell subsets at high frequencies (discussed in the accompanying article by Souquette et al.). ... CMV immune evasion and manipulation of the immune system with aging Article Full-text available Jun 2017 Sarah Jackson Anke Redeker Ramon Arens Mark R. Wills Human cytomegalovirus (HCMV) encodes numerous proteins and microRNAs that function to evade the immune response and allow the virus to replicate and disseminate in the face of a competent innate and acquired immune system. The establishment of a latent infection by CMV, which if completely quiescent at the level of viral gene expression would represent an ultimate in immune evasion strategies, is not sufficient for lifelong persistence and dissemination of the virus. CMV needs to reactivate and replicate in a lytic cycle of infection in order to disseminate further, which occurs in the face of a fully primed secondary immune response. Without reactivation, latency itself would be redundant for the virus. It is also becoming clear that latency is not a totally quiescent state, but is characterized by limited viral gene expression. Therefore, the virus also needs immune evasion strategies during latency. An effective immune response to CMV is required or viral replication will cause morbidity and ultimately mortality in the host. There is clearly a complex balance between virus immune evasion and host immune recognition over a lifetime. This poses the important question of whether long-term evasion or manipulation of the immune response driven by CMV is detrimental to health. In this meeting report, three groups used the murine model of CMV (MCMV) to examine if the contribution of the virus to immune senescence is set by the (i) initial viral inoculum, (ii) inflation of T cell responses, (iii) or the balance between functionally distinct effector CD4+ T cells. The work of other groups studying the CMV response in humans is discussed. Their work asks whether the ability to make immune responses to new antigens is compromised by (i) age and HCMV carriage, (ii) long-term exposure to HCMV giving rise to an overall immunosuppressive environment and increased levels of latent virus, or (iii) adapted virus mutants (used as potential vaccines) that have the capacity to elicit conventional and unconventional T cell responses. View Show abstract ... More accurate CMV risk stratification may additionally be performed not only by dissecting donor/recipients serology subgroups-for example after lung transplantation, 34 % of D+/R+ and only 6 % of D−/R+ developed infection or disease after 6 months of prophylaxis [79]-but also by specifically assaying host ability to respond to infection. In particular, hypogammaglobulinemia, as long as negative CMV serology, is a risk factor for infections and transfer of passive immunity by CMV hyperimmune sera may be effective in reducing CMV disease and its adverse graft-related consequences [80, 81] . Lack of recovery of T-cell mediated CMV-specific immunity represents an additional risk factor for infection and disease potentially benefitting of more aggressive CMV prevention strategies [82]. ... Current Perspectives on Cytomegalovirus in Heart Transplantation Article Full-text available Dec 2016 Marco Masetti Antonio Russo Francesco Grigioni Luciano Potena Cytomegalovirus (CMV) is the most common infectious agent in heart transplantation. Its peculiar features, including a complex interplay with host’s immune system and lifelong latency, have been associated with reduced graft survival by the development of manifestations of graft rejection, such as cardiac allograft vasculopathy. Modern antiviral agents have been effective in reducing the clinical impact of acute CMV syndromes, thus questioning the current role of the virus on chronic graft dysfunction. In this setting, prolonged universal antiviral prophylaxis may appear an easy and effective strategy to abolish the CMV issue in clinical practice. Nevertheless, several unanswered questions are still open: is CMV still involved in CAV development despite ganciclovir use? Do we have evidence that benefits from universal antiviral prophylaxis overcome costs and toxicities? Do we have anything new in the pipeline? In this article, we review the latest evidences unraveling the answers to these burning questions. View Show abstract ... In solid organ transplant patients, global suppression of innate and adaptive immunity by immunosuppressive agents limits immunological defense, including inhibition of natural killer cell activity with ongoing lowering of IG levels and CMV-specific antibody titers. CMVIG preparations have the potential to exert immunomodulatory effects as well as providing passive immunization: the role of CMVIG in reconstituting specific anti-CMV antibodies may be enhanced by some degree of modulation of the innate and adaptive immune responses, which could help to control some of the direct and indirect effects of CMV infection [56] . In this context, it should be noted that postoperative hypogammaglobulinemia, which is a common finding after thoracic transplantation, is associated with a marked increase in the risk for CMV infection [57], and that preemptive CMVIG treatment in patients with moderate or severe hypogammaglobulinemia showed a trend to fewer rejection episodes in a randomized trial [58]. ... Treatment and prevention of cytomegalovirus infection in heart and lung transplantation: an update Article Jun 2016 Luciano Potena Paolo Solidoro Filippo Patrucco L. Borgese Introduction: Heart and lung transplantation are standard therapeutic strategies to improve survival and quality of life in selected patients with end-stage heart or lung diseases. Cytomegalovirus (CMV) is one the most clinically relevant and frequent post-transplant infectious agents, which may cause direct acute syndromes, and chronic indirect graft-related injury. Despite effective antiviral drugs being available to prevent and treat CMV infection, due to the immunosuppression burden and the specific characteristics of thoracic grafts, CMV infection remains a major clinical problem in heart and lung transplant recipients. Areas covered: We performed an extensive literature search focused on studies specifically including heart or lung transplantation, when available, or kidney transplant recipients when data on thoracic transplants were not available. We discuss the pros and cons supporting the use of currently available drugs and strategies for CMV prevention and treatment, highlighting current unmet needs. Expert opinion: While (Val)Ganciclovir remains the cornerstone of anti-CMV therapy, prolonged universal prophylaxis may expose a large number of patients to an excess of drug toxicity. Additional drugs with lower toxicity may be available in the context of anti-CMV prophylaxis, and effective CMV-risk stratification, by means of novel immune monitoring assays, which may help to customize the therapeutic approach. View Show abstract Keep on tailoring CMV management in lung transplantation: 24 vs 12-month CMV hyperimmune globulins regimen effects in combined universal prophylaxis Article Sep 2022 PANMINERVA MED Paolo Solidoro Filippo Patrucco Carlo Albera Cristina Costa Background: In lung transplantation (LTx) cytomegalovirus (CMV) management is based on prophylaxis or pre-emptive therapy; CMV Hyperimmune Globulins (CMV IG) added to prophylactic antiviral agents reduce CMV manifestations and acute rejection. The length of prophylaxis regimens is variable among studies with different results. Methods: We conduced, after demonstrating efficacy of 12 months prophylaxis on acute rejections and CMV pneumonia, a single centre retrospective study comparing, during the second year after LTx, clinical effects of a long (24 months) versus short (12 months) course of combined CMV prophylaxis scheme, based on antiviral agents and CMV IG. Results: We included 120 patients, 70 received a long (24 months) and 50 a short (12 months) prophylaxis. The long prophylaxis group, at 18th month, had a lower rate of neutrophilic alveolitis in BAL (63.6% vs 94.4%, p=0.029). No other statistically significant differences were observed among the two groups of patients although we observed a reduction in both CMV (56,4% vs 76.0% p=n.s.) and bacterial infections (23,7% vs 32.0%, p=n.s.) during the 18th month of follow-up. We did not observe differences among two groups in acute rejection rate on transbronchial lung biopsies. Conclusions: The combined long prophylaxis course based on antiviral agents and CMV IG provides a reduction trend in CMV or bacterial infections even if not statistically significant; the significant reduction in neutrophilia in BAL compared to the cohort undergoing prophylaxis for 12 months should be carefully interpreted. An 18-month prophylaxis could be a good suggestion to be tested by other larger prospective studies. View Show abstract Management of ganciclovir resistance Cytomegalovirus infection with CMV hyperimmune globulin and leflunomide in 7 cardiothoracic transplant recipients and literature review Article Sep 2021 Kapil Iyer Rajamaiyer Venkateswaran Karthik Santhanakrishnan Nizar Yonan CMV disease caused by genetically resistant cytomegalovirus (CMV) poses a major challenge in solid organ transplant recipients, the development of resistance is associated with increased morbidity and mortality. Antiviral resistance affects 5–12% of patients following ganciclovir (GCV) therapy, but is more common in individuals with specific underlying risk factors. These include the CMV D+R- serostatus, type of transplanted organ, dose and duration of (Val)ganciclovir ([V]GCV) prophylaxis, peak viral loads and the intensity of immunosuppressive therapy. Guideline recommendations for the management of GCV resistance (GanR) in solid organ transplant recipients are based on expert opinion as there is a lack of data from controlled trials. Second-line options to treat GanR include foscarnet (FOS) and cidofovir (CDV), but these drugs are often poorly tolerated due to high rates of toxicity, such as renal dysfunction and neutropenia. Here we report 7 Cardiothoracic Transplant recipients with Ganciclovir Resistance CMV Infection from our centre treated with CMV immunoglobulin (CMVIG) +/- Leflunomide (LEF), and reviewed the literature on the use of these agents in this therapeutic setting. This article is protected by copyright. All rights reserved View Show abstract Cytomegalovirus reactivation after hematopoietic stem cell transplant with CMV-IG prophylaxis: A monocentric retrospective analysis Article Feb 2021 J MED VIROL Andrea Gilioli Andrea Messerotti Paola Bresciani Franco Narni Human Cytomegalovirus (CMV) represents the most common viral infection after hematopoietic stem cell transplant (HSCT), mainly occurring as reactivation from latency in seropositive patients, with a different prevalence based on the extent and timing of seroconversion in a specific population. Here, we retrospectively analyzed a cohort of patients who underwent HSCT at our Institution between 2013 and 2018, all of which were prophylactically treated with CMV‐Ig (Megalotect Biotest ®), to define the incidence and clinical outcomes of CMV reactivation and clinically significant infection. CMV infection occurred in 69% of our patient series, mainly resulted from reactivation, and CMV clinically significant infection (CS‐CMVi) occurred in 48% of prophylactically treated patients. CMV infection and CS‐CMVi impacted neither on relapse incidence, nor on overall survival (OS) nor on relapse free survival (RFS). Moreover, a very low incidence of CMV end‐organ disease was documented. CMV‐Ig used alone as prophylactic therapy after HSCT do not effectively prevent CMV reactivation. This article is protected by copyright. All rights reserved. View Show abstract The Risk Factor Analysis and Treatment Experience in Pneumocystis Jirovecii Pneumonia After Kidney Transplantation Article Dec 2020 Pengfeng Yang Xiaoqiu Zhu Weixiang Liang Ruiming Cai Background Pneumocystis jirovecii pneumonia (PJP) is an opportunistic infection among solid organ transplantation. The occurrence of PJP is dangerous and fatal if there is no early identification and sufficient treatment. Objective The aim of this study was to evaluate the risk factors and provide appropriate strategies of prophylaxis and treatment for PJP after kidney transplantation in our center. Patients/Methods From January 2009 to December 2018, a total of 167 kidney transplantation recipients with pneumonia were enrolled, including 47 PJP patients as PJP group and120 non‐PJP patients as control group. The clinical characteristics of the two groups were analyzed retrospectively. Results Multivariate analysis showed that high total dosage of ATG [OR, 2.03; 95% CI, 1.12‐3.68] and cytomegalovirus (CMV) infection were independent risk factors for PJP. Trimethoprim‐sulfamethoxazole (TMP‐SMX) (1.44g q6h) based treatment was used for 2 weeks and its dosage and course were adjusted according to the therapeutic effect and side effects. Forty‐five cases were recovered after 3 months follow‐up, and two patients died of respiratory failure. TMP‐SMX (0.48 g/d) prophylaxis was used for 3‐6 months and prolonged to 7‐8 months after treatment for acute rejection, which reduced the incidence of PJP compared with those without prophylaxis. Conclusion Our study suggests that the high total dosage of ATG and CMV infection indicate the increased risk of PJP. The strategies of prophylaxis and treatment for PJP after kidney transplantation in our center were effective. View Show abstract Progress and Challenges in the Prevention, Diagnosis, and Management of Cytomegalovirus Infection in Transplantation Article Oct 2020 Ajit P Limaye Tara M. Babu Michael Boeckh Hosts with compromised or naive immune systems, such as individuals living with HIV/AIDS, transplant recipients, and fetuses, are at the highest risk for complications from cytomegalovirus (CMV) infection. Despite substantial progress in prevention, diagnostics, and treatment, CMV continues to negatively impact both solid-organ transplant (SOT) and hematologic cell transplant (HCT) recipients. In this article, we summarize important developments in the field over the past 10 years and highlight new approaches and remaining challenges to the optimal control of CMV infection and disease in transplant settings. View Show abstract The development and implementation of stewardship initiatives to optimize the prevention and treatment of cytomegalovirus infection in solid-organ transplant recipients Article May 2020 Margaret R. Jorgenson Jillian L. Descourouez Lucas Schulz Didier A Mandelbrot Classical stewardship efforts have targeted immunocompetent patients; however, appropriate use of antimicrobials in the immunocompromised host has become a target of interest. Cytomegalovirus (CMV) infection is one of the most common and significant complications after solid-organ transplant (SOT). The treatment of CMV requires a dual approach of antiviral drug therapy and reduction of immunosuppression for optimal outcomes. This dual approach to CMV management increases complexity and requires individualization of therapy to balance antiviral efficacy with the risk of allograft rejection. In this review, we focus on the development and implementation of CMV stewardship initiatives, as a component of antimicrobial stewardship in the immunocompromised host, to optimize the management of prevention and treatment of CMV in SOT recipients. These initiatives have the potential not only to improve judicious use of antivirals and prevent resistance but also to improve patient and graft survival given the interconnection between CMV infection and allograft function. View Show abstract The use of CMVIg rescue therapy in cardiothoracic transplantation: A single-center experience over 6 years (2011-2017) Article Jun 2019 Karthik Santhanakrishnan Nizar Yonan Paul Callan Rajamiyer Venkateswaran Cytomegalovirus (CMV) is a common human pathogen that infects between 30 and 97% of the population in different geographic areas 2. In immunocompetent hosts, the virus is normally latent but can undergo reactivation leading to a range of clinical sequelae, especially in immunocompromised patients. Solid organ transplant (SOT) recipients are at high risk of CMV infection as they often receive intense and prolonged immunosuppression to minimise graft rejection. In such patients, CMV infection may occur either as a primary infection, when a CMV‐seronegative SOT recipient receives an allograft from a CMV‐seropositive donor (D+R‐) or as a secondary infection, following reactivation of endogenous latent virus in the recipient. Without CMV prevention strategies, CMV infection and disease typically occur during the first 3 months after SOT, but can also occur after prophylactic antiviral therapy has ended 2. This article is protected by copyright. All rights reserved. View Show abstract A Lymphoma Patient with Cytomegalovirus Retinitis and Post‐Autologous Hematopoietic Cell Transplantation Immune Reconstitution Uveitis: A Case Report and Review of the Literature Article Apr 2019 Risa Fuller Danielle Strauss Amir Steinberg Samantha E. Jacobs Cytomegalovirus (CMV) retinitis in hematologic malignancies in the absence of hematopoietic cell transplant (HCT) is uncommon. We report a case of a 54 year‐old woman with peripheral T cell lymphoma who develops CMV retinitis and subsequently undergoes an autologous HCT, with eventual development of immune reconstitution uveitis. We further reviewed the PubMed literature on CMV retinitis in patients with lymphoma. We describe that CMV retinitis in patients with lymphoma has variable clinical presentations, may occur at any time during the course of the disease and chemotherapy, and is associated with significant morbidity. This article is protected by copyright. All rights reserved. View Show abstract Cytomegalovirus and Epstein-Barr Virus Infection and Disease Chapter Apr 2019 Emanuele Nicastro Lorenzo D'Antiga Epstein-Barr virus (EBV) and cytomegalovirus (CMV) infections represent the two main infectious threats following paediatric liver transplantation. In particular, EBV primary infection is observed in 60–80% of seronegative children within 3 months of solid organ transplant, either from the oropharyngeal route or via donor passenger lymphocytes in the transplanted organ from a seropositive donor. Symptomatic EBV infection occurs in 8–22% of the cases and its most threatening complication, post-transplant lymphoproliferative disease (PTLD), in about 5% of all paediatric liver transplant recipients. PTLD is a potentially fatal condition that requires aggressive diagnostic approach and multidisciplinary management. In this chapter we present a tactic to treat PTLD through a clinical algorithm, based on stratification according to the disease risk. View Show abstract Indications for and outcomes of therapeutic plasma exchange after cardiac transplantation: A single center retrospective study Article Mar 2018 Oluwatoyosi A. Onwuemene Steven C. Grambow Chetan B. Patel Thomas L. Ortel Introduction: Limited data are available describing indications for and outcomes of therapeutic plasma exchange (TPE) in cardiac transplantation. Methods: In a retrospective study of patients who underwent cardiac transplantation at Duke University Medical Center from 2010 to 2014, we reviewed 3 TPE treatment patterns: a Single TPE procedure within 24 h of transplant; Multiple TPE procedures initiated within 24 h of transplant; and 1 or more TPE procedures beginning >24 h post-transplant. Primary and secondary outcomes were overall survival (OS) and TPE survival (TS), respectively. Results: Of 313 patients meeting study criteria, 109 (35%) underwent TPE. TPE was initiated in 82 patients within 24 h, 40 (37%) receiving a single procedure (Single TPE), and 42 (38%) multiple procedures (Multiple TPE). Twenty-seven (25%) began TPE >24 h after transplant (Delayed TPE). The most common TPE indication was elevated/positive panel reactive or human leukocyte antigen antibodies (32%). With a median follow-up of 49 months, the non-TPE treated and Single TPE cohorts had similar OS (HR 1.08 [CI, 0.54, 2.14], P = .84), while the Multiple and Delayed TPE cohorts had worse OS (HR 2.62 [CI, 1.53, 4.49] and HR 1.98 [CI, 1.02, 3.83], respectively). The Multiple and Delayed TPE cohorts also had worse TS (HR 2.59 [CI, 1.31, 5.14] and HR 3.18 [CI, 1.56, 6.50], respectively). Infection rates did not differ between groups but was independently associated with OS (HR 2.31 [CI, 1.50, 3.54]). Conclusions: TPE is an important therapeutic modality in cardiac transplant patients. Prospective studies are needed to better define TPE's different roles in this patient population. View Show abstract Potential Immunomodulatory Role of Specific Anticytomegalovirus Intravenous Immunoglobulin in Heart Recipients Article Nov 2016 TRANSPL P Javier Carbone A. Gallego J. Fernandez Yañez Elizabeth Sarmiento Background: Specific anticytomegalovirus (anti-CMV) intravenous immunoglobulin (IVIG) has the potential to influence the immune response, but its complex mode of action has not been well evaluated. Methods: An immunologic study of 6 CMV-seronegative heart transplant patients receiving anti-CMV prophylaxis with the use of ganciclovir and CMV-IVIG (150 mg/kg within 24 hours after transplantation and 100 mg/kg on days 2, 7, 14, 22, 35, 56, and 77 after transplantation) was performed in a single center. Lymphocyte subsets were evaluated by means of 4-color flow cytometry at the time of inclusion in the waiting list and at 3 months after transplantation. Results: High-risk heart recipients receiving CMV-IVIG showed a clear reduction in the frequency of activated CD4+CD38+DR+ T-helper cells at 3 months after transplantation compared with a group of 27 untreated control subjects who received only anti-CMV prophylaxis with the use of ganciclovir. In this study, an increase of CD19+CD27-IgM+IgD+ naïve B cells was also observed in seronegative recipients after prophylaxis with the use of CMV-IVIG but not in control subjects. None of the CMV-IVIG-treated recipients developed acute cellular rejection during the 1st 6 months after transplantation. Conclusions: The immune modulation of activated CD4+ lymphocyte and of naïve B-cell subsets after CMV-IVIG use should be further evaluated in future prospective studies with higher numbers of patients. View Show abstract CMV tratment Expert Opinion on Pharmaco Therapy 2016 Data Oct 2016 Luciano Potena Paolo Solidoro Filippo Patrucco L. Borgese View Functional Properties of Human Cytomegalovirus Hyperimmunoglobulin and Standard Immunoglobulin Preparations Article Sep 2016 ANN TRANSPL Matthias Germer Peter Herbener Joerg Schuettrumpf Background: Cytomegalovirus hyperimmunoglobulin (CMV-HIG) preparations reduce mortality after solid organ transplantation. Polyspecific intravenous immunoglobulin (IVIg) products are also used prophylactically by some centers. Since direct comparative characterizations of the preparations are scarce, it is challenging to compare different clinical studies. Material/Methods: The functionality of 2 CMV-HIG preparations (Cytotect® CP, Cytogam®) and 2 IVIg preparations (Ig Vena®, Flebogamma®) were compared in terms of: (i) CMV-specific immunoglobulin G (IgG) antibody levels determined by enzyme-linked immunoabsorbent assay (ELISA), (ii) avidity index using a CMV IgG avidity enzyme immunoassay, (iii) immunoblot assay against CMV-specific antigens, and (iv) anti-CMV microneutralization assay. Results: Median CMV-specific IgG antibody concentration was similar in the 2 CMV-HIG preparations (Cytotect® CP 101.8 PEIU/ml, Cytogam® 112.5 PEIU/ml) but markedly lower in the IVIg preparations (13.5 PEIU/ml and 21.3 PEIU/ml). CMV binding avidity was virtually identical for both CMV-HIG products (~90%). Immunoblot assay showed consistently high binding of both CMV-HIG preparations against all antigenic CMV glycoproteins tested. Recognition of some CMV-specific antigens (IE1, CM2, and p65) was weaker for the 2 IVIg products. Median CMV neutralizing antibody titers were identical for both CMV-HIG preparations (1:256), and 4-fold lower (1:64) for the IVIg products. CMV IgG antibody concentration correlated with the CMV neutralization titer. Conclusions: Compared to the polyspecific IVIg products tested here, CMV-HIG preparations showed higher CMV binding activity and wider recognition of tested CMV-specific glycoprotein antigens, with markedly higher neutralizing activity. There do not appear to be any relevant distinctions between the Cytotect® CP and Cytogam® CMV-HIG products in terms of functional activity. View Show abstract Mannose-Binding Lectin Serum Levels and Pre-Tranplant Genotypes for Personalized Anti-CMV Prophylaxis in Heart Recipients Article Full-text available Apr 2015 J HEART LUNG TRANSPL Mauricio Arraya Francisco Lozano Javier Carbone Elizabeth Sarmiento View A multicenter, randomized, open-labeled study to steer immunosuppressive and antiviral therapy by measurement of virus (CMV, ADV, HSV)-specific T cells in addition to determination of trough levels of immunosuppressants in pediatric kidney allograft recipients (IVIST01-trial): Study protocol for a randomized controlled trial Article Full-text available Aug 2014 TRIALS Thurid Ahlenstiel-Grunow Armin Koch Grosshennig Anika Lars Pape Background After kidney transplantation, immunosuppressive therapy causes impaired cellular immune defense leading to an increased risk of viral complications. Trough level monitoring of immunosuppressants is insufficient to estimate the individual intensity of immunosuppression. We have already shown that virus-specific T cells (Tvis) correlate with control of virus replication as well as with the intensity of immunosuppression. The multicentre IVIST01-trial should prove that additional steering of immunosuppressive and antiviral therapy by Tvis levels leads to better graft function by avoidance of over-immunosuppression (for example, viral infections) and drug toxicity (for example, nephrotoxicity). Methods/design The IVIST-trial starts 4 weeks after transplantation. Sixty-four pediatric kidney recipients are randomized either to a non-intervention group that is only treated conservatively or to an intervention group with additional monitoring by Tvis. The randomization is stratified by centre and cytomegalovirus (CMV) prophylaxis. In both groups the immunosuppressive medication (cyclosporine A and everolimus) is adopted in the same target range of trough levels. In the non-intervention group the immunosuppressive therapy (cyclosporine A and everolimus) is only steered by classical trough level monitoring and the antiviral therapy of a CMV infection is performed according to a standard protocol. In contrast, in the intervention group the dose of immunosuppressants is individually adopted according to Tvis levels as a direct measure of the intensity of immunosuppression in addition to classical trough level monitoring. In case of CMV infection or reactivation the antiviral management is based on the individual CMV-specific immune defense assessed by the CMV-Tvis level. Primary endpoint of the study is the glomerular filtration rate 2 years after transplantation; secondary endpoints are the number and severity of viral infections and the incidence of side effects of immunosuppressive and antiviral drugs. Discussion This IVIST01-trial will answer the question whether the new concept of steering immunosuppressive and antiviral therapy by Tvis levels leads to better future graft function. In terms of an effect-related drug monitoring, the study design aims to realize a personalization of immunosuppressive and antiviral management after transplantation. Based on the IVIST01-trial, immunomonitoring by Tvis might be incorporated into routine care after kidney transplantation. Trial Registration EudraCT No: 2009-012436-32, ISRCTN89806912 (17 June 2009). View Show abstract Preformed Frequencies of Cytomegalovirus (CMV)-Specific Memory T and B Cells Identify Protected CMV-Sensitized Individuals Among Seronegative Kidney Transplant Recipients Article Full-text available Jul 2014 Joan Torras Ambros Elena Crespo Edoardo Melilli Oriol Bestard Background: Cytomegalovirus (CMV) infection remains a major complication after kidney transplantation. Baseline CMV risk is typically determined by the serological presence of preformed CMV-specific immunoglobulin (Ig) G antibodies, even though T-cell responses to major viral antigens are crucial when controlling viral replication. Some IgG-seronegative patients who receive an IgG-seropositive allograft do not develop CMV infection despite not receiving prophylaxis. We hypothesized that a more precise evaluation of pretransplant CMV-specific immune-sensitization using the B and T-cell enzyme-linked immunospot assays may identify CMV-sensitized individuals more accurately, regardless of serological evidence of CMV-specific IgG titers. Methods: We compared the presence of preformed CMV-specific memory B and T cells in kidney transplant recipients between 43 CMV IgG-seronegative (sR(-)) and 86 CMV IgG-seropositive (sR(+)) patients. Clinical outcome was evaluated in both groups. Results: All sR(+) patients showed a wide range of CMV-specific memory T- and B-cell responses. High memory T- and B-cell frequencies were also clearly detected in 30% of sR(-) patients, and those with high CMV-specific T-cell frequencies had a significantly lower incidence of late CMV infection after prophylactic therapy. Receiver operating characteristic curve analysis for predicting CMV viremia and disease showed a high area under the receiver operating characteristic curve (>0.8), which translated into a high sensitivity and negative predictive value of the test. Conclusions: Assessment of CMV-specific memory T- and B-cell responses before kidney transplantation among sR(-) recipients may help identify immunized individuals more precisely, being ultimately at lower risk for CMV infection. View Show abstract Neutrophils Recruited by IL-22 in Peripheral Tissues Function as TRAIL-Dependent Antiviral Effectors against MCMV Article Full-text available Apr 2014 Maria A Oliver Morgan Marsden Tu Anh Pham N Ian R Humphreys During primary infection, murine cytomegalovirus (MCMV) spreads systemically, resulting in virus replication and pathology in multiple organs. This disseminated infection is ultimately controlled, but the underlying immune defense mechanisms are unclear. Investigating the role of the cytokine IL-22 in MCMV infection, we discovered an unanticipated function for neutrophils as potent antiviral effector cells that restrict viral replication and associated pathogenesis in peripheral organs. NK-, NKT-, and T cell-secreted IL-22 orchestrated antiviral neutrophil-mediated responses via induction in stromal nonhematopoietic tissue of the neutrophil-recruiting chemokine CXCL1. The antiviral effector properties of infiltrating neutrophils were directly linked to the expression of TNF-related apoptosis-inducing ligand (TRAIL). Our data identify a role for neutrophils in antiviral defense, and establish a functional link between IL-22 and the control of antiviral neutrophil responses that prevents pathogenic herpesvirus infection in peripheral organs. View Show abstract Assessment of Cytomegalovirus-Specific Cell-Mediated Immunity for the Prediction of Cytomegalovirus Disease in High-Risk Solid-Organ Transplant Recipients: A Multicenter Cohort Study Article Full-text available Nov 2012 CLIN INFECT DIS Oriol Manuel Shahid Husain Deepali Kumar Atul Humar Background. Cytomegalovirus (CMV) disease remains an important problem in solid-organ transplant recipients, with the greatest risk among donor CMV-seropositive, recipient-seronegative (D⁺/R⁻) patients. CMV-specific cell-mediated immunity may be able to predict which patients will develop CMV disease. Methods. We prospectively included D⁺/R⁻ patients who received antiviral prophylaxis. We used the Quantiferon-CMV assay to measure interferon-γ levels following in vitro stimulation with CMV antigens. The test was performed at the end of prophylaxis and 1 and 2 months later. The primary outcome was the incidence of CMV disease at 12 months after transplant. We calculated positive and negative predictive values of the assay for protection from CMV disease. Results. Overall, 28 of 127 (22%) patients developed CMV disease. Of 124 evaluable patients, 31 (25%) had a positive result, 81 (65.3%) had a negative result, and 12 (9.7%) had an indeterminate result (negative mitogen and CMV antigen) with the Quantiferon-CMV assay. At 12 months, patients with a positive result had a subsequent lower incidence of CMV disease than patients with a negative and an indeterminate result (6.4% vs 22.2% vs 58.3%, respectively; P < .001). Positive and negative predictive values of the assay for protection from CMV disease were 0.90 (95% confidence interval [CI], .74–.98) and 0.27 (95% CI, .18–.37), respectively. Conclusions. This assay may be useful to predict if patients are at low, intermediate, or high risk for the development of subsequent CMV disease after prophylaxis. Clinical Trials Registration. NCT00817908. View Show abstract Antibodies against the gH/gL/UL128/UL130/UL131 Complex Comprise the Majority of the Anti-Cytomegalovirus (Anti-CMV) Neutralizing Antibody Response in CMV Hyperimmune Globulin Article Full-text available Apr 2012 J VIROL Ashley Fouts Pamela Chan Jean-Philippe Stephan Becket Feierbach Anti-cytomegalovirus (anti-CMV) hyperimmune globulin (HIG) has demonstrated efficacy in preventing CMV disease in solid-organ transplant patients as well as congenital disease when administered to pregnant women. To identify the neutralizing component of cytomegalovirus hyperimmune globulin (CMV-HIG), we performed serial depletions of CMV-HIG on cell-surface-expressed CMV antigens as well as purified antigens. Using this approach, we demonstrate that the major neutralizing antibody response is directed at the gH/gL/UL128/UL130/UL131 complex, suggesting little role for anti-gB antibodies in CMV-HIG neutralization. View Show abstract Human Cytomegalovirus-Specific T-Cell Immune Reconstitution in Preemptively Treated Heart Transplant Recipients Identifies Subjects at Critical Risk for Infection Article Full-text available Mar 2012 J CLIN MICROBIOL Davide Abate Marta Fiscon Alda Saldan Giorgio Palu Human cytomegalovirus (CMV) infection represents a major threat for heart transplant recipients (HTXs). CMV-specific T cells effectively control virus infection, and thus, assessment of antiviral immune recovery may have clinical utility in identifying HTXs at risk of infection. In this study, 10 CMV-seropositive (R(+)) pretransplant patients and 48 preemptively treated R(+) HTXs were examined before and after 100 days posttransplant. Preemptive treatment is supposed to favor the immune recovery. CMV DNAemia and gamma interferon enzyme-linked immunosorbent spot (ELISPOT) assay were employed to assess the viremia and immune reconstitution. HTXs could be categorized into three groups characterized by high (>100), medium (50 to 100), and low (<50) spot levels. Early-identified high responders efficiently controlled the infection and also maintained high immunity levels after 100 days after transplant. No episodes of grade ≥2R rejection occurred in the high responders. Midresponders were identified as a group with heterogeneous trends of immune reconstitution. Low responders were 41% and 21% of HTXs before and after 100 days posttransplant, respectively. Low responders were associated with a higher incidence of infection. The effect of viremia on immune recovery was investigated: a statistically significant inverse correlation between magnitude of viremia and immune recovery emerged; in particular, each 10-fold increase in viremia (>4 log(10) DNAemia/ml) was associated with a 36% decrease of the ELISPOT assay spot levels. All episodes of high viremia (>4 log(10) DNAemia/ml) occurred from 1 to 60 days after transplant. Thus, the concomitant evaluation of viremia and CMV immune reconstitution has clinical utility in identifying HTXs at risk of infection and may represent a helpful guide in making therapeutic choices. View Show abstract Simultaneous Monitoring of Cytomegalovirus-Specific Antibody and T-cell levels in Seropositive Heart Transplant Recipients Article Full-text available Feb 2012 J CLIN IMMUNOL Javier Carbone Nallibe Lanio A. Gallego Elizabeth Sarmiento Human cytomegalovirus (CMV) active infection (CMV infection) poses serious risks to CMV-seropositive heart transplant recipients. We evaluated the usefulness of simultaneous assessment of CMV-specific values for parameters of the humoral (antibodies) and cellular (CD4+ and CD8+ T-cells) immune responses in the identification of heart recipients at risk of developing CMV infection after transplantation. We prospectively studied 38 CMV-seropositive heart recipients. Anti-CMV antibody titers were assessed using enzyme-linked immunosorbent assays. CD4+ and CD8+ T-cell responses to overlapping peptide pools of the CMV proteins pp65 and immediate early protein-1 (IE1) were evaluated by flow cytometry. Immunological studies were performed before transplantation and at 30 days after transplantation. Patients with CMV infection were compared with heart recipients without CMV infection. During the 6-month follow-up period, 13 (34.2%) patients developed CMV infection. At baseline, the mean anti-CMV-IgG antibody titer was lower in patients who developed CMV infection. This difference remained at 30 days after transplantation. One month after transplantation, the mean percentage of IE1-specific CD8+ T cells that are IFNg-positive (CD8/IFNg + IE1) was lower in CMV-infected patients. The predictive value of these variables at 30 days was increased when they were combined. Cox regression analysis revealed an association between the risk of developing CMV infection and the combination marker (low anti-CMV titer [<16,100] and low CD8/IFNg + IE1 percentages [<0.40%], relative hazard, 6.07; p = 0.019). The combination marker remained significant after adjustment for clinical variables. This novel approach of a simultaneous assessment of specific anti-CMV antibody titers and CD8/IFNg + IE1 percentages might help identify heart transplant recipients with an increased risk of developing CMV infection. View Show abstract Homozygosity for the Toll-Like Receptor 2 R753Q Single-Nucleotide Polymorphism Is a Risk Factor for Cytomegalovirus Disease After Liver Transplantation Article Full-text available Jan 2012 J INFECT DIS Seung H Kang Abdelhakim Chillali Robert A Brown Raymund R Razonable Immunity against cytomegalovirus (CMV) is initiated after its recognition by Toll-like receptor 2 (TLR2). We assessed the association between a single-nucleotide polymorphism (SNP) that impairs TLR2 function and CMV disease in a cohort of 737 liver recipients. Ninety-two of 737 patients (7.1%, 10.9%, 12.3%, and 12.5% by 3, 6, 12, and 24 months, respectively) developed CMV disease. Kaplan-Meier estimation demonstrated an association between TLR2 R753Q SNP homozygosity and CMV disease (P = .044), especially tissue-invasive CMV disease (P = .001). A multivariate Cox proportional hazard model that accounted for other significant predictors demonstrated a significant association between TLR2 R753Q SNP homozygosity and tissue-invasive CMV disease (hazard ratio, 3.407; 95% confidence interval, 1.518–7.644; P = .0029). In conclusion, homozygosity for TLR2 R753Q SNP is a marker for CMV disease risk, especially for tissue-invasive disease, after liver transplantation. This observation supports the critical role of TLR2 in the pathogenesis of CMV disease in humans. View Show abstract Expansion of a unique CD57(+) NKG2C(hi) natural killer cell subset during acute human cytomegalovirus infection Article Full-text available Aug 2011 P NATL ACAD SCI USA Sandra L López-Vergès Jeffrey M Milush Brian S Schwartz Lewis L Lanier During human CMV infection, there is a preferential expansion of natural killer (NK) cells expressing the activating CD94-NKG2C receptor complex, implicating this receptor in the recognition of CMV-infected cells. We hypothesized that NK cells expanded in response to pathogens will be marked by expression of CD57, a carbohydrate antigen expressed on highly mature cells within the CD56(dim)CD16(+) NK cell compartment. Here we demonstrate the preferential expansion of a unique subset of NK cells coexpressing the activating CD94-NKG2C receptor and CD57 in CMV(+) donors. These CD57(+)NKG2C(hi) NK cells degranulated in response to stimulation through their NKG2C receptor. Furthermore, CD57(+)NKG2C(hi) NK cells preferentially lack expression of the inhibitory NKG2A receptor and the inhibitory KIR3DL1 receptor in individuals expressing its HLA-Bw4 ligand. Moreover, in solid-organ transplant recipients with active CMV infection, the percentage of CD57(+)NKG2C(hi) NK cells in the total NK cell population preferentially increased. During acute CMV infection, the NKG2C(+) NK cells proliferated, became NKG2C(hi), and finally acquired CD57. Thus, we propose that CD57 might provide a marker of "memory" NK cells that have been expanded in response to infection. View Show abstract Cytomegalovirus Incidence Between Everolimus Versus Mycophenolate in De Novo Renal Transplants: Pooled Analysis of Three Clinical Trials Article Full-text available Aug 2011 AM J TRANSPLANT Daniel C. Brennan C Legendre D. Patel Fuad Shihab Everolimus (EVR) in heart and renal transplant (RTx) recipients may be associated with a decreased incidence of cytomegalovirus (CMV). A detailed analysis of the association between EVR versus mycophenolic acid (MPA) and CMV events has not been reported. CMV data from 2004 de novo RTx recipients from three-randomized, prospective, EVR studies A2309 (N = 833), B201 (N = 588) and B251 (N = 583) were retrospectively analyzed to identify differences between two EVR dosing groups and MPA. EVR groups received 1.5 mg/day, or 3 mg/day with either standard (SD-CsA) or reduced dose cyclosporine (RD-CsA). Controls received MPA with SD-CsA. CMV prophylaxis was as per center practice. CMV incidence (infection/syndrome, disease, viremia) was captured per local center evaluations. Kaplan-Meier analyses demonstrated that freedom from CMV viremia and infection/syndrome was significantly greater for EVR versus MPA for recipients without CMV prophylaxis. Among recipients who received prophylaxis, freedom from viremia was greater for EVR 3.0 mg; freedom from infection/syndrome was greater for EVR 3.0 and 1.5 mg. Although freedom from organ involvement was numerically greater for EVR, it was not statistically significant. This analysis documents significant reductions in the incidence of CMV infection/syndrome and viremia in EVR-treated de novo RTx recipients, especially those who did not receive CMV prophylaxis versus MPA. View Show abstract Incidence, Clinical Characteristics and Risk Factors of Late Infection in Solid Organ Transplant Recipients: Data from the RESITRA Study Group Article Full-text available Apr 2007 Rafael San Juan José María Aguado C Lumbreras J Torre-Cisneros Information describing the incidence and clinical characteristics of late infection (LI) in solid organ transplantation (SOT) is scarce. The aim of this study was to define the incidence, clinical characteristics and risk factors for LI (>6 months) as compared with infection in the early period (<6 months) after SOT. By the online database of the Spanish Network of Infection in Transplantation (RESITRA) we prospectively analyzed 2702 SOT recipients from September 2003 to February 2005. Univariate and multivariate analysis using logistic regression were performed to calculate the risk factors associated with the development of LI. A total of 131 patients developed 176 LI episodes (8%). Global incidence of LI was 0.4 per 1000 transplant-days, ranging from 0.3/1000 in kidney transplants to 1.4 in lung transplants. Independent risk factors for LI in were: acute rejection in the early period (OR 1.5; CI 95%: 1.1-2.3), chronic graft malfunction (OR 2; CI 95%: 1.4-3), re-operation (OR 1.9; CI 95%: 1.3-2.8) relapsing viral infection apart from CMV (OR 1.9; CI 95%: 1.1-3.5), previous bacterial infection (OR 1.8; CI 95%: 1.2-2.6) and lung transplantation (OR 4.5; CI 95%: 2.6-7.8). Severe LI occurs in a subgroup of high-risk SOT recipients who deserve a more careful follow-up and could benefit from prolonged prophylactic measures similar to that performed in the early period after transplantation. View Show abstract High Frequency of Cytomegalovirus‐Specific Cytotoxic T‐Effector Cells in HLA‐A*0201–Positive Subjects during Multiple Viral Coinfections Article Full-text available Jan 2000 Jun-Jie Xu Marie-Ange Demoitie Sean M. Donahoe Douglas F. Nixon How the cellular immune response copes with diverse antigenic competition is poorly understood. Responses of virus-specific cytotoxic T lymphocytes (CTL) were examined longitudinally in an individual coinfected with human immunodeficiency virus type 1 (HIV-1), Epstein-Barr virus (EBV), and cytomegalovirus (CMV). CTL responses to all 3 viruses were quantified by limiting dilution analysis and staining with HLA-A*0201 tetrameric complexes folded with HIV-1, EBV, and CMV peptides. A predominance of CMV-pp65-speciflc CTL was found, with a much lower frequency of CTL to HIV-1 Gag and Pol and to EBV-BMLF1 and LMP2. The high frequency of CMV-speciflc CTL, compared with HIV-1- and EBV-specific CTL, was confirmed in an additional 16 HLA-A*0201-positive virus-coinfected subjects. Therefore, the human immune system can mount CTL responses to multiple viral antigens simultaneously, albeit with different strengths. View Show abstract Recombinant glycoprotein B vaccine formulation with Toll-like receptor 9 agonist and immune-stimulating complex induces specific immunity against multiple strains of cytomegalovirus Article Full-text available Feb 2011 J GEN VIROL Vijayendra Dasari Corey Smith Jie Zhong Rajiv Khanna Natural human cytomegalovirus (CMV) infection is characterized by a strain-specific neutralizing antibody response. This is particularly relevant in clinical settings such as transplantation and pregnancy where reinfection with heterologous strains occurs and the immune system does not mount an effective response against the infecting strain due to underlying immunosuppression. There is an emerging argument that a CMV vaccine that induces high titres of cross-neutralizing antibodies will be more effective in protecting individuals from infection with antigenically different CMV strains. In addition, induction of cell-mediated immunity offers the additional advantage of targeting virus-infected cells. This study presents a novel formulation of a CMV vaccine that, by combining recombinant soluble gB protein with a Toll-like receptor 9 agonist (CpG ODN1826) and immune-stimulating complexes (AbISCO 100), was able to elicit strong polyfunctional CMV-specific cellular and cross-neutralizing humoral immune responses. These data demonstrated that prime-boost immunization of human leukocyte antigen (HLA)-A2 mice with gB protein in combination with CpG ODN1826 and AbISCO 100 induced long-lasting CMV-specific CD4(+) and CD8(+) T-cell and humoral responses. Furthermore, these responses neutralized infection with multiple strains of CMV expressing different gB genotypes and afforded protection against challenge with recombinant vaccinia virus encoding the gB protein. These observations argue that this novel vaccine strategy, if applied to humans, should facilitate the generation of a robust, pluripotent immune response, which may be more effective in preventing infection with multiple strains of CMV. View Show abstract Differential kinetics of effector and regulatory T cells in patients on calcineurin inhibitor-based drug regimens Article Full-text available Jul 2009 KIDNEY INT Daniela Presser Janine Mohrbach Urban Sester Martina Sester Besides iatrogenic immunosuppression, endogenous suppression by regulatory T cells (Tregs) may also mediate inhibition of effector T cells after transplantation. Here we determined the effect of common immunosuppressive drug regimens on both Treg and effector T cells. Tregs and cytomegalovirus (CMV)-specific T cells were quantified in 88 renal transplant recipients, 58 hemodialysis patients, and 22 controls. T cell dynamics were longitudinally assessed within 20 weeks after transplantation. The number of Tregs was quantified by measurement of CD25 and/or FOXP3-positive cells and by functional assays. CMV-specific T cells were quantified by stimulation-induced intracellular cytokine analysis. Treg frequencies in transplant recipients were significantly lower compared to those in hemodialysis patients and controls. These lower Treg levels were associated with a less pronounced suppression of effector function. Treg levels decreased within the first weeks after transplantation and remained low in the long term. In contrast, although decreased at early post-transplant, long-term levels of CMV-specific T cells normalized to levels found in hemodialysis patients and controls. These studies suggest that there is an initial decrease of Tregs and effector T cells as a consequence of a direct inhibitory effect of immunosuppressive drugs. In the long term, persistently low Treg levels may favor normalization of effector T cells to ensure sufficient pathogen control. View Show abstract Neutralization of Cytomegalovirus Virions: The Role of Complement Article Full-text available Sep 1997 Owen Brad Spiller S. Melanie Hanna Dana Devine Frank Tufaro Complement provides a key immunologic defense against invading pathogens; thus, a clear understanding of the interactions between cytomegalovirus (CMV) and complement may permit the development of strategies to enhance CMV neutralization. In the presence of specific anti-CMV antibodies, complement enhanced the neutralizing ability of serum by 2- to 3-fold. However, in the absence of specific anti-CMV antibodies, complement was ineffective in neutralizing CMV virions by plaque assay. Although complement alone did not mediate any neutralizing effect, CMV consumed complement activity from seronegative serum, resulting in the deposition of C3 on the virion. However, only in the presence of specific anti-CMV antibody did complement activation continue to the deposition of C9 on the virions. These results strongly suggest complement regulation by CMV virions that is modulated by anti-CMV antibody; this regulation may be attributed to three host complement regulators on the virions: CD55, CD46, and CD59. View Show abstract Circulating cytomegalovirus (CMV) neutralizing activity in bone marrow transplant recipients: comparison of passive immunity in a randomized study of four intravenous IgG products administered to CMV-seronegative patients Article Nov 1992 Alexandra H Filipovich M H Peltier M K Bechtel J A Englund Forty-two cytomegalovirus (CMV)-seronegative bone marrow transplant (BMT) recipients were randomized in a double-blind fashion to receive one of four commercially available intravenous Ig (IVIgG) products (Gamimmune N, Immune Globulin Intravenous, Gammagard, or Sandoglobulin) at a dose of 500 mg/kg every other week. The four treatment groups were similar in distribution of patient ages, weights, autologous versus allogeneic donor type, and underlying diseases. Every other week administration of IVIgG provided total serum IgG levels within the physiologic range for age. CMV titers by latex agglutination were stable (average geometric mean titer of 18.4 after the second IVIgG dose), with no statistically significant differences among the four product groups. CMV neutralizing activity (CMVNA) and CMV enzyme-linked immunosorbent assay (ELISA) titers were determined on a subset of sera from 27 study patients representing the four product groups. Patient serum samples obtained before IVIgG infusions and 2 weeks after the second IVIgG dose (ie, 3 weeks post-BMT) were assayed for CMVNA and CMV ELISA titers. Geometric mean titers of CMVNA and CMV ELISA varied among the product groups. The highest mean 50% CMVNA was 1:43 for product B, whereas the lowest mean 50% CMVNA was 1:14 for product A; two of the IVIgG product groups showed intermediate 50% mean titers of 1:27 (product C) and 1:26 (product D) for an overall P = .02. CMV ELISA titers (expressed as Paul Ehrlich International units [PEI U]) also showed the highest mean of 2.95 PEI U/mL for product B and the lowest mean of 1.34 PEI U/mL for product A. Intermediate mean values of 2.27 PEI U/mL and 2.03 PEI U/mL were obtained with products C and D, respectively (overall P = .003). The CMV ELISA titers show a minimal correlation (r = .566) to the observed CMVNA titers. We conclude that commercially available IVIgG products provide passive CMVNA, and that the level of circulating CMVNA is affected by the IVIgG product used. View Show abstract Increased in vitro Immunosuppressive Action of Anti-CMV and Anti-HBs Intravenous Immunoglobulins due to Higher Amounts of Interferon-Gamma Specific Neutralizing Antibodies Article Jan 1997 E C Dupont C Denys Michel Toungouz View Pharmacokinetics of an Anti-Cytomegalovirus Hyperimmunoglobulin after Single Intravenous Administration to Healthy Volunteers Article Jan 1991 A Millendorfer M Scriba Helmut Bachmayer E. Gaudera View Updated International Consensus Guidelines on the Management of Cytomegalovirus in Solid-Organ Transplantation Article Aug 2013 Camille Kotton Angela M Caliendo on behalf of The Transplantation Society International CMV Consensus Group Deepali Kumar Cytomegalovirus (CMV) continues to be one of the most common infections after solid-organ transplantation, resulting in significant morbidity, graft loss, and adverse outcomes. Management of CMV varies considerably among transplant centers but has been become more standardized by publication of consensus guidelines by the Infectious Diseases Section of The Transplantation Society. An international panel of experts was reconvened in October 2012 to revise and expand evidence and expert opinion-based consensus guidelines on CMV management, including diagnostics, immunology, prevention, treatment, drug resistance, and pediatric issues. The following report summarizes the recommendations. View Show abstract Broadly targeted human cytomegalovirus-specific CD4(+) and CD8(+) T-cells dominate the memory compartments of exposed subjects (vol 202, pg 673, 2005) Article Nov 2005 Andrew W Sylwester BL Mitchell John B Edgar Louis J Picker View Humoral Immunity Profiles to Identify CMV-seropositive Heart Recipients at Risk of CMV Disease: A Prospective Multicenter Study Article Apr 2014 J HEART LUNG TRANSPL Maria Isabel Jaramillo Elizabeth Sarmiento Jordi Navarro Javier Carbone View CMV-Seronegative Heart Recipients Disclose a Lower Immunocompetence Status Before Transplantation in Comparison With Seropositive Patients Article Apr 2014 J HEART LUNG TRANSPL J. Rodriguez-Molina Javier Carbone Jordi Navarro Elizabeth Sarmiento View Antibody-Mediated Response of NKG2Cbright NK Cells against Human Cytomegalovirus Article Feb 2015 J IMMUNOL Marcel Costa-Garcia Andrea Vera Manuela Moraru Aura Muntasell Human CMV (HCMV) infection promotes a variable and persistent expansion of functionally mature NKG2C(bright) NK cells. We analyzed NKG2C(bright) NK cell responses triggered by Abs from HCMV(+) sera against HCMV-infected MRC5 fibroblasts. Specific Abs promoted the degranulation (i.e., CD107a expression) and the production of cytokines (TNF-α and IFN-γ) by a significant fraction of NK cells, exceeding the low natural cytotoxicity against HCMV-infected targets. NK cell-mediated Ab-dependent cell-mediated cytotoxicity was limited by viral Ag availability and HLA class I expression on infected cells early postinfection and increased at late stages, overcoming viral immunoevasion strategies. Moreover, the presence of specific IgG triggered the activation of NK cells against Ab-opsonized cell-free HCMV virions. As compared with NKG2A(+) NK cells, a significant proportion of NKG2C(bright) NK cells was FcεR γ-chain defective and highly responsive to Ab-driven activation, being particularly efficient in the production of antiviral cytokines, mainly TNF-α. Remarkably, the expansion of NKG2C(bright) NK cells in HCMV(+) subjects was related to the overall magnitude of TNF-α and IFN-γ cytokine secretion upon Ab-dependent and -independent activation. We show the power and sensitivity of the anti-HCMV response resulting from the cooperation between specific Abs and the NKG2C(bright) NK-cell subset. Furthermore, we disclose the proinflammatory potential of NKG2C(bright) NK cells, a variable that could influence the individual responses to other pathogens and tumors. Copyright © 2015 by The American Association of Immunologists, Inc. View Show abstract Cytomegalovirus Enhances Macrophage TLR Expression and MyD88-Mediated Signal Transduction To Potentiate Inducible Inflammatory Responses Article Oct 2014 Phillip D Smith Masako Shimamura Lois C Musgrove Lesley E. Smythies Circulating monocytes carrying human CMV (HCMV) migrate into tissues, where they differentiate into HCMV-infected resident macrophages that upon interaction with bacterial products may potentiate tissue inflammation. In this study, we investigated the mechanism by which HCMV promotes macrophage-orchestrated inflammation using a clinical isolate of HCMV (TR) and macrophages derived from primary human monocytes. HCMV infection of the macrophages, which was associated with viral DNA replication, significantly enhanced TNF-α, IL-6, and IL-8 gene expression and protein production in response to TLR4 ligand (LPS) stimulation compared with mock-infected LPS-stimulated macrophages during a 6-d in vitro infection. HCMV infection also potentiated TLR5 ligand-stimulated cytokine production. To elucidate the mechanism by which HCMV infection potentiated inducible macrophage responses, we show that infection by HCMV promoted the maintenance of surface CD14 and TLR4 and TLR5, which declined over time in mock-infected macrophages, and enhanced both the intracellular expression of adaptor protein MyD88 and the inducible phosphorylation of IκBα and NF-κB. These findings provide additional information toward elucidating the mechanism by which HCMV potentiates bacteria-induced NF-κB-mediated macrophage inflammatory responses, thereby enhancing organ inflammation in HCMV-infected tissues. View Show abstract Differential kinetics of human cytomegalovirus (HCMV) load and antibody responses in primary infection of the immunocompetent and immunocompromised host. Article Oct 2014 J GEN VIROL Daniele Lilleri Chiara Fornara Maria Grazia Revello Giuseppe Gerna The comparative long-term kinetics of human cytomegalovirus (HCMV) load and HCMV-specific antibody responses in the immunocompetent and immunocompromised solid-organ transplanted host during primary HCMV infection was investigated. On the whole, 40 immunocompetent subjects and 17 transplanted patients were examined for viral load as well as for IgG antibody responses to HCMV glycoproteins gH/gL/pUL128L, gH/gL and gB, and neutralizing antibodies in ARPE-19 epithelial cells and human fibroblasts. In parallel, the CD4+ and CD8+ HCMV-specific T-cell responses were determined by cytokine flow cytometry. Transplanted patients reached significantly higher viral DNA peaks, which persisted longer than in immunocompetent subjects. The ELISA-IgG responses to the pentamer, gH/gL and gB were significantly higher in primary infections of the immunocompetent until six months after onset, then the two antibody levels overlapped from six to 12 months. Antibody levels neutralizing infection of epithelial cells were significantly higher in transplanted patients after six months, persisting up to a year after transplantation. This trend was not observed for antibodies neutralizing infection of human fibroblasts, which showed higher titers in the immunocompetent over the entire 1-year follow-up. In conclusion, in immunocompromised patients the viral load peak was much higher, while the neutralizing antibody response exceeded that detected in the immunocompetent host starting six months after onset of follow-up, often concomitantly with a lack of specific CD4+ T-cells. In these setting, the elevated antibody response occurred in the presence of differentiated follicular helper T-cells in blood, which decreased in number as did antibody titers upon reappearance of HCMV-specific CD4+ T-cells. View Show abstract Evaluation of an immunological score to assess the risk of severe infection in heart recipients Article Aug 2014 TRANSPL INFECT DIS Jordi Navarro Juan Fernández-Yáñez Elizabeth Sarmiento Javier Carbone Background We previously reported how specific humoral and cellular immunological markers that are readily available in clinical practice can be used to identify heart transplant recipients (HTR) at risk of developing severe infections. In this study, we perform an extended analysis to identify immunological profiles that could prove to be superior to individual markers in assessing the risk of infection early after heart transplantation.Methods In a prospective follow-up study, we evaluated 100 HTR at 1 week after transplantation. Laboratory tests included determination of immunoglobulin (Ig) levels (IgG, IgA, IgM), complement factors (C3 and C4), and lymphocyte subsets (CD3+, CD4+, CD8+ T cells, B cells, and natural killer [NK] cells). The prevalence of infection during the first 3 months was registered at scheduled visits after transplantation. Severe infections were defined as all infections requiring hospitalization and intravenous antimicrobial therapy.ResultsDuring follow-up, 33 patients (33%) developed severe infections. The individual risk factors of severe infection, according to the Cox regression analysis, were as follows: IgG <600 mg/dL (hazard ratio [HR], 2.41; 95% confidence interval [CI], 1.21–4.78; P = 0.012), C3 <80 mg/dL (HR, 4.65; 95% CI, 2.31–9.38; P < 0.0001), C4 <18 mg/dL (HR 2.30, 95% CI, 1.15–4.59; P = 0.018), NK count <30 cells/μL (HR 4.07, 95% CI, 1.76–9.38; P = 0.001), and CD4 count <350 cells/μL (HR, 3.04; 95% CI, 1.47–6.28; P = 0.0027). An immunological score was created. HRs were used to determine the number of points assigned to each of the 5 previously mentioned individual risk factors. The score was obtained from the sum of these factors. In the multivariate Cox regression analysis, the immunological score was useful for identifying patients at risk of infection and was the only variable that maintained a significant association with the development of infection, after adjustment for the 5 individual factors.Conclusion Patients with an immunological score ≥13 were at the highest risk of severe infections (HR, 9.29; 95% CI, 4.57–18.90; P < 0.0001). This score remained significantly associated with the risk of severe infection after adjustment for clinical risk factors of infection. An immunological score was useful for identifying HTR at risk of developing severe infections. If this score is validated in multicenter studies, it could be easily introduced into clinical practice. View Show abstract Impact of Antiviral Preventive Strategies on the Incidence and Outcomes of Cytomegalovirus Disease in Solid Organ Transplant Recipients Article Aug 2013 AM J TRANSPLANT Georg Kralidis Nicolas J Mueller Pascal Meylan Oriol Manuel We assessed the impact of antiviral prophylaxis and preemptive therapy on the incidence and outcomes of cytomegalovirus (CMV) disease in a nationwide prospective cohort of solid organ transplant recipients. Risk factors associated with CMV disease and graft failure-free survival were analyzed using Cox regression models. One thousand two hundred thirty-nine patients transplanted from May 2008 until March 2011 were included; 466 (38%) patients received CMV prophylaxis and 522 (42%) patients were managed preemptively. Overall incidence of CMV disease was 6.05% and was linked to CMV serostatus (D+/R- vs. R+, hazard ratio [HR] 5.36 [95% CI 3.14-9.14], p < 0.001). No difference in the incidence of CMV disease was observed in patients receiving antiviral prophylaxis as compared to the preemptive approach (HR 1.16 [95% CI 0.63-2.17], p = 0.63). CMV disease was not associated with a lower graft failure-free survival (HR 1.27 [95% CI 0.64-2.53], p = 0.50). Nevertheless, patients followed by the preemptive approach had an inferior graft failure-free survival after a median of 1.05 years of follow-up (HR 1.63 [95% CI 1.01-2.64], p = 0.044). The incidence of CMV disease in this cohort was low and not influenced by the preventive strategy used. However, patients on CMV prophylaxis were more likely to be free from graft failure. View Show abstract T-cell inhibitory capacity of hyperimmunoglobulins is influenced by the production process Article Jan 2014 Rogier van Gent Haziz Jaadar Angela Tjon Jaap Kwekkeboom Intravenous immunoglobulin (IVIg) preparations are widely used for anti-inflammatory therapy of autoimmune and systemic inflammatory diseases. Hyperimmunoglobulins enriched in neutralizing antibodies against viruses can, in addition to their virus-neutralizing activity, also exert immunomodulatory activity. Previously, we observed that Cytotect®, an anti-CMV hyperimmunoglobulin, was less effective in suppressing human T-cell responses in vitro compared to Hepatect® CP, an anti-HBV hyperimmunoglobulin. We hypothesized that the poor immunomodulatory activity of Cytotect® results from treatment with β-propiolactone during the manufacturing process. The manufacturer of these hyperimmunoglobulins has now introduced a new anti-CMV hyperimmunoglobulin, called Cytotect® CP, in which β-propiolactone treatment is omitted. Here we show that Cytotect® CP inhibits PHA-driven T-cell proliferation and cytokine production with similar efficacy as Hepatect® CP, whereas the former Cytotect® does not. In addition, Cytotect® CP inhibits allogeneic T-cell responses better than Cytotect®. Our results advocate the use of hyperimmunoglobulins that have not been exposed to β-propiolactone in order to benefit from their immunomodulatory properties. View Show abstract PD-1 Analysis on CD28(-)CD27(-) CD4 T Cells Allows Stimulation-Independent Assessment of CMV Viremic Episodes in Transplant Recipients Article Oct 2013 AM J TRANSPLANT Jan Dirks H. Tas T. Schmidt Martina Sester Expression of the inhibitory receptor programmed death 1 (PD-1) on cytomegalovirus (CMV)-specific CD4 T cells defines a phenotype associated with CMV viremia in transplant recipients. Moreover, CD28(-) CD27(-) double negativity is known as a typical phenotype of CMV-specific CD4 T cells. Therefore, the co-expression of inhibitory receptors on CD28(-) CD27(-) CD4 T cells was assessed as a rapid, stimulation-independent parameter for monitoring CMV complications after transplantation. Ninety-three controls, 67 hemodialysis patients and 81 renal transplant recipients were recruited in a cross-sectional and longitudinal manner. CMV-specific CD4 T cell levels quantified after stimulation were compared to levels of CD28(-) CD27(-) CD4 T cells. PD-1 and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) expression on CD28(-) CD27(-) CD4 T cells were related to viremia. A percentage of ≥0.44% CD28(-) CD27(-) CD4 T cells defined CMV seropositivity (93.3% sensitivity, 97.1% specificity), and their frequencies correlated strongly with CMV-specific CD4 T cell levels after stimulation (r = 0.73, p < 0.0001). Highest PD-1 expression levels on CD28(-) CD27(-) CD4 T cells were observed in patients with primary CMV viremia and reactivation (p < 0.0001), whereas CTLA-4 expression was only elevated during primary CMV viremia (p < 0.05). Longitudinal analysis showed a significant increase in PD-1 expression in relation to viremia (p < 0.001), whereas changes in nonviremic patients were nonsignificant. In conclusion, increased PD-1 expression on CD28(-) CD27(-) CD4 T cells correlates with CMV viremia in transplant recipients and may serve as a specific, stimulation-independent parameter to guide duration of antiviral therapy. View Show abstract What Is the Impact of Hypogammaglobulinemia on the Rate of Infections and Survival in Solid Organ Transplantation? A Meta-Analysis Article Aug 2013 AM J TRANSPLANT Diana Florescu Andre C Kalil Fang Qiu Uriel Sandkovsky Hypogammaglobulinemia has been described after solid organ transplantation and has been associated with increased risk of infections. The aim of the study was to evaluate the rate of severe hypogammaglobulinemia and its relationship with the risk of infections during the first year posttransplantation. Eighteen studies (1756 patients) that evaluated hypogammaglobulinemia and posttransplant infections were included. The data were pooled using the DerSimonian and Laird random-effects model. Q statistic method was used to assess statistical heterogeneity. Within the first year posttransplantation, the rate of hypogammaglobulinemia (IgG < 700 mg/dL) was 45% (95% CI: 0.34-0.55; Q = 330.1, p < 0.0001), the rate of mild hypogammaglobulinemia (IgG = 400-700 mg/dL) was 39% (95% CI: 0.22-0.56; Q = 210.09, p < 0.0001) and the rate of severe hypogammaglobulinemia (IgG < 400 mg/dL) was 15% (95% CI: 0.08-0.22; Q = 50.15, p < 0.0001). The rate of hypogammaglobulinemia by allograft type: heart 49% (21%-78%; Q = 131.16, p < 0.0001); kidney 40% (30%-49%; Q = 24.55, p = 0.0002); liver 16% (0.001%-35%; Q = 14.31, p = 0.0002) and lung 63% (53%-74%; Q = 6.85, p = 0.08). The odds of respiratory infection (OR = 4.83; 95% CI: 1.66-14.05; p = 0.004; I(2) = 0%), CMV (OR = 2.40; 95% CI: 1.16-4.96; p = 0.02; I(2) = 26.66%), Aspergillus (OR = 8.19; 95% CI: 2.38-28.21; p = 0.0009; I(2) = 17.02%) and other fungal infections (OR = 3.69; 95% CI: 1.11-12.33; p = 0.03; I(2) = 0%) for patients with IgG <400 mg/dL were higher than the odds for patients with IgG >400 mg/dL. The odds for 1-year all-cause mortality for severe hypogammaglobulinemia group was 21.91 times higher than those for IgG >400 mg/dL group (95% CI: 2.49-192.55; p = 0.005; I(2) = 0%). Severe hypogammaglobulinemia during the first year posttransplantation significantly increased the risk of CMV, fungal and respiratory infections, and was associated with higher 1-year all-cause mortality. View Show abstract Neutralizing antibodies are unable to inhibit direct viral cell-to-cell spread of human cytomegalovirus Article Jul 2013 VIROLOGY Christian L Jacob Louie Lamorte Eliud Sepulveda Michael Franti Infection with human cytomegalovirus (CMV) during pregnancy is the most common cause of congenital disorders, and can lead to severe life-long disabilities with associated high cost of care. Since there is no vaccine or effective treatment, current efforts are focused on identifying potent neutralizing antibodies. A panel of CMV monoclonal antibodies identified from patent applications, was synthesized and expressed in order to reproduce data from the literature showing that anti-glycoprotein B antibodies neutralized virus entry into all cell types and that anti-pentameric complex antibodies are highly potent in preventing virus entry into epithelial cells. It had not been established whether antibodies could prevent subsequent rounds of infection that are mediated primarily by direct cell-to-cell transmission. A thorough validation of a plaque reduction assay to monitor cell-to-cell spread led to the conclusion that neutralizing antibodies do not significantly inhibit plaque formation or reduce plaque size when they are added post-infection. View Show abstract Cytomegalovirus Infection Rate among Heart Transplant Patients in Relation to Antithymocyte Immunoglobulin Induction Therapy Article Jan 1994 Kim Krogsgaard Søren Boesgaard J Aldershvile G Petterson During a 2-year period, 49 patients underwent heart transplantation at Rigshospitalet, Copenhagen. Nine (18%) were females and the mean age for all patients was 44 years (range 14–56 years). Immunosuppressive therapy included cyclosporin, azathioprine and steroids in all patients. 43 patients received in addition short-term (approx. 4 days) induction treatment with antithymocyte immunoglobulin (ATG). 17 patients received ATG Fresenius®, 2.5mg/kg/day or ATGAM®, 12.5 mg/kg/day, whereas the remaining 26 patients received ATG Merieux®, 2.5 mg/kg/day. Prophylactic antimicrobial chemotherapy included ceftriaxone, acyclovir (1 g daily), nystatin, and pyrimethamine in toxoplasmosis mismatch patients. Serological assays for cytomegalovirus (CMV), Epstein-Barr virus, varicella-zoster virus, herpes simplex virus, legionella and toxoplasmosis as well as CMV and bacterial culturing were carried out before transplantation, at regular intervals and when clinically indicated. Five patients developed septicaemia. Nine had pulmonary bacterial infections, including 2 cases of legionella pneumonia. Two had Clostridium difficile diarrhoea. Three patients had Pneumocystis carinii pneumonitis. 24 patients (49%) had evidence of CMV infection/reactivation. Seven out of 10 CMV mismatch (pos donor/neg recipient) patients and 3 out of 12 CMV match (pos donor/pos recipient) patients developed clinical CMV disease. The rate of CMV infection/reactivation was significantly higher among patients who had CMV-positive donors (p < 0.01) and among patients receiving ATG Merieux®, induction treatment (p < 0.0001). Logistic regression analysis showed that both positive CMV donor status and ATG Merieux® induction treatment were significant independent predictors of CMV infection. Six patients (12%) died. Two out of 4 infection related deaths could be ascribed to CMV disease. View Show abstract Everolimus is associated with a reduced incidence of cytomegalovirus infection following de novo cardiac transplantation Article Sep 2012 TRANSPL INFECT DIS J. Kobashigawa Heather Ross Christoph Bara Luciano Potena Cytomegalovirus (CMV) causes several complications following cardiac transplantation including cardiac allograft vasculopathy. Previous studies suggested that immunosuppressive treatment based on everolimus might reduce CMV infection. Aiming to better characterize the action of everolimus on CMV and its interplay with patient/recipient serology and anti-CMV prophylaxis, we analyzed data from 3 large randomized studies comparing various everolimus regimens with azathioprine (AZA)- and mycophenolate mofetil (MMF)-based regimens. CMV data were analyzed from 1009 patients in 3 trials of de novo cardiac transplant recipients who were randomized to everolimus 1.5 mg/day, everolimus 3 mg/day, or AZA 1–3 mg/kg/day, plus standard-dose (SD) cyclosporine (CsA; study B253, n = 634); everolimus 1.5 mg/day plus SD- or reduced-dose (RD)-CsA (study A2403, n = 199); and everolimus 1.5 mg/day plus RD-CsA or MMF plus SD-CsA (study A2411, n = 176). In study B253, patients allocated to everolimus experienced almost a 70% reduction in odds of experiencing CMV infection compared with AZA (P < 0.001). In study A2403, CMV infection was low in both everolimus arms, irrespective of CsA dosing, and in study A2411, patients allocated to everolimus experienced an 80% reduction in odds of experiencing CMV infection, compared with MMF (P < 0.001). CMV syndrome/disease was rare and less frequent in everolimus-treated patients. Subgroup analyses showed that the benefit everolimus provides, in terms of CMV events, is retained in CMV-naïve recipients and is independent of anti-CMV prophylaxis or preemptive approaches. Everolimus is associated with a lower incidence of CMV infection compared with AZA and MMF, which combined with its immunosuppressive efficacy and antiproliferative effects may positively impact long-term outcomes. View Show abstract Decreased levels of serum complement C3 and natural killer cells add to the predictive value of total immunoglobulin G for severe infection in heart transplant recipients Article Oct 2012 TRANSPL INFECT DIS N. Del Pozo Elizabeth Sarmiento A. Gallego Javier Carbone Infection remains a source of mortality in heart recipients. We previously reported that post-transplant immunoglobulin G (IgG) quantification can help identify the risk for infection. We assessed whether other standardized parameters of humoral and cellular immunity could prove useful when identifying patients at risk of infection. We prospectively studied 133 heart recipients over a 12-month period. Forty-eight patients had at least one episode of severe infection. An event was defined as an infection requiring intravenous antimicrobial therapy. Cox regression analysis revealed an association between the risk of developing infection and the following: lower IgG2 subclass levels (day 7: relative hazard [RH] 1.71; day 30: RH 1.76), lower IgA levels (day 7: RH 1.61; day 30: RH 1.91), lower complement C3 values (day 7: RH 1.25), lower CD3 absolute counts (day 30: RH 1.10), lower absolute natural killer [NK] cell count (day 7: RH 1.24), and lower IgG concentrations (day 7: RH 1.31; day 30: RH 1.36). Cox regression bivariate analysis revealed that lower day 7 C3 levels, IgG2 concentration, and absolute NK cell count remained significant after adjustment for total IgG levels. Data suggest that early immune monitoring including C3, IgG2, and NK cell testing in addition to IgG concentrations is useful when attempting to identify the risk of infection in heart transplant recipients. View Show abstract Immune response to CMV in solid organ transplant recipients: Current concepts and future directions Article May 2012 Expet Rev Clin Immunol Richard R Watkins Tracy L Lemonovich Raymund R Razonable Despite advances in immunosuppression and antiviral therapy, CMV continues to be a significant opportunistic pathogen adversely affecting the outcome of solid organ transplantation (SOT) recipients. While a significant proportion of CMV disease is caused by reactivation of latent virus, the risk is highest among CMV donor+ and recipient- SOT patients. CMV is responsible for both direct (e.g., pneumonitis, colitis) and indirect (e.g., rejection, atherosclerosis) morbidity and mortality. Healthy CMV-seropositive individuals have a high frequency of CMV-specific CD4(+) and CD8(+) T cells that provide immune protection by limiting CMV reactivation and replication. Changes to the innate and adaptive immune system from immunosuppressive therapy following SOT contribute to CMV disease pathogenesis. CMV disease after SOT is associated with poorer outcomes, thus novel strategies to prevent it are an area of active research. In this article, we review the current state of knowledge on the immune response to CMV following SOT. View Show abstract Neutralizing Activity of Immunoglobulin Preparations Article Feb 2012 Stuart P Adler View Serum antibody response to the gH/gL/pUL128-131 five-protein complex of human cytomegalovirus (HCMV) in primary and reactivated HCMV infections Article Aug 2011 Emilia Genini Elena Percivalle Antonella Sarasini Giuseppe Gerna Recently, a new human cytomegalovirus (HCMV) glycoprotein complex has been identified and potentially proposed as a vaccine. The aim of this study was to determine whether the HCMV gH/gL/pUL128-pUL130-pUL131 (gH/gL/pUL128-131) 5-protein (pentameric) complex (which has been recently found to be indispensable for the infection of endothelial and epithelial cells) is able to elicit a consistent antibody response in both primary and reactivated HCMV infections. The antibody response was determined by both indirect immunofluorescence (IFA) and ELISA, using fixed (IFA) or lysed (ELISA) epithelial (ARPE-19) cells infected with one or more adenoviral vectors, each carrying one HCMV gene and, in parallel, with a control adenovirus vector. The specificity of results was determined by the reactivity of human neutralizing mAbs recognizing two, three, or four proteins of the complex. In 14 cases of primary infection, an IgG antibody seroconversion to the UL128-131 gene products was consistently detected within 2-4 weeks after onset of infection, while antibodies persisted for at least 12 months. The IgG antibody response to UL128-131 gene products was generally superior to the response to gH and appeared to follow the neutralizing antibody response (as determined in epithelial cells). In reactivated infections, the antibody response showed a trend reminiscent of a booster response. IgG antibodies were detected in HCMV-seropositive healthy adult controls, but not in HCMV-seronegative individuals. The IgG antibody response to the pentameric complex could be a major target for the evaluation of the antibody response to a pentamer-based vaccine. View Show abstract Cytomegalovirus Neutralization by Hyperimmune and Standard Intravenous Immunoglobulin Preparations Article Jun 2011 Christina B Planitzer Marcus D Saemann Hartwig Gajek Thomas R. Kreil Cytomegalovirus (CMV) remains one of the most important pathogens after transplantation, potentially leading to CMV disease, allograft dysfunction, acute, and chronic rejection and opportunistic infections. Immunoglobulin G (IgG) preparations with high antibody titers against CMV are a valuable adjunctive prevention and treatment option for clinicians and apart from standard intravenous immunoglobulin (IVIG), CMV hyperimmune preparations are available. The CMV antibody titer of these preparations is typically determined by Enzyme-linked immunosorbent assay (ELISA), also used for the selection of high titer plasma donors for the production of the CMV Hyperimmune product. However, CMV ELISA titers do not necessarily correlate with CMV antibody function which is determined by virus neutralization tests. CMV antibody titers were determined by both ELISA and virus neutralization assay and the IgG subclass distribution was compared between a CMV hyperimmune licensed in Europe and standard IVIG preparations. Although the expected high CMV IgG ELISA antibody titers were confirmed for three lots of a CMV hyperimmune preparation, the functionally more relevant CMV neutralizing antibody titers were significantly higher for 31 lots of standard IVIG preparations. Moreover, considerably lower IgG3 levels were found for the CMV hyperimmune preparation compared with standard IVIG preparations. The higher functional CMV neutralization titers of standard IVIG preparations and the better availability of these preparations, suggest that these products could be a valuable alternative to the CMV hyperimmune preparation. View Show abstract Lack of antibodies against the antigen domain 2 epitope of cytomegalovirus (CMV) glycoprotein B is associated with CMV disease after renal transplantation in recipients having the same glycoprotein H serotypes as their donors Article Jun 2011 TRANSPL INFECT DIS K. Ishibashi Tadahiko Tokumoto H Shirakawa Tatsuo Suzutani Cytomegalovirus (CMV) reinfection of seropositive individuals has been associated with adverse outcomes in organ transplantation and is a frequent cause of congenital infection. Previously we demonstrated that mismatching of CMV glycoprotein H (gH) serotypes was associated with CMV disease after renal transplantation. Because the antigen domain 2 (AD2) epitope of glycoprotein B (gB) is conserved among CMV isolates and is one of the known targets of neutralizing antibodies, in this study we investigated whether antibodies against the epitope contribute to protection from CMV reinfection in renal transplantation, irrespective of gH serological matching. For this purpose, the gB and gH serology and clinical outcomes were analyzed retrospectively for 77 transplant recipients in the donor positive/recipient positive setting, who were managed by preemptive strategy. We found that there was a good negative correlation between the numbers of antigenemia-positive cells and the levels of antibodies against gB AD2 in the CMV-gH antibody matched group, but not in the CMV-gH antibody mismatched group. None of the recipients with antibodies against both gB AD2 and strain-specific epitopes of gH have experienced CMV disease during 6 month after transplantation, while 28% of those who lacked either/both antibody response needed preemptive therapy. Because the outcome was statistically significant, antibodies against gB AD2 can be a useful indicator to predict emergence of CMV disease for preemptive therapy, in addition to antibodies against the mismatched gH types. View Show abstract Recombinant human monoclonal antibodies to human cytomegalovirus glycoprotein B neutralize virus in a complement-dependent manner Article Aug 2009 Akane Ohta Ayano Fujita Tsugiya Murayama Yoshizo Asano Human antibodies specific for HCMV are currently considered as potential anti-HCMV therapeutic agents. In this study, we used a combinatorial human antibody library to isolate and characterize complete human monoclonal antibodies that effectively neutralize HCMV in a complement-dependent manner. One hundred and six clones were isolated in two independent screens using HCMV virions and recombinant glycoprotein B, gB654, as antigens. All of the clones recognized the same molecule gB and were classified into 14 groups based on the amino acid sequence of the V(H) region. Seven representative clones from these 14 groups had a strong gB654 binding affinity by surface plasmon resonance (SPR). A pairwise binding competition analysis suggested that there were three groups based on differences in the gB recognition sites. Although Fab fragments of the seven groups showed strong affinity for gB, none of the Fab fragments neutralized HCMV infectivity in vitro. In contrast, complete human IgG(1) antibodies of at least three groups neutralized HCMV in a complement-dependent manner. These data suggest that potent therapeutic antibodies can be obtained from a human antibody library, including most of the functional antibodies that mediate humoral immunity to the selected pathogen. View Show abstract Immune monitoring of anti cytomegalovirus antibodies and risk of cytomegalovirus disease in heart transplantation Article Oct 2008 Elizabeth Sarmiento Nallibe Lanio A. Gallego Javier Carbone We sought to determine whether quantitative assessment of anti-cytomegalovirus (CMV) antibodies could be useful to identify patients at risk of cytomegalovirus (CMV) disease after heart transplantation (HT). 75 patients who underwent HT at a single health care center were prospectively studied. Induction therapy included 2 doses of daclizumab and maintenance tacrolimus (n=42) or cyclosporine (n=29), mycophenolate mofetil and prednisone. All patients received prophylaxis with gancyclovir or valganciclovir. Anti-CMV intravenous immunoglobulin (CMV-IG) was added in high risk patients (CMV D+/R- serostatus). Serial determinations of anti-CMV antibodies, immunoglobulins (IgG, IgA, IgM) and IgG-subclasses were analysed. CMV infection was based on detection of the virus by antigenemia. CMV disease consisted of detection of signs or symptoms attributable to this microorganism. Ten patients (13.3%) developed CMV disease. Mean time of development of CMV disease was 3.4+/-1.6 months. In Cox regression analysis, patients with low baseline anti-CMV titers (<4.26 natural logarithm of titer, RH: 8.1, 95%CI: 1.93-34.1, p=0.004) and recipients with 1-month post-HT IgG hypogammaglobulinemia (IgG<500 mg/dl, RH: 4.49, 95%CI: 1.26-15.94, p=0.02) were at higher risk of having CMV disease. Despite use of prophylactic CMV-IG, D+/R- patients showed significantly lower titers of anti-CMV antibodies at 7 d, 30 d and 90 d post HT as compared with HT recipients without infections. Four out of 6 of these patients developed late CMV disease. Monitoring of specific anti-CMV antibodies on the bedside warrants further evaluation as a potential tool to identify heart transplant recipients at higher risk of CMV disease. View Show abstract Combined Cytomegalovirus Prophylaxis in Lung Transplantation: Effects on Acute Rejection, Lymphocytic Bronchitis/Bronchiolitis, and Herpesvirus Infections Article Jul 2008 TRANSPL P Paolo Solidoro Daniela Libertucci Luisa Delsedime Sergio Baldi Lung transplantation recipients are at high risk for herpesvirus infections. We evaluated the effect of combined cytomegalovirus (CMV) prophylaxis on CMV pneumonia, acute rejection episodes (ARE), lymphocytic bronchitis/bronchiolitis (LB), and obliterans bronchiolitis (OB) diagnosed in 180 transbronchial biopsies (TBB) of lung transplant recipients. At our center, 25 patients (control group; 1999-2002) received acyclovir for 12 months and 21 recipients (study group; 2003-2007) received combined CMV prophylaxis consisting of CMV-IG (Cytotect Biotest) for 12 months and ganciclovir or valganciclovir from postoperative day 21 for 3 weeks. Among the study group (since 2005), CMV shell vial viral culture and Epstein-Barr virus (EBV), human herpesvirus-6 (HHV-6), and HHV-7 DNA were determined on BAL specimens. In the study group, the number of LB was significantly lower than in the control group (2% vs 11%; P= .04). Similar results were obtained for ARE (6% vs 17%; P= .04). No difference was observed in OB (5% vs 5%; P= .53, NS). A reduction trend was found in CMV pneumonia (2% vs 7%; P= .23, NS). Logistic regression analysis showed a relationship between prophylaxis and a reduced prevalence of ARE (odds ratio [OR] 3.25, confidence interval [CI] 1.12-9.40; P= .03). Finally, in the study group, BAL EBV-DNA positivity and EBV-CMV coinfections were low (6% and 0%, respectively) compared with other herpesviruses and with the literature. Our data suggested the efficacy of combined CMV prophylaxis to prevent ARE and LB, 2 risk factors for chronic rejection, and a possible role to reduce the trend toward CMV pneumonia and EBV infections. View Show abstract Circulating Cytomegalovirus (CMV) neutralizing activity in bone marrow transplant recipients: Comparison of passive immunity in a randomized study of four intravenous IgG products administered to CMV-seronegative patients Article Dec 1992 Alexandra H Filipovich M H Peltier M K Bechtel J A Englund Forty-two cytomegalovirus (CMV)-seronegative bone marrow transplant (BMT) recipients were randomized in a double-blind fashion to receive one of four commercially available intravenous Ig (IVIgG) products (Gamimmune N, Immune Globulin Intravenous, Gammagard, or Sandoglobulin) at a dose of 500 mg/kg every other week. The four treatment groups were similar in distribution of patient ages, weights, autologous versus allogeneic donor type, and underlying diseases. Every other week administration of IVIgG provided total serum IgG levels within the physiologic range for age. CMV titers by latex agglutination were stable (average geometric mean titer of 18.4 after the second IVIgG dose), with no statistically significant differences among the four product groups. CMV neutralizing activity (CMVNA) and CMV enzyme-linked immunosorbent assay (ELISA) titers were determined on a subset of sera from 27 study patients representing the four product groups. Patient serum samples obtained before IVIgG infusions and 2 weeks after the second IVIgG dose (ie, 3 weeks post-BMT) were assayed for CMVNA and CMV ELISA titers. Geometric mean titers of CMVNA and CMV ELISA varied among the product groups. The highest mean 50% CMVNA was 1:43 for product B, whereas the lowest mean 50% CMVNA was 1:14 for product A; two of the IVIgG product groups showed intermediate 50% mean titers of 1:27 (product C) and 1:26 (product D) for an overall P = .02. CMV ELISA titers (expressed as Paul Ehrlich International units [PEI U]) also showed the highest mean of 2.95 PEI U/mL for product B and the lowest mean of 1.34 PEI U/mL for product A. Intermediate mean values of 2.27 PEI U/mL and 2.03 PEI U/mL were obtained with products C and D, respectively (overall P = .003). The CMV ELISA titers show a minimal correlation (r = .566) to the observed CMVNA titers. We conclude that commercially available IVIgG products provide passive CMVNA, and that the level of circulating CMVNA is affected by the IVIgG product used. View Show abstract Pharmacokinetics of an Anti-Cytomegalovirus Hyperimmunoglobulin after Single Intravenous Administration to Healthy Volunteers Article Feb 1991 U Ganzinger Jacqueline J. Martindale E. Gaudera Helmut Bachmayer By selecting blood donors with high cytomegalovirus (CMV) antibody titres, a plasma pool was obtained which was used to produce an IgG hyperimmunoglobulin by means of pepsin fractionation. After administration of approximately 100 mg/kg body weight to healthy subjects, the time course both of anti-CMV IgG antibody titres by ELISA and of virus neutralisation (VN) titres was followed for 15 days. Seronegative subjects showed an increase in CMV-IgG antibodies as well as a significant enhancement of VN. The time course of both titres was non-uniform. The decline of both titres was biphasic: CMV-IgG antibodies fell slowly during the first week and remained unchanged thereafter, whereas VN titres decreased markedly faster in the first than in the second week. In seropositive subjects, on the other hand, VN remained unchanged. CMV-IgG antibodies increased by approximately 3 times, followed by a similar biphasic decline as seen in seronegative subjects. Due to the differences between seronegative and seropositive subjects and to the non-uniform time course, no calculations of the elimination rate were feasible. View Show abstract Cytomegalovirus infection rate among heart transplant patients in relation to anti-thymocyte immunoglobulin induction therapy. Copenhagen Heart Transplant Group Article Jan 1994 K Krogsgaard Søren Boesgaard J Aldershvile G Petterson During a 2-year period, 49 patients underwent heart transplantation at Rigshospitalet, Copenhagen. Nine (18%) were females and the mean age for all patients was 44 years (range 14-56 years). Immunosuppressive therapy included cyclosporin, azathioprine and steroids in all patients. 43 patients received in addition short-term (approx. 4 days) induction treatment with antithymocyte immunoglobulin (ATG). 17 patients received ATG Fresenius, 2.5 mg/kg/day or ATGAM, 12.5 mg/kg/day, whereas the remaining 26 patients received ATG Merieux, 2.5 mg/kg/day. Prophylactic antimicrobial chemotherapy included ceftriaxone, acyclovir (1 g daily), nystatin, and pyrimethamine in toxoplasmosis mismatch patients. Serological assays for cytomegalovirus (CMV), Epstein-Barr virus, varicella-zoster virus, herpes simplex virus, legionella and toxoplasmosis as well as CMV and bacterial culturing were carried out before transplantation, at regular intervals and when clinically indicated. Five patients developed septicaemia. Nine had pulmonary bacterial infections, including 2 cases of legionella pneumonia. Two had Clostridium difficile diarrhoea. Three patients had Pneumocystis carinii pneumonitis. 24 patients (49%) had evidence of CMV infection/reactivation. Seven out of 10 CMV mismatch (pos donor/neg recipient) patients and 3 out of 12 CMV match (pos donor/pos recipient) patients developed clinical CMV disease. The rate of CMV infection/reactivation was significantly higher among patients who had CMV-positive donors (p < 0.01) and among patients receiving ATG Merieux induction treatment (p < 0.0001). Logistic regression analysis showed that both positive CMV donor status and ATG Merieux induction treatment were significant independent predictors of CMV infection. Six patients (12%) died. Two out of 4 infection related deaths could be ascribed to CMV disease. View Show abstract Immune Modulatory Effects of Immunoglobulins on Cell-Mediated Immune Responses In Vitro Article Dec 1993 Sven Klaesson Olle Ringdén L Markling Inger Lundkvist Intravenous Immunoglobulin (IVIG) at a concentration of 5 mg/ml, significantly inhibited mitogenic responses to phytohaemagglutinin (PHA), concanavalin A (conA) and pokeweed mitogen (PWM) by peripheral blood cells from healthy donors. No difference in inhibition by IVIG was seen when stimulating different T-lymphocyte cell subsets. Inhibition by IVIG was dose-dependent. An increased response was observed when IVIG was added more than 12 h after PHA compared to adding 1 h before [p= 0·05]. Intravenous immunoglobulin added to mixed lymphocyte cultures (MLC), reduced the median response by more than 60% (range 14–89%; P= 0·03) and almost completely abrogated the lymphocyte response to Staphylococcus aureus protein A (SPA), whose median inhibition was 94% (range 90–99%; P= 0·02). View Show abstract Incidence of rejection and infection after liver transplantation as a function of the primary disease: Possible influence of alcohol and polyclonal immunoglobulins Article Mar 1996 O Farges Faouzi Saliba H Farhamant Henri Bismuth A retrospective analysis was undertaken to determine if the incidence, timing, and severity of acute and chronic rejection were influenced by the primary disease necessitating transplantation. Of the 875 liver transplantations performed between 1984 and 1992, 768 were primary transplantations and 107 were retransplantations. Among the former, 330 patients that were liver transplant recipients for a chronic liver disease without cancer in the native liver received an ABO-compatible and cross-match-negative graft and were given a cyclosporine- or tacrolimus-based immunosuppression. These included primary biliary cirrhosis (PBC, 66 patients), primary sclerosing cholangitis (PSC, 23 patients), alcoholic cirrhosis (ALC, 21 patients), autoimmune cirrhosis (AIC, 17 patients), hepatitis B virus-induced cirrhosis (HBV-C, 116 patients) and hepatitis C virus-induced cirrhosis (HCV-C, 87 patients). The incidence of acute (48% +/- 3% [SE] at 1 year) and chronic rejection (10% +/- 2% at 3 years) was comparable in patients who have undergone transplantation for PBC, PSC, AIC, and HCV-C. However, the incidence of acute (but not chronic) rejection was significantly lower in patients who have undergone transplantation for ALC (29% at 1 year). This reduced incidence of acute rejection was associated with an increased incidence of bacterial infections. In patients who have undergone transplantation for HBV-C (the majority of whom had received long-term anti-hepatitis B surface antigen [HBs] immunoglobulins), the incidence of both acute (21% at 1 year) and chronic rejection (0% at 3 years) was significantly lower, whereas the incidence of septic complications was comparable with that in the other groups. The incidence of acute rejection in patients who have undergone transplantation for nonviral disease receiving polyclonal human anti-cytomegalovirus (CMV) immunoglobulins was also significantly lower than that of patients who did not receive the immunoglobulins (19% vs. 48% at 3 months; P = .01), and this was identical to that of patients who have undergone transplantation for viral disease receiving polyclonal human anti-HBs immunoglobulins (19% at 3 months). These results show that the risk of rejection is unequal among patients, being lower in patients who have undergone transplantation for ALC (probably as a result of a state of nonspecific hyporesponsiveness) and in patients who have undergone transplantation for HBV-C (possibly as a result of long-term administration of polyclonal human immunoglobulins). View Show abstract Increased in vitro Immunosuppressive Action of Anti-CMV and Anti-HBs Intravenous Immunoglobulins due to Higher Amounts of Interferon-Gamma Specific Neutralizing Antibodies Article Feb 1997 C Denys Michel Toungouz Etienne Dupont We previously found that interferon-gamma (IFN-gamma) antibodies in intravenous immunoglobulins (IVIG) can block not only IFN-gamma production and tumor necrosis factor-alpha secretion, but also T-cell proliferation. Since the presence of IFN-gamma antibodies has been attributed to previous viral infection, we hypothesized that the viral status of the plasma donors used for IVIG pools might be a decisive factor in controlling the immunosuppressive capacity of IVIG. We tested three different pooled, human IVIG preparations for the presence of IFN-gamma antibodies by ELISA. Comparison of the immunomodulatory activity of polyvalent IVIG with that of specific CMV and HBs IVIG showed that the latter-had higher levels of IFN-gamma antibodies and an increased capacity to block mixed lymphocyte reaction and cytokine production. We propose that these in vitro assays constitute a basis for the selection of plasma intended for manufacturing IVIG aimed at immunosuppression in the transplant setting. View Show abstract Levels of Virus-Specific Cd4 T Cells Correlate With Cytomegalovirus Control and Predict Virus-Induced Disease After Renal Transplantation1 Article Jun 2001 TRANSPLANTATION Martina Sester Urban Sester Barbara C Gärtner Hans Köhler Immunosuppressive treatment in transplant patients frequently causes infectious complications with cytomegalovirus (CMV). The extent of CMV replication can be followed by a number of diagnostic methods. There is, however, no simple diagnostic tool to assess the quality of the cellular antiviral immune response of an individual patient. This would be of particular importance for therapy decisions, as patients with detectable virus load do not necessarily develop CMV-related disease. Using a rapid whole blood assay, the frequencies of CMV-reactive CD4 and CD8 T cells were followed after renal transplantation to characterize their relative contribution in the containment of CMV infection. T cells from transplant patients ands healthy control persons were stimulated with CMV antigen in vitro. Based on specific cellular activation and induction of intracellular cytokines, the frequency of CMV-reactive CD4 and CD8 T cells was determined using flow cytometry. Viral load quantified using the "hybrid-capture" assay. The absence of CMV complications in long-term transplant recipients is reflected by stable virus-specific T-cell frequencies, which do not differ from healthy CMV-positive controls. In contrast, during the first months after transplantation, clinical symptoms are preceded by a decrease in CMV-reactive CD4 T-cell frequencies and an increase in CMV load. The individual immune response and CMV replication are critically balanced and can be characterized by assesing both viral load and antiviral T cells. Our experimental design allows the identification of patients with sufficient, insufficient, or absent T-cell activity and can serve as diagnostic tool to facilitate decisions on antiviral therapy. View Show abstract Show more Recommendations Discover more Article A prospective cross-sectional study of BK virus infection in non-renal solid organ transplant recipi... June 2006 · Transplant Infectious Disease Todd D Barton Emily Blumberg Alden M Doyle [...] A.P. Limaye Polyomavirus (primarily BK virus [BKV]) infection is an important cause of chronic renal dysfunction in renal transplant recipients, but its possible contribution to chronic renal dysfunction in non-renal solid organ transplant (NRSOT) recipients has not been fully explored. We performed a prospective, cross-sectional study of consecutive NRSOT recipients with unexplained chronic renal ... [Show full abstract] dysfunction of at least a 3 months duration. Medical records were reviewed, and polymerase chain reaction was used to amplify BKV-specific sequences from serum and urine samples. The potential associations between various demographic and transplant variables and BKV infection were assessed. Thirty-four consecutive NRSOT recipients (23 lung, 8 liver, 2 heart, 1 heart-lung) with chronic renal dysfunction were enrolled at a median of 3.5 years (range 0.3-12.5 years) post transplantation. Five of the 34 (15%) patients had BKV viruria (range 1040-1.8 x 10(6) copies/mL), but none had BKV viremia. BK viruria was associated with mycophenolate mofetil use (5 of 19 [26%] vs. 0 of 15, P = 0.03) and a history of cytomegalovirus disease (3 of 4 [75%] vs. 2 of 30 [7%], P < 0.01). However, the mean estimated creatinine clearance was similar in patients with or without BKV viruria (49 vs. 47 mL/min). BKV viruria was present in a proportion of NRSOT patients with otherwise unexplained chronic renal dysfunction. The possibility that BKV infection might contribute to chronic renal dysfunction in this setting warrants further investigation. Read more Article Full-text available CMV in Hematopoietic Stem Cell Transplantation June 2016 · Mediterranean Journal of Hematology and Infectious Diseases Rafael de la Camara Due to its negative impact on the outcome of stem cell transplant (SCT) and solid organ transplant patients (SOT) CMV has been called "the troll of transplantation". One of the greatest advances in the management of SCT has been the introduction of the preemptive strategy. Since its introduction, the incidence of the viremia, as expected, remains unchanged but there has been a marked decline in ... [Show full abstract] the incidence of early CMV disease. However, in spite of the advances in prevention of CMV disease, CMV is still today an important cause of morbidity and mortality. Late CMV disease is still occurring in a significant proportion of patients and the so-called indirect effects of CMV are causing significant morbidity and mortality. Fortunately there have been several advances in the development of new antivirals, adoptive immunotherapy and DNA-CMV vaccines that might transform the management of CMV in the near future. View full-text Article Identifying the patient at risk for post-transplant lymphoproliferative disorder July 2001 · Transplant Infectious Disease S M Cockfield Post-transplant lymphoproliferative disorders (PTLD) are a recognized complication of the immunosuppression required to prevent allograft rejection, occurring in 1-20% of recipients of solid organ transplants. Several factors greatly increase the risk of developing PTLD early post-transplant in any individual recipient. Epstein-Barr virus (EBV) infection is critical in the pathogenesis of the ... [Show full abstract] majority of these cases. Pre-transplant EBV seronegativity increases the incidence of PTLD 10- to 75-fold over that of EBV-seropositive recipients. Other risk factors include very young recipient age, cytomegalovirus infection or mismatching (donor positive-recipient negative), aggressive immunosuppression with conventional biologic agents, and the type of organ transplanted. In contrast, the risk of developing PTLD late in the post-transplant course does not appear to be influenced by the type of immunosuppressive agents employed, but rather by the duration of any immunosuppression. The role of EBV in late PTLD is also less certain, as a greater proportion of lesions are not associated with evidence of EBV infection. As the understanding of these risk factors has expanded, opportunities exist to target those populations at highest risk for the development of PTLD for aggressive monitoring and pre-emptive or prophylactic therapy. It is hoped that implementation of such strategies will render early PTLD a preventable complication of transplantation. Read more Article Unraveling the Biologic Basis of Late-Onset Cytomegalovirus Disease in High-Risk Organ Transplant Re... March 2011 · Transplantation Nina Singh Read more Last Updated: 13 Jan 2023 Looking for the full-text? You can request the full-text of this article directly from the authors on ResearchGate. Request full-text Already a member? Log in ResearchGate iOS App Get it from the App Store now. Install Keep up with your stats and more Access scientific knowledge from anywhere or Discover by subject area Recruit researchers Join for free Login Email Tip: Most researchers use their institutional email address as their ResearchGate login Password Forgot password? Keep me logged in Log in or Continue with Google Welcome back! Please log in. 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https://www.researchgate.net/publication/295399308_The_Immunology_of_Posttransplant_CMV_Infection

[RFC v4 0/8] Proposal for a GPU cgroup controller * [RFC v4 0/8] Proposal for a GPU cgroup controller @ 2022-03-28 3:59 T.J. Mercier 2022-03-28 3:59 ` [RFC v4 1/8] gpu: rfc: " T.J. Mercier ` (7 more replies) 0 siblings, 8 replies; 22+ messages in thread From: T.J. Mercier @ 2022-03-28 3:59 UTC ( permalink / raw ) To: tjmercier, David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan Cc: kaleshsingh, Kenny.Ho, mkoutny, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest This patch series revisits the proposal for a GPU cgroup controller to track and limit memory allocations by various device/allocator subsystems. The patch series also contains a simple prototype to illustrate how Android intends to implement DMA-BUF allocator attribution using the GPU cgroup controller. The prototype does not include resource limit enforcements. Changelog: v4: Skip test if not run as root per Shuah Khan Add better test logging for abnormal child termination per Shuah Khan Adjust ordering of charge/uncharge during transfer to avoid potentially hitting cgroup limit per Michal Koutný Adjust gpucg_try_charge critical section for charge transfer functionality Fix uninitialized return code error for dmabuf_try_charge error case v3: Remove Upstreaming Plan from gpu-cgroup.rst per John Stultz Use more common dual author commit message format per John Stultz Remove android from binder changes title per Todd Kjos Add a kselftest for this new behavior per Greg Kroah-Hartman Include details on behavior for all combinations of kernel/userspace versions in changelog (thanks Suren Baghdasaryan) per Greg Kroah-Hartman. Fix pid and uid types in binder UAPI header v2: See the previous revision of this change submitted by Hridya Valsaraju at: https://lore.kernel.org/all/[email protected]/ Move dma-buf cgroup charge transfer from a dma_buf_op defined by every heap to a single dma-buf function for all heaps per Daniel Vetter and Christian König. Pointers to struct gpucg and struct gpucg_device tracking the current associations were added to the dma_buf struct to achieve this. Fix incorrect Kconfig help section indentation per Randy Dunlap. History of the GPU cgroup controller ==================================== The GPU/DRM cgroup controller came into being when a consensus[1] was reached that the resources it tracked were unsuitable to be integrated into memcg. Originally, the proposed controller was specific to the DRM subsystem and was intended to track GEM buffers and GPU-specific resources[2]. In order to help establish a unified memory accounting model for all GPU and all related subsystems, Daniel Vetter put forth a suggestion to move it out of the DRM subsystem so that it can be used by other DMA-BUF exporters as well[3]. This RFC proposes an interface that does the same. [1]: https://patchwork.kernel.org/project/dri-devel/cover/[email protected]/#22624705 [2]: https://lore.kernel.org/amd-gfx/[email protected]/ [3]: https://lore.kernel.org/amd-gfx/YCVOl8%[email protected]/ Hridya Valsaraju (5): gpu: rfc: Proposal for a GPU cgroup controller cgroup: gpu: Add a cgroup controller for allocator attribution of GPU memory dmabuf: heaps: export system_heap buffers with GPU cgroup charging dmabuf: Add gpu cgroup charge transfer function binder: Add a buffer flag to relinquish ownership of fds T.J. Mercier (3): dmabuf: Use the GPU cgroup charge/uncharge APIs binder: use __kernel_pid_t and __kernel_uid_t for userspace selftests: Add binder cgroup gpu memory transfer test Documentation/gpu/rfc/gpu-cgroup.rst | 183 +++++++ Documentation/gpu/rfc/index.rst | 4 + drivers/android/binder.c | 26 + drivers/dma-buf/dma-buf.c | 107 ++++ drivers/dma-buf/dma-heap.c | 27 + drivers/dma-buf/heaps/system_heap.c | 3 + include/linux/cgroup_gpu.h | 139 +++++ include/linux/cgroup_subsys.h | 4 + include/linux/dma-buf.h | 22 +- include/linux/dma-heap.h | 11 + include/uapi/linux/android/binder.h | 5 +- init/Kconfig | 7 + kernel/cgroup/Makefile | 1 + kernel/cgroup/gpu.c | 362 +++++++++++++ .../selftests/drivers/android/binder/Makefile | 8 + .../drivers/android/binder/binder_util.c | 254 +++++++++ .../drivers/android/binder/binder_util.h | 32 ++ .../selftests/drivers/android/binder/config | 4 + .../binder/test_dmabuf_cgroup_transfer.c | 484 ++++++++++++++++++ 19 files changed, 1679 insertions(+), 4 deletions(-) create mode 100644 Documentation/gpu/rfc/gpu-cgroup.rst create mode 100644 include/linux/cgroup_gpu.h create mode 100644 kernel/cgroup/gpu.c create mode 100644 tools/testing/selftests/drivers/android/binder/Makefile create mode 100644 tools/testing/selftests/drivers/android/binder/binder_util.c create mode 100644 tools/testing/selftests/drivers/android/binder/binder_util.h create mode 100644 tools/testing/selftests/drivers/android/binder/config create mode 100644 tools/testing/selftests/drivers/android/binder/test_dmabuf_cgroup_transfer.c -- 2.35.1.1021.g381101b075-goog ^ permalink raw reply [ flat | nested ] 22+ messages in thread * [RFC v4 1/8] gpu: rfc: Proposal for a GPU cgroup controller 2022-03-28 3:59 [RFC v4 0/8] Proposal for a GPU cgroup controller T.J. Mercier @ 2022-03-28 3:59 ` T.J. Mercier 2022-03-28 3:59 ` [RFC v4 2/8] cgroup: gpu: Add a cgroup controller for allocator attribution of GPU memory T.J. Mercier ` (6 subsequent siblings) 7 siblings, 0 replies; 22+ messages in thread From: T.J. Mercier @ 2022-03-28 3:59 UTC ( permalink / raw ) To: tjmercier, David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan Cc: kaleshsingh, Kenny.Ho, mkoutny, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest From: Hridya Valsaraju <[email protected]> This patch adds a proposal for a new GPU cgroup controller for accounting/limiting GPU and GPU-related memory allocations. The proposed controller is based on the DRM cgroup controller[1] and follows the design of the RDMA cgroup controller. The new cgroup controller would: * Allow setting per-cgroup limits on the total size of buffers charged to it. * Allow setting per-device limits on the total size of buffers allocated by device within a cgroup. * Expose a per-device/allocator breakdown of the buffers charged to a cgroup. The prototype in the following patches is only for memory accounting using the GPU cgroup controller and does not implement limit setting. [1]: https://lore.kernel.org/amd-gfx/[email protected]/ Signed-off-by: Hridya Valsaraju <[email protected]> Signed-off-by: T.J. Mercier <[email protected]> --- v3 changes Remove Upstreaming Plan from gpu-cgroup.rst per John Stultz. Use more common dual author commit message format per John Stultz. --- Documentation/gpu/rfc/gpu-cgroup.rst | 183 +++++++++++++++++++++++++++ Documentation/gpu/rfc/index.rst | 4 + 2 files changed , 187 insertions(+) create mode 100644 Documentation/gpu/rfc/gpu-cgroup.rst diff --git a/Documentation/gpu/rfc/gpu-cgroup.rst b/Documentation/gpu/rfc/gpu-cgroup.rst new file mode 100644 index 000000000000..5b40d5518a5e --- /dev/null +++ b/Documentation/gpu/rfc/gpu-cgroup.rst @@ -0,0 +1,183 @@ +=================================== +GPU cgroup controller +=================================== + +Goals +===== +This document intends to outline a plan to create a cgroup v2 controller subsystem +for the per-cgroup accounting of device and system memory allocated by the GPU +and related subsystems. + +The new cgroup controller would: + +* Allow setting per-cgroup limits on the total size of buffers charged to it. + +* Allow setting per-device limits on the total size of buffers allocated by a + device/allocator within a cgroup. + +* Expose a per-device/allocator breakdown of the buffers charged to a cgroup. + +Alternatives Considered +======================= + +The following alternatives were considered: + +The memory cgroup controller +____________________________ + +1. As was noted in [1], memory accounting provided by the GPU cgroup +controller is not a good fit for integration into memcg due to the +differences in how accounting is performed. It implements a mechanism +for the allocator attribution of GPU and GPU-related memory by +charging each buffer to the cgroup of the process on behalf of which +the memory was allocated. The buffer stays charged to the cgroup until +it is freed regardless of whether the process retains any references +to it. On the other hand, the memory cgroup controller offers a more +fine-grained charging and uncharging behavior depending on the kind of +page being accounted. + +2. Memcg performs accounting in units of pages. In the DMA-BUF buffer sharing model, +a process takes a reference to the entire buffer(hence keeping it alive) even if +it is only accessing parts of it. Therefore, per-page memory tracking for DMA-BUF +memory accounting would only introduce additional overhead without any benefits. + +[1]: https://patchwork.kernel.org/project/dri-devel/cover/[email protected]/#22624705 + +Userspace service to keep track of buffer allocations and releases +__________________________________________________________________ + +1. There is no way for a userspace service to intercept all allocations and releases. +2. In case the process gets killed or restarted, we lose all accounting so far. + +UAPI +==== +When enabled, the new cgroup controller would create the following files in every cgroup. + +:: + + gpu.memory.current (R) + gpu.memory.max (R/W) + +gpu.memory.current is a read-only file and would contain per-device memory allocations +in a key-value format where key is a string representing the device name +and the value is the size of memory charged to the device in the cgroup in bytes. + +For example: + +:: + + cat /sys/kernel/fs/cgroup1/gpu.memory.current + dev1 4194304 + dev2 4194304 + +The string key for each device is set by the device driver when the device registers +with the GPU cgroup controller to participate in resource accounting(see section +'Design and Implementation' for more details). + +gpu.memory.max is a read/write file. It would show the current total +size limits on memory usage for the cgroup and the limits on total memory usage +for each allocator/device. + +Setting a total limit for a cgroup can be done as follows: + +:: + + echo “total 41943040” > /sys/kernel/fs/cgroup1/gpu.memory.max + +Setting a total limit for a particular device/allocator can be done as follows: + +:: + + echo “dev1 4194304” > /sys/kernel/fs/cgroup1/gpu.memory.max + +In this example, 'dev1' is the string key set by the device driver during +registration. + +Design and Implementation +========================= + +The cgroup controller would closely follow the design of the RDMA cgroup controller +subsystem where each cgroup maintains a list of resource pools. +Each resource pool contains a struct device and the counter to track current total, +and the maximum limit set for the device. + +The below code block is a preliminary estimation on how the core kernel data structures +and APIs would look like. + +.. code-block:: c + + /** + * The GPU cgroup controller data structure. + */ + struct gpucg { + struct cgroup_subsys_state css; + + /* list of all resource pools that belong to this cgroup */ + struct list_head rpools; + }; + + struct gpucg_device { + /* + * list of various resource pools in various cgroups that the device is + * part of. + */ + struct list_head rpools; + + /* list of all devices registered for GPU cgroup accounting */ + struct list_head dev_node; + + /* name to be used as identifier for accounting and limit setting */ + const char *name; + }; + + struct gpucg_resource_pool { + /* The device whose resource usage is tracked by this resource pool */ + struct gpucg_device *device; + + /* list of all resource pools for the cgroup */ + struct list_head cg_node; + + /* + * list maintained by the gpucg_device to keep track of its + * resource pools + */ + struct list_head dev_node; + + /* tracks memory usage of the resource pool */ + struct page_counter total; + }; + + /** + * gpucg_register_device - Registers a device for memory accounting using the + * GPU cgroup controller. + * + * @device: The device to register for memory accounting. Must remain valid + * after registration. + * @name: Pointer to a string literal to denote the name of the device. + */ + void gpucg_register_device(struct gpucg_device *gpucg_dev, const char *name); + + /** + * gpucg_try_charge - charge memory to the specified gpucg and gpucg_device. + * + * @gpucg: The gpu cgroup to charge the memory to. + * @device: The device to charge the memory to. + * @usage: size of memory to charge in bytes. + * + * Return: returns 0 if the charging is successful and otherwise returns an + * error code. + */ + int gpucg_try_charge(struct gpucg *gpucg, struct gpucg_device *device, u64 usage); + + /** + * gpucg_uncharge - uncharge memory from the specified gpucg and gpucg_device. + * + * @gpucg: The gpu cgroup to uncharge the memory from. + * @device: The device to charge the memory from. + * @usage: size of memory to uncharge in bytes. + */ + void gpucg_uncharge(struct gpucg *gpucg, struct gpucg_device *device, u64 usage); + +Future Work +=========== +Additional GPU resources can be supported by adding new controller files. diff --git a/Documentation/gpu/rfc/index.rst b/Documentation/gpu/rfc/index.rst index 91e93a705230..0a9bcd94e95d 100644 --- a/Documentation/gpu/rfc/index.rst +++ b/Documentation/gpu/rfc/index.rst @@ -23,3 +23,7 @@ host such documentation: .. toctree:: i915_scheduler.rst + +.. toctree:: + + gpu-cgroup.rst -- 2.35.1.1021.g381101b075-goog ^ permalink raw reply related [ flat | nested ] 22+ messages in thread * [RFC v4 2/8] cgroup: gpu: Add a cgroup controller for allocator attribution of GPU memory 2022-03-28 3:59 [RFC v4 0/8] Proposal for a GPU cgroup controller T.J. Mercier 2022-03-28 3:59 ` [RFC v4 1/8] gpu: rfc: " T.J. Mercier @ 2022-03-28 3:59 ` T.J. Mercier 2022-03-29 16:59 ` Tejun Heo 2022-03-28 3:59 ` [RFC v4 3/8] dmabuf: Use the GPU cgroup charge/uncharge APIs T.J. Mercier ` (5 subsequent siblings) 7 siblings, 1 reply; 22+ messages in thread From: T.J. Mercier @ 2022-03-28 3:59 UTC ( permalink / raw ) To: tjmercier, David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan Cc: kaleshsingh, Kenny.Ho, mkoutny, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest From: Hridya Valsaraju <[email protected]> The cgroup controller provides accounting for GPU and GPU-related memory allocations. The memory being accounted can be device memory or memory allocated from pools dedicated to serve GPU-related tasks. This patch adds APIs to: -allow a device to register for memory accounting using the GPU cgroup controller. -charge and uncharge allocated memory to a cgroup. When the cgroup controller is enabled, it would expose information about the memory allocated by each device(registered for GPU cgroup memory accounting) for each cgroup. The API/UAPI can be extended to set per-device/total allocation limits in the future. The cgroup controller has been named following the discussion in [1]. [1]: https://lore.kernel.org/amd-gfx/YCJp%2F%[email protected]/ Signed-off-by: Hridya Valsaraju <[email protected]> Signed-off-by: T.J. Mercier <[email protected]> --- v4 changes Adjust gpucg_try_charge critical section for future charge transfer functionality. v3 changes Use more common dual author commit message format per John Stultz. v2 changes Fix incorrect Kconfig help section indentation per Randy Dunlap. --- include/linux/cgroup_gpu.h | 127 ++++++++++++++ include/linux/cgroup_subsys.h | 4 + init/Kconfig | 7 + kernel/cgroup/Makefile | 1 + kernel/cgroup/gpu.c | 303 ++++++++++++++++++++++++++++++++++ 5 files changed , 442 insertions(+) create mode 100644 include/linux/cgroup_gpu.h create mode 100644 kernel/cgroup/gpu.c diff --git a/include/linux/cgroup_gpu.h b/include/linux/cgroup_gpu.h new file mode 100644 index 000000000000..c90069719022 --- /dev/null +++ b/include/linux/cgroup_gpu.h @@ -0,0 +1,127 @@ +/* SPDX-License-Identifier: MIT + * Copyright 2019 Advanced Micro Devices, Inc. + * Copyright (C) 2022 Google LLC. + */ +#ifndef _CGROUP_GPU_H +#define _CGROUP_GPU_H + +#include <linux/cgroup.h> +#include <linux/page_counter.h> + +#ifdef CONFIG_CGROUP_GPU + /* The GPU cgroup controller data structure */ +struct gpucg { + struct cgroup_subsys_state css; + + /* list of all resource pools that belong to this cgroup */ + struct list_head rpools; +}; + +struct gpucg_device { + /* + * list of various resource pools in various cgroups that the device is + * part of. + */ + struct list_head rpools; + + /* list of all devices registered for GPU cgroup accounting */ + struct list_head dev_node; + + /* + * pointer to string literal to be used as identifier for accounting and + * limit setting + */ + const char *name; +}; + +/** + * css_to_gpucg - get the corresponding gpucg ref from a cgroup_subsys_state + * @css: the target cgroup_subsys_state + * + * Returns: gpu cgroup that contains the @css + */ +static inline struct gpucg *css_to_gpucg(struct cgroup_subsys_state *css) +{ + return css ? container_of(css, struct gpucg, css) : NULL; +} + +/** + * gpucg_get - get the gpucg reference that a task belongs to + * @task: the target task + * + * This increases the reference count of the css that the @task belongs to. + * + * Returns: reference to the gpu cgroup the task belongs to. + */ +static inline struct gpucg *gpucg_get(struct task_struct *task) +{ + if (!cgroup_subsys_enabled(gpu_cgrp_subsys)) + return NULL; + return css_to_gpucg(task_get_css(task, gpu_cgrp_id)); +} + +/** + * gpucg_put - put a gpucg reference + * @gpucg: the target gpucg + * + * Put a reference obtained via gpucg_get + */ +static inline void gpucg_put(struct gpucg *gpucg) +{ + if (gpucg) + css_put(&gpucg->css); +} + +/** + * gpucg_parent - find the parent of a gpu cgroup + * @cg: the target gpucg + * + * This does not increase the reference count of the parent cgroup + * + * Returns: parent gpu cgroup of @cg + */ +static inline struct gpucg *gpucg_parent(struct gpucg *cg) +{ + return css_to_gpucg(cg->css.parent); +} + +int gpucg_try_charge(struct gpucg *gpucg, struct gpucg_device *device, u64 usage); +void gpucg_uncharge(struct gpucg *gpucg, struct gpucg_device *device, u64 usage); +void gpucg_register_device(struct gpucg_device *gpucg_dev, const char *name); +#else /* CONFIG_CGROUP_GPU */ + +struct gpucg; +struct gpucg_device; + +static inline struct gpucg *css_to_gpucg(struct cgroup_subsys_state *css) +{ + return NULL; +} + +static inline struct gpucg *gpucg_get(struct task_struct *task) +{ + return NULL; +} + +static inline void gpucg_put(struct gpucg *gpucg) {} + +static inline struct gpucg *gpucg_parent(struct gpucg *cg) +{ + return NULL; +} + +static inline int gpucg_try_charge(struct gpucg *gpucg, + struct gpucg_device *device, + u64 usage) +{ + return 0; +} + +static inline void gpucg_uncharge(struct gpucg *gpucg, + struct gpucg_device *device, + u64 usage) {} + +static inline void gpucg_register_device(struct gpucg_device *gpucg_dev, + const char *name) {} +#endif /* CONFIG_CGROUP_GPU */ +#endif /* _CGROUP_GPU_H */ diff --git a/include/linux/cgroup_subsys.h b/include/linux/cgroup_subsys.h index 445235487230..46a2a7b93c41 100644 --- a/include/linux/cgroup_subsys.h +++ b/include/linux/cgroup_subsys.h @@ -65,6 +65,10 @@ SUBSYS(rdma) SUBSYS(misc) #endif +#if IS_ENABLED(CONFIG_CGROUP_GPU) +SUBSYS(gpu) +#endif + /* * The following subsystems are not supported on the default hierarchy. */ diff --git a/init/Kconfig b/init/Kconfig index e9119bf54b1f..43568472930a 100644 --- a/init/Kconfig +++ b/init/Kconfig @@ -980,6 +980,13 @@ config BLK_CGROUP See Documentation/admin-guide/cgroup-v1/blkio-controller.rst for more information. +config CGROUP_GPU + bool "gpu cgroup controller (EXPERIMENTAL)" + select PAGE_COUNTER + help + Provides accounting and limit setting for memory allocations by the GPU and + GPU-related subsystems. + config CGROUP_WRITEBACK bool depends on MEMCG && BLK_CGROUP diff --git a/kernel/cgroup/Makefile b/kernel/cgroup/Makefile index 12f8457ad1f9..be95a5a532fc 100644 --- a/kernel/cgroup/Makefile +++ b/kernel/cgroup/Makefile @@ -7,3 +7,4 @@ obj-$(CONFIG_CGROUP_RDMA) += rdma.o obj-$(CONFIG_CPUSETS) += cpuset.o obj-$(CONFIG_CGROUP_MISC) += misc.o obj-$(CONFIG_CGROUP_DEBUG) += debug.o +obj-$(CONFIG_CGROUP_GPU) += gpu.o diff --git a/kernel/cgroup/gpu.c b/kernel/cgroup/gpu.c new file mode 100644 index 000000000000..ac4c470914b5 --- /dev/null +++ b/kernel/cgroup/gpu.c @@ -0,0 +1,303 @@ +// SPDX-License-Identifier: MIT +// Copyright 2019 Advanced Micro Devices, Inc. +// Copyright (C) 2022 Google LLC. + +#include <linux/cgroup.h> +#include <linux/cgroup_gpu.h> +#include <linux/mm.h> +#include <linux/page_counter.h> +#include <linux/seq_file.h> +#include <linux/slab.h> + +static struct gpucg *root_gpucg __read_mostly; + +/* + * Protects list of resource pools maintained on per cgroup basis + * and list of devices registered for memory accounting using the GPU cgroup + * controller. + */ +static DEFINE_MUTEX(gpucg_mutex); +static LIST_HEAD(gpucg_devices); + +struct gpucg_resource_pool { + /* The device whose resource usage is tracked by this resource pool */ + struct gpucg_device *device; + + /* list of all resource pools for the cgroup */ + struct list_head cg_node; + + /* list maintained by the gpucg_device to keep track of its resource pools */ + struct list_head dev_node; + + /* tracks memory usage of the resource pool */ + struct page_counter total; +}; + +static void free_cg_rpool_locked(struct gpucg_resource_pool *rpool) +{ + lockdep_assert_held(&gpucg_mutex); + + list_del(&rpool->cg_node); + list_del(&rpool->dev_node); + kfree(rpool); +} + +static void gpucg_css_free(struct cgroup_subsys_state *css) +{ + struct gpucg_resource_pool *rpool, *tmp; + struct gpucg *gpucg = css_to_gpucg(css); + + // delete all resource pools + mutex_lock(&gpucg_mutex); + list_for_each_entry_safe(rpool, tmp, &gpucg->rpools, cg_node) + free_cg_rpool_locked(rpool); + mutex_unlock(&gpucg_mutex); + + kfree(gpucg); +} + +static struct cgroup_subsys_state * +gpucg_css_alloc(struct cgroup_subsys_state *parent_css) +{ + struct gpucg *gpucg, *parent; + + gpucg = kzalloc(sizeof(struct gpucg), GFP_KERNEL); + if (!gpucg) + return ERR_PTR(-ENOMEM); + + parent = css_to_gpucg(parent_css); + if (!parent) + root_gpucg = gpucg; + + INIT_LIST_HEAD(&gpucg->rpools); + + return &gpucg->css; +} + +static struct gpucg_resource_pool *find_cg_rpool_locked( + struct gpucg *cg, + struct gpucg_device *device) +{ + struct gpucg_resource_pool *pool; + + lockdep_assert_held(&gpucg_mutex); + + list_for_each_entry(pool, &cg->rpools, cg_node) + if (pool->device == device) + return pool; + + return NULL; +} + +static struct gpucg_resource_pool *init_cg_rpool(struct gpucg *cg, + struct gpucg_device *device) +{ + struct gpucg_resource_pool *rpool = kzalloc(sizeof(*rpool), + GFP_KERNEL); + if (!rpool) + return ERR_PTR(-ENOMEM); + + rpool->device = device; + + page_counter_init(&rpool->total, NULL); + INIT_LIST_HEAD(&rpool->cg_node); + INIT_LIST_HEAD(&rpool->dev_node); + list_add_tail(&rpool->cg_node, &cg->rpools); + list_add_tail(&rpool->dev_node, &device->rpools); + + return rpool; +} + +/** + * get_cg_rpool_locked - find the resource pool for the specified device and + * specified cgroup. If the resource pool does not exist for the cg, it is + * created in a hierarchical manner in the cgroup and its ancestor cgroups who + * do not already have a resource pool entry for the device. + * + * @cg: The cgroup to find the resource pool for. + * @device: The device associated with the returned resource pool. + * + * Return: return resource pool entry corresponding to the specified device in + * the specified cgroup (hierarchically creating them if not existing already). + * + */ +static struct gpucg_resource_pool * +get_cg_rpool_locked(struct gpucg *cg, struct gpucg_device *device) +{ + struct gpucg *parent_cg, *p, *stop_cg; + struct gpucg_resource_pool *rpool, *tmp_rpool; + struct gpucg_resource_pool *parent_rpool = NULL, *leaf_rpool = NULL; + + rpool = find_cg_rpool_locked(cg, device); + if (rpool) + return rpool; + + stop_cg = cg; + do { + rpool = init_cg_rpool(stop_cg, device); + if (IS_ERR(rpool)) + goto err; + + if (!leaf_rpool) + leaf_rpool = rpool; + + stop_cg = gpucg_parent(stop_cg); + if (!stop_cg) + break; + + rpool = find_cg_rpool_locked(stop_cg, device); + } while (!rpool); + + /* + * Re-initialize page counters of all rpools created in this invocation + * to enable hierarchical charging. + * stop_cg is the first ancestor cg who already had a resource pool for + * the device. It can also be NULL if no ancestors had a pre-existing + * resource pool for the device before this invocation. + */ + rpool = leaf_rpool; + for (p = cg; p != stop_cg; p = parent_cg) { + parent_cg = gpucg_parent(p); + if (!parent_cg) + break; + parent_rpool = find_cg_rpool_locked(parent_cg, device); + page_counter_init(&rpool->total, &parent_rpool->total); + + rpool = parent_rpool; + } + + return leaf_rpool; +err: + for (p = cg; p != stop_cg; p = gpucg_parent(p)) { + tmp_rpool = find_cg_rpool_locked(p, device); + free_cg_rpool_locked(tmp_rpool); + } + return rpool; +} + +/** + * gpucg_try_charge - charge memory to the specified gpucg and gpucg_device. + * Caller must hold a reference to @gpucg obtained through gpucg_get(). The size + * of the memory is rounded up to be a multiple of the page size. + * + * @gpucg: The gpu cgroup to charge the memory to. + * @device: The device to charge the memory to. + * @usage: size of memory to charge in bytes. + * + * Return: returns 0 if the charging is successful and otherwise returns an + * error code. + */ +int gpucg_try_charge(struct gpucg *gpucg, struct gpucg_device *device, u64 usage) +{ + struct page_counter *counter; + u64 nr_pages; + struct gpucg_resource_pool *rp; + int ret = 0; + + nr_pages = PAGE_ALIGN(usage) >> PAGE_SHIFT; + + mutex_lock(&gpucg_mutex); + rp = get_cg_rpool_locked(gpucg, device); + /* + * Continue to hold gpucg_mutex because we use it to block charges + * while transfers are in progress. + */ + if (IS_ERR(rp)) { + mutex_unlock(&gpucg_mutex); + return PTR_ERR(rp); + } + + if (page_counter_try_charge(&rp->total, nr_pages, &counter)) + css_get_many(&gpucg->css, nr_pages); + else + ret = -ENOMEM; + mutex_unlock(&gpucg_mutex); + + return ret; +} + +/** + * gpucg_uncharge - uncharge memory from the specified gpucg and gpucg_device. + * The caller must hold a reference to @gpucg obtained through gpucg_get(). + * + * @gpucg: The gpu cgroup to uncharge the memory from. + * @device: The device to uncharge the memory from. + * @usage: size of memory to uncharge in bytes. + */ +void gpucg_uncharge(struct gpucg *gpucg, struct gpucg_device *device, u64 usage) +{ + u64 nr_pages; + struct gpucg_resource_pool *rp; + + mutex_lock(&gpucg_mutex); + rp = find_cg_rpool_locked(gpucg, device); + /* + * gpucg_mutex can be unlocked here, rp will stay valid until gpucg is freed and there are + * active refs on gpucg. Uncharges are fine while transfers are in progress. + */ + mutex_unlock(&gpucg_mutex); + + if (unlikely(!rp)) { + pr_err("Resource pool not found, incorrect charge/uncharge ordering?\n"); + return; + } + + nr_pages = PAGE_ALIGN(usage) >> PAGE_SHIFT; + page_counter_uncharge(&rp->total, nr_pages); + css_put_many(&gpucg->css, nr_pages); +} + +/** + * gpucg_register_device - Registers a device for memory accounting using the + * GPU cgroup controller. + * + * @device: The device to register for memory accounting. + * @name: Pointer to a string literal to denote the name of the device. + * + * Both @device andd @name must remain valid. + */ +void gpucg_register_device(struct gpucg_device *device, const char *name) +{ + if (!device) + return; + + INIT_LIST_HEAD(&device->dev_node); + INIT_LIST_HEAD(&device->rpools); + + mutex_lock(&gpucg_mutex); + list_add_tail(&device->dev_node, &gpucg_devices); + mutex_unlock(&gpucg_mutex); + + device->name = name; +} + +static int gpucg_resource_show(struct seq_file *sf, void *v) +{ + struct gpucg_resource_pool *rpool; + struct gpucg *cg = css_to_gpucg(seq_css(sf)); + + mutex_lock(&gpucg_mutex); + list_for_each_entry(rpool, &cg->rpools, cg_node) { + seq_printf(sf, "%s %lu\n", rpool->device->name, + page_counter_read(&rpool->total) * PAGE_SIZE); + } + mutex_unlock(&gpucg_mutex); + + return 0; +} + +struct cftype files[] = { + { + .name = "memory.current", + .seq_show = gpucg_resource_show, + }, + { } /* terminate */ +}; + +struct cgroup_subsys gpu_cgrp_subsys = { + .css_alloc = gpucg_css_alloc, + .css_free = gpucg_css_free, + .early_init = false, + .legacy_cftypes = files, + .dfl_cftypes = files, +}; -- 2.35.1.1021.g381101b075-goog ^ permalink raw reply related [ flat | nested ] 22+ messages in thread * [RFC v4 3/8] dmabuf: Use the GPU cgroup charge/uncharge APIs 2022-03-28 3:59 [RFC v4 0/8] Proposal for a GPU cgroup controller T.J. Mercier 2022-03-28 3:59 ` [RFC v4 1/8] gpu: rfc: " T.J. Mercier 2022-03-28 3:59 ` [RFC v4 2/8] cgroup: gpu: Add a cgroup controller for allocator attribution of GPU memory T.J. Mercier @ 2022-03-28 3:59 ` T.J. Mercier 2022-03-28 3:59 ` [RFC v4 4/8] dmabuf: heaps: export system_heap buffers with GPU cgroup charging T.J. Mercier ` (4 subsequent siblings) 7 siblings, 0 replies; 22+ messages in thread From: T.J. Mercier @ 2022-03-28 3:59 UTC ( permalink / raw ) To: tjmercier, David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan Cc: kaleshsingh, Kenny.Ho, mkoutny, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest This patch uses the GPU cgroup charge/uncharge APIs to charge buffers allocated by any DMA-BUF exporter that exports a buffer with a GPU cgroup device association. By doing so, it becomes possible to track who allocated/exported a DMA-BUF even after the allocating process drops all references to a buffer. Originally-by: Hridya Valsaraju <[email protected]> Signed-off-by: T.J. Mercier <[email protected]> --- v4 changes Fix uninitialized return code error for dmabuf_try_charge error case. v3 changes Use more common dual author commit message format per John Stultz. v2 changes Move dma-buf cgroup charging/uncharging from a dma_buf_op defined by every heap to a single dma-buf function for all heaps per Daniel Vetter and Christian König. --- drivers/dma-buf/dma-buf.c | 58 +++++++++++++++++++++++++++++++++++++++ include/linux/dma-buf.h | 20 ++++++++++++-- 2 files changed , 76 insertions(+), 2 deletions(-) diff --git a/drivers/dma-buf/dma-buf.c b/drivers/dma-buf/dma-buf.c index 602b12d7470d..1ee5c60d3d6d 100644 --- a/drivers/dma-buf/dma-buf.c +++ b/drivers/dma-buf/dma-buf.c @@ -56,6 +56,53 @@ static char *dmabuffs_dname(struct dentry *dentry, char *buffer, int buflen) dentry->d_name.name, ret > 0 ? name : ""); } +#ifdef CONFIG_CGROUP_GPU +static inline struct gpucg_device * +exp_info_gpucg_dev(const struct dma_buf_export_info *exp_info) +{ + return exp_info->gpucg_dev; +} + +static int dmabuf_try_charge(struct dma_buf *dmabuf, + struct gpucg_device *gpucg_dev) +{ + int ret; + + dmabuf->gpucg = gpucg_get(current); + dmabuf->gpucg_dev = gpucg_dev; + + ret = gpucg_try_charge(dmabuf->gpucg, dmabuf->gpucg_dev, dmabuf->size); + if (ret) { + gpucg_put(dmabuf->gpucg); + dmabuf->gpucg = NULL; + dmabuf->gpucg_dev = NULL; + } + return ret; +} + +static void dmabuf_uncharge(struct dma_buf *dmabuf) +{ + if (dmabuf->gpucg && dmabuf->gpucg_dev) { + gpucg_uncharge(dmabuf->gpucg, dmabuf->gpucg_dev, dmabuf->size); + gpucg_put(dmabuf->gpucg); + } +} +#else /* CONFIG_CGROUP_GPU */ +static inline struct gpucg_device *exp_info_gpucg_dev( +const struct dma_buf_export_info *exp_info) +{ + return NULL; +} + +static inline int dmabuf_try_charge(struct dma_buf *dmabuf, + struct gpucg_device *gpucg_dev)) +{ + return 0; +} + +static inline void dmabuf_uncharge(struct dma_buf *dmabuf) {} +#endif /* CONFIG_CGROUP_GPU */ + static void dma_buf_release(struct dentry *dentry) { struct dma_buf *dmabuf; @@ -79,6 +126,8 @@ static void dma_buf_release(struct dentry *dentry) if (dmabuf->resv == (struct dma_resv *)&dmabuf[1]) dma_resv_fini(dmabuf->resv); + dmabuf_uncharge(dmabuf); + WARN_ON(!list_empty(&dmabuf->attachments)); module_put(dmabuf->owner); kfree(dmabuf->name); @@ -484,6 +533,7 @@ struct dma_buf *dma_buf_export(const struct dma_buf_export_info *exp_info) { struct dma_buf *dmabuf; struct dma_resv *resv = exp_info->resv; + struct gpucg_device *gpucg_dev = exp_info_gpucg_dev(exp_info); struct file *file; size_t alloc_size = sizeof(struct dma_buf); int ret; @@ -534,6 +584,12 @@ struct dma_buf *dma_buf_export(const struct dma_buf_export_info *exp_info) } dmabuf->resv = resv; + if (gpucg_dev) { + ret = dmabuf_try_charge(dmabuf, gpucg_dev); + if (ret) + goto err_charge; + } + file = dma_buf_getfile(dmabuf, exp_info->flags); if (IS_ERR(file)) { ret = PTR_ERR(file); @@ -565,6 +621,8 @@ struct dma_buf *dma_buf_export(const struct dma_buf_export_info *exp_info) file->f_path.dentry->d_fsdata = NULL; fput(file); err_dmabuf: + dmabuf_uncharge(dmabuf); +err_charge: kfree(dmabuf); err_module: module_put(exp_info->owner); diff --git a/include/linux/dma-buf.h b/include/linux/dma-buf.h index 7ab50076e7a6..742f29c3daaf 100644 --- a/include/linux/dma-buf.h +++ b/include/linux/dma-buf.h @@ -13,6 +13,7 @@ #ifndef __DMA_BUF_H__ #define __DMA_BUF_H__ +#include <linux/cgroup_gpu.h> #include <linux/dma-buf-map.h> #include <linux/file.h> #include <linux/err.h> @@ -303,7 +304,7 @@ struct dma_buf { /** * @size: * - * Size of the buffer; invariant over the lifetime of the buffer. + * Size of the buffer in bytes; invariant over the lifetime of the buffer. */ size_t size; @@ -453,6 +454,17 @@ struct dma_buf { struct dma_buf *dmabuf; } *sysfs_entry; #endif + +#ifdef CONFIG_CGROUP_GPU + /** @gpucg: Pointer to the cgroup this buffer currently belongs to. */ + struct gpucg *gpucg; + + /** @gpucg_dev: + * + * Pointer to the cgroup GPU device whence this buffer originates. + */ + struct gpucg_device *gpucg_dev; +#endif }; /** @@ -529,9 +541,10 @@ struct dma_buf_attachment { * @exp_name: name of the exporter - useful for debugging. * @owner: pointer to exporter module - used for refcounting kernel module * @ops: Attach allocator-defined dma buf ops to the new buffer - * @size: Size of the buffer - invariant over the lifetime of the buffer + * @size: Size of the buffer in bytes - invariant over the lifetime of the buffer * @flags: mode flags for the file * @resv: reservation-object, NULL to allocate default one + * @gpucg_dev: pointer to the gpu cgroup device this buffer belongs to * @priv: Attach private data of allocator to this buffer * * This structure holds the information required to export the buffer. Used @@ -544,6 +557,9 @@ struct dma_buf_export_info { size_t size; int flags; struct dma_resv *resv; +#ifdef CONFIG_CGROUP_GPU + struct gpucg_device *gpucg_dev; +#endif void *priv; }; -- 2.35.1.1021.g381101b075-goog ^ permalink raw reply related [ flat | nested ] 22+ messages in thread * [RFC v4 4/8] dmabuf: heaps: export system_heap buffers with GPU cgroup charging 2022-03-28 3:59 [RFC v4 0/8] Proposal for a GPU cgroup controller T.J. Mercier ` (2 preceding siblings ...) 2022-03-28 3:59 ` [RFC v4 3/8] dmabuf: Use the GPU cgroup charge/uncharge APIs T.J. Mercier @ 2022-03-28 3:59 ` T.J. Mercier 2022-03-28 14:36 ` Daniel Vetter 2022-03-28 3:59 ` [RFC v4 5/8] dmabuf: Add gpu cgroup charge transfer function T.J. Mercier ` (3 subsequent siblings) 7 siblings, 1 reply; 22+ messages in thread From: T.J. Mercier @ 2022-03-28 3:59 UTC ( permalink / raw ) To: tjmercier, David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan Cc: kaleshsingh, Kenny.Ho, mkoutny, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest From: Hridya Valsaraju <[email protected]> All DMA heaps now register a new GPU cgroup device upon creation, and the system_heap now exports buffers associated with its GPU cgroup device for tracking purposes. Signed-off-by: Hridya Valsaraju <[email protected]> Signed-off-by: T.J. Mercier <[email protected]> --- v3 changes Use more common dual author commit message format per John Stultz. v2 changes Move dma-buf cgroup charge transfer from a dma_buf_op defined by every heap to a single dma-buf function for all heaps per Daniel Vetter and Christian König. --- drivers/dma-buf/dma-heap.c | 27 +++++++++++++++++++++++++++ drivers/dma-buf/heaps/system_heap.c | 3 +++ include/linux/dma-heap.h | 11 +++++++++++ 3 files changed , 41 insertions(+) diff --git a/drivers/dma-buf/dma-heap.c b/drivers/dma-buf/dma-heap.c index 8f5848aa144f..885072427775 100644 --- a/drivers/dma-buf/dma-heap.c +++ b/drivers/dma-buf/dma-heap.c @@ -7,6 +7,7 @@ */ #include <linux/cdev.h> +#include <linux/cgroup_gpu.h> #include <linux/debugfs.h> #include <linux/device.h> #include <linux/dma-buf.h> @@ -31,6 +32,7 @@ * @heap_devt heap device node * @list list head connecting to list of heaps * @heap_cdev heap char device + * @gpucg_dev gpu cgroup device for memory accounting * * Represents a heap of memory from which buffers can be made. */ @@ -41,6 +43,9 @@ struct dma_heap { dev_t heap_devt; struct list_head list; struct cdev heap_cdev; +#ifdef CONFIG_CGROUP_GPU + struct gpucg_device gpucg_dev; +#endif }; static LIST_HEAD(heap_list); @@ -216,6 +221,26 @@ const char *dma_heap_get_name(struct dma_heap *heap) return heap->name; } +#ifdef CONFIG_CGROUP_GPU +/** + * dma_heap_get_gpucg_dev() - get struct gpucg_device for the heap. + * @heap: DMA-Heap to get the gpucg_device struct for. + * + * Returns: + * The gpucg_device struct for the heap. NULL if the GPU cgroup controller is + * not enabled. + */ +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap) +{ + return &heap->gpucg_dev; +} +#else /* CONFIG_CGROUP_GPU */ +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap) +{ + return NULL; +} +#endif /* CONFIG_CGROUP_GPU */ + struct dma_heap *dma_heap_add(const struct dma_heap_export_info *exp_info) { struct dma_heap *heap, *h, *err_ret; @@ -288,6 +313,8 @@ struct dma_heap *dma_heap_add(const struct dma_heap_export_info *exp_info) list_add(&heap->list, &heap_list); mutex_unlock(&heap_list_lock); + gpucg_register_device(dma_heap_get_gpucg_dev(heap), exp_info->name); + return heap; err2: diff --git a/drivers/dma-buf/heaps/system_heap.c b/drivers/dma-buf/heaps/system_heap.c index ab7fd896d2c4..752a05c3cfe2 100644 --- a/drivers/dma-buf/heaps/system_heap.c +++ b/drivers/dma-buf/heaps/system_heap.c @@ -395,6 +395,9 @@ static struct dma_buf *system_heap_allocate(struct dma_heap *heap, exp_info.ops = &system_heap_buf_ops; exp_info.size = buffer->len; exp_info.flags = fd_flags; +#ifdef CONFIG_CGROUP_GPU + exp_info.gpucg_dev = dma_heap_get_gpucg_dev(heap); +#endif exp_info.priv = buffer; dmabuf = dma_buf_export(&exp_info); if (IS_ERR(dmabuf)) { diff --git a/include/linux/dma-heap.h b/include/linux/dma-heap.h index 0c05561cad6e..e447a61d054e 100644 --- a/include/linux/dma-heap.h +++ b/include/linux/dma-heap.h @@ -10,6 +10,7 @@ #define _DMA_HEAPS_H #include <linux/cdev.h> +#include <linux/cgroup_gpu.h> #include <linux/types.h> struct dma_heap; @@ -59,6 +60,16 @@ void *dma_heap_get_drvdata(struct dma_heap *heap); */ const char *dma_heap_get_name(struct dma_heap *heap); +/** + * dma_heap_get_gpucg_dev() - get a pointer to the struct gpucg_device for the + * heap. + * @heap: DMA-Heap to retrieve gpucg_device for. + * + * Returns: + * The gpucg_device struct for the heap. + */ +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap); + /** * dma_heap_add - adds a heap to dmabuf heaps * @exp_info: information needed to register this heap -- 2.35.1.1021.g381101b075-goog ^ permalink raw reply related [ flat | nested ] 22+ messages in thread * [RFC v4 5/8] dmabuf: Add gpu cgroup charge transfer function 2022-03-28 3:59 [RFC v4 0/8] Proposal for a GPU cgroup controller T.J. Mercier ` (3 preceding siblings ...) 2022-03-28 3:59 ` [RFC v4 4/8] dmabuf: heaps: export system_heap buffers with GPU cgroup charging T.J. Mercier @ 2022-03-28 3:59 ` T.J. Mercier 2022-03-29 15:21 ` Michal Koutný 2022-03-28 3:59 ` [RFC v4 6/8] binder: Add a buffer flag to relinquish ownership of fds T.J. Mercier ` (2 subsequent siblings) 7 siblings, 1 reply; 22+ messages in thread From: T.J. Mercier @ 2022-03-28 3:59 UTC ( permalink / raw ) To: tjmercier, David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan Cc: kaleshsingh, Kenny.Ho, mkoutny, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest From: Hridya Valsaraju <[email protected]> The dma_buf_charge_transfer function provides a way for processes to transfer charge of a buffer to a different process. This is essential for the cases where a central allocator process does allocations for various subsystems, hands over the fd to the client who requested the memory and drops all references to the allocated memory. Signed-off-by: Hridya Valsaraju <[email protected]> Signed-off-by: T.J. Mercier <[email protected]> --- v4 changes Adjust ordering of charge/uncharge during transfer to avoid potentially hitting cgroup limit per Michal Koutný. v3 changes Use more common dual author commit message format per John Stultz. v2 changes Move dma-buf cgroup charge transfer from a dma_buf_op defined by every heap to a single dma-buf function for all heaps per Daniel Vetter and Christian König. --- drivers/dma-buf/dma-buf.c | 49 +++++++++++++++++++++++++++++++ include/linux/cgroup_gpu.h | 12 ++++++++ include/linux/dma-buf.h | 2 ++ kernel/cgroup/gpu.c | 59 ++++++++++++++++++++++++++++++++++++++ 4 files changed , 122 insertions(+) diff --git a/drivers/dma-buf/dma-buf.c b/drivers/dma-buf/dma-buf.c index 1ee5c60d3d6d..7748c3453b91 100644 --- a/drivers/dma-buf/dma-buf.c +++ b/drivers/dma-buf/dma-buf.c @@ -1380,6 +1380,55 @@ void dma_buf_vunmap(struct dma_buf *dmabuf, struct dma_buf_map *map) } EXPORT_SYMBOL_NS_GPL(dma_buf_vunmap, DMA_BUF); +/** + * dma_buf_transfer_charge - Change the GPU cgroup to which the provided dma_buf is charged. + * @dmabuf: [in] buffer whose charge will be migrated to a different GPU cgroup + * @gpucg: [in] the destination GPU cgroup for dmabuf's charge + * + * Only tasks that belong to the same cgroup the buffer is currently charged to + * may call this function, otherwise it will return -EPERM. + * + * Returns 0 on success, or a negative errno code otherwise. + */ +int dma_buf_transfer_charge(struct dma_buf *dmabuf, struct gpucg *gpucg) +{ +#ifdef CONFIG_CGROUP_GPU + struct gpucg *current_gpucg; + int ret; + + /* If the source and destination cgroups are the same, don't do anything. */ + current_gpucg = gpucg_get(current); + if (current_gpucg == gpucg) { + ret = 0; + goto skip_transfer; + } + + /* + * Verify that the cgroup of the process requesting the transfer is the + * same as the one the buffer is currently charged to. + */ + current_gpucg = gpucg_get(current); + mutex_lock(&dmabuf->lock); + if (current_gpucg != dmabuf->gpucg) { + ret = -EPERM; + goto err; + } + + ret = gpucg_transfer_charge(current_gpucg, gpucg, dmabuf->gpucg_dev, dmabuf->size); + if (ret) + goto err; + dmabuf->gpucg = gpucg; +err: + mutex_unlock(&dmabuf->lock); +skip_transfer: + gpucg_put(current_gpucg); + return ret; +#else + return 0; +#endif /* CONFIG_CGROUP_GPU */ +} +EXPORT_SYMBOL_NS_GPL(dma_buf_transfer_charge, DMA_BUF); + #ifdef CONFIG_DEBUG_FS static int dma_buf_debug_show(struct seq_file *s, void *unused) { diff --git a/include/linux/cgroup_gpu.h b/include/linux/cgroup_gpu.h index c90069719022..e30f15d5e9be 100644 --- a/include/linux/cgroup_gpu.h +++ b/include/linux/cgroup_gpu.h @@ -87,6 +87,10 @@ static inline struct gpucg *gpucg_parent(struct gpucg *cg) int gpucg_try_charge(struct gpucg *gpucg, struct gpucg_device *device, u64 usage); void gpucg_uncharge(struct gpucg *gpucg, struct gpucg_device *device, u64 usage); +int gpucg_transfer_charge(struct gpucg *source, + struct gpucg *dest, + struct gpucg_device *device, + u64 usage); void gpucg_register_device(struct gpucg_device *gpucg_dev, const char *name); #else /* CONFIG_CGROUP_GPU */ @@ -121,6 +125,14 @@ static inline void gpucg_uncharge(struct gpucg *gpucg, struct gpucg_device *device, u64 usage) {} +static inline int gpucg_transfer_charge(struct gpucg *source, + struct gpucg *dest, + struct gpucg_device *device, + u64 usage) +{ + return 0; +} + static inline void gpucg_register_device(struct gpucg_device *gpucg_dev, const char *name) {} #endif /* CONFIG_CGROUP_GPU */ diff --git a/include/linux/dma-buf.h b/include/linux/dma-buf.h index 742f29c3daaf..646827156213 100644 --- a/include/linux/dma-buf.h +++ b/include/linux/dma-buf.h @@ -646,4 +646,6 @@ int dma_buf_mmap(struct dma_buf *, struct vm_area_struct *, unsigned long); int dma_buf_vmap(struct dma_buf *dmabuf, struct dma_buf_map *map); void dma_buf_vunmap(struct dma_buf *dmabuf, struct dma_buf_map *map); + +int dma_buf_transfer_charge(struct dma_buf *dmabuf, struct gpucg *gpucg); #endif /* __DMA_BUF_H__ */ diff --git a/kernel/cgroup/gpu.c b/kernel/cgroup/gpu.c index ac4c470914b5..40531323d6da 100644 --- a/kernel/cgroup/gpu.c +++ b/kernel/cgroup/gpu.c @@ -247,6 +247,65 @@ void gpucg_uncharge(struct gpucg *gpucg, struct gpucg_device *device, u64 usage) css_put_many(&gpucg->css, nr_pages); } +/** + * gpucg_transfer_charge - Transfer a GPU charge from one cgroup to another. + * @source: [in] The GPU cgroup the charge will be transferred from. + * @dest: [in] The GPU cgroup the charge will be transferred to. + * @device: [in] The GPU cgroup device corresponding to the charge. + * @usage: [in] The size of the memory in bytes. + * + * Returns 0 on success, or a negative errno code otherwise. + */ +int gpucg_transfer_charge(struct gpucg *source, + struct gpucg *dest, + struct gpucg_device *device, + u64 usage) +{ + struct page_counter *counter; + u64 nr_pages; + struct gpucg_resource_pool *rp_source, *rp_dest; + int ret = 0; + + nr_pages = PAGE_ALIGN(usage) >> PAGE_SHIFT; + + mutex_lock(&gpucg_mutex); + rp_source = find_cg_rpool_locked(source, device); + if (unlikely(!rp_source)) { + ret = -ENOENT; + goto exit_early; + } + + rp_dest = get_cg_rpool_locked(dest, device); + if (IS_ERR(rp_dest)) { + ret = PTR_ERR(rp_dest); + goto exit_early; + } + + /* + * First uncharge from the pool it's currently charged to. This ordering avoids double + * charging while the transfer is in progress, which could cause us to hit a limit. + * If the try_charge fails for this transfer, we need to be able to reverse this uncharge, + * so we continue to hold the gpucg_mutex here. + */ + page_counter_uncharge(&rp_source->total, nr_pages); + css_put_many(&source->css, nr_pages); + + /* Now attempt the new charge */ + if (page_counter_try_charge(&rp_dest->total, nr_pages, &counter)) { + css_get_many(&dest->css, nr_pages); + } else { + /* + * The new charge failed, so reverse the uncharge from above. This should always + * succeed since charges on source are blocked by gpucg_mutex. + */ + WARN_ON(!page_counter_try_charge(&rp_source->total, nr_pages, &counter)); + css_get_many(&source->css, nr_pages); + } +exit_early: + mutex_unlock(&gpucg_mutex); + return ret; +} + /** * gpucg_register_device - Registers a device for memory accounting using the * GPU cgroup controller. -- 2.35.1.1021.g381101b075-goog ^ permalink raw reply related [ flat | nested ] 22+ messages in thread * [RFC v4 6/8] binder: Add a buffer flag to relinquish ownership of fds 2022-03-28 3:59 [RFC v4 0/8] Proposal for a GPU cgroup controller T.J. Mercier ` (4 preceding siblings ...) 2022-03-28 3:59 ` [RFC v4 5/8] dmabuf: Add gpu cgroup charge transfer function T.J. Mercier @ 2022-03-28 3:59 ` T.J. Mercier 2022-03-28 3:59 ` [RFC v4 7/8] binder: use __kernel_pid_t and __kernel_uid_t for userspace T.J. Mercier 2022-03-28 3:59 ` [RFC v4 8/8] selftests: Add binder cgroup gpu memory transfer test T.J. Mercier 7 siblings, 0 replies; 22+ messages in thread From: T.J. Mercier @ 2022-03-28 3:59 UTC ( permalink / raw ) To: tjmercier, David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan Cc: kaleshsingh, Kenny.Ho, mkoutny, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest From: Hridya Valsaraju <[email protected]> This patch introduces a buffer flag BINDER_BUFFER_FLAG_SENDER_NO_NEED that a process sending an fd array to another process over binder IPC can set to relinquish ownership of the fds being sent for memory accounting purposes. If the flag is found to be set during the fd array translation and the fd is for a DMA-BUF, the buffer is uncharged from the sender's cgroup and charged to the receiving process's cgroup instead. It is up to the sending process to ensure that it closes the fds regardless of whether the transfer failed or succeeded. Most graphics shared memory allocations in Android are done by the graphics allocator HAL process. On requests from clients, the HAL process allocates memory and sends the fds to the clients over binder IPC. The graphics allocator HAL will not retain any references to the buffers. When the HAL sets the BINDER_BUFFER_FLAG_SENDER_NO_NEED for fd arrays holding DMA-BUF fds, the gpu cgroup controller will be able to correctly charge the buffers to the client processes instead of the graphics allocator HAL. Since this is a new feature exposed to userspace, the kernel and userspace must be compatible for the accounting to work for transfers. In all cases the allocation and transport of DMA buffers via binder will succeed, but only when both the kernel supports, and userspace depends on this feature will the transfer accounting work. The possible scenarios are detailed below: 1. new kernel + old userspace The kernel supports the feature but userspace does not use it. The old userspace won't mount the new cgroup controller, accounting is not performed, charge is not transferred. 2. old kernel + new userspace The new cgroup controller is not supported by the kernel, accounting is not performed, charge is not transferred. 3. old kernel + old userspace Same as #2 4. new kernel + new userspace Cgroup is mounted, feature is supported and used. Signed-off-by: Hridya Valsaraju <[email protected]> Signed-off-by: T.J. Mercier <[email protected]> --- v3 changes Remove android from title per Todd Kjos. Use more common dual author commit message format per John Stultz. Include details on behavior for all combinations of kernel/userspace versions in changelog (thanks Suren Baghdasaryan) per Greg Kroah-Hartman. v2 changes Move dma-buf cgroup charge transfer from a dma_buf_op defined by every heap to a single dma-buf function for all heaps per Daniel Vetter and Christian König. --- drivers/android/binder.c | 26 ++++++++++++++++++++++++++ include/uapi/linux/android/binder.h | 1 + 2 files changed , 27 insertions(+) diff --git a/drivers/android/binder.c b/drivers/android/binder.c index 8351c5638880..4357d2efc8e1 100644 --- a/drivers/android/binder.c +++ b/drivers/android/binder.c @@ -42,6 +42,7 @@ #define pr_fmt(fmt) KBUILD_MODNAME ": " fmt +#include <linux/dma-buf.h> #include <linux/fdtable.h> #include <linux/file.h> #include <linux/freezer.h> @@ -2482,8 +2483,10 @@ static int binder_translate_fd_array(struct list_head *pf_head, { binder_size_t fdi, fd_buf_size; binder_size_t fda_offset; + bool transfer_gpu_charge = false; const void __user *sender_ufda_base; struct binder_proc *proc = thread->proc; + struct binder_proc *target_proc = t->to_proc; int ret; fd_buf_size = sizeof(u32) * fda->num_fds; @@ -2521,8 +2524,15 @@ static int binder_translate_fd_array(struct list_head *pf_head, if (ret) return ret; + if (IS_ENABLED(CONFIG_CGROUP_GPU) && + parent->flags & BINDER_BUFFER_FLAG_SENDER_NO_NEED) + transfer_gpu_charge = true; + for (fdi = 0; fdi < fda->num_fds; fdi++) { u32 fd; + struct dma_buf *dmabuf; + struct gpucg *gpucg; + binder_size_t offset = fda_offset + fdi * sizeof(fd); binder_size_t sender_uoffset = fdi * sizeof(fd); @@ -2532,6 +2542,22 @@ static int binder_translate_fd_array(struct list_head *pf_head, in_reply_to); if (ret) return ret > 0 ? -EINVAL : ret; + + if (!transfer_gpu_charge) + continue; + + dmabuf = dma_buf_get(fd); + if (IS_ERR(dmabuf)) + continue; + + gpucg = gpucg_get(target_proc->tsk); + ret = dma_buf_transfer_charge(dmabuf, gpucg); + if (ret) { + pr_warn("%d:%d Unable to transfer DMA-BUF fd charge to %d", + proc->pid, thread->pid, target_proc->pid); + gpucg_put(gpucg); + } + dma_buf_put(dmabuf); } return 0; } diff --git a/include/uapi/linux/android/binder.h b/include/uapi/linux/android/binder.h index 3246f2c74696..169fd5069a1a 100644 --- a/include/uapi/linux/android/binder.h +++ b/include/uapi/linux/android/binder.h @@ -137,6 +137,7 @@ struct binder_buffer_object { enum { BINDER_BUFFER_FLAG_HAS_PARENT = 0x01, + BINDER_BUFFER_FLAG_SENDER_NO_NEED = 0x02, }; /* struct binder_fd_array_object - object describing an array of fds in a buffer -- 2.35.1.1021.g381101b075-goog ^ permalink raw reply related [ flat | nested ] 22+ messages in thread [RFC v4 0/8] Proposal for a GPU cgroup controller [RFC v4 6/8] binder: Add a buffer flag to relinquish ownership of fds ) To: tjmercier, David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan Cc: kaleshsingh, Kenny.Ho, mkoutny, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest The kernel interface should use types that the kernel defines instead of pid_t and uid_t, whose definiton is owned by libc. This fixes the header so that it can be included without first including sys/types.h. Signed-off-by: T.J. Mercier <[email protected]> --- include/uapi/linux/android/binder.h | 4 ++-- 1 file changed , 2 insertions(+), 2 deletions(-) diff --git a/include/uapi/linux/android/binder.h b/include/uapi/linux/android/binder.h index 169fd5069a1a..aa28454dbca3 100644 --- a/include/uapi/linux/android/binder.h +++ b/include/uapi/linux/android/binder.h @@ -289,8 +289,8 @@ struct binder_transaction_data { /* General information about the transaction. */ __u32 flags; - pid_t sender_pid; - uid_t sender_euid; + __kernel_pid_t sender_pid; + __kernel_uid_t sender_euid; binder_size_t data_size; /* number of bytes of data */ binder_size_t offsets_size; /* number of bytes of offsets */ -- 2.35.1.1021.g381101b075-goog * [RFC v4 8/8] selftests: Add binder cgroup gpu memory transfer test 2022-03-28 3:59 [RFC v4 0/8] Proposal for a GPU cgroup controller T.J. Mercier ` (6 preceding siblings ...) 2022-03-28 3:59 ` [RFC v4 7/8] binder: use __kernel_pid_t and __kernel_uid_t for userspace T.J. Mercier @ 2022-03-28 3:59 ` T.J. Mercier 7 siblings, 0 replies; 22+ messages in thread From: T.J. Mercier @ 2022-03-28 3:59 UTC ( permalink / raw ) To: tjmercier, David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan Cc: kaleshsingh, Kenny.Ho, mkoutny, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest This test verifies that the cgroup GPU memory charge is transferred correctly when a dmabuf is passed between processes in two different cgroups and the sender specifies BINDER_BUFFER_FLAG_SENDER_NO_NEED in the binder transaction data containing the dmabuf file descriptor. Signed-off-by: T.J. Mercier <[email protected]> --- v4 changes Skip test if not run as root per Shuah Khan. Add better logging for abnormal child termination per Shuah Khan. --- .../selftests/drivers/android/binder/Makefile | 8 + .../drivers/android/binder/binder_util.c | 254 +++++++++ .../drivers/android/binder/binder_util.h | 32 ++ .../selftests/drivers/android/binder/config | 4 + .../binder/test_dmabuf_cgroup_transfer.c | 484 ++++++++++++++++++ 5 files changed , 782 insertions(+) create mode 100644 tools/testing/selftests/drivers/android/binder/Makefile create mode 100644 tools/testing/selftests/drivers/android/binder/binder_util.c create mode 100644 tools/testing/selftests/drivers/android/binder/binder_util.h create mode 100644 tools/testing/selftests/drivers/android/binder/config create mode 100644 tools/testing/selftests/drivers/android/binder/test_dmabuf_cgroup_transfer.c diff --git a/tools/testing/selftests/drivers/android/binder/Makefile b/tools/testing/selftests/drivers/android/binder/Makefile new file mode 100644 index 000000000000..726439d10675 --- /dev/null +++ b/tools/testing/selftests/drivers/android/binder/Makefile @@ -0,0 +1,8 @@ +# SPDX-License-Identifier: GPL-2.0 +CFLAGS += -Wall + +TEST_GEN_PROGS = test_dmabuf_cgroup_transfer + +include ../../../lib.mk + +$(OUTPUT)/test_dmabuf_cgroup_transfer: ../../../cgroup/cgroup_util.c binder_util.c diff --git a/tools/testing/selftests/drivers/android/binder/binder_util.c b/tools/testing/selftests/drivers/android/binder/binder_util.c new file mode 100644 index 000000000000..c9dcf5b9d42b --- /dev/null +++ b/tools/testing/selftests/drivers/android/binder/binder_util.c @@ -0,0 +1,254 @@ +// SPDX-License-Identifier: GPL-2.0 + +#include "binder_util.h" + +#include <errno.h> +#include <fcntl.h> +#include <stdio.h> +#include <stdlib.h> +#include <string.h> +#include <unistd.h> +#include <sys/ioctl.h> +#include <sys/mman.h> +#include <sys/mount.h> + +#include <linux/limits.h> +#include <linux/android/binder.h> +#include <linux/android/binderfs.h> + +static const size_t BINDER_MMAP_SIZE = 64 * 1024; + +static void binderfs_unmount(const char *mountpoint) +{ + if (umount2(mountpoint, MNT_DETACH)) + fprintf(stderr, "Failed to unmount binderfs at %s: %s\n", + mountpoint, strerror(errno)); + else + fprintf(stderr, "Binderfs unmounted: %s\n", mountpoint); + + if (rmdir(mountpoint)) + fprintf(stderr, "Failed to remove binderfs mount %s: %s\n", + mountpoint, strerror(errno)); + else + fprintf(stderr, "Binderfs mountpoint destroyed: %s\n", mountpoint); +} + +struct binderfs_ctx create_binderfs(const char *name) +{ + int fd, ret, saved_errno; + struct binderfs_device device = { 0 }; + struct binderfs_ctx ctx = { 0 }; + + /* + * P_tmpdir is set to "/tmp/" on Android platforms where Binder is most + * commonly used, but this path does not actually exist on Android. We + * will first try using "/data/local/tmp" and fallback to P_tmpdir if + * that fails for non-Android platforms. + */ + static const char tmpdir[] = "/data/local/tmp"; + static const size_t MAX_TMPDIR_SIZE = + sizeof(tmpdir) > sizeof(P_tmpdir) ? + sizeof(tmpdir) : sizeof(P_tmpdir); + static const char template[] = "/binderfs_XXXXXX"; + + char *mkdtemp_result; + char binderfs_mntpt[MAX_TMPDIR_SIZE + sizeof(template)]; + char device_path[MAX_TMPDIR_SIZE + sizeof(template) + BINDERFS_MAX_NAME]; + + snprintf(binderfs_mntpt, sizeof(binderfs_mntpt), "%s%s", tmpdir, template); + + mkdtemp_result = mkdtemp(binderfs_mntpt); + if (mkdtemp_result == NULL) { + fprintf(stderr, "Failed to create binderfs mountpoint at %s: %s.\n", + binderfs_mntpt, strerror(errno)); + fprintf(stderr, "Trying fallback mountpoint...\n"); + snprintf(binderfs_mntpt, sizeof(binderfs_mntpt), "%s%s", P_tmpdir, template); + if (mkdtemp(binderfs_mntpt) == NULL) { + fprintf(stderr, "Failed to create binderfs mountpoint at %s: %s\n", + binderfs_mntpt, strerror(errno)); + return ctx; + } + } + fprintf(stderr, "Binderfs mountpoint created at %s\n", binderfs_mntpt); + + if (mount(NULL, binderfs_mntpt, "binder", 0, 0)) { + perror("Could not mount binderfs"); + rmdir(binderfs_mntpt); + return ctx; + } + fprintf(stderr, "Binderfs mounted at %s\n", binderfs_mntpt); + + strncpy(device.name, name, sizeof(device.name)); + snprintf(device_path, sizeof(device_path), "%s/binder-control", binderfs_mntpt); + fd = open(device_path, O_RDONLY | O_CLOEXEC); + if (!fd) { + perror("Failed to open binder-control device"); + binderfs_unmount(binderfs_mntpt); + return ctx; + } + + ret = ioctl(fd, BINDER_CTL_ADD, &device); + saved_errno = errno; + close(fd); + errno = saved_errno; + if (ret) { + perror("Failed to allocate new binder device"); + binderfs_unmount(binderfs_mntpt); + return ctx; + } + + fprintf(stderr, "Allocated new binder device with major %d, minor %d, and name %s at %s\n", + device.major, device.minor, device.name, binderfs_mntpt); + + ctx.name = strdup(name); + ctx.mountpoint = strdup(binderfs_mntpt); + return ctx; +} + +void destroy_binderfs(struct binderfs_ctx *ctx) +{ + char path[PATH_MAX]; + + snprintf(path, sizeof(path), "%s/%s", ctx->mountpoint, ctx->name); + + if (unlink(path)) + fprintf(stderr, "Failed to unlink binder device %s: %s\n", path, strerror(errno)); + else + fprintf(stderr, "Destroyed binder %s at %s\n", ctx->name, ctx->mountpoint); + + binderfs_unmount(ctx->mountpoint); + + free(ctx->name); + free(ctx->mountpoint); +} + +struct binder_ctx open_binder(struct binderfs_ctx *bfs_ctx) +{ + struct binder_ctx ctx = {.fd = -1, .memory = NULL}; + char path[PATH_MAX]; + + snprintf(path, sizeof(path), "%s/%s", bfs_ctx->mountpoint, bfs_ctx->name); + ctx.fd = open(path, O_RDWR | O_NONBLOCK | O_CLOEXEC); + if (ctx.fd < 0) { + fprintf(stderr, "Error opening binder device %s: %s\n", path, strerror(errno)); + return ctx; + } + + ctx.memory = mmap(NULL, BINDER_MMAP_SIZE, PROT_READ, MAP_SHARED, ctx.fd, 0); + if (ctx.memory == NULL) { + perror("Error mapping binder memory"); + close(ctx.fd); + ctx.fd = -1; + } + + return ctx; +} + +void close_binder(struct binder_ctx *ctx) +{ + if (munmap(ctx->memory, BINDER_MMAP_SIZE)) + perror("Failed to unmap binder memory"); + ctx->memory = NULL; + + if (close(ctx->fd)) + perror("Failed to close binder"); + ctx->fd = -1; +} + +int become_binder_context_manager(int binder_fd) +{ + return ioctl(binder_fd, BINDER_SET_CONTEXT_MGR, 0); +} + +int do_binder_write_read(int binder_fd, void *writebuf, binder_size_t writesize, + void *readbuf, binder_size_t readsize) +{ + int err; + struct binder_write_read bwr = { + .write_buffer = (binder_uintptr_t)writebuf, + .write_size = writesize, + .read_buffer = (binder_uintptr_t)readbuf, + .read_size = readsize + }; + + do { + if (ioctl(binder_fd, BINDER_WRITE_READ, &bwr) >= 0) + err = 0; + else + err = -errno; + } while (err == -EINTR); + + if (err < 0) { + perror("BINDER_WRITE_READ"); + return -1; + } + + if (bwr.write_consumed < writesize) { + fprintf(stderr, "Binder did not consume full write buffer %llu %llu\n", + bwr.write_consumed, writesize); + return -1; + } + + return bwr.read_consumed; +} + +static const char *reply_string(int cmd) +{ + switch (cmd) { + case BR_ERROR: + return("BR_ERROR"); + case BR_OK: + return("BR_OK"); + case BR_TRANSACTION_SEC_CTX: + return("BR_TRANSACTION_SEC_CTX"); + case BR_TRANSACTION: + return("BR_TRANSACTION"); + case BR_REPLY: + return("BR_REPLY"); + case BR_ACQUIRE_RESULT: + return("BR_ACQUIRE_RESULT"); + case BR_DEAD_REPLY: + return("BR_DEAD_REPLY"); + case BR_TRANSACTION_COMPLETE: + return("BR_TRANSACTION_COMPLETE"); + case BR_INCREFS: + return("BR_INCREFS"); + case BR_ACQUIRE: + return("BR_ACQUIRE"); + case BR_RELEASE: + return("BR_RELEASE"); + case BR_DECREFS: + return("BR_DECREFS"); + case BR_ATTEMPT_ACQUIRE: + return("BR_ATTEMPT_ACQUIRE"); + case BR_NOOP: + return("BR_NOOP"); + case BR_SPAWN_LOOPER: + return("BR_SPAWN_LOOPER"); + case BR_FINISHED: + return("BR_FINISHED"); + case BR_DEAD_BINDER: + return("BR_DEAD_BINDER"); + case BR_CLEAR_DEATH_NOTIFICATION_DONE: + return("BR_CLEAR_DEATH_NOTIFICATION_DONE"); + case BR_FAILED_REPLY: + return("BR_FAILED_REPLY"); + case BR_FROZEN_REPLY: + return("BR_FROZEN_REPLY"); + case BR_ONEWAY_SPAM_SUSPECT: + return("BR_ONEWAY_SPAM_SUSPECT"); + default: + return("Unknown"); + }; +} + +int expect_binder_reply(int32_t actual, int32_t expected) +{ + if (actual != expected) { + fprintf(stderr, "Expected %s but received %s\n", + reply_string(expected), reply_string(actual)); + return -1; + } + return 0; +} + diff --git a/tools/testing/selftests/drivers/android/binder/binder_util.h b/tools/testing/selftests/drivers/android/binder/binder_util.h new file mode 100644 index 000000000000..807f5abe987e --- /dev/null +++ b/tools/testing/selftests/drivers/android/binder/binder_util.h @@ -0,0 +1,32 @@ +/* SPDX-License-Identifier: GPL-2.0 */ + +#ifndef SELFTEST_BINDER_UTIL_H +#define SELFTEST_BINDER_UTIL_H + +#include <stdint.h> + +#include <linux/android/binder.h> + +struct binderfs_ctx { + char *name; + char *mountpoint; +}; + +struct binder_ctx { + int fd; + void *memory; +}; + +struct binderfs_ctx create_binderfs(const char *name); +void destroy_binderfs(struct binderfs_ctx *ctx); + +struct binder_ctx open_binder(struct binderfs_ctx *bfs_ctx); +void close_binder(struct binder_ctx *ctx); + +int become_binder_context_manager(int binder_fd); + +int do_binder_write_read(int binder_fd, void *writebuf, binder_size_t writesize, + void *readbuf, binder_size_t readsize); + +int expect_binder_reply(int32_t actual, int32_t expected); +#endif diff --git a/tools/testing/selftests/drivers/android/binder/config b/tools/testing/selftests/drivers/android/binder/config new file mode 100644 index 000000000000..fcc5f8f693b3 --- /dev/null +++ b/tools/testing/selftests/drivers/android/binder/config @@ -0,0 +1,4 @@ +CONFIG_CGROUP_GPU=y +CONFIG_ANDROID=y +CONFIG_ANDROID_BINDERFS=y +CONFIG_ANDROID_BINDER_IPC=y diff --git a/tools/testing/selftests/drivers/android/binder/test_dmabuf_cgroup_transfer.c b/tools/testing/selftests/drivers/android/binder/test_dmabuf_cgroup_transfer.c new file mode 100644 index 000000000000..586465d7bf6d --- /dev/null +++ b/tools/testing/selftests/drivers/android/binder/test_dmabuf_cgroup_transfer.c @@ -0,0 +1,484 @@ +// SPDX-License-Identifier: GPL-2.0 + +/* + * This test verifies that the cgroup GPU memory charge is transferred correctly + * when a dmabuf is passed between processes in two different cgroups and the + * sender specifies BINDER_BUFFER_FLAG_SENDER_NO_NEED in the binder transaction + * data containing the dmabuf file descriptor. + * + * The gpu_cgroup_dmabuf_transfer test function becomes the binder context + * manager, then forks a child who initiates a transaction with the context + * manager by specifying a target of 0. The context manager reply contains a + * dmabuf file descriptor which was allocated by the gpu_cgroup_dmabuf_transfer + * test function, but should be charged to the child cgroup after the binder + * transaction. + */ + +#include <errno.h> +#include <fcntl.h> +#include <stddef.h> +#include <stdint.h> +#include <stdio.h> +#include <stdlib.h> +#include <string.h> +#include <unistd.h> +#include <sys/epoll.h> +#include <sys/ioctl.h> +#include <sys/types.h> +#include <sys/wait.h> + +#include "binder_util.h" +#include "../../../cgroup/cgroup_util.h" +#include "../../../kselftest.h" +#include "../../../kselftest_harness.h" + +#include <linux/limits.h> +#include <linux/dma-heap.h> +#include <linux/android/binder.h> + +#define UNUSED(x) ((void)(x)) + +static const unsigned int BINDER_CODE = 8675309; /* Any number will work here */ + +struct cgroup_ctx { + char *root; + char *source; + char *dest; +}; + +void destroy_cgroups(struct __test_metadata *_metadata, struct cgroup_ctx *ctx) +{ + if (ctx->source != NULL) { + TH_LOG("Destroying cgroup: %s", ctx->source); + rmdir(ctx->source); + free(ctx->source); + } + + if (ctx->dest != NULL) { + TH_LOG("Destroying cgroup: %s", ctx->dest); + rmdir(ctx->dest); + free(ctx->dest); + } + + free(ctx->root); + ctx->root = ctx->source = ctx->dest = NULL; +} + +struct cgroup_ctx create_cgroups(struct __test_metadata *_metadata) +{ + struct cgroup_ctx ctx = {0}; + char root[PATH_MAX], *tmp; + static const char template[] = "/gpucg_XXXXXX"; + + if (cg_find_unified_root(root, sizeof(root))) { + TH_LOG("Could not find cgroups root"); + return ctx; + } + + if (cg_read_strstr(root, "cgroup.controllers", "gpu")) { + TH_LOG("Could not find GPU controller"); + return ctx; + } + + if (cg_write(root, "cgroup.subtree_control", "+gpu")) { + TH_LOG("Could not enable GPU controller"); + return ctx; + } + + ctx.root = strdup(root); + + snprintf(root, sizeof(root), "%s/%s", ctx.root, template); + tmp = mkdtemp(root); + if (tmp == NULL) { + TH_LOG("%s - Could not create source cgroup", strerror(errno)); + destroy_cgroups(_metadata, &ctx); + return ctx; + } + ctx.source = strdup(tmp); + + snprintf(root, sizeof(root), "%s/%s", ctx.root, template); + tmp = mkdtemp(root); + if (tmp == NULL) { + TH_LOG("%s - Could not create destination cgroup", strerror(errno)); + destroy_cgroups(_metadata, &ctx); + return ctx; + } + ctx.dest = strdup(tmp); + + TH_LOG("Created cgroups: %s %s", ctx.source, ctx.dest); + + return ctx; +} + +int dmabuf_heap_alloc(int fd, size_t len, int *dmabuf_fd) +{ + struct dma_heap_allocation_data data = { + .len = len, + .fd = 0, + .fd_flags = O_RDONLY | O_CLOEXEC, + .heap_flags = 0, + }; + int ret; + + if (!dmabuf_fd) + return -EINVAL; + + ret = ioctl(fd, DMA_HEAP_IOCTL_ALLOC, &data); + if (ret < 0) + return ret; + *dmabuf_fd = (int)data.fd; + return ret; +} + +/* The system heap is known to export dmabufs with support for cgroup tracking */ +int alloc_dmabuf_from_system_heap(struct __test_metadata *_metadata, size_t bytes) +{ + int heap_fd = -1, dmabuf_fd = -1; + static const char * const heap_path = "/dev/dma_heap/system"; + + heap_fd = open(heap_path, O_RDONLY); + if (heap_fd < 0) { + TH_LOG("%s - open %s failed!\n", strerror(errno), heap_path); + return -1; + } + + if (dmabuf_heap_alloc(heap_fd, bytes, &dmabuf_fd)) + TH_LOG("dmabuf allocation failed! - %s", strerror(errno)); + close(heap_fd); + + return dmabuf_fd; +} + +int binder_request_dmabuf(int binder_fd) +{ + int ret; + + /* + * We just send an empty binder_buffer_object to initiate a transaction + * with the context manager, who should respond with a single dmabuf + * inside a binder_fd_array_object. + */ + + struct binder_buffer_object bbo = { + .hdr.type = BINDER_TYPE_PTR, + .flags = 0, + .buffer = 0, + .length = 0, + .parent = 0, /* No parent */ + .parent_offset = 0 /* No parent */ + }; + + binder_size_t offsets[] = {0}; + + struct { + int32_t cmd; + struct binder_transaction_data btd; + } __attribute__((packed)) bc = { + .cmd = BC_TRANSACTION, + .btd = { + .target = { 0 }, + .cookie = 0, + .code = BINDER_CODE, + .flags = TF_ACCEPT_FDS, /* We expect a FDA in the reply */ + .data_size = sizeof(bbo), + .offsets_size = sizeof(offsets), + .data.ptr = { + (binder_uintptr_t)&bbo, + (binder_uintptr_t)offsets + } + }, + }; + + struct { + int32_t reply_noop; + } __attribute__((packed)) br; + + ret = do_binder_write_read(binder_fd, &bc, sizeof(bc), &br, sizeof(br)); + if (ret >= sizeof(br) && expect_binder_reply(br.reply_noop, BR_NOOP)) { + return -1; + } else if (ret < sizeof(br)) { + fprintf(stderr, "Not enough bytes in binder reply %d\n", ret); + return -1; + } + return 0; +} + +int send_dmabuf_reply(int binder_fd, struct binder_transaction_data *tr, int dmabuf_fd) +{ + int ret; + /* + * The trailing 0 is to achieve the necessary alignment for the binder + * buffer_size. + */ + int fdarray[] = { dmabuf_fd, 0 }; + + struct binder_buffer_object bbo = { + .hdr.type = BINDER_TYPE_PTR, + .flags = BINDER_BUFFER_FLAG_SENDER_NO_NEED, + .buffer = (binder_uintptr_t)fdarray, + .length = sizeof(fdarray), + .parent = 0, /* No parent */ + .parent_offset = 0 /* No parent */ + }; + + struct binder_fd_array_object bfdao = { + .hdr.type = BINDER_TYPE_FDA, + .num_fds = 1, + .parent = 0, /* The binder_buffer_object */ + .parent_offset = 0 /* FDs follow immediately */ + }; + + uint64_t sz = sizeof(fdarray); + uint8_t data[sizeof(sz) + sizeof(bbo) + sizeof(bfdao)]; + binder_size_t offsets[] = {sizeof(sz), sizeof(sz)+sizeof(bbo)}; + + memcpy(data, &sz, sizeof(sz)); + memcpy(data + sizeof(sz), &bbo, sizeof(bbo)); + memcpy(data + sizeof(sz) + sizeof(bbo), &bfdao, sizeof(bfdao)); + + struct { + int32_t cmd; + struct binder_transaction_data_sg btd; + } __attribute__((packed)) bc = { + .cmd = BC_REPLY_SG, + .btd.transaction_data = { + .target = { tr->target.handle }, + .cookie = tr->cookie, + .code = BINDER_CODE, + .flags = 0, + .data_size = sizeof(data), + .offsets_size = sizeof(offsets), + .data.ptr = { + (binder_uintptr_t)data, + (binder_uintptr_t)offsets + } + }, + .btd.buffers_size = sizeof(fdarray) + }; + + struct { + int32_t reply_noop; + } __attribute__((packed)) br; + + ret = do_binder_write_read(binder_fd, &bc, sizeof(bc), &br, sizeof(br)); + if (ret >= sizeof(br) && expect_binder_reply(br.reply_noop, BR_NOOP)) { + return -1; + } else if (ret < sizeof(br)) { + fprintf(stderr, "Not enough bytes in binder reply %d\n", ret); + return -1; + } + return 0; +} + +struct binder_transaction_data *binder_wait_for_transaction(int binder_fd, + uint32_t *readbuf, + size_t readsize) +{ + static const int MAX_EVENTS = 1, EPOLL_WAIT_TIME_MS = 3 * 1000; + struct binder_reply { + int32_t reply0; + int32_t reply1; + struct binder_transaction_data btd; + } *br; + struct binder_transaction_data *ret = NULL; + struct epoll_event events[MAX_EVENTS]; + int epoll_fd, num_events, readcount; + uint32_t bc[] = { BC_ENTER_LOOPER }; + + do_binder_write_read(binder_fd, &bc, sizeof(bc), NULL, 0); + + epoll_fd = epoll_create1(EPOLL_CLOEXEC); + if (epoll_fd == -1) { + perror("epoll_create"); + return NULL; + } + + events[0].events = EPOLLIN; + if (epoll_ctl(epoll_fd, EPOLL_CTL_ADD, binder_fd, &events[0])) { + perror("epoll_ctl add"); + goto err_close; + } + + num_events = epoll_wait(epoll_fd, events, MAX_EVENTS, EPOLL_WAIT_TIME_MS); + if (num_events < 0) { + perror("epoll_wait"); + goto err_ctl; + } else if (num_events == 0) { + fprintf(stderr, "No events\n"); + goto err_ctl; + } + + readcount = do_binder_write_read(binder_fd, NULL, 0, readbuf, readsize); + fprintf(stderr, "Read %d bytes from binder\n", readcount); + + if (readcount < (int)sizeof(struct binder_reply)) { + fprintf(stderr, "read_consumed not large enough\n"); + goto err_ctl; + } + + br = (struct binder_reply *)readbuf; + if (expect_binder_reply(br->reply0, BR_NOOP)) + goto err_ctl; + + if (br->reply1 == BR_TRANSACTION) { + if (br->btd.code == BINDER_CODE) + ret = &br->btd; + else + fprintf(stderr, "Received transaction with unexpected code: %u\n", + br->btd.code); + } else { + expect_binder_reply(br->reply1, BR_TRANSACTION_COMPLETE); + } + +err_ctl: + if (epoll_ctl(epoll_fd, EPOLL_CTL_DEL, binder_fd, NULL)) + perror("epoll_ctl del"); +err_close: + close(epoll_fd); + return ret; +} + +static int child_request_dmabuf_transfer(const char *cgroup, void *arg) +{ + UNUSED(cgroup); + int ret = -1; + uint32_t readbuf[32]; + struct binderfs_ctx bfs_ctx = *(struct binderfs_ctx *)arg; + struct binder_ctx b_ctx; + + fprintf(stderr, "Child PID: %d\n", getpid()); + + b_ctx = open_binder(&bfs_ctx); + if (b_ctx.fd < 0) { + fprintf(stderr, "Child unable to open binder\n"); + return -1; + } + + if (binder_request_dmabuf(b_ctx.fd)) + goto err; + + /* The child must stay alive until the binder reply is received */ + if (binder_wait_for_transaction(b_ctx.fd, readbuf, sizeof(readbuf)) == NULL) + ret = 0; + + /* + * We don't close the received dmabuf here so that the parent can + * inspect the cgroup gpu memory charges to verify the charge transfer + * completed successfully. + */ +err: + close_binder(&b_ctx); + fprintf(stderr, "Child done\n"); + return ret; +} + +TEST(gpu_cgroup_dmabuf_transfer) +{ + static const char * const GPUMEM_FILENAME = "gpu.memory.current"; + static const size_t ONE_MiB = 1024 * 1024; + + int ret, dmabuf_fd; + uint32_t readbuf[32]; + long memsize; + pid_t child_pid; + struct binderfs_ctx bfs_ctx; + struct binder_ctx b_ctx; + struct cgroup_ctx cg_ctx; + struct binder_transaction_data *tr; + struct flat_binder_object *fbo; + struct binder_buffer_object *bbo; + + if (geteuid() != 0) + ksft_exit_skip("Need to be root to mount binderfs\n"); + + bfs_ctx = create_binderfs("testbinder"); + if (bfs_ctx.name == NULL) + ksft_exit_skip("The Android binderfs filesystem is not available\n"); + + cg_ctx = create_cgroups(_metadata); + if (cg_ctx.root == NULL) { + destroy_binderfs(&bfs_ctx); + ksft_exit_skip("cgroup v2 isn't mounted\n"); + } + + ASSERT_EQ(cg_enter_current(cg_ctx.source), 0) { + TH_LOG("Could not move parent to cgroup: %s", cg_ctx.source); + goto err_cg; + } + + dmabuf_fd = alloc_dmabuf_from_system_heap(_metadata, ONE_MiB); + ASSERT_GE(dmabuf_fd, 0) { + goto err_cg; + } + TH_LOG("Allocated dmabuf"); + + memsize = cg_read_key_long(cg_ctx.source, GPUMEM_FILENAME, "system"); + ASSERT_EQ(memsize, ONE_MiB) { + TH_LOG("GPU memory used after allocation: %ld but it should be %lu", + memsize, (unsigned long)ONE_MiB); + goto err_dmabuf; + } + + b_ctx = open_binder(&bfs_ctx); + ASSERT_GE(b_ctx.fd, 0) { + TH_LOG("Parent unable to open binder"); + goto err_dmabuf; + } + TH_LOG("Opened binder at %s/%s", bfs_ctx.mountpoint, bfs_ctx.name); + + ASSERT_EQ(become_binder_context_manager(b_ctx.fd), 0) { + TH_LOG("Cannot become context manager: %s", strerror(errno)); + goto err_binder; + } + + child_pid = cg_run_nowait(cg_ctx.dest, child_request_dmabuf_transfer, &bfs_ctx); + ASSERT_GT(child_pid, 0) { + TH_LOG("Error forking: %s", strerror(errno)); + goto err_binder; + } + + tr = binder_wait_for_transaction(b_ctx.fd, readbuf, sizeof(readbuf)); + ASSERT_NE(tr, NULL) { + TH_LOG("Error receiving transaction request from child"); + goto err_child; + } + fbo = (struct flat_binder_object *)tr->data.ptr.buffer; + ASSERT_EQ(fbo->hdr.type, BINDER_TYPE_PTR) { + TH_LOG("Did not receive a buffer object from child"); + goto err_child; + } + bbo = (struct binder_buffer_object *)fbo; + ASSERT_EQ(bbo->length, 0) { + TH_LOG("Did not receive an empty buffer object from child"); + goto err_child; + } + + TH_LOG("Received transaction from child"); + send_dmabuf_reply(b_ctx.fd, tr, dmabuf_fd); + + ASSERT_EQ(cg_read_key_long(cg_ctx.dest, GPUMEM_FILENAME, "system"), ONE_MiB) { + TH_LOG("Destination cgroup does not have system charge!"); + goto err_child; + } + ASSERT_EQ(cg_read_key_long(cg_ctx.source, GPUMEM_FILENAME, "system"), 0) { + TH_LOG("Source cgroup still has system charge!"); + goto err_child; + } + TH_LOG("Charge transfer succeeded!"); + +err_child: + waitpid(child_pid, &ret, 0); + if (WIFEXITED(ret)) + TH_LOG("Child %d terminated normally with %d", child_pid, WEXITSTATUS(ret)); + else if (WIFSIGNALED(ret)) + TH_LOG("Child %d terminated with signal %d\n", child_pid, WTERMSIG(ret)); +err_binder: + close_binder(&b_ctx); +err_dmabuf: + close(dmabuf_fd); +err_cg: + destroy_cgroups(_metadata, &cg_ctx); + destroy_binderfs(&bfs_ctx); +} + +TEST_HARNESS_MAIN -- 2.35.1.1021.g381101b075-goog ^ permalink raw reply related [ flat | nested ] 22+ messages in thread * Re: [RFC v4 4/8] dmabuf: heaps: export system_heap buffers with GPU cgroup charging 2022-03-28 3:59 ` [RFC v4 4/8] dmabuf: heaps: export system_heap buffers with GPU cgroup charging T.J. Mercier @ 2022-03-28 14:36 ` Daniel Vetter 2022-03-28 18:28 ` T.J. Mercier 0 siblings, 1 reply; 22+ messages in thread From: Daniel Vetter @ 2022-03-28 14:36 UTC ( permalink / raw ) To: T.J. Mercier Cc: David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan, kaleshsingh, Kenny.Ho, mkoutny, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest On Mon, Mar 28, 2022 at 03:59:43AM +0000, T.J. Mercier wrote: > From: Hridya Valsaraju <[email protected]> > > All DMA heaps now register a new GPU cgroup device upon creation, and the > system_heap now exports buffers associated with its GPU cgroup device for > tracking purposes. > > Signed-off-by: Hridya Valsaraju <[email protected]> > Signed-off-by: T.J. Mercier <[email protected]> > > --- > v3 changes > Use more common dual author commit message format per John Stultz. > > v2 changes > Move dma-buf cgroup charge transfer from a dma_buf_op defined by every > heap to a single dma-buf function for all heaps per Daniel Vetter and > Christian König. Apologies for being out of the loop quite a bit. I scrolled through this all and I think it looks good to get going. The only thing I have is whether we should move the cgroup controllers out of dma-buf heaps, since that's rather android centric. E.g. - a system gpucg_device which is used by all the various single page allocators (dma-buf heap but also shmem helpers and really anything else) - same for cma, again both for dma-buf heaps and also for the gem cma helpers in drm Otherwise this will only work on non-upstream android where gpu drivers allocate everything from dma-buf heap. If you use something like the x86 android project with mesa drivers, then driver-internal buffers will be allocated through gem and not through dma-buf heaps. Or at least I think that's how it works. But also meh, we can fix this fairly easily later on by adding these standard gpucg_dev somwehere with a bit of kerneldoc. Anyway has my all my ack, but don't count this as my in-depth review :-) -Daniel > --- > drivers/dma-buf/dma-heap.c | 27 +++++++++++++++++++++++++++ > drivers/dma-buf/heaps/system_heap.c | 3 +++ > include/linux/dma-heap.h | 11 +++++++++++ > 3 files changed, 41 insertions(+) > > diff --git a/drivers/dma-buf/dma-heap.c b/drivers/dma-buf/dma-heap.c > index 8f5848aa144f..885072427775 100644 > --- a/drivers/dma-buf/dma-heap.c > +++ b/drivers/dma-buf/dma-heap.c > @@ -7,6 +7,7 @@ > */ > > #include <linux/cdev.h> > +#include <linux/cgroup_gpu.h> > #include <linux/debugfs.h> > #include <linux/device.h> > #include <linux/dma-buf.h> > @@ -31,6 +32,7 @@ > * @heap_devt heap device node > * @list list head connecting to list of heaps > * @heap_cdev heap char device > + * @gpucg_dev gpu cgroup device for memory accounting > * > * Represents a heap of memory from which buffers can be made. > */ > @@ -41,6 +43,9 @@ struct dma_heap { > dev_t heap_devt; > struct list_head list; > struct cdev heap_cdev; > +#ifdef CONFIG_CGROUP_GPU > + struct gpucg_device gpucg_dev; > +#endif > }; > > static LIST_HEAD(heap_list); > @@ -216,6 +221,26 @@ const char *dma_heap_get_name(struct dma_heap *heap) > return heap->name; > } > > +#ifdef CONFIG_CGROUP_GPU > +/** > + * dma_heap_get_gpucg_dev() - get struct gpucg_device for the heap. > + * @heap: DMA-Heap to get the gpucg_device struct for. > + * > + * Returns: > + * The gpucg_device struct for the heap. NULL if the GPU cgroup controller is > + * not enabled. > + */ > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap) > +{ > + return &heap->gpucg_dev; > +} > +#else /* CONFIG_CGROUP_GPU */ > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap) > +{ > + return NULL; > +} > +#endif /* CONFIG_CGROUP_GPU */ > + > struct dma_heap *dma_heap_add(const struct dma_heap_export_info *exp_info) > { > struct dma_heap *heap, *h, *err_ret; > @@ -288,6 +313,8 @@ struct dma_heap *dma_heap_add(const struct dma_heap_export_info *exp_info) > list_add(&heap->list, &heap_list); > mutex_unlock(&heap_list_lock); > > + gpucg_register_device(dma_heap_get_gpucg_dev(heap), exp_info->name); > + > return heap; > > err2: > diff --git a/drivers/dma-buf/heaps/system_heap.c b/drivers/dma-buf/heaps/system_heap.c > index ab7fd896d2c4..752a05c3cfe2 100644 > --- a/drivers/dma-buf/heaps/system_heap.c > +++ b/drivers/dma-buf/heaps/system_heap.c > @@ -395,6 +395,9 @@ static struct dma_buf *system_heap_allocate(struct dma_heap *heap, > exp_info.ops = &system_heap_buf_ops; > exp_info.size = buffer->len; > exp_info.flags = fd_flags; > +#ifdef CONFIG_CGROUP_GPU > + exp_info.gpucg_dev = dma_heap_get_gpucg_dev(heap); > +#endif > exp_info.priv = buffer; > dmabuf = dma_buf_export(&exp_info); > if (IS_ERR(dmabuf)) { > diff --git a/include/linux/dma-heap.h b/include/linux/dma-heap.h > index 0c05561cad6e..e447a61d054e 100644 > --- a/include/linux/dma-heap.h > +++ b/include/linux/dma-heap.h > @@ -10,6 +10,7 @@ > #define _DMA_HEAPS_H > > #include <linux/cdev.h> > +#include <linux/cgroup_gpu.h> > #include <linux/types.h> > > struct dma_heap; > @@ -59,6 +60,16 @@ void *dma_heap_get_drvdata(struct dma_heap *heap); > */ > const char *dma_heap_get_name(struct dma_heap *heap); > > +/** > + * dma_heap_get_gpucg_dev() - get a pointer to the struct gpucg_device for the > + * heap. > + * @heap: DMA-Heap to retrieve gpucg_device for. > + * > + * Returns: > + * The gpucg_device struct for the heap. > + */ > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap); > + > /** > * dma_heap_add - adds a heap to dmabuf heaps > * @exp_info: information needed to register this heap > -- > 2.35.1.1021.g381101b075-goog > -- Daniel Vetter Software Engineer, Intel Corporation http://blog.ffwll.ch ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 4/8] dmabuf: heaps: export system_heap buffers with GPU cgroup charging 2022-03-28 14:36 ` Daniel Vetter @ 2022-03-28 18:28 ` T.J. Mercier 2022-03-29 8:42 ` Daniel Vetter 0 siblings, 1 reply; 22+ messages in thread From: T.J. Mercier @ 2022-03-28 18:28 UTC ( permalink / raw ) To: T.J. Mercier, David Airlie, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan, Kalesh Singh, Kenny.Ho, Michal Koutný, Shuah Khan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest Cc: Daniel Vetter On Mon, Mar 28, 2022 at 7:36 AM Daniel Vetter <[email protected]> wrote: > > On Mon, Mar 28, 2022 at 03:59:43AM +0000, T.J. Mercier wrote: > > From: Hridya Valsaraju <[email protected]> > > > > All DMA heaps now register a new GPU cgroup device upon creation, and the > > system_heap now exports buffers associated with its GPU cgroup device for > > tracking purposes. > > > > Signed-off-by: Hridya Valsaraju <[email protected]> > > Signed-off-by: T.J. Mercier <[email protected]> > > > > --- > > v3 changes > > Use more common dual author commit message format per John Stultz. > > > > v2 changes > > Move dma-buf cgroup charge transfer from a dma_buf_op defined by every > > heap to a single dma-buf function for all heaps per Daniel Vetter and > > Christian König. > > Apologies for being out of the loop quite a bit. I scrolled through this > all and I think it looks good to get going. > > The only thing I have is whether we should move the cgroup controllers out > of dma-buf heaps, since that's rather android centric. E.g. > - a system gpucg_device which is used by all the various single page > allocators (dma-buf heap but also shmem helpers and really anything > else) > - same for cma, again both for dma-buf heaps and also for the gem cma > helpers in drm Thanks Daniel, in general that makes sense to me as an approach to making this more universal. However for the Android case I'm not sure if the part about a single system gpucg_device would be sufficient, because there are at least 12 different graphics related heaps that could potentially be accounted/limited differently. [1] So that raises the question of how fine grained we want this to be... I tend towards separating them all, but I haven't formed a strong opinion about this at the moment. It sounds like you are in favor of a smaller, more rigidly defined set of them? Either way, we need to add code for accounting at points where we know memory is specifically for graphics use and not something else right? (I.E. Whether it is a dma-buf heap or somewhere like drm_gem_object_init.) So IIUC the only question is what to use for the gpucg_device(s) at these locations. [1] https://cs.android.com/android/platform/superproject/+/master:hardware/google/graphics/common/libion/ion.cpp;l=39-50 > > Otherwise this will only work on non-upstream android where gpu drivers > allocate everything from dma-buf heap. If you use something like the x86 > android project with mesa drivers, then driver-internal buffers will be > allocated through gem and not through dma-buf heaps. Or at least I think > that's how it works. > > But also meh, we can fix this fairly easily later on by adding these > standard gpucg_dev somwehere with a bit of kerneldoc. This is what I was thinking would happen next, but IDK if anyone sees a more central place to do this type of use-specific accounting. > > Anyway has my all my ack, but don't count this as my in-depth review :-) > -Daniel Thanks again for taking a look! > > > --- > > drivers/dma-buf/dma-heap.c | 27 +++++++++++++++++++++++++++ > > drivers/dma-buf/heaps/system_heap.c | 3 +++ > > include/linux/dma-heap.h | 11 +++++++++++ > > 3 files changed, 41 insertions(+) > > > > diff --git a/drivers/dma-buf/dma-heap.c b/drivers/dma-buf/dma-heap.c > > index 8f5848aa144f..885072427775 100644 > > --- a/drivers/dma-buf/dma-heap.c > > +++ b/drivers/dma-buf/dma-heap.c > > @@ -7,6 +7,7 @@ > > */ > > > > #include <linux/cdev.h> > > +#include <linux/cgroup_gpu.h> > > #include <linux/debugfs.h> > > #include <linux/device.h> > > #include <linux/dma-buf.h> > > @@ -31,6 +32,7 @@ > > * @heap_devt heap device node > > * @list list head connecting to list of heaps > > * @heap_cdev heap char device > > + * @gpucg_dev gpu cgroup device for memory accounting > > * > > * Represents a heap of memory from which buffers can be made. > > */ > > @@ -41,6 +43,9 @@ struct dma_heap { > > dev_t heap_devt; > > struct list_head list; > > struct cdev heap_cdev; > > +#ifdef CONFIG_CGROUP_GPU > > + struct gpucg_device gpucg_dev; > > +#endif > > }; > > > > static LIST_HEAD(heap_list); > > @@ -216,6 +221,26 @@ const char *dma_heap_get_name(struct dma_heap *heap) > > return heap->name; > > } > > > > +#ifdef CONFIG_CGROUP_GPU > > +/** > > + * dma_heap_get_gpucg_dev() - get struct gpucg_device for the heap. > > + * @heap: DMA-Heap to get the gpucg_device struct for. > > + * > > + * Returns: > > + * The gpucg_device struct for the heap. NULL if the GPU cgroup controller is > > + * not enabled. > > + */ > > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap) > > +{ > > + return &heap->gpucg_dev; > > +} > > +#else /* CONFIG_CGROUP_GPU */ > > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap) > > +{ > > + return NULL; > > +} > > +#endif /* CONFIG_CGROUP_GPU */ > > + > > struct dma_heap *dma_heap_add(const struct dma_heap_export_info *exp_info) > > { > > struct dma_heap *heap, *h, *err_ret; > > @@ -288,6 +313,8 @@ struct dma_heap *dma_heap_add(const struct dma_heap_export_info *exp_info) > > list_add(&heap->list, &heap_list); > > mutex_unlock(&heap_list_lock); > > > > + gpucg_register_device(dma_heap_get_gpucg_dev(heap), exp_info->name); > > + > > return heap; > > > > err2: > > diff --git a/drivers/dma-buf/heaps/system_heap.c b/drivers/dma-buf/heaps/system_heap.c > > index ab7fd896d2c4..752a05c3cfe2 100644 > > --- a/drivers/dma-buf/heaps/system_heap.c > > +++ b/drivers/dma-buf/heaps/system_heap.c > > @@ -395,6 +395,9 @@ static struct dma_buf *system_heap_allocate(struct dma_heap *heap, > > exp_info.ops = &system_heap_buf_ops; > > exp_info.size = buffer->len; > > exp_info.flags = fd_flags; > > +#ifdef CONFIG_CGROUP_GPU > > + exp_info.gpucg_dev = dma_heap_get_gpucg_dev(heap); > > +#endif > > exp_info.priv = buffer; > > dmabuf = dma_buf_export(&exp_info); > > if (IS_ERR(dmabuf)) { > > diff --git a/include/linux/dma-heap.h b/include/linux/dma-heap.h > > index 0c05561cad6e..e447a61d054e 100644 > > --- a/include/linux/dma-heap.h > > +++ b/include/linux/dma-heap.h > > @@ -10,6 +10,7 @@ > > #define _DMA_HEAPS_H > > > > #include <linux/cdev.h> > > +#include <linux/cgroup_gpu.h> > > #include <linux/types.h> > > > > struct dma_heap; > > @@ -59,6 +60,16 @@ void *dma_heap_get_drvdata(struct dma_heap *heap); > > */ > > const char *dma_heap_get_name(struct dma_heap *heap); > > > > +/** > > + * dma_heap_get_gpucg_dev() - get a pointer to the struct gpucg_device for the > > + * heap. > > + * @heap: DMA-Heap to retrieve gpucg_device for. > > + * > > + * Returns: > > + * The gpucg_device struct for the heap. > > + */ > > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap); > > + > > /** > > * dma_heap_add - adds a heap to dmabuf heaps > > * @exp_info: information needed to register this heap > > -- > > 2.35.1.1021.g381101b075-goog > > > > -- > Daniel Vetter > Software Engineer, Intel Corporation > http://blog.ffwll.ch ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 4/8] dmabuf: heaps: export system_heap buffers with GPU cgroup charging 2022-03-28 18:28 ` T.J. Mercier @ 2022-03-29 8:42 ` Daniel Vetter 2022-03-29 16:50 ` Tejun Heo 2022-03-29 17:52 ` T.J. Mercier 0 siblings, 2 replies; 22+ messages in thread From: Daniel Vetter @ 2022-03-29 8:42 UTC ( permalink / raw ) To: T.J. Mercier Cc: David Airlie, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan, Kalesh Singh, Kenny.Ho, Michal Koutný, Shuah Khan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest, Daniel Vetter On Mon, Mar 28, 2022 at 11:28:24AM -0700, T.J. Mercier wrote: > On Mon, Mar 28, 2022 at 7:36 AM Daniel Vetter <[email protected]> wrote: > > > > On Mon, Mar 28, 2022 at 03:59:43AM +0000, T.J. Mercier wrote: > > > From: Hridya Valsaraju <[email protected]> > > > > > > All DMA heaps now register a new GPU cgroup device upon creation, and the > > > system_heap now exports buffers associated with its GPU cgroup device for > > > tracking purposes. > > > > > > Signed-off-by: Hridya Valsaraju <[email protected]> > > > Signed-off-by: T.J. Mercier <[email protected]> > > > > > > --- > > > v3 changes > > > Use more common dual author commit message format per John Stultz. > > > > > > v2 changes > > > Move dma-buf cgroup charge transfer from a dma_buf_op defined by every > > > heap to a single dma-buf function for all heaps per Daniel Vetter and > > > Christian König. > > > > Apologies for being out of the loop quite a bit. I scrolled through this > > all and I think it looks good to get going. > > > > The only thing I have is whether we should move the cgroup controllers out > > of dma-buf heaps, since that's rather android centric. E.g. > > - a system gpucg_device which is used by all the various single page > > allocators (dma-buf heap but also shmem helpers and really anything > > else) > > - same for cma, again both for dma-buf heaps and also for the gem cma > > helpers in drm > > Thanks Daniel, in general that makes sense to me as an approach to > making this more universal. However for the Android case I'm not sure > if the part about a single system gpucg_device would be sufficient, > because there are at least 12 different graphics related heaps that > could potentially be accounted/limited differently. [1] So that > raises the question of how fine grained we want this to be... I tend > towards separating them all, but I haven't formed a strong opinion > about this at the moment. It sounds like you are in favor of a > smaller, more rigidly defined set of them? Either way, we need to add > code for accounting at points where we know memory is specifically for > graphics use and not something else right? (I.E. Whether it is a > dma-buf heap or somewhere like drm_gem_object_init.) So IIUC the only > question is what to use for the gpucg_device(s) at these locations. We don't have 12 in upstream, so this is a lot easier here :-) I'm not exactly sure why you have such a huge pile of them. For gem buffers it would be fairly similar to what you've done for dma-buf heaps I think, with the various helper libraries (drivers stopped hand-rolling their gem buffer) setting the right accounting group. And yeah for system memory I think we'd need to have standard ones, for driver specific ones it's kinda different. > [1] https://cs.android.com/android/platform/superproject/+/master:hardware/google/graphics/common/libion/ion.cpp;l=39-50 > > > > > Otherwise this will only work on non-upstream android where gpu drivers > > allocate everything from dma-buf heap. If you use something like the x86 > > android project with mesa drivers, then driver-internal buffers will be > > allocated through gem and not through dma-buf heaps. Or at least I think > > that's how it works. > > > > But also meh, we can fix this fairly easily later on by adding these > > standard gpucg_dev somwehere with a bit of kerneldoc. > > This is what I was thinking would happen next, but IDK if anyone sees > a more central place to do this type of use-specific accounting. Hm I just realized ... are the names in the groups abi? If yes then I think we need to fix this before we merge anything. -Daniel > > > > > Anyway has my all my ack, but don't count this as my in-depth review :-) > > -Daniel > > Thanks again for taking a look! > > > > > --- > > > drivers/dma-buf/dma-heap.c | 27 +++++++++++++++++++++++++++ > > > drivers/dma-buf/heaps/system_heap.c | 3 +++ > > > include/linux/dma-heap.h | 11 +++++++++++ > > > 3 files changed, 41 insertions(+) > > > > > > diff --git a/drivers/dma-buf/dma-heap.c b/drivers/dma-buf/dma-heap.c > > > index 8f5848aa144f..885072427775 100644 > > > --- a/drivers/dma-buf/dma-heap.c > > > +++ b/drivers/dma-buf/dma-heap.c > > > @@ -7,6 +7,7 @@ > > > */ > > > > > > #include <linux/cdev.h> > > > +#include <linux/cgroup_gpu.h> > > > #include <linux/debugfs.h> > > > #include <linux/device.h> > > > #include <linux/dma-buf.h> > > > @@ -31,6 +32,7 @@ > > > * @heap_devt heap device node > > > * @list list head connecting to list of heaps > > > * @heap_cdev heap char device > > > + * @gpucg_dev gpu cgroup device for memory accounting > > > * > > > * Represents a heap of memory from which buffers can be made. > > > */ > > > @@ -41,6 +43,9 @@ struct dma_heap { > > > dev_t heap_devt; > > > struct list_head list; > > > struct cdev heap_cdev; > > > +#ifdef CONFIG_CGROUP_GPU > > > + struct gpucg_device gpucg_dev; > > > +#endif > > > }; > > > > > > static LIST_HEAD(heap_list); > > > @@ -216,6 +221,26 @@ const char *dma_heap_get_name(struct dma_heap *heap) > > > return heap->name; > > > } > > > > > > +#ifdef CONFIG_CGROUP_GPU > > > +/** > > > + * dma_heap_get_gpucg_dev() - get struct gpucg_device for the heap. > > > + * @heap: DMA-Heap to get the gpucg_device struct for. > > > + * > > > + * Returns: > > > + * The gpucg_device struct for the heap. NULL if the GPU cgroup controller is > > > + * not enabled. > > > + */ > > > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap) > > > +{ > > > + return &heap->gpucg_dev; > > > +} > > > +#else /* CONFIG_CGROUP_GPU */ > > > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap) > > > +{ > > > + return NULL; > > > +} > > > +#endif /* CONFIG_CGROUP_GPU */ > > > + > > > struct dma_heap *dma_heap_add(const struct dma_heap_export_info *exp_info) > > > { > > > struct dma_heap *heap, *h, *err_ret; > > > @@ -288,6 +313,8 @@ struct dma_heap *dma_heap_add(const struct dma_heap_export_info *exp_info) > > > list_add(&heap->list, &heap_list); > > > mutex_unlock(&heap_list_lock); > > > > > > + gpucg_register_device(dma_heap_get_gpucg_dev(heap), exp_info->name); > > > + > > > return heap; > > > > > > err2: > > > diff --git a/drivers/dma-buf/heaps/system_heap.c b/drivers/dma-buf/heaps/system_heap.c > > > index ab7fd896d2c4..752a05c3cfe2 100644 > > > --- a/drivers/dma-buf/heaps/system_heap.c > > > +++ b/drivers/dma-buf/heaps/system_heap.c > > > @@ -395,6 +395,9 @@ static struct dma_buf *system_heap_allocate(struct dma_heap *heap, > > > exp_info.ops = &system_heap_buf_ops; > > > exp_info.size = buffer->len; > > > exp_info.flags = fd_flags; > > > +#ifdef CONFIG_CGROUP_GPU > > > + exp_info.gpucg_dev = dma_heap_get_gpucg_dev(heap); > > > +#endif > > > exp_info.priv = buffer; > > > dmabuf = dma_buf_export(&exp_info); > > > if (IS_ERR(dmabuf)) { > > > diff --git a/include/linux/dma-heap.h b/include/linux/dma-heap.h > > > index 0c05561cad6e..e447a61d054e 100644 > > > --- a/include/linux/dma-heap.h > > > +++ b/include/linux/dma-heap.h > > > @@ -10,6 +10,7 @@ > > > #define _DMA_HEAPS_H > > > > > > #include <linux/cdev.h> > > > +#include <linux/cgroup_gpu.h> > > > #include <linux/types.h> > > > > > > struct dma_heap; > > > @@ -59,6 +60,16 @@ void *dma_heap_get_drvdata(struct dma_heap *heap); > > > */ > > > const char *dma_heap_get_name(struct dma_heap *heap); > > > > > > +/** > > > + * dma_heap_get_gpucg_dev() - get a pointer to the struct gpucg_device for the > > > + * heap. > > > + * @heap: DMA-Heap to retrieve gpucg_device for. > > > + * > > > + * Returns: > > > + * The gpucg_device struct for the heap. > > > + */ > > > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap); > > > + > > > /** > > > * dma_heap_add - adds a heap to dmabuf heaps > > > * @exp_info: information needed to register this heap > > > -- > > > 2.35.1.1021.g381101b075-goog > > > > > > > -- > > Daniel Vetter > > Software Engineer, Intel Corporation > > http://blog.ffwll.ch -- Daniel Vetter Software Engineer, Intel Corporation http://blog.ffwll.ch ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 5/8] dmabuf: Add gpu cgroup charge transfer function 2022-03-28 3:59 ` [RFC v4 5/8] dmabuf: Add gpu cgroup charge transfer function T.J. Mercier @ 2022-03-29 15:21 ` Michal Koutný 2022-04-01 18:41 ` T.J. Mercier 0 siblings, 1 reply; 22+ messages in thread From: Michal Koutný @ 2022-03-29 15:21 UTC ( permalink / raw ) To: T.J. Mercier Cc: David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan, kaleshsingh, Kenny.Ho, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest Hi. On Mon, Mar 28, 2022 at 03:59:44AM +0000, "T.J. Mercier" <[email protected]> wrote: > From: Hridya Valsaraju <[email protected]> > > The dma_buf_charge_transfer function provides a way for processes to (s/dma_bug_charge_transfer/dma_bug_transfer_charge/) > transfer charge of a buffer to a different process. This is essential > for the cases where a central allocator process does allocations for > various subsystems, hands over the fd to the client who requested the > memory and drops all references to the allocated memory. I understood from [1] some buffers are backed by regular RAM. How are these charges going to be transferred (if so)? Thanks, Michal [1] https://lore.kernel.org/r/CABdmKX2NSAKMC6rReMYfo2SSVNxEXcS466hk3qF6YFt-j-+_NQ@mail.gmail.com ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 4/8] dmabuf: heaps: export system_heap buffers with GPU cgroup charging 2022-03-29 8:42 ` Daniel Vetter @ 2022-03-29 16:50 ` Tejun Heo 2022-03-29 17:52 ` T.J. Mercier 1 sibling, 0 replies; 22+ messages in thread From: Tejun Heo @ 2022-03-29 16:50 UTC ( permalink / raw ) To: Daniel Vetter Cc: T.J. Mercier, David Airlie, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Zefan Li, Johannes Weiner, Shuah Khan, Kalesh Singh, Kenny.Ho, Michal Koutný, Shuah Khan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest On Tue, Mar 29, 2022 at 10:42:20AM +0200, Daniel Vetter wrote: > Hm I just realized ... are the names in the groups abi? If yes then I > think we need to fix this before we merge anything. Yes. Thanks. -- tejun ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 2/8] cgroup: gpu: Add a cgroup controller for allocator attribution of GPU memory 2022-03-28 3:59 ` [RFC v4 2/8] cgroup: gpu: Add a cgroup controller for allocator attribution of GPU memory T.J. Mercier @ 2022-03-29 16:59 ` Tejun Heo 2022-03-30 20:56 ` T.J. Mercier 0 siblings, 1 reply; 22+ messages in thread From: Tejun Heo @ 2022-03-29 16:59 UTC ( permalink / raw ) To: T.J. Mercier Cc: David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Zefan Li, Johannes Weiner, Shuah Khan, kaleshsingh, Kenny.Ho, mkoutny, skhan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest Hello, On Mon, Mar 28, 2022 at 03:59:41AM +0000, T.J. Mercier wrote: > The API/UAPI can be extended to set per-device/total allocation limits > in the future. This total thing kinda bothers me. Can you please provide some concrete examples of how this and per-device limits would be used? Thanks. -- tejun ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 4/8] dmabuf: heaps: export system_heap buffers with GPU cgroup charging 2022-03-29 8:42 ` Daniel Vetter 2022-03-29 16:50 ` Tejun Heo @ 2022-03-29 17:52 ` T.J. Mercier 1 sibling, 0 replies; 22+ messages in thread From: T.J. Mercier @ 2022-03-29 17:52 UTC ( permalink / raw ) To: T.J. Mercier, David Airlie, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan, Kalesh Singh, Kenny.Ho, Michal Koutný, Shuah Khan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest Cc: Daniel Vetter On Tue, Mar 29, 2022 at 1:42 AM Daniel Vetter <[email protected]> wrote: > > On Mon, Mar 28, 2022 at 11:28:24AM -0700, T.J. Mercier wrote: > > On Mon, Mar 28, 2022 at 7:36 AM Daniel Vetter <[email protected]> wrote: > > > > > > On Mon, Mar 28, 2022 at 03:59:43AM +0000, T.J. Mercier wrote: > > > > From: Hridya Valsaraju <[email protected]> > > > > > > > > All DMA heaps now register a new GPU cgroup device upon creation, and the > > > > system_heap now exports buffers associated with its GPU cgroup device for > > > > tracking purposes. > > > > > > > > Signed-off-by: Hridya Valsaraju <[email protected]> > > > > Signed-off-by: T.J. Mercier <[email protected]> > > > > > > > > --- > > > > v3 changes > > > > Use more common dual author commit message format per John Stultz. > > > > > > > > v2 changes > > > > Move dma-buf cgroup charge transfer from a dma_buf_op defined by every > > > > heap to a single dma-buf function for all heaps per Daniel Vetter and > > > > Christian König. > > > > > > Apologies for being out of the loop quite a bit. I scrolled through this > > > all and I think it looks good to get going. > > > > > > The only thing I have is whether we should move the cgroup controllers out > > > of dma-buf heaps, since that's rather android centric. E.g. > > > - a system gpucg_device which is used by all the various single page > > > allocators (dma-buf heap but also shmem helpers and really anything > > > else) > > > - same for cma, again both for dma-buf heaps and also for the gem cma > > > helpers in drm > > > > Thanks Daniel, in general that makes sense to me as an approach to > > making this more universal. However for the Android case I'm not sure > > if the part about a single system gpucg_device would be sufficient, > > because there are at least 12 different graphics related heaps that > > could potentially be accounted/limited differently. [1] So that > > raises the question of how fine grained we want this to be... I tend > > towards separating them all, but I haven't formed a strong opinion > > about this at the moment. It sounds like you are in favor of a > > smaller, more rigidly defined set of them? Either way, we need to add > > code for accounting at points where we know memory is specifically for > > graphics use and not something else right? (I.E. Whether it is a > > dma-buf heap or somewhere like drm_gem_object_init.) So IIUC the only > > question is what to use for the gpucg_device(s) at these locations. > > We don't have 12 in upstream, so this is a lot easier here :-) > > I'm not exactly sure why you have such a huge pile of them. > > For gem buffers it would be fairly similar to what you've done for dma-buf > heaps I think, with the various helper libraries (drivers stopped > hand-rolling their gem buffer) setting the right accounting group. And > yeah for system memory I think we'd need to have standard ones, for driver > specific ones it's kinda different. > > > [1] https://cs.android.com/android/platform/superproject/+/master:hardware/google/graphics/common/libion/ion.cpp;l=39-50 > > > > > > > > Otherwise this will only work on non-upstream android where gpu drivers > > > allocate everything from dma-buf heap. If you use something like the x86 > > > android project with mesa drivers, then driver-internal buffers will be > > > allocated through gem and not through dma-buf heaps. Or at least I think > > > that's how it works. > > > > > > But also meh, we can fix this fairly easily later on by adding these > > > standard gpucg_dev somwehere with a bit of kerneldoc. > > > > This is what I was thinking would happen next, but IDK if anyone sees > > a more central place to do this type of use-specific accounting. > > Hm I just realized ... are the names in the groups abi? If yes then I > think we need to fix this before we merge anything. > -Daniel Do you mean the set of possible names being part of the ABI for GPU cgroups? I'm not exactly sure what you mean here. The name is a settable string inside the gpucg_device struct, and right now the docs say it must be from a string literal but this can be changed. The only one this patchset adds is "system", which comes from the name field in its dma_heap_export_info struct when it's first created (and that string is hardcoded). The heap gpucg_devices are all registered from one spot in dma-heap.c, so maybe we should append "-heap" to the gpucg_device names there so "system" is available for a standardized name. Let me make those two changes, and I will also rebase this series before resending. > > > > > > > > > Anyway has my all my ack, but don't count this as my in-depth review :-) > > > -Daniel > > > > Thanks again for taking a look! > > > > > > > --- > > > > drivers/dma-buf/dma-heap.c | 27 +++++++++++++++++++++++++++ > > > > drivers/dma-buf/heaps/system_heap.c | 3 +++ > > > > include/linux/dma-heap.h | 11 +++++++++++ > > > > 3 files changed, 41 insertions(+) > > > > > > > > diff --git a/drivers/dma-buf/dma-heap.c b/drivers/dma-buf/dma-heap.c > > > > index 8f5848aa144f..885072427775 100644 > > > > --- a/drivers/dma-buf/dma-heap.c > > > > +++ b/drivers/dma-buf/dma-heap.c > > > > @@ -7,6 +7,7 @@ > > > > */ > > > > > > > > #include <linux/cdev.h> > > > > +#include <linux/cgroup_gpu.h> > > > > #include <linux/debugfs.h> > > > > #include <linux/device.h> > > > > #include <linux/dma-buf.h> > > > > @@ -31,6 +32,7 @@ > > > > * @heap_devt heap device node > > > > * @list list head connecting to list of heaps > > > > * @heap_cdev heap char device > > > > + * @gpucg_dev gpu cgroup device for memory accounting > > > > * > > > > * Represents a heap of memory from which buffers can be made. > > > > */ > > > > @@ -41,6 +43,9 @@ struct dma_heap { > > > > dev_t heap_devt; > > > > struct list_head list; > > > > struct cdev heap_cdev; > > > > +#ifdef CONFIG_CGROUP_GPU > > > > + struct gpucg_device gpucg_dev; > > > > +#endif > > > > }; > > > > > > > > static LIST_HEAD(heap_list); > > > > @@ -216,6 +221,26 @@ const char *dma_heap_get_name(struct dma_heap *heap) > > > > return heap->name; > > > > } > > > > > > > > +#ifdef CONFIG_CGROUP_GPU > > > > +/** > > > > + * dma_heap_get_gpucg_dev() - get struct gpucg_device for the heap. > > > > + * @heap: DMA-Heap to get the gpucg_device struct for. > > > > + * > > > > + * Returns: > > > > + * The gpucg_device struct for the heap. NULL if the GPU cgroup controller is > > > > + * not enabled. > > > > + */ > > > > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap) > > > > +{ > > > > + return &heap->gpucg_dev; > > > > +} > > > > +#else /* CONFIG_CGROUP_GPU */ > > > > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap) > > > > +{ > > > > + return NULL; > > > > +} > > > > +#endif /* CONFIG_CGROUP_GPU */ > > > > + > > > > struct dma_heap *dma_heap_add(const struct dma_heap_export_info *exp_info) > > > > { > > > > struct dma_heap *heap, *h, *err_ret; > > > > @@ -288,6 +313,8 @@ struct dma_heap *dma_heap_add(const struct dma_heap_export_info *exp_info) > > > > list_add(&heap->list, &heap_list); > > > > mutex_unlock(&heap_list_lock); > > > > > > > > + gpucg_register_device(dma_heap_get_gpucg_dev(heap), exp_info->name); > > > > + > > > > return heap; > > > > > > > > err2: > > > > diff --git a/drivers/dma-buf/heaps/system_heap.c b/drivers/dma-buf/heaps/system_heap.c > > > > index ab7fd896d2c4..752a05c3cfe2 100644 > > > > --- a/drivers/dma-buf/heaps/system_heap.c > > > > +++ b/drivers/dma-buf/heaps/system_heap.c > > > > @@ -395,6 +395,9 @@ static struct dma_buf *system_heap_allocate(struct dma_heap *heap, > > > > exp_info.ops = &system_heap_buf_ops; > > > > exp_info.size = buffer->len; > > > > exp_info.flags = fd_flags; > > > > +#ifdef CONFIG_CGROUP_GPU > > > > + exp_info.gpucg_dev = dma_heap_get_gpucg_dev(heap); > > > > +#endif > > > > exp_info.priv = buffer; > > > > dmabuf = dma_buf_export(&exp_info); > > > > if (IS_ERR(dmabuf)) { > > > > diff --git a/include/linux/dma-heap.h b/include/linux/dma-heap.h > > > > index 0c05561cad6e..e447a61d054e 100644 > > > > --- a/include/linux/dma-heap.h > > > > +++ b/include/linux/dma-heap.h > > > > @@ -10,6 +10,7 @@ > > > > #define _DMA_HEAPS_H > > > > > > > > #include <linux/cdev.h> > > > > +#include <linux/cgroup_gpu.h> > > > > #include <linux/types.h> > > > > > > > > struct dma_heap; > > > > @@ -59,6 +60,16 @@ void *dma_heap_get_drvdata(struct dma_heap *heap); > > > > */ > > > > const char *dma_heap_get_name(struct dma_heap *heap); > > > > > > > > +/** > > > > + * dma_heap_get_gpucg_dev() - get a pointer to the struct gpucg_device for the > > > > + * heap. > > > > + * @heap: DMA-Heap to retrieve gpucg_device for. > > > > + * > > > > + * Returns: > > > > + * The gpucg_device struct for the heap. > > > > + */ > > > > +struct gpucg_device *dma_heap_get_gpucg_dev(struct dma_heap *heap); > > > > + > > > > /** > > > > * dma_heap_add - adds a heap to dmabuf heaps > > > > * @exp_info: information needed to register this heap > > > > -- > > > > 2.35.1.1021.g381101b075-goog > > > > > > > > > > -- > > > Daniel Vetter > > > Software Engineer, Intel Corporation > > > http://blog.ffwll.ch > > -- > Daniel Vetter > Software Engineer, Intel Corporation > http://blog.ffwll.ch ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 2/8] cgroup: gpu: Add a cgroup controller for allocator attribution of GPU memory 2022-03-29 16:59 ` Tejun Heo @ 2022-03-30 20:56 ` T.J. Mercier 2022-04-04 17:41 ` Tejun Heo 0 siblings, 1 reply; 22+ messages in thread From: T.J. Mercier @ 2022-03-30 20:56 UTC ( permalink / raw ) To: Tejun Heo Cc: David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Zefan Li, Johannes Weiner, Shuah Khan, Kalesh Singh, Kenny.Ho, Michal Koutný, Shuah Khan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest On Tue, Mar 29, 2022 at 9:59 AM Tejun Heo <[email protected]> wrote: > > Hello, I'm sorry for the delay Tejun, my test device stopped working and my attention has been occupied with that. > > On Mon, Mar 28, 2022 at 03:59:41AM +0000, T.J. Mercier wrote: > > The API/UAPI can be extended to set per-device/total allocation limits > > in the future. > > This total thing kinda bothers me. Can you please provide some concrete > examples of how this and per-device limits would be used? The use case we have for accounting the total (separate from the individual devices) is to include the value as part of bugreports, for understanding the system-wide amount of dmabuf allocations. I'm not aware of an existing need to limit the total. Admittedly this is just the sum over the devices, but we currently maintain out of tree code to do this sort of thing today. [1] The per-device limits would be used to restrict the amount of each type of allocation charged to an individual application to prevent hogging or to completely prevent access. This limitation is not something we have implemented today, but it is on our roadmap. [1] https://android-review.googlesource.com/c/kernel/common/+/1566704/3/drivers/dma-buf/dma-heap.c > > Thanks. > > -- > tejun ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 5/8] dmabuf: Add gpu cgroup charge transfer function 2022-03-29 15:21 ` Michal Koutný @ 2022-04-01 18:41 ` T.J. Mercier 2022-04-05 12:12 ` Michal Koutný 0 siblings, 1 reply; 22+ messages in thread From: T.J. Mercier @ 2022-04-01 18:41 UTC ( permalink / raw ) To: Michal Koutný Cc: David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan, Kalesh Singh, Kenny.Ho, Shuah Khan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest On Tue, Mar 29, 2022 at 8:21 AM Michal Koutný <[email protected]> wrote: > > Hi. > > On Mon, Mar 28, 2022 at 03:59:44AM +0000, "T.J. Mercier" <[email protected]> wrote: > > From: Hridya Valsaraju <[email protected]> > > > > The dma_buf_charge_transfer function provides a way for processes to > > (s/dma_bug_charge_transfer/dma_bug_transfer_charge/) > Doh! Thanks. > > transfer charge of a buffer to a different process. This is essential > > for the cases where a central allocator process does allocations for > > various subsystems, hands over the fd to the client who requested the > > memory and drops all references to the allocated memory. > > I understood from [1] some buffers are backed by regular RAM. How are > these charges going to be transferred (if so)? > This link doesn't work for me, but I think you're referring to the discussion about your "RAM_backed_buffers" comment from March 23rd. I wanted to do a simple test to confirm my own understanding here, but that got delayed due to some problems on my end. Anyway the test I did goes like this: enable memcg and gpu cgoups tracking and run a process that allocates 100MiB of dmabufs. Observe memcg and gpu accounting values before and after the allocation. Before # cat memory.current gpu.memory.current 14909440 system 0 <Test program does the allocation of 100MiB of dmabufs> After # cat memory.current gpu.memory.current 48025600 system 104857600 So the memcg value increases by about 30 MiB while the gpu values increases by 100 MiB. This is with kmem enabled, and the /proc/maps file for this process indicates that the majority of that 30 MiB is kernel memory. I think this result shows that neither the kernel nor process memory overlap with the gpu cgroup tracking of these allocations. So despite the fact that these buffers are in main memory, they are allocated in a way that does not result in memcg attribution. (It looks to me like __GFP_ACCOUNT is not set for these.) > > Thanks, > Michal > > [1] > https://lore.kernel.org/r/CABdmKX2NSAKMC6rReMYfo2SSVNxEXcS466hk3qF6YFt-j-+_NQ@mail.gmail.com ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 2/8] cgroup: gpu: Add a cgroup controller for allocator attribution of GPU memory 2022-03-30 20:56 ` T.J. Mercier @ 2022-04-04 17:41 ` Tejun Heo 2022-04-04 17:49 ` T.J. Mercier 0 siblings, 1 reply; 22+ messages in thread From: Tejun Heo @ 2022-04-04 17:41 UTC ( permalink / raw ) To: T.J. Mercier Cc: David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Zefan Li, Johannes Weiner, Shuah Khan, Kalesh Singh, Kenny.Ho, Michal Koutný, Shuah Khan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest Hello, On Wed, Mar 30, 2022 at 01:56:09PM -0700, T.J. Mercier wrote: > The use case we have for accounting the total (separate from the > individual devices) is to include the value as part of bugreports, for > understanding the system-wide amount of dmabuf allocations. I'm not > aware of an existing need to limit the total. Admittedly this is just > the sum over the devices, but we currently maintain out of tree code > to do this sort of thing today. [1] So, drop this part? Thanks. -- tejun ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 2/8] cgroup: gpu: Add a cgroup controller for allocator attribution of GPU memory 2022-04-04 17:41 ` Tejun Heo @ 2022-04-04 17:49 ` T.J. Mercier 0 siblings, 0 replies; 22+ messages in thread From: T.J. Mercier @ 2022-04-04 17:49 UTC ( permalink / raw ) To: Tejun Heo Cc: David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Zefan Li, Johannes Weiner, Shuah Khan, Kalesh Singh, Kenny.Ho, Michal Koutný, Shuah Khan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest On Mon, Apr 4, 2022 at 10:42 AM Tejun Heo <[email protected]> wrote: > > Hello, > > On Wed, Mar 30, 2022 at 01:56:09PM -0700, T.J. Mercier wrote: > > The use case we have for accounting the total (separate from the > > individual devices) is to include the value as part of bugreports, for > > understanding the system-wide amount of dmabuf allocations. I'm not > > aware of an existing need to limit the total. Admittedly this is just > > the sum over the devices, but we currently maintain out of tree code > > to do this sort of thing today. [1] > > So, drop this part? Ok, will do - I'll drop the "total" limitation text from the series. > > Thanks. > > -- > tejun ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 5/8] dmabuf: Add gpu cgroup charge transfer function 2022-04-01 18:41 ` T.J. Mercier @ 2022-04-05 12:12 ` Michal Koutný 2022-04-05 17:48 ` T.J. Mercier 0 siblings, 1 reply; 22+ messages in thread From: Michal Koutný @ 2022-04-05 12:12 UTC ( permalink / raw ) To: T.J. Mercier Cc: David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan, Kalesh Singh, Kenny.Ho, Shuah Khan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest On Fri, Apr 01, 2022 at 11:41:36AM -0700, "T.J. Mercier" <[email protected]> wrote: > This link doesn't work for me, but I think you're referring to the > discussion about your "RAM_backed_buffers" comment from March 23rd. (Oops, it's a non-public message. But yes, you guessed it right ;-)) > Anyway the test I did goes like this: enable memcg and gpu cgoups > tracking and run a process that allocates 100MiB of dmabufs. Observe > memcg and gpu accounting values before and after the allocation. Thanks for this measurement/dem/demoo. > Before > # cat memory.current gpu.memory.current > 14909440 > system 0 > > <Test program does the allocation of 100MiB of dmabufs> > > After > # cat memory.current gpu.memory.current > 48025600 > system 104857600 > > So the memcg value increases by about 30 MiB while the gpu values > increases by 100 MiB. > This is with kmem enabled, and the /proc/maps > file for this process indicates that the majority of that 30 MiB is > kernel memory. > I think this result shows that neither the kernel nor process memory > overlap with the gpu cgroup tracking of these allocations. It depends how the semantics of the 'system' entry is defined, no? As I grasped from other thread, the 'total' is going to be removed, so 'system' represents exclusively device memory? > So despite the fact that these buffers are in main memory, they are > allocated in a way that does not result in memcg attribution. (It > looks to me like __GFP_ACCOUNT is not set for these.) (I thought you knew what dmabufs your program used :-p) So, the goal is to do the tracking and migrations only via the gpu cg layer, regardless how memcg charges it (or not). (I have no opinion on that, I'm just summing it so that we're on the same page.) Michal ^ permalink raw reply [ flat | nested ] 22+ messages in thread * Re: [RFC v4 5/8] dmabuf: Add gpu cgroup charge transfer function 2022-04-05 12:12 ` Michal Koutný @ 2022-04-05 17:48 ` T.J. Mercier 0 siblings, 0 replies; 22+ messages in thread From: T.J. Mercier @ 2022-04-05 17:48 UTC ( permalink / raw ) To: Michal Koutný Cc: David Airlie, Daniel Vetter, Maarten Lankhorst, Maxime Ripard, Thomas Zimmermann, Jonathan Corbet, Greg Kroah-Hartman, Arve Hjønnevåg, Todd Kjos, Martijn Coenen, Joel Fernandes, Christian Brauner, Hridya Valsaraju, Suren Baghdasaryan, Sumit Semwal, Christian König, Benjamin Gaignard, Liam Mark, Laura Abbott, Brian Starkey, John Stultz, Tejun Heo, Zefan Li, Johannes Weiner, Shuah Khan, Kalesh Singh, Kenny.Ho, Shuah Khan, dri-devel, linux-doc, linux-kernel, linux-media, linaro-mm-sig, cgroups, linux-kselftest On Tue, Apr 5, 2022 at 5:12 AM Michal Koutný <[email protected]> wrote: > > On Fri, Apr 01, 2022 at 11:41:36AM -0700, "T.J. Mercier" <[email protected]> wrote: > > This link doesn't work for me, but I think you're referring to the > > discussion about your "RAM_backed_buffers" comment from March 23rd. > > (Oops, it's a non-public message. But yes, you guessed it right ;-)) > > > Anyway the test I did goes like this: enable memcg and gpu cgoups > > tracking and run a process that allocates 100MiB of dmabufs. Observe > > memcg and gpu accounting values before and after the allocation. > > Thanks for this measurement/dem/demoo. > > > Before > > # cat memory.current gpu.memory.current > > 14909440 > > system 0 > > > > <Test program does the allocation of 100MiB of dmabufs> > > > > After > > # cat memory.current gpu.memory.current > > 48025600 > > system 104857600 > > > > So the memcg value increases by about 30 MiB while the gpu values > > increases by 100 MiB. > > > This is with kmem enabled, and the /proc/maps > > file for this process indicates that the majority of that 30 MiB is > > kernel memory. > > > I think this result shows that neither the kernel nor process memory > > overlap with the gpu cgroup tracking of these allocations. > > It depends how the semantics of the 'system' entry is defined, no? > As I grasped from other thread, the 'total' is going to be removed, so > 'system' represents exclusively device memory? > That's right. The system charges (soon to be renamed "system-heap") result only from an allocator (in this case the system heap) deciding to call gpucg_try_charge for the buffer which is entirely device memory. > > > So despite the fact that these buffers are in main memory, they are > > allocated in a way that does not result in memcg attribution. (It > > looks to me like __GFP_ACCOUNT is not set for these.) > > (I thought you knew what dmabufs your program used :-p) > I'm coming up to speed on a lot of new-to-me code here. :) Just for completeness, these buffers were allocated with libdmabufheap's AllocSystem. > So, the goal is to do the tracking and migrations only via the gpu cg > layer, regardless how memcg charges it (or not). > > (I have no opinion on that, I'm just summing it so that we're on the > same page.) > Yes, this reflects my intention and current state of the code in this series. > Michal Thanks, T.J. ^ permalink raw reply [ flat | nested ] 22+ messages in thread This is a public inbox, see mirroring instructions for how to clone and mirror all data and code used for this inbox; as well as URLs for NNTP newsgroup(s).

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Lipad - August 2, 1963 - Browse the Canadian House of Commons August 2, 1963 LIB George James McIlraith (Minister of Transport) Liberal Hon. G. J. Mcllraiih (Minister of Transport): Mr. Speaker, I am familiar with this situation and I do not think it is proper to say that the air line was not living up to its commitment. The air line took the precaution to have its senior representative fly to Toronto, when these people were going overseas, to hand them individually their one way tickets for the express purpose of having the opportunity to advise and warn each of them that they did not have return tickets, and that the air line would not return them to Canada unless it were paid the fare. That having been done, I do not know there is much that can properly be done by the department at the taxpayer's expense. I must say I have a great deal of sympathy for the plight of these people, and I think it most unfortunate that after what I hope was a very happy and pleasant holiday on the other side they now find themselves without return tickets. But I think it would be a pretty dangerous precedent for the government to take taxpayers' money and expend it to provide return tickets when the individuals, if I am correctly informed as I believe I am, were warned before they went that they did not have return tickets and that the air line would not fly them back unless return tickets were paid for. Topic: AIR TRANSPORT Subtopic: ASSISTANCE TO PASSENGERS STRANDED IN ENGLAND John Angus MacLean Progressive Conservative Hon. A. J. MacLean (Queens): I have a supplementary question, Mr. Speaker. I would like to ask the Minister of Transport whether consideration has been given to having the Department of External Affairs advance money, as is sometimes done, to those would-be passengers who are Canadian citizens, to allow them to return to their homes and jobs. Topic: AIR TRANSPORT Subtopic: ASSISTANCE TO PASSENGERS STRANDED IN ENGLAND LIB George James McIlraith (Minister of Transport) Liberal Mr. Mcllrailh: Mr. Speaker, so far I understand there has been no request from any of them for an advance of money. (Translation): Topic: AIR TRANSPORT Subtopic: ASSISTANCE TO PASSENGERS STRANDED IN ENGLAND NORTHERN AFFAIRS INQUIRY AS TO TRANSFER OF JURISDICTION On the orders of the day: PC Paul Raymond Martineau Progressive Conservative Hon. Paul Marlineau (Ponliac-Temisca-mingue): Mr. Speaker, I have a question for the Prime Minister. Is he considering the possibility of direct negotiations with Mr. Lesage concerning the transfer of jurisdiction over New Quebec to the government of the province of Quebec? (Text): Topic: NORTHERN AFFAIRS Subtopic: INQUIRY AS TO TRANSFER OF JURISDICTION LIB Lester Bowles Pearson (Prime Minister) Liberal Right Hon. L. B. Pearson (Prime Minister): Mr. Speaker, that matter is already under study, as the house knows, by at least two members of the cabinet. Topic: NORTHERN AFFAIRS Subtopic: INQUIRY AS TO TRANSFER OF JURISDICTION PUBLIC WORKS CLINTON, ONT.-INQUIRY AS TO NEW POST OFFICE On the orders of the day: Lewis Elston Cardiff Progressive Conservative Mr. L. E. Cardiff (Huron): Mr. Speaker, I have a question for the Minister of Public Works. I sent him notice of the question yesterday. Could the minister tell me whether any progress has been made with respect to a start on the new post office building at Clinton, Ontario? Topic: PUBLIC WORKS Subtopic: CLINTON, ONT.-INQUIRY AS TO NEW POST OFFICE LIB Hubert Badanai (Parliamentary Secretary to the Minister of Public Works) Liberal Mr. Hubert Badanai (Parliamentary Secretary to Minister of Public Works): Mr. Speaker, on behalf of the minister I am very happy to inform the hon. member that the Inquiries of the Ministry architect has submitted his preliminary sketches in which some revisions have been made, and he is now preparing the working drawings and specifications. It is planned that tenders will be called in October, for a completion date in September, 1964. Topic: PUBLIC WORKS Subtopic: CLINTON, ONT.-INQUIRY AS TO NEW POST OFFICE INDIAN AFFAIRS INQUIRY AS TO SETTING UP COURT OF CLAIMS On the orders of the day: CCF Thomas Speakman Barnett Co-operative Commonwealth Federation (C.C.F.) Mr. T. S. Barnett (Comox-Alberni): Mr. Speaker, I have a question for the Minister of Citizenship and Immigration. I would like to ask the minister whether we may expect that there will be prepared for submission to parliament when we return from the recess, draft legislation in respect of the establishment of a court of Indian claims, which is a matter of great concern to many residents of this country. Topic: INDIAN AFFAIRS Subtopic: INQUIRY AS TO SETTING UP COURT OF CLAIMS LIB Guy Favreau (Minister of Citizenship and Immigration) Liberal Hon. Guy Favreau (Minisler of Citizenship and Immigration): Mr. Speaker, I must say this is a question which has already been thoroughly reviewed, and it is my intention to submit to cabinet a recommendation for the establishment of such a court upon the return of the house. (Translation): Topic: INDIAN AFFAIRS Subtopic: INQUIRY AS TO SETTING UP COURT OF CLAIMS ROYAL CANADIAN MOUNTED POLICE REQUEST FOR INVESTIGATION INTO UNDERCOVER ACTIVITIES

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Biomedicines | Free Full-Text | Association of Polymorphisms in FSHR, INHA, ESR1, and BMP15 with Recurrent Implantation Failure Recurrent implantation failure (RIF) refers to two or more unsuccessful in vitro fertilization embryo transfers in the same individual. Embryonic characteristics, immunological factors, and coagulation factors are known to be the causes of RIF. Genetic factors have also been reported to be involved in the occurrence of RIF, and some single nucleotide polymorphisms (SNPs) may contribute to RIF. We examined SNPs in FSHR, INHA, ESR1, and BMP15, which have been associated with primary ovarian failure. A cohort of 133 RIF patients and 317 healthy controls consisting of all Korean women was included. Genotyping was performed by Taq-Man genotyping assays to determine the frequency of the following polymorphisms: FSHR rs6165, INHA rs11893842 and rs35118453, ESR1 rs9340799 and rs2234693, and BMP15 rs17003221 and rs3810682. The differences in these SNPs were compared between the patient and control groups. Our results demonstrate a decreased prevalence of RIF in subjects with the FSHR rs6165 A>G polymorphism [AA vs. AG adjusted odds ratio (AOR) = 0.432; confidence interval (CI) = 0.206–0.908; p = 0.027, AA+AG vs. GG AOR = 0.434; CI = 0.213–0.885; p = 0.022]. Based on a genotype combination analysis, the GG/AA (FSHR rs6165/ESR1 rs9340799: OR = 0.250; CI = 0.072–0.874; p = 0.030) and GG-CC (FSHR rs6165/BMP15 rs3810682: OR = 0.466; CI = 0.220–0.987; p = 0.046) alleles were also associated with a decreased RIF risk. Additionally, the FSHR rs6165GG and BMP15 rs17003221TT+TC genotype combination was associated with a decreased RIF risk (OR = 0.430; CI = 0.210–0.877; p = 0.020) and increased FSH levels, as assessed by an analysis of variance. The FSHR rs6165 polymorphism and genotype combinations are significantly associated with RIF development in Korean women. Association of Polymorphisms in FSHR , INHA , ESR1 , and BMP15 with Recurrent Implantation Failure by Eun-Ju Ko 1,† , Ji-Eun Shin 2,† , Jung-Yong Lee 1 , Chang-Soo Ryu 1 , Ji-Young Hwang 3 , Young-Ran Kim 2 , Eun-Hee Ahn 2 , Ji-Hyang Kim 2,* and Nam-Keun Kim 1,* 1 Department of Biomedical Science, College of Life Science, CHA University, Seongnam 13488, Republic of Korea 2 Department of Obstetrics and Gynecology, Fertility Center of CHA Bundang Medical Center, CHA University, Seongnam 13520, Republic of Korea 3 Department of Obstetrics and Gynecology, Fertility Center of CHA Gangnam Medical Center, CHA University, Seoul 06125, Republic of Korea * Authors to whom correspondence should be addressed. † These authors contributed equally to this work. Biomedicines 2023 , 11 (5), 1374; https://doi.org/10.3390/biomedicines11051374 Received: 31 March 2023 / Revised: 29 April 2023 / Accepted: 3 May 2023 / Published: 5 May 2023 Abstract Recurrent implantation failure (RIF) refers to two or more unsuccessful in vitro fertilization embryo transfers in the same individual. Embryonic characteristics, immunological factors, and coagulation factors are known to be the causes of RIF. Genetic factors have also been reported to be involved in the occurrence of RIF, and some single nucleotide polymorphisms (SNPs) may contribute to RIF. We examined SNPs in FSHR , INHA , ESR1 , and BMP15 , which have been associated with primary ovarian failure. A cohort of 133 RIF patients and 317 healthy controls consisting of all Korean women was included. Genotyping was performed by Taq-Man genotyping assays to determine the frequency of the following polymorphisms: FSHR rs6165, INHA rs11893842 and rs35118453, ESR1 rs9340799 and rs2234693, and BMP15 rs17003221 and rs3810682. The differences in these SNPs were compared between the patient and control groups. Our results demonstrate a decreased prevalence of RIF in subjects with the FSHR rs6165 A>G polymorphism [AA vs. AG adjusted odds ratio (AOR) = 0.432; confidence interval (CI) = 0.206–0.908; p = 0.027, AA+AG vs. GG AOR = 0.434; CI = 0.213–0.885; p = 0.022]. Based on a genotype combination analysis, the GG/AA ( FSHR rs6165/ESR1 rs9340799: OR = 0.250; CI = 0.072–0.874; p = 0.030) and GG-CC ( FSHR rs6165/ BMP15 rs3810682: OR = 0.466; CI = 0.220–0.987; p = 0.046) alleles were also associated with a decreased RIF risk. Additionally, the FSHR rs6165GG and BMP15 rs17003221TT+TC genotype combination was associated with a decreased RIF risk (OR = 0.430; CI = 0.210–0.877; p = 0.020) and increased FSH levels, as assessed by an analysis of variance. The FSHR rs6165 polymorphism and genotype combinations are significantly associated with RIF development in Korean women. Keywords: recurrent implantation failure ; single-nucleotide polymorphism ; genotype combination ; female hormones 1. Introduction Implantation is a process by which an embryo attaches to the lumen surface of the endometrium and then migrates into the deep layer of the endometrium [ 1 ]. Successful embryo implantation requires success at each event, such as sperm and oocyte quality, development of the early embryo and endometrium, and the interaction between the blastocyst and endometrium [ 2 , 3 ]. Recurrent implantation failure (RIF) is defined as repeated embryo implantation failure after four high-quality embryo transfers in women. Despite advances in in vitro fertilization (IVF) technology, success rates remain stable, and approximately 10% of women undergoing IVF treatment suffer from RIF [ 4 ]. Implantation failure can be a consequence of immunological, uterine, male, embryo, or coagulation factors and genetics [ 5 , 6 ]. Among them, embryogenesis and development are some of the most important processes in pregnancy [ 7 ]. In the IVF procedure, only embryos classified as healthy according to specific criteria are selected and transplanted into the uterus. However, a good or “transplantable” class of embryos may increase the success rate of transplantation and pregnancy [ 8 , 9 ]. The morphological criteria do not necessarily correlate with implantation success rates, and approximately 50% of healthy embryos fail to implant [ 10 ]. Additionally, several factors are involved in embryogenesis and development [ 11 , 12 , 13 , 14 , 15 , 16 ]. Maternal hormones comprise an important factor for successful implantation during pregnancy. In particular, ovarian hormones interact with signaling molecules, including cytokines and growth factors, to assist embryo implantation [ 17 ]. Additionally, hormone receptors are associated with the implantation process and pregnancy result, and changes in the hormone receptor expression and function may affect the pregnancy outcome [ 18 , 19 , 20 ]. One of the most important hormones is follicle-stimulating hormone (FSH). FSH binds to the FSH receptor (FSHR) expressed in follicular granulosa cells to induce estrogen production and secretion, which stimulates the growth and maturation of ovarian follicles and subsequently improves the quality and recovery rate of oocytes [ 21 ]. Previously, the FSHR was thought to be expressed only in the ovaries, but a recent study showed that it was expressed in extra-gonadal tissues including the cervix, endometrium, myometrium, vascular smooth muscle, and vascular endothelium [ 22 ]. One study confirmed that FSHR expression was upregulated during decidualization and in the myometrium during pregnancy. In addition, a study in mice showed that FSHR was expressed in the placental blood vessels and was related to fetal vascular formation, and deletion of the FSHR gene affected pregnancy [ 23 , 24 ]. Extraovarian FSHRs play an important role in establishing and maintaining successful pregnancies in humans [ 22 ]. Previously reported studies showed that FSHR was expressed in fetal vascular endothelium [ 23 ], and it was identified that the FSH–FSHR signaling system promotes the angiogenesis of vascular endothelial cells [ 25 ]. FSHR is also expressed in the uterine myometrium and plays an important role in regulating uterine muscle contraction [ 26 ]. The INHA gene encodes inhibin α, which suppresses FSHR expression, thereby inhibiting the action of FSH [ 27 ]. Inhibin α is expressed in the luminal epithelium, glandular epithelium, stromal tissue, and vascular endothelium throughout the menstrual cycle [ 28 ]. Inhibin α is secreted by mature follicles and reflects follicular maturity. One study showed that serum inhibin α is a predictor for determining oocyte maturation [ 29 ]. Furthermore, concentrations of inhibin α in follicular fluid were higher in the pregnancy group than in the non-pregnancy group, and a positive correlation was found between the number of oocytes retrieved and the fertility rate [ 30 ]. Estrogen receptors, ERα and ERβ, mediate well-characterized effects on follicle growth, maturation, oocyte release, and endometrial preparation for implantation by binding to estrogen [ 31 ]. Among them, ERα is encoded by estrogen receptor 1 ( ESR1 ) and plays an essential role in regulating decidualization [ 32 ]. Bone morphogenetic protein ( BMP ) 15 is a TGFβ family member secreted by oocytes during follicle formation and is expressed in granulocytes and follicles as well as in oocytes. BMP15 plays an important role in follicular development during primordial follicle recruitment, ovulation, and corpus luteum formation and is closely related to fertilization, embryonic quality, and pregnancy outcomes. Therefore, it can be considered a new molecular marker for predicting follicular development potential [ 33 ]. In addition, one study showed that the concentration of BMP15 in women with an intracytoplasmic sperm injection was related to the fertility level, and an increased follicular concentration of BMP15 indicated an increased fertility level [ 34 ]. Single-nucleotide polymorphisms (SNPs) have been associated with reproductive diseases [ 35 , 36 , 37 ]. Additionally, RIF has previously been associated with SNPs, and many studies have been published exploring the relevance of these associations [ 38 , 39 ]. Polymorphisms in regulatory regions (promoter, 5′, 3′ UTR) or gene-coding regions may alter gene expression [ 40 , 41 ]. Maternal hormones play an important role in maintaining pregnancy, and hormone levels may be altered by certain polymorphisms [ 42 , 43 ]. Therefore, four genes ( FSHR , INHA , ESR1 , and BMP15 ) related to hormones were selected, and polymorphisms located in gene regulatory or coding regions were selected. Finally, a total of seven mutations were selected: coding region ( FSHR rs6165, ESR1 rs2234693, and BMP15 rs17003221), promoter region ( INHA rs11893842, rs35118453, and ESR1 rs9340799), and 5′UTR region ( BMP15 rs3810682). The occurrence of FSHR , INHA , ESR1 , and BMP15 gene polymorphisms has also been reported to be associated with various reproductive diseases, including primary ovarian insufficiency (POI), pregnancy loss, and preeclampsia [ 44 , 45 , 46 , 47 ]. However, few studies have examined the associations between RIF and FSHR , INHA , ESR1 , and BMP15 gene polymorphisms. In this study, we investigated whether FSHR , INHA , ESR1 , and BMP15 polymorphisms are associated with RIF and whether these polymorphisms affect the levels of clinical factors in Korean women. To reveal the relationship between RIF and FSHR , INHA , ESR1 , and BMP15 gene polymorphisms, we assessed the differences between RIF patients and healthy controls by examining the known FSHR (rs6165), INHA (rs11893842, rs35118453), ESR1 (rs9340799, rs2234693), and BMP15 (rs17003221, rs3810682) gene polymorphisms. 2. Materials and Methods 2.1. Study Population Blood samples were obtained from women with RIF treated at the Department of Obstetrics and Gynecology and the Fertility Center of CHA Bundang Medical Center in Seongnam, South Korea, between March 2010 and December 2022. In total, we obtained blood samples from 133 patients with RIF and 317 control participants. All patients and controls were Korean. The institutional review board of CHA Bundang Medical Center approved the study, and all patients provided written informed consent (reference no. CHAMC2009-12-120). All embryos were examined by an embryologist prior to transfer, and embryos that showed good quality were transferred. Implantation failure refers to cases where the level of human chorionic gonadotropin measured on the 14th day of embryo transfer is <5 U/mL [ 48 ]. In the study group, subjects diagnosed with implantation failure due to anatomical, chromosomal, hormonal, infectious, autoimmune, or thrombotic causes were excluded. Uterine anatomical abnormalities in RIF patients were confirmed by hysterosalpingography, hysteroscopy, uterine sonography, computed tomography, or magnetic resonance imaging. A karyotype analysis was conducted to confirm chromosomal abnormalities, and the karyotype analysis followed the standard protocol. Hormonal causes of RIF such as hyperproactinemia, lutein insufficiency, and thyroid disease were identified by measuring prolactin, thyroid stimulating hormone (TSH), free T4, FSH, luteinizing hormone (LH), and progesterone levels in peripheral blood. Lupus anticoagulants and anti-cardiolipin antibodies were tested to confirm the autoimmune disease lupus and antiphospholipid syndrome, respectively. A deficiency of protein C and protein S and the presence of anti-β2 glycoprotein antibodies were diagnosed as thrombosis. As a control group, women with regular menstrual cycles, pregnancy history of at least one naturally conceived pregnancy, no history of pregnancy loss, and karyotype of 46, XX were recruited from CHA Bundang Medical Center. 2.2. Estimation of Homocysteine, Folate, Total Cholesterol, Uric Acid, Blood Urea Nitrogen, Creatinine, and Blood Coagulation Status Blood samples were collected from RIF subjects after 12 h of fasting. We performed a fluorescence polarization immunoassay using the Abbott IMx analyzer (Abbott Laboratories, Abbott Park, IL, USA) to measure the homocysteine level. Folate was measured via a competitive immunoassay using the ACS 180Plus automated chemiluminescence system (Bayer Diagnostics, Tarrytown, NY, USA). Total cholesterol, uric acid, blood urea nitrogen, and creatinine were measured using commercially available enzymatic colorimetric assays (Roche Diagnostics, GmbH, Mannheim, Germany). The platelet, white blood cell, and hemoglobin levels were obtained using the Sysmex XE 2100 automated hematology system (Sysmex Corporation, Kobe, Japan). The prothrombin time (PT) and activated partial thromboplastin time (aPTT) were measured with an ACL TOP automated photo-optical coagulometer (LSI Medience, Tokyo, Japan). 2.3. Flow Cytometry Analysis of Immune Cell Proportion Immune cell measurement was performed by flow cytometry using CellQuest software (BD FACS Calibur; BD Biosciences, Seoul, Republic of Korea). Fluorescently-labeled [fluorescein isothiocyanate, phycoerythrin (PE), peridinin chlorophyll protein, and allophycocyanin] monoclonal antibodies specific to CD3, CD4, CD8, CD19, CD16, and CD56 were purchased from BD Biosciences. Anti-NKG2A-PE antibodies were obtained from Beckman Coulter (Fullerton, CA, USA). To determine cell surface antigen expression, peripheral blood mononuclear cells (2.5 × 105) were stained for 30 min at 4 °C in the dark, washed twice with 2% phosphate-buffered saline containing 1% bovine serum albumin and 0.01% sodium azide (FACS wash buffer), and then fixed with 1% formaldehyde solution (Sigma-Aldrich, St. Louis, MO, USA) prior to sorting, as previously described [ 49 ]. 2.4. Hormone Assays On the second to third days of the women’s menstrual cycle, blood samples were collected and measured using serum samples. Following the manufacturer’s instructions, the estradiol (E2) and TSH levels were measured using radioimmunoassays (Beckman Coulter), and the FSH and LH levels were measured using enzyme immunoassays (Siemens, Munich, Germany). 2.5. SNP Selection and Genetic Analysis We selected FSHR , INHA , ESR1 , and BMP15 , which are hormone-related genes associated with pregnancy. To select polymorphisms of the FSHR , INHA , ESR1 , and BMP15 genes, studies on the association between pregnancy-related diseases (recurrent pregnancy loss, recurrent implantation failure, preeclampsia, premature ovarian failure, and poor ovarian response) and polymorphisms were investigated [ 46 , 50 , 51 , 52 , 53 , 54 , 55 ]. Finally, a total of seven polymorphisms in FSHR (rs6165), INHA (rs11893842 and rs35118453), ESR1 (rs9340799 and rs2234693), and BMP15 (rs17003221 and rs3810682) were selected and studied. Genomic DNA was extracted from anticoagulated peripheral blood using a G-DEX blood extraction kit (Intron, Seongnam, Republic of Korea). All genetic polymorphisms were identified by a real-time polymerase chain reaction (PCR) using the TaqMan SNP Genotyping Assay Kit (Applied Biosystems, Foster City, CA, USA). To validate the real-time analysis, DNA sequencing was performed on approximately 10~15% of the samples by random selection using an ABI 3730XL DNA Analyzer (Applied Biosystems). The concordance of the quality control samples was 100%. 2.6. Statistical Analysis Regarding the clinical characteristics of participants, the differences between the categorical variables were analyzed using a chi-square test and continuous variables using an independent sample t -test. The data were presented as mean and standard deviations for continuous variables, and frequency and percentage for categorical variables. The statistical normality of continuous variables was confirmed using the Kolmogorov–Smirnov test. For continuous variables showing a non-normal distribution ( p < 0.05 in the Kolmogorov–Smirnov test), group difference analyses were performed using the Mann–Whitney test. Allele frequencies were determined to confirm deviations from the Hardy–Weinberg equilibrium. To select the best inheritance model for a specific polymorphism, Akaike’s information criterion was calculated. The associations between FSHR , INHA , ESR1 , and BMP15 polymorphisms and RIF incidence were calculated using adjusted odds ratios (AORs) and 95% confidence intervals (95% CIs) obtained from a multivariate logistic regression analysis adjusted for age. False discovery rate correction was used to adjust multiple comparison tests and provide a measure of the expected proportion of false positives among the data. The open-source MDR software package (v.2.0, www.epistasis.org accessed on 1 April 2022) was used to perform the genetic interaction analysis. Using this MDR analysis, all possible genotype combinations for gene–gene interactions were identified and analyzed. The associations between each FSHR , INHA , ESR1 , and BMP15 gene polymorphism and each clinical value (platelets, PT, aPTT, homocysteine, folate, natural killer cells, uric acid, and total cholesterol) for RIF patients were assessed using ANOVA and Kruskal–Wallis tests. For the overall statistical analysis, the level of statistical significance was set as p < 0.05. 3. Results Table 1 presents the clinical variables of the 133 RIF patients and 317 control subjects. No significant differences in age distribution were observed between the RIF and control groups, indicating that our age frequency matching was satisfactory. The blood urea nitrogen, creatinine, PT, TSH, E2, LH, BMI, total cholesterol, and white blood cell levels were different between the patient and the control group. The blood urea nitrogen ( p < 0.0001), creatinine ( p < 0.0001), PT ( p < 0.0001), TSH ( p = 0.0001), E2 ( p = 0.0002), and LH ( p < 0.0001) levels increased significantly in the patient group, and on the contrary, BMI ( p = 0.047), total cholesterol ( p < 0.0001), and white blood cell ( p = 0.005) values increased significantly in the control group. Table 1. Clinical profiles between RIF patient and control subjects. We investigated the distribution of FSHR (rs6165), INHA (rs11893842, rs35118453), ESR1 (rs9340799, rs2234693), and BMP15 (rs17003221, rs3810682) polymorphisms in RIF patients and the control group ( Table 2 ). The AOR with respect to age was calculated from the logistic regression analysis. The frequency of each genotype in the control group was consistent with the Hardy–Weinberg equilibrium. Table 2. Comparison of genotype frequencies of FSHR , INHA , ESR1 , and BMP15 polymorphisms between the RIF and control subjects. We investigated the genotype frequencies according to the number of implantation failures among RIF patients ( Table 2 ). For FSHR rs6165A>G, GG homozygous genotypes and the recessive model were found to exert a protective effect against RIF [AA vs. GG: AOR, 0.463; 95% CI, 0.218–0.980; p = 0.044 and AA+AG vs. GG (recessive model): AOR, 0.430; 95% CI, 0.210–0.877; p = 0.020]. The other gene polymorphisms ( INHA rs11893842 and rs35118453, ESR1 rs9340799 and rs2234693, and BMP15 rs17003221 and rs3810682) did not show significant differences between the control and patient groups. We also identified the genotype frequencies among patients according to the number of RIFs ( Table 3 ). FSHR rs6165A>G GG genotypes and the recessive model showed protective effects in patients with more than three implantation failures (AA vs. GG: AOR, 0.412; 95% CI, 0.183–0.931; p = 0.033 and recessive model: AOR, 0.392; 95% CI, 0.156–0.982; p = 0.046), and INHA rs3511845C>T CT genotypes and CT+TT (dominant model) showed risk effects in patients with more than four implantation failures (CC vs. CT: AOR, 3.793; 95% CI,1.171–12.285; p = 0.026 and dominant model: AOR, 3.745; 95% CI, 1.176–11.921; p = 0.025). Table 3. Comparison of genotype frequencies of FSHR , INHA , ESR1 , and BMP15 polymorphisms between the RIF and control subjects. We performed a genotype combination analysis of RIF patients and control subjects ( Table 4 ). In the genotype combination type of FSHR rs6165 A>G and ESR1 rs9340799 A>G, the GG/AA (OR, 0.250; 95% CI, 0.072–0.874; p = 0.030) and GG/AA+AG (OR, 0.373; 95% CI, 0.171–0.816; p = 0.014) combinations were significantly associated with RIF risk. Additionally, in the FSHR rs6165A>G/ INHA rs35118453C>T combination genotype, GG/CC+CT (OR, 0.399; 95% CI, 0.190–0.841; p = 0.016) showed a significantly decreased OR. In contrast, in the genotype combination analysis of INHA rs35118453 and C>T/ ESR1 rs2234693 T>C, TT/TC+CC (OR, 7.001; 95% CI, 1.298–37.776; p = 0.024) was associated with an increased RIF risk. Additionally, in the INHA rs11893842 A>G and ESR1 rs9340799 A>G genotype combination, AA+AG/GG (OR, 3.065; 95% CI, 1.081–8.690; p = 0.035) was associated with an increased RIF risk. In the genotype combination analysis of ESR1 rs9340799 A>G/ ESR1 rs2234693 T>C, GG/TT+TC (OR, 12.930; 95% CI, 1.492–112.052; p = 0.020) was associated with an increased RIF risk. Table 4. Combined genotype analysis for the FSHR , INHA , ESR1 , and BMP15 polymorphisms in RIF patients and controls. We performed an analysis of the differences in clinical factors according to the genotype of the FSHR (rs6165), INHA (rs11893842, rs35118453), ESR1 (rs9340799, rs2234693), and BMP15 (rs17003221, rs3810682) polymorphisms using a one-way analysis of variance (ANOVA). We found that increased LH levels were associated with the INHA rs35118453 polymorphism among all subjects ( Table 5 , p < 0.05). In RIF patients, increased E2 levels were associated with INHA rs11893842, and decreased CD3 (pan T cell) levels were associated with ESR1 rs2234693 ( Table S1 , p < 0.05). TSH levels in the control group showed an increasing trend according to the FSHR rs6165 genotype ( Table S2 , p < 0.05). Table 5. Clinical variables in RIF patients and controls stratified by FSHR , INHA , ESR1 , and BMP15 polymorphisms status by ANOVA and Kruskal–Wallis test. We then analyzed the synergistic effects of the FSHR (rs6165), INHA (rs11893842, rs35118453), ESR1 (rs9340799, rs2234693), and BMP15 (rs17003221, rs3810682) polymorphisms and clinical factors (i.e., folate, homocysteine, E2, FSH, and LH) on RIF risk ( Table S3 ). In Table S3 , we set the reference values for homocysteine, E2, FSH, and LH levels by determining the threshold for the highest 25% of levels (homocysteine 7.94 µmol/L, E2 37.6 pg/mL, FSH 9.6 U/L, and LH 5.12 U/L), and the folate level was determined by the lowest 25% of levels (8.12 ng/mL) in RIF patients and controls. FSHR rs6165 and high E2 and FSH levels had a synergistic effect on an increased susceptibility to RIF (AA+AG with high E2 levels, AOR = 11.411, p < 0.0001; AA+AG with high FSH levels, AOR = 3.009, p = 0.025, Figure 1 ). Additionally, INHA rs11893842, high homocysteine levels, and low folate levels had a synergistic effect on an increasing susceptibility to RIF (AG+GG with high homocysteine levels, AOR = 6.180, p = 0.004; AG+GG with low folate levels, AOR = 8.090, p = 0.003, Figure 1 ). Moreover, combining the ESR1 rs9340799 AG+GG type with E2 (AOR = 6.538, p = 0.0001), FSH (AOR = 5.511, p = 0.003), and LH (AOR = 12.305, p = 0.002) resulted in an increased RIF risk ( Figure 2 ). Furthermore, the AORs of the ESR1 rs2234693 TC+CC group when combined with each female hormone factor including ≥37.6 pg/mL E2, ≥9.6 U/L FSH, and ≥5.12 U/L LH were 6.997 ( p < 0.0001), 3.371 ( p = 0.003), and 6.161 ( p < 0.0001), respectively. Figure 1. Synergistic effect of FSHR rs6165 and INHA rs11893842 polymorphisms with clinical parameters. ( A – D ) panels show AOR of FSHR rs6165 and INHA rs11893842 with clinical parameters including E2 ( A ), FSH ( B ), homocysteine ( C ), and folate ( D ). Figure 2. Synergistic effect of ESR1 rs9340799 and rs2234693 polymorphisms with clinical parameters. ( A – F ) panels show AOR of ESR1 rs9340799 and rs2234693 with clinical parameters including E2 ( A , D ), FSH ( B , E ), and LH ( C , F ). 4. Discussion The formation and quality of gametocytes and embryo development are important factors for successful pregnancy progression, and many infertile couples have experienced recurrent failure due to the poor quality or quantity of embryos before transplantation. Currently, morphological indicators are used for embryo selection, and there is no other predictive indicator that can evaluate the quality of embryo cells. Recently, many studies have elucidated molecular and genetic factors to assess embryo quality and select developmentally competent embryos. Previous studies have reported several genes associated with the pathogenesis of oocyte maturation arrest ( TUBB8 , PATL2 ) and fertilization failures ( TLE6 , WEE2 ). [ 56 ]. In this study, seven polymorphisms were selected from four hormone-related genes ( FSHR rs6165, INHA rs11893842 and rs35118453, ESR1 rs9340799 and rs2234693, BMP15 rs1700321, and rs3810682), and the association between RIF occurrence and polymorphisms was analyzed. We examined the association between RIF occurrence and SNPs in the FSHR , INHA , ESR1 , and BMP15 genes. Our results showed that the genotype frequency of the FSHR rs6165 SNP was significantly different in the control and RIF patient groups. A genotype combination analysis of the seven genetic markers revealed that the GG/AA ( p = 0.030) and GG/AA+AG ( p = 0.014) combinations significantly reduced the RIF risk in the FSHR rs6165 and ESR1 rs9340799 combination type. Additionally, in the genotype combination analysis of FSHR rs6165A>G/ INHA rs35118453C>T, GG/CC+CT ( p = 0.016) showed a significantly decreased risk of RIF. In contrast, the INHA rs35118453 TT and ESR1 rs2234693 TC+CC combination were associated with increased RIF risk ( p = 0.024). Moreover, the INHA rs11893842 A>G/ ESR1 rs9340799 A>G, AA+AG/GG combination ( p = 0.035) was associated with an increased RIF risk. In the genotype combination of ESR1 rs9340799 A>G/ ESR1 rs2234693 T>C, GG/TT+TC ( p = 0.020) was associated with an increased RIF risk. Furthermore, our gene–environment combinatorial analysis data revealed statistically significant relationships between the FSHR rs6165, INHA rs11893842, ESR1 rs9340799, and rs2234693 genotypes and clinical factors (folate, homocysteine, E2, FSH, and LH) in Korean RIF patients. When combined with clinical parameters, the RIF risk increased by 2- to 11-fold. Previously, FSRH , INHA , ESR1 , and BMP15 gene polymorphisms have been studied with regard to the conditions of the female reproductive system, including polycystic ovary syndrome, poor ovarian response, POI, breast cancer, and endometriosis, as well as male infertility [ 50 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 ]. The FSHR rs6165 polymorphism is located in exon 10 of the FSHR gene and is in the transmembrane region of the FSHR protein. The rs6165 polymorphism produces a change of A to G in position 919 and changes codon 307 from threonine (ACT) to alanine (GCT) [ 57 ]. According to Ganesh et al. [ 50 ], the FSHR rs6165 polymorphism was associated with unsuccessful IVF outcomes, and a higher frequency of the heterozygous AG genotype was observed in the infertile group than in the control group. For FSHR rs6165 AA carriers, the number of oocytes retrieved was significantly higher and ovarian stimulation was significantly shorter than those in GG and AG carriers [ 58 ]. Additionally, Rod et al. [ 59 ] found that FSHR rs6165 was associated with controlled ovarian stimulation and was 3-fold higher in poor responders than in good responders. Moreover, one study showed that the rs6165 AG genotype was associated with an increased risk of male infertility [ 60 ]. INHA rs11893842 and rs35119453 are located in the promoter region of the INHA gene and have been studied with regard to POI, male infertility (sperm parameters), and adrenocortical cancer [ 44 , 61 , 62 ]. Neither variant was associated with POI. Rafaqat et al. [ 48 ] found that the GG genotype frequency was increased in male infertility patients and showed a significant association with male infertility in the Pakistani population. Furthermore, rs11893842 minor alleles showed a low frequency in adrenocortical cancer patients and the rs11893842 AA genotype was associated with decreased INHA mRNA levels [ 62 ]. ESR1 rs9340799 and rs2234693 are located in intron 1, 351 and 1397 bp upstream of exon 2, respectively. The two polymorphisms are associated with cancer in females, including breast cancer and endometrial cancer [ 63 , 64 ], and are associated with POI [ 65 ]. The ESR1 rs9340799 GA and rs2234693 TC genotypes were associated with a decreased risk of POI, and the ESR1 rs9340799 AA and rs2234693 TT genotypes were associated with an increased risk of POI [ 65 ]. Additionally, the ESR1 rs9340799 GG genotype was associated with a 4-fold increased risk of endometriosis and a 3-fold increased risk of IVF failure in infertile patients [ 66 ]. Furthermore, the FSHR rs6165 GG, rs6165 AA, ESR1 rs9340799 GA, and rs2234693 TC gene combination enhanced the protective effect of FSHR gene variants and was associated with a reduced risk of fibrocystic mastopathy in infertile women [ 67 ]. Several clinical factors are involved in embryo implantation and pregnancy maintenance, including folate, homocysteine, and female hormones (E2, FSH, LH). Gaskins et al. [ 68 ] found that, among women receiving assisted reproductive technology, women with high serum folate levels (>26.3 ng/mL) had a 1.62-fold higher live birth rate than women with low folate levels (<16.6 ng/mL). Additionally, Ocal et al. [ 69 ] found that high homocysteine levels in follicular fluid resulted in reduced cell division and increased fragmentation in embryo cultures, which was associated with decreased oocyte and embryo quality. Another study showed that exposure to high E2 concentrations is detrimental to blastocyst implantation and early post-implantation development. Moreover, during clinically-assisted reproductive technology, high serum E2 levels not only affected the endometrium but also directly affected the blastocyst during implantation [ 70 ]. Our study revealed that patients with several clinical factors (low folate levels and high homocysteine, E2, FSH, and LH levels) and FSHR rs6165, INHA rs11893842, ESR1 rs9340799, and rs2234693 polymorphisms had an approximately 3- to 12-fold increased risk of RIF. This study has several limitations. First, how FSHR , INHA , ESR1 , and BMP15 polymorphisms influence RIF development is still unclear. The effect of these SNPs should also be confirmed through in vitro and in vivo studies. Second, additional environmental risk factors to RIFs need to be evaluated. Finally, the size of the RIF patient and control group was small, and this study group included only Koreans. To determine whether the studied genetic polymorphism can be used as a predictor of RIF, our results should be validated using larger sample sizes and other ethnic groups. 5. Conclusions We investigated the association between the FSHR rs6165, INHA rs11893842 and rs35118453, ESR1 rs9340799 and rs2234693, and BMP15 rs17003221 and rs3810682 polymorphisms and the risk of RIF in Korean women. We found the frequency of the GG genotype of FSHR rs6165 was lower among RIF patients than among controls, suggesting this genotype may be associated with a reduced risk of RIF. Additionally, the interaction of the FSHR rs6165, INHA rs11893842, and ESR1 rs9340799 and rs2234693 polymorphisms with some clinical factors may increase the risk of RIF. Supplementary Materials The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/biomedicines11051374/s1 , Table S1: Clinical variables in RIF patients stratified by FSHR , INHA , ESR1 , and BMP15 polymorphisms status by ANOVA and Kruskal–Wallis tests; Table S2: Clinical variables in controls stratified by FSHR , INHA , ESR1 , and BMP15 polymorphisms status by ANOVA and Kruskal–Wallis tests; Table S3: Synergic effect of FSHR , INHA , ESR1 , and BMP15 polymorphisms with clinical risk factor. Author Contributions Conceptualization, E.-J.K. and J.-E.S.; methodology, J.-Y.L.; validation, C.-S.R.; investigation, E.-J.K.; resources, Y.-R.K. and E.-H.A.; data curation, E.-J.K., J.-E.S. and J.-Y.H.; writing—original draft preparation, E.-J.K.; writing—review and editing, E.-J.K. and J.-E.S.; supervision, J.-H.K. and N.-K.K.; project administration, J.-H.K. and N.-K.K.; funding acquisition, Y.-R.K., E.-H.A., J.-H.K. and N.-K.K. All authors have read and agreed to the published version of the manuscript. Funding This research was supported by the National Research Foundation of Korea (NRF) grants funded by the Korean Government (MSIT; grant number 2022R1F1A1064169, 2022R1F1A1074986, and 2020R1F1A1074525). This research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health and Welfare, Republic of Korea (grant number: HR22C1605). Informed Consent Statement The institutional review board of CHA Bundang Medical Center approved the study, and all patients provided written informed consent (reference no. CHAMC2009-12-120). Data Availability Statement The data presented in this study are available on request from the corresponding author. Conflicts of Interest The authors declare no conflict of interest. References Achache, H.; Revel, A. Endometrial receptivity markers, the journey to successful embryo implantation. Hum. Reprod. Update 2006 , 12 , 731–746. [ Google Scholar ] [ CrossRef ] [ PubMed ] Simon, A.; Laufer, N. Assessment and treatment of repeated implantation failure (RIF). J. Assist. Reprod. Genet. 2012 , 29 , 1227–1239. [ Google Scholar ] [ CrossRef ] Zhou, W.; Dimitriadis, E. Secreted microRNA to predict embryo implantation outcome: From research to clinical diagnostic application. Front. Cell. Dev. Biol. 2020 , 8 , 586510. [ Google Scholar ] [ CrossRef ] Busnelli, A.; Reschini, M.; Cardellicchio, L.; Vegetti, W.; Somigliana, E.; Vercellini, P. How common is real repeated implantation failure? An indirect estimate of the prevalence. Reprod. Biomed. Online 2020 , 40 , 91–97. [ Google Scholar ] [ CrossRef ] Bashiri, A.; Halper, K.I.; Orvieto, R. Recurrent implantation failure-update overview on etiology, diagnosis, treatment and future directions. Reprod. Biol. Endocrinol. 2018 , 16 , 121. [ Google Scholar ] [ CrossRef ] Tan, B.K.; Vandekerckhove, P.; Kennedy, R.; Keay, S.D. Investigation and current management of recurrent IVF treatment failure in the UK. BJOG 2005 , 112 , 773–780. [ Google Scholar ] [ CrossRef ] [ PubMed ] Zhai, J.; Xiao, Z.; Wang, Y.; Wang, H. Human embryonic development: From peri-implantation to gastrulation. Trends. Cell. Biol. 2022 , 32 , 18–29. [ Google Scholar ] [ CrossRef ] [ PubMed ] Giorgetti, C.; Terriou, P.; Auquier, P.; Hans, E.; Spach, J.L.; Salzmann, J.; Roulier, R. Embryo score to predict implantation after in-vitro fertilization: Based on 957 single embryo transfers. Hum. Reprod. 1995 , 10 , 2427–2431. [ Google Scholar ] [ CrossRef ] [ PubMed ] Gardner, D.K.; Lane, M.; Stevens, J.; Schlenker, T.; Schoolcraft, W.B. Blastocyst score affects implantation and pregnancy outcome: Towards a single blastocyst transfer. Fertil. Steril. 2000 , 73 , 1155–1158. [ Google Scholar ] [ CrossRef ] Gardner, D.K.; Balaban, B. Assessment of human embryo development using morphological criteria in an era of time-lapse, algorithms and ‘OMICS’: Is looking good still important? Mol. Hum. Reprod. 2016 , 22 , 704–718. [ Google Scholar ] [ CrossRef ] Hartshorne, G.M. Factors controlling embryo viability. Hum. Fertil. 2001 , 4 , 225–234. [ Google Scholar ] [ CrossRef ] Assou, S.; Boumela, I.; Haouzi, D.; Anahory, T.; Dechaud, H.; De Vos, J.; Hamamah, S. Dynamic changes in gene expression during human early embryo development: From fundamental aspects to clinical applications. Hum. Reprod. Update 2011 , 17 , 272–290. [ Google Scholar ] [ CrossRef ] Paonessa, M.; Borini, A.; Coticchio, G. Genetic causes of preimplantation embryo developmental failure. Mol. Reprod. Dev. 2021 , 88 , 338–348. [ Google Scholar ] [ CrossRef ] [ PubMed ] Yi, H.; Xue, L.; Guo, M.X.; Ma, J.; Zeng, Y.; Wang, W.; Cai, J.Y.; Hu, H.M.; Shu, H.B.; Shi, Y.B.; et al. Gene expression atlas for human embryogenesis. FASEB J. 2010 , 24 , 3341–3350. [ Google Scholar ] [ CrossRef ] Wale, P.L.; Gardner, D.K. The effects of chemical and physical factors on mammalian embryo culture and their importance for the practice of assisted human reproduction. Hum. Reprod. Update 2016 , 22 , 2–22. [ Google Scholar ] [ CrossRef ] [ PubMed ] Zhang, X.; Chen, Y.; Wang, X.; Zhang, Z.; Wang, J.; Shen, Y.; Hu, Y.; Wu, X. NINJ1 triggers extravillous trophoblast cell dysfunction through blocking the STAT3 signaling pathway. Genes Genom. 2022 , 44 , 1385–1397. [ Google Scholar ] [ CrossRef ] Dey, S.K.; Lim, H.; Das, S.K.; Reese, J.; Paria, B.C.; Daikoku, T.; Wang, H. Molecular cues to implantation. Endocr. Rev. 2004 , 25 , 341–373. [ Google Scholar ] [ CrossRef ] Robinson, D.P.; Klein, S.L. Pregnancy and pregnancy-associated hormones alter immune responses and disease pathogenesis. Horm. Behav. 2012 , 62 , 263–271. [ Google Scholar ] [ CrossRef ] Large, M.J.; DeMayo, F.J. The regulation of embryo implantation and endometrial decidualization by progesterone receptor signaling. Mol. Cell. Endocrinol. 2012 , 358 , 155–165. [ Google Scholar ] [ CrossRef ] Klonos, E.; Katopodis, P.; Karteris, E.; Papanikolaou, E.; Tarlatzis, B.; Pados, G. Endometrial changes in estrogen and progesterone receptor expression during implantation in an oocyte donation program. Exp. Ther. Med. 2020 , 20 , 178. [ Google Scholar ] [ CrossRef ] [ PubMed ] Park, S.R.; Kim, S.R.; Kim, S.K.; Park, J.R.; Hong, I.S. A novel role of follicle-stimulating hormone (FSH) in various regeneration-related functions of endometrial stem cells. Exp. Mol. Med. 2022 , 54 , 1524–1535. [ Google Scholar ] [ CrossRef ] Stilley, J.A.W.; Segaloff, D.L. FSH actions and pregnancy: Looking beyond ovarian FSH receptors. Endocrinology 2018 , 159 , 4033–4042. [ Google Scholar ] [ CrossRef ] [ PubMed ] Stilley, J.A.W.; Christensen, D.E.; Dahlem, K.B.; Guan, R.; Santillan, D.A.; England, S.K.; Al-Hendy, A.; Kirby, P.A.; Segaloff, D.L. FSH receptor (FSHR) expression in human extragonadal reproductive tissues and the developing placenta, and the impact of its deletion on pregnancy in mice. Biol. Reprod. 2014 , 91 , 74. [ Google Scholar ] [ CrossRef ] Stilley, J.A.W.; Segaloff, D.L. Deletion of fetoplacental Fshr inhibits fetal vessel angiogenesis in the mouse placenta. Mol. Cell. Endocrinol. 2018 , 476 , 79–83. [ Google Scholar ] [ CrossRef ] Stelmaszewska, J.; Chrusciel, M.; Doroszko, M.; Akerfelt, M.; Ponikwicka-Tyszko, D.; Nees, M.; Frentsch, M.; Li, X.; Kero, J.; Huhtaniemi, I.; et al. Revisiting the expression and function of follicle-stimulation hormone receptor in human umbilical vein endothelial cells. Sci. Rep. 2016 , 6 , 37095. [ Google Scholar ] [ CrossRef ] Stilley, J.A.; Guan, R.; Santillan, D.A.; Mitchell, B.F.; Lamping, K.G.; Segaloff, D.L. Differential regulation of human and mouse myometrial contractile activity by FSH as a function of FSH receptor density. Biol. Reprod. 2016 , 95 , 36. [ Google Scholar ] [ CrossRef ] Lu, C.; Yang, W.; Chen, M.; Liu, T.; Yang, J.; Tan, P.; Li, L.; Hu, X.; Fan, C.; Hu, Z.; et al. Inhibin A inhibits follicle-stimulating hormone (FSH) action by suppressing its receptor expression in cultured rat granulosa cells. Mol. Cell. Endocrinol. 2009 , 298 , 48–56. [ Google Scholar ] [ CrossRef ] Leung, P.H.; Salamonsen, L.A.; Findlay, J.K. Immunolocalization of inhibin and activin subunits in human endometrium across the menstrual cycle. Hum. Reprod. 1998 , 13 , 3469–3477. [ Google Scholar ] [ CrossRef ] [ PubMed ] Lawrenz, B.; Depret Bixio, L.; Coughlan, C.; Andersen, C.Y.; Melado, L.; Kalra, B.; Savjani, G.; Fatemi, H.M.; Kumar, A. Inhibin A—A promising predictive parameter for determination of final oocyte maturation in ovarian stimulation for IVF/ICSI. Front. Endocrinol. 2020 , 11 , 307. [ Google Scholar ] [ CrossRef ] Ocal, P.; Aydin, S.; Cepni, I.; Idil, S.; Idil, M.; Uzun, H.; Benian, A. Follicular fluid concentrations of vascular endothelial growth factor, inhibin A and inhibin B in IVF cycles: Are they markers for ovarian response and pregnancy outcome? Eur. J. Obstet. Gynecol. Reprod. Biol. 2004 , 115 , 194–199. [ Google Scholar ] [ CrossRef ] [ PubMed ] Goldenberg, R.L.; Vaitukaitis, J.L.; Ross, G.T. Estrogen and follicle stimulation hormone interactions on follicle growth in rats. Endocrinology 1972 , 90 , 1492–1498. [ Google Scholar ] [ CrossRef ] [ PubMed ] Pawar, S.; Laws, M.J.; Bagchi, I.C.; Bagchi, M.K. Uterine epithelial estrogen receptor-alpha controls decidualization via a paracrine mechanism. Mol. Endocrinol. 2015 , 29 , 1362–1374. [ Google Scholar ] [ CrossRef ] Li, Y.; Li, R.Q.; Ou, S.B.; Zhang, N.F.; Ren, L.; Wei, L.N.; Zhang, Q.X.; Yang, D.Z. Increased GDF9 and BMP15 mRNA levels in cumulus granulosa cells correlate with oocyte maturation, fertilization, and embryo quality in humans. Reprod. Biol. Endocrinol. 2014 , 12 , 81. [ Google Scholar ] [ CrossRef ] Wu, Y.T.; Tang, L.; Cai, J.; Lu, X.E.; Xu, J.; Zhu, X.M.; Luo, Q.; Huang, H.F. High bone morphogenetic protein-15 level in follicular fluid is associated with high quality oocyte and subsequent embryonic development. Hum. Reprod. 2007 , 22 , 1526–1531. [ Google Scholar ] [ CrossRef ] Barbitoff, Y.A.; Tsarev, A.A.; Vashukova, E.S.; Maksiutenko, E.M.; Kovalenko, L.V.; Belotserkovtseva, L.D.; Glotov, A.S. A data-driven review of the genetic factors of pregnancy complications. Int. J. Mol. Sci. 2020 , 21 , 3384. [ Google Scholar ] [ CrossRef ] [ PubMed ] Ryu, C.S.; Sakong, J.H.; Ahn, E.H.; Kim, J.O.; Ko, D.; Kim, J.H.; Lee, W.S.; Kim, N.K. Association study of the three functional polymorphisms (TAS2R46G>A, OR4C16G>A, and OR4X1A>T) with recurrent pregnancy loss. Genes. Genom. 2019 , 41 , 61–70. [ Google Scholar ] [ CrossRef ] Kwon, B.N.; Lee, N.R.; Kim, H.J.; Kang, Y.D.; Kim, J.S.; Park, J.W.; Jin, H.J. Folate metabolizing gene polymorphisms and genetic vulnerability to preterm birth in Korean women. Genes Genom. 2021 , 43 , 937–945. [ Google Scholar ] [ CrossRef ] Lee, J.Y.; Ahn, E.H.; Park, H.W.; Kim, J.H.; Kim, Y.R.; Lee, W.S.; Kim, N.K. Association between HOX transcript antisense RNA single-nucleotide variants and recurrent implantation failure. Int. J. Mol. Sci. 2021 , 22 , 3021. [ Google Scholar ] [ CrossRef ] [ PubMed ] Salazar Garcia, M.D.; Sung, N.; Mullenix, T.M.; Dambaeva, S.; Beaman, K.; Gilman-Sachs, A.; Kwak-Kim, J. Plasminogen activator inhibitor-1 4G/5G polymorphism is associated with reproductive failure: Metabolic, hormonal, and immune profiles. Am. J. Reprod. Immunol. 2016 , 76 , 70–81. [ Google Scholar ] [ CrossRef ] [ PubMed ] Zhang, Z.; Miteva, M.A.; Wang, L.; Alexov, E. Analyzing effects of naturally occurring missense mutations. Comput. Math Methods Med. 2012 , 2012 , 805827. [ Google Scholar ] [ CrossRef ] Guo, Y.; Jamison, D.C. The distribution of SNPs in human gene regulatory regions. BMC Genom. 2005 , 6 , 140. [ Google Scholar ] [ CrossRef ] [ PubMed ] Borgbo, T.; Jeppesen, J.V.; Lindgren, I.; Lundberg Giwercman, Y.; Hansen, L.L.; Yding Andersen, C. Effect of the FSH receptor single nucleotide polymorphisms (FSHR 307/680) on the follicular fluid hormone profile and the granulosa cell gene expression in human small antral follicles. Mol. Hum. Reprod. 2015 , 21 , 255–261. [ Google Scholar ] [ CrossRef ] [ PubMed ] Eldafira, E.; Prasasty, V.D.; Abinawanto, A.; Syahfirdi, L.; Pujianto, D.A. Polymorphisms of estrogen receptor-alpha and estrogen receptor-beta genes and its expression in endometriosis. Turk. J. Pharm. Sci. 2021 , 18 , 91–95. [ Google Scholar ] [ CrossRef ] [ PubMed ] Pu, D.; Xing, Y.; Gao, Y.; Gu, L.; Wu, J. Gene variation and premature ovarian failure: A meta-analysis. Eur. J. Obstet. Gynecol. Reprod. Biol. 2014 , 182 , 226–237. [ Google Scholar ] [ CrossRef ] Meireles, A.J.C.; Bilibio, J.P.; Lorenzzoni, P.L.; Conto, E.; Nascimento, F.C.D.; Cunha-Filho, J.S.D. Association of FSHR, LH, LHR, BMP15, GDF9, AMH, and AMHR polymorphisms with poor ovarian response in patients undergoing in vitro fertilization. JBRA Assist. Reprod. 2021 , 25 , 439–446. [ Google Scholar ] [ CrossRef ] Jalilvand, A.; Yari, K.; Heydarpour, F. Role of polymorphisms on the recurrent pregnancy loss: A systematic review, meta-analysis and bioinformatic analysis. Gene 2022 , 844 , 146804. [ Google Scholar ] [ CrossRef ] [ PubMed ] Ke, Y.; Bin, L.; Lin, L.; MingRong, X. ESR1 polymorphisms and risk of preeclampsia. J. Matern. Fetal. Neonatal. Med. 2022 , 35 , 402–409. [ Google Scholar ] [ CrossRef ] Choi, Y.; Kim, J.O.; Shim, S.H.; Lee, Y.; Kim, J.H.; Jeon, Y.J.; Ko, J.J.; Lee, W.S.; Kim, N.K. Genetic variation of methylenetetrahydrofolate reductase (MTHFR) and thymidylate synthase (TS) genes is associated with idiopathic recurrent implantation failure. PLoS ONE 2016 , 11 , e01608842016. [ Google Scholar ] [ CrossRef ] Kim, E.S.; Kim, J.O.; An, H.J.; Sakong, J.H.; Lee, H.A.; Kim, J.H.; Ahn, E.H.; Kim, Y.R.; Lee, W.S.; Kim, N.K. MTHFR 3′-untranslated region polymorphisms contribute to recurrent pregnancy loss risk and alterations in peripheral natural killer cell proportions. Clin. Exp. Reprod. Med. 2017 , 44 , 152–158. [ Google Scholar ] [ CrossRef ] Ganesh, V.; Venkatesan, V.; Koshy, T.; Reddy, S.N.; Muthumuthiah, S.; Paul, S.F.D. Association of estrogen, progesterone and follicle stimulating hormone receptor polymorphisms with in vitro fertilization outcomes. Syst. Biol. Reprod. Med. 2018 , 64 , 260–265. [ Google Scholar ] [ CrossRef ] Santos, M.; Cordts, E.B.; Peluso, C.; Dornas, M.; Neto, F.H.V.; Bianco, B.; Barbosa, C.P.; Christofolini, D.M. Association of BMP15 and GDF9 variants to premature ovarian insufficiency. J. Assist. Reprod. Genet. 2019 , 36 , 2163–2169. [ Google Scholar ] [ CrossRef ] Rah, H.; Jeon, Y.J.; Ko, J.J.; Kim, J.H.; Kim, Y.R.; Cha, S.H.; Choi, Y.; Lee, W.S.; Kim, N.K. Association of inhibin alpha gene promoter polymorphisms with risk of idiopathic primary ovarian insufficiency in Korean women. Maturitas 2014 , 77 , 163–167. [ Google Scholar ] [ CrossRef ] Cristina Dos Santos Lopes, A.; Perucci, L.O.; Gontijo Evangelista, F.C.; Godoi, L.C.; de Paula Sabino, A.; Gomes, K.B.; Talvani, A.; Dusse, L.M.S.; Alpoim, P.N. Association among ACE, ESR1 polymorphisms and preeclampsia in Brazilian pregnant women. Mol. Cell. Probes. 2019 , 45 , 43–47. [ Google Scholar ] [ CrossRef ] Neves, A.R.; Vuong, N.L.; Blockeel, C.; Garcia, S.; Alviggi, C.; Spits, C.; Ma, P.Q.M.; Ho, M.T.; Tournaye, H.; Polyzos, N.P. The effect of polymorphisms in FSHR gene on late follicular phase progesterone and estradiol serum levels in predicted normoresponders. Hum. Reprod. 2022 , 37 , 2646–2654. [ Google Scholar ] [ CrossRef ] Sindiani, A.M.; Batiha, O.; Al-Zoubi, E.; Khadrawi, S.; Alsoukhni, G.; Alkofahi, A.; Alahmad, N.A.; Shaaban, S.; Alshdaifat, E.; Abu-Halima, M. Association of single-nucleotide polymorphisms in the ESR2 and FSHR genes with poor ovarian response in infertile Jordanian women. Clin. Exp. Reprod. Med. 2021 , 48 , 69–79. [ Google Scholar ] [ CrossRef ] Solovova, O.A.; Chernykh, V.B. Genetics of oocyte maturation defects and early embryo development arrest. Genes 2022 , 13 , 1920. [ Google Scholar ] [ CrossRef ] [ PubMed ] Aittomaki, K.; Lucena, J.L.; Pakarinen, P.; Sistonen, P.; Tapanainen, J.; Gromoll, J.; Kaskikari, R.; Sankila, E.M.; Lehvaslaiho, H.; Engel, A.R.; et al. Mutation in the follicle-stimulating hormone receptor gene causes hereditary hypergonadotropic ovarian failure. Cell 1995 , 82 , 959–968. [ Google Scholar ] [ CrossRef ] Alviggi, C.; Conforti, A.; Santi, D.; Esteves, S.C.; Andersen, C.Y.; Humaidan, P.; Chiodini, P.; De Placido, G.; Simoni, M. Clinical relevance of genetic variants of gonadotrophins and their receptors in controlled ovarian stimulation: A systematic review and meta-analysis. Hum. Reprod. Update 2018 , 24 , 599–614. [ Google Scholar ] [ CrossRef ] [ PubMed ] Rod, A.; Jarzabek, K.; Wolczynski, S.; Benhaim, A.; Reznik, Y.; Denoual-Ziad, C.; Herlicoviez, M.; Mittre, H. ESR1 and FSHR gene polymorphisms influence ovarian response to FSH in poor responder women with normal FSH levels. Endocrinol. Metab. Synd. 2014 , 3 , 1–5. [ Google Scholar ] Wu, Q.; Zhang, J.; Zhu, P.; Jiang, W.; Liu, S.; Ni, M.; Zhang, M.; Li, W.; Zhou, Q.; Cui, Y.; et al. The susceptibility of FSHB -211G > T and FSHR G-29A, 919A > G, 2039A > G polymorphisms to men infertility: An association study and meta-analysis. BMC Med. Genet. 2017 , 18 , 81. [ Google Scholar ] [ CrossRef ] Rafaqat, W.; Kayani, M.R.; Fatima, T.; Shaharyar, S.; Khan, S.; Ashraf, M.; Afzal, U.; Rehman, R. Association of polymorphism c.-124G>A and c.-16 C>T in the promoter region of human INHA gene with altered sperm parameters; a pilot study. Int. J. Clin. Pract. 2020 , 74 , e135952020. [ Google Scholar ] [ CrossRef ] [ PubMed ] Hofland, J.; Steenbergen, J.; Voorsluijs, J.M.; Verbiest, M.M.; de Krijger, R.R.; Hofland, L.J.; de Herder, W.W.; Uitterlinden, A.G.; Feelders, R.A.; de Jong, F.H. Inhibin alpha-subunit (INHA) expression in adrenocortical cancer is linked to genetic and epigenetic INHA promoter variation. PLoS ONE 2014 , 9 , e1049442014. [ Google Scholar ] [ CrossRef ] Tan, S.C.; Low, T.Y.; Mohamad Hanif, E.A.; Sharzehan, M.A.K.; Kord-Varkaneh, H.; Islam, M.A. The rs9340799 polymorphism of the estrogen receptor alpha (ESR1) gene and its association with breast cancer susceptibility. Sci. Rep. 2021 , 11 , 18619. [ Google Scholar ] [ CrossRef ] Ashton, K.A.; Proietto, A.; Otton, G.; Symonds, I.; McEvoy, M.; Attia, J.; Gilbert, M.; Hamann, U.; Scott, R.J. Estrogen receptor polymorphisms and the risk of endometrial cancer. BJOG 2009 , 116 , 1053–1061. [ Google Scholar ] [ CrossRef ] Sadat Eshaghi, F.; Dehghan Tezerjani, M.; Ghasemi, N.; Dehghani, M. Association study of ESR1 rs9340799, rs2234693, and MMP2 rs243865 variants in Iranian women with premature ovarian insufficiency: A case-control study. Int. J. Reprod. Biomed. 2022 , 20 , 841–850. [ Google Scholar ] [ CrossRef ] Paskulin, D.D.; Cunha-Filho, J.S.; Paskulin, L.D.; Souza, C.A.; Ashton-Prolla, P. ESR1 rs9340799 is associated with endometriosis-related infertility and in vitro fertilization failure. Dis. Markers 2013 , 35 , 907–913. [ Google Scholar ] [ CrossRef ] Kornatska, A.G.; Rossokha, Z.I.; Fishchuk, L.Y.; Dubenko, O.D.; Medvedieva, N.L.; Flaksemberg, M.A.; Chubei, G.V.; Popova, O.F.; Gorovenko, N.G. ESR1 gene variants affect FSHR-depended risk of fibrocystic mastopathy in infertile women. Exp. Oncol. 2021 , 43 , 266–269. [ Google Scholar ] [ PubMed ] Gaskins, A.J.; Chiu, Y.H.; Williams, P.L.; Ford, J.B.; Toth, T.L.; Hauser, R.; Chavarro, J.E.; Team, E.S. Association between serum folate and vitamin B-12 and outcomes of assisted reproductive technologies. Am. J. Clin. Nutr. 2015 , 102 , 943–950. [ Google Scholar ] [ CrossRef ] [ PubMed ] Ocal, P.; Ersoylu, B.; Cepni, I.; Guralp, O.; Atakul, N.; Irez, T.; Idil, M. The association between homocysteine in the follicular fluid with embryo quality and pregnancy rate in assisted reproductive techniques. J. Assist. Reprod. Genet. 2012 , 29 , 299–304. [ Google Scholar ] [ CrossRef ] Chang, K.T.; Su, Y.T.; Tsai, Y.R.; Lan, K.C.; Hsuuw, Y.D.; Kang, H.Y.; Chan, W.H.; Huang, F.J. High levels estradiol affect blastocyst implantation and post-implantation development directly in mice. Biomed. J. 2022 , 45 , 179–189. [ Google Scholar ] [ CrossRef ] Figure 1. Synergistic effect of FSHR rs6165 and INHA rs11893842 polymorphisms with clinical parameters. ( A – D ) panels show AOR of FSHR rs6165 and INHA rs11893842 with clinical parameters including E2 ( A ), FSH ( B ), homocysteine ( C ), and folate ( D ). Figure 2. Synergistic effect of ESR1 rs9340799 and rs2234693 polymorphisms with clinical parameters. ( A – F ) panels show AOR of ESR1 rs9340799 and rs2234693 with clinical parameters including E2 ( A , D ), FSH ( B , E ), and LH ( C , F ). Table 1. Clinical profiles between RIF patient and control subjects. Characteristics Controls ( n = 317) RIF ( n = 133) p a Age (years, mean ± SD) 33.5 ± 3.4 33.6 ± 2.9 0.579 BMI (kg/m 2 , mean ± SD) 22.0 ± 3.4 21.1 ± 3.2 0.047 Previous implantation failure ( n , mean ± SD) - 4.7 ± 2.0 N/A Live births ( n , mean ± SD) 1.6 ± 0.6 - N/A Mean gestational age (weeks, mean ± SD) 39.3 ± 1.6 - N/A Homocysteine (μmol/L mean ± SD) 6.4 ± 3.0 6.9 ± 1.8 0.402 Folate (ng/mL, mean ± SD) 14.0 ± 7.5 15.3 ± 10.7 0.896 BUN (mg/dL, mean ± SD) 8.8 ± 2.8 10.4 ± 2.9 <0.0001 Creatinine (mg/dL, mean ± SD) 0.6 ± 0.2 0.8 ± 0.1 <0.0001 Uric acid (mg/dL, mean ± SD 3.9 ± 1.0 4.0 ± 1.0 0.266 Total cholesterol (mg/dL) 215.3 ± 57.2 185.2 ± 42.6 <0.0001 WBC(10 3 /μL, mean ± SD 7.9 ± 2.2 7.4 ± 3.0 0.005 Hgb (g/dL, mean ± SD) 12.3 ± 1.2 12.4 ± 1.5 0.080 PLT (10 3 /μL) 230.0 ± 63.1 238.7 ± 67.0 0.465 PT (sec) 10.7 ± 1.6 11.3 ± 0.6 <0.0001 aPTT (sec) 29.1 ± 3.5 29.6 ± 3.4 0.164 CD3 (pan T) (%, mean ± SD) - 66.9 ± 11.2 N/A CD4 (helper T) (%, mean ± SD) - 34.4 ± 8.8 N/A CD8 (suppressor) (%, mean ± SD) - 28.9 ± 7.7 N/A CD19 (B cell) (%, mean ± SD) - 11.7 ± 4.7 N/A CD56 (NK cell) (%, mean ± SD) - 18.1 ± 9.4 N/A TSH (mU/L, mean ± SD) 1.6 ± 0.9 2.3 ± 1.5 0.0001 E2 (pg/mL, mean ± SD) 26.6 ± 14.4 33.3 ± 15.4 0.0002 FSH (U/L, mean ± SD) 8.1 ± 2.8 8.9 ± 4.5 0.599 LH (U/L, mean ± SD) 3.7 ± 2.6 4.8 ± 2.1 <0.0001 Note: RIF, recurrent implantation failure; BMI, body mass index; BUN, blood urea nitrogen; WBC, white blood cell; Hgb, hemoglobin; PLT, platelet count; PT, prothrombin time; aPTT, activated partial thromboplastin time; TSH, thyroid stimulating hormone; E2, estradiol; FSH, follicle stimulating hormone; LH, luteinizing hormone; N/A, not applicable. a Mann-Whitney test. Table 2. Comparison of genotype frequencies of FSHR , INHA , ESR1 , and BMP15 polymorphisms between the RIF and control subjects. Genotypes Controls ( n = 317) RIF ( n = 133) COR (95% CI) p FDR- p AIC AOR (95% CI) p FDR- p AIC FSHR rs6165 A>G AA 134 (42.3) 62 (46.6) 1.000 (reference) 1.000 (reference) AG 133 (42.0) 61 (45.9) 0.991 (0.647–1.520) 0.968 0.968 430.36 0.996 (0.650–1.528) 0.986 0.986 428.34 GG 50 (15.8) 10 (7.5) 0.432 (0.206–0.908) 0.027 0.135 282.93 0.463 (0.218–0.980) 0.044 0.165 280.03 Dominant (AA vs. AG+GG) 0.839 (0.558–1.260) 0.397 0.702 542.20 0.838 (0.558–1.260) 0.396 0.707 540.16 Recessive (AA+AG vs. GG) 0.434 (0.213–0.885) 0.022 0.11 540.73 0.430 (0.210–0.877) 0.020 0.100 538.69 HWE- P 0.083 0.340 INHA rs11893842 A>G AA 106 (33.4) 42 (31.6) 1.000 (reference) 1.000 (reference) AG 156 (49.2) 61 (45.9) 0.987 (0.621–1.570) 0.956 0.968 518.96 0.988 (0.621–1.573) 0.961 0.986 518.96 GG 55 (17.4) 30 (22.6) 1.377 (0.778–2.436) 0.272 0.334 363.05 1.358 (0.767–2.407) 0.294 0.350 362.07 Dominant (AA vs. AG+GG) 1.089 (0.705–1.680) 0.702 0.702 540.03 1.086 (0.703–1.677) 0.710 0.710 539.99 Recessive (AA+AG vs. GG) 1.388 (0.842–2.287) 0.199 0.249 536.91 1.386 (0.841–2.286) 0.200 0.250 536.87 HWE- P 0.853 0.386 INHA rs35118453 C>T CC 209 (65.9) 84 (63.2) 1.000 (reference) 1.000 (reference) CT 102 (32.2) 42 (31.6) 1.025 (0.660–1.590) 0.914 0.968 480.21 1.025 (0.660–1.592) 0.913 0.986 479.51 TT 6 (1.9) 7 (5.3) 2.903 (0.948–8.892) 0.062 0.155 292.70 2.865 (0.934–8.786) 0.066 0.165 286.68 Dominant (CC vs. CT+TT) 1.129 (0.740–1.722) 0.574 0.702 539.63 1.126 (0.738–1.718) 0.583 0.710 539.58 Recessive (CC+CT vs. TT) 2.880 (0.949–8.737) 0.062 0.155 534.27 2.875 (0.947–8.724) 0.062 0.155 534.09 HWE- P 0.106 0.564 ESR1 rs9340799 A>G AA 212 (66.9) 83 (62.4) 1.000 (reference) 1.000 (reference) AG 94 (29.7) 41 (30.8) 1.114 (0.713–1.740) 0.635 0.968 510.37 1.117 (0.715–1.745) 0.627 0.986 510.09 GG 11 (3.5) 9 (6.8) 2.090 (0.836–5.227) 0.115 0.192 247.08 2.107 (0.841–5.278) 0.112 0.187 240.74 Dominant (AA vs. AG+GG) 1.216 (0.798–1.854) 0.363 0.702 539.52 1.218 (0.799–1.857) 0.360 0.710 539.46 Recessive (AA+AG vs. GG) 2.019 (0.817–4.993) 0.128 0.213 538.12 2.023 (0.818–5.003) 0.127 0.212 538.05 HWE- P 0.884 0.217 ESR1 rs2234693 T>C TT 123 (38.8) 44 (33.1) 1.000 (reference) 1.000 (reference) TC 144 (45.4) 65 (48.9) 1.262 (0.803–1.983) 0.313 0.968 445.72 1.258 (0.800–1.978) 0.320 0.986 445.09 CC 50 (15.8) 24 (18.0) 1.342 (0.739–2.436) 0.334 0.334 279.86 1.330 (0.732–2.417) 0.350 0.350 279.60 Dominant (TT vs. TC+CC) 1.283 (0.838–1.964) 0.252 0.702 539.02 1.281 (0.836–1.962) 0.255 0.710 538.98 Recessive (TT+TC vs. CC) 1.176 (0.688–2.008) 0.553 0.553 540.00 1.173 (0.687–2.005) 0.559 0.559 539.96 HWE- P 0.470 0.999 BMP15 rs17003221 C>T CC 288 (90.9) 124 (93.2) 1.000 (reference) 1.000 (reference) CT 29 (9.1) 9 (6.8) 0.721 (0.331–1.568) 0.409 0.968 539.63 0.718 (0.330–1.563) 0.404 0.986 539.56 TT 0 (0.0) 0 (0.0) N/A N/A N/A N/A N/A N/A N/A N/A Dominant (CC vs. CT+TT) 0.721 (0.331–1.568) 0.409 0.702 539.63 0.718 (0.330–1.563) 0.404 0.710 539.56 Recessive (CC+CT vs. TT) N/A N/A N/A N/A N/A N/A N/A N/A HWE- P 0.393 0.686 BMP15 rs3810682 C>G CC 305 (96.2) 129 (97.0) 1.000 (reference) 1.000 (reference) CG 12 (3.8) 4 (3.0) 0.788 (0.250–2.490) 0.685 0.968 540.18 0.780 (0.246–2.470) 0.672 0.986 540.11 GG 0 (0.0) 0 (0.0) N/A N/A N/A N/A N/A N/A N/A N/A Dominant (CC vs. CG+GG) 0.788 (0.250–2.490) 0.685 0.702 540.18 0.780 (0.246–2.470) 0.672 0.710 540.11 Recessive (CC+CG vs. GG) N/A N/A N/A N/A N/A N/A N/A N/A HWE- P 0.731 0.860 Note: AOR was adjusted by age. RIF, recurrent implantation failure; COR, crude odds ratio; AOR, adjusted odds ratio; 95% CI, 95% confidence interval; FDR; false discovery rate; AIC, Akaike information criterion; HWE, Hardy-Weinberg equilibrium, N/A, not applicable. Table 3. Comparison of genotype frequencies of FSHR , INHA , ESR1 , and BMP15 polymorphisms between the RIF and control subjects. Genotypes Controls RIF ≥ 3 AOR (95% CI) p FDR- p AIC RIF ≥ 4 AOR (95% CI) p FDR- p AIC ( n = 317) ( n = 119) ( n = 89) FSHR rs6165 A>G AA 106 (33.4) 36 (30.3) 1.000 (reference) 28 (31.5) 1.000 (reference) AG 156 (49.2) 54 (45.4) 1.037 (0.665–1.617) 0.871 0.967 393.87 42 (47.2) 0.955 (0.582–1.566) 0.854 0.969 335.77 GG 55 (17.4) 29 (24.4) 0.412 (0.183–0.931) 0.033 0.165 261.48 19 (21.3) 0.392 (0.156–0.982) 0.046 0.115 213.89 Dominant (AA vs. AG+GG) 0.852 (0.558–1.303) 0.461 0.706 504.07 0.787 (0.491–1.261) 0.319 0.809 420.25 Recessive (AA+AG vs. GG) 0.373 (0.171–0.815) 0.013 0.065 501.78 0.372 (0.154–0.901) 0.029 0.073 419.62 INHA rs11893842 A>G AA 209 (65.9) 75 (63.0) 1.000 (reference) 55 (61.8) 1.000 (reference) AG 102 (32.2) 38 (31.9) 1.010 (0.619–1.649) 0.967 0.967 485.21 28 (31.5) 1.011 (0.589–1.734) 0.969 0.969 399.29 GG 6 (1.9) 6 (5.0) 1.534 (0.851–2.767) 0.155 0.194 335.63 6 (6.7) 1.284 (0.657–2.511) 0.465 0.465 277.85 Dominant (AA vs. AG+GG) 1.147 (0.727–1.811) 0.555 0.706 504.15 1.082 (0.652–1.794) 0.761 0.809 419.89 Recessive (AA+AG vs. GG) 1.536 (0.922–2.557) 0.099 0.165 501.57 1.292 (0.720–2.319) 0.391 0.391 415.63 INHA rs35118453 C>T CC 134 (42.3) 55 (46.2) 1.000 (reference) 43 (48.3) 1.000 (reference) CT 133 (42.0) 56 (47.1) 1.029 (0.651–1.626) 0.903 0.967 449.91 40 (44.9) 1.033 (0.617–1.727) 0.903 0.969 376.03 TT 50 (15.8) 8 (6.7) 2.771 (0.863–8.895) 0.087 0.194 265.38 6 (6.7) 3.793 (1.171–12.285) 0.026 0.115 226.80 Dominant (CC vs. CT+TT) 1.123 (0.724–1.744) 0.605 0.706 503.88 1.182 (0.725–1.925) 0.503 0.809 419.35 Recessive (CC+CT vs. TT) 2.749 (0.868–8.704) 0.086 0.165 497.08 3.745 (1.176–11.921) 0.025 0.073 414.45 ESR1 rs9340799 A>G AA 212 (66.9) 76 (63.9) 1.000 (reference) 56 (62.9) 1.000 (reference) AG 94 (29.7) 35 (29.4) 1.042 (0.652–1.666) 0.863 0.967 475.82 26 (29.2) 1.054 (0.623–1.783) 0.844 0.969 392.70 GG 11 (3.5) 8 (6.7) 2.060 (0.796–5.328) 0.136 0.194 351.20 7 (7.9) 2.459 (0.907–6.664) 0.077 0.128 291.35 Dominant (AA vs. AG+GG) 1.146 (0.737–1.782) 0.546 0.706 504.06 1.196 (0.732–1.952) 0.475 0.809 419.83 Recessive (AA+AG vs. GG) 2.025 (0.793–5.173) 0.140 0.175 502.35 2.400 (0.900–6.397) 0.080 0.133 417.51 ESR1 rs2234693 T>C TT 123 (38.8) 38 (31.9) 1.000 (reference) 29 (32.6) TC 144 (45.4) 59 (49.6) 1.318 (0.820–2.118) 0.254 0.967 413.24 42 (47.2) 1.227 (0.720–2.089) 0.452 0.969 338.94 CC 50 (15.8) 22 (18.5) 1.393 (0.748–2.594) 0.297 0.297 257.72 18 (20.2) 1.498 (0.762–2.945) 0.241 0.301 219.97 Dominant (TT vs. TC+CC) 1.342 (0.858–2.100) 0.197 0.706 502.73 1.303 (0.792–2.144) 0.298 0.809 419.24 Recessive (TT+TC vs. CC) 1.197 (0.688–2.082) 0.524 0.524 504.02 1.340 (0.736–2.442) 0.339 0.391 419.45 BMP15 rs17003221 C>T CC 288 (90.9) 110 (92.4) 1.000 (reference) 82 (92.1) 1.000 (reference) CT 29 (9.1) 9 (7.6) 0.804 (0.369–1.755) 0.584 0.967 504.11 7 (7.9) 0.834 (0.352–1.977) 0.681 0.969 420.16 TT 0 (0.0) 0 (0.0) N/A N/A N/A N/A 0 (0.0) N/A N/A N/A N/A Dominant (CC vs. CT+TT) 0.804 (0.369–1.755) 0.584 0.706 504.11 0.834 (0.352–1.977) 0.681 0.809 420.16 Recessive (CC+CT vs. TT) N/A N/A N/A N/A N/A N/A N/A N/A BMP15 rs3810682 C>G CC 305 (96.2) 115 (96.6) 1.000 (reference) 86 (96.6) 1.000 (reference) CG 12 (3.8) 4 (3.4) 0.853 (0.269–2.709) 0.788 0.967 504.35 3 (3.4) 0.853 (0.234–3.104) 0.809 0.969 420.28 GG 0 (0.0) 0 (0.0) N/A N/A N/A N/A 0 (0.0) N/A N/A N/A N/A Dominant (CC vs. CG+GG) 0.853 (0.269–2.709) 0.788 0.788 504.35 0.853 (0.234–3.104) 0.809 0.809 420.28 Recessive (CC+CG vs. GG) N/A N/A N/A N/A N/A N/A N/A N/A Note: AOR was adjusted by age. RIF, recurrent implantation failure, AOR, adjusted odds ratio; 95% CI, 95% confidence interval; AIC, Akaike information criterion; N/A, not applicable. Table 4. Combined genotype analysis for the FSHR , INHA , ESR1 , and BMP15 polymorphisms in RIF patients and controls. Genotype Combinations Controls RIF AOR (95% CI) p ( n = 317) ( n = 133) FSHR rs6165 A>G/ ESR1 rs9340799 A>G AA/AA 92 (29.0) 38 (28.6) 1.000 (reference) AA/AG 37 (11.7) 20 (15.0) 1.312 (0.673–2.556) 0.425 AA/GG 5 (1.6) 4 (3.0) 2.017 (0.487–8.352) 0.333 AG/AA 88 (27.8) 42 (31.6) 1.163 (0.685–1.974) 0.576 AG/AG 42 (13.2) 16 (12.0) 1.767 (0.278–11.232) 0.546 AG/GG 3 (0.9) 3 (2.3) 0.807 (0.270–2.410) 0.701 GG/AA 32 (10.1) 3 (2.3) 0.250 (0.072–0.874) 0.030 GG/AG 15 (4.7) 5 (3.8) 0.807 (0.270–2.410) 0.701 GG/GG 3 (0.9) 2 (1.5) 1.767 (0.278–11.232) 0.546 INHA rs35118453 C>T/ ESR1 rs2234693 T>C CC+CT/TT 119 (37.5) 42 (31.6) 1.000 (reference) CC+CT/TC+CC 192 (60.6) 84 (63.2) 1.240 (0.802–1.917) 0.333 TT/TT 4 (1.3) 2 (1.5) 1.403 (0.247–7.976) 0.703 TT/TC+CC 2 (0.6) 5 (3.8) 7.001 (1.298–37.776) 0.024 FSHR rs6165 A>G/ ESR1 rs9340799 A>G AA+AG/AA 180 (56.8) 80 (60.2) 1.000 (reference) AA+AG/AG+GG 87 (27.4) 43 (32.3) 1.115 (0.710–1.750) 0.636 GG/AA 32 (10.1) 3 (2.3) 0.199 (0.059–0.670) 0.009 GG/AG+GG 18 (5.7) 7 (5.3) 0.867 (0.348–2.160) 0.758 FSHR rs6165 A>G/ BMP15 rs17003221 C>T AA+AG/CC 241 (76.0) 114 (85.7) 1.000 (reference) AA+AG/CT+TT 26 (8.2) 9 (6.8) 0.723 (0.328–1.594) 0.421 GG/CC 47 (14.8) 10 (7.5) 0.447 (0.218–0.918) 0.028 GG/CT+TT 3 (0.9) 0 (0.0) N/A N/A FSHR rs6165 A>G/ BMP15 rs3810682 C>G AA+AG/CC 257 (81.1) 119 (89.5) 1.000 (reference) AA+AG/CG+GG 10 (3.2) 4 (3.0) 0.842 (0.258–2.745) 0.775 GG/CC 48 (15.1) 10 (7.5) 0.446 (0.218–0.914) 0.027 GG/CG+GG 2 (0.6) 0 (0.0) N/A N/A INHA rs11893842 A>G/ ESR1 rs9340799 A>G AA+AG/AA+AG 255 (80.4) 95 (71.4) 1.000 (reference) AA+AG/GG 7 (2.2) 8 (6.0) 3.065 (1.081–8.690) 0.035 GG/AA+AG 51 (16.1) 29 (21.8) 1.526 (0.913–2.549) 0.107 GG/GG 4 (1.3) 1 (0.8) 0.670 (0.074–6.073) 0.722 FSHR rs6165A>G/ INHA rs35118453C>T AA+AG/CC+CT 261 (82.3) 117 (88.0) AA+AG/TT 6 (1.9) 6 (4.5) 2.203 (0.695–6.979) 0.180 GG/CC+CT 50 (15.8) 9 (6.8) 0.399 (0.190–0.841) 0.016 GG/TT 0 (0.0) 1 (0.8) N/A N/A FSHR rs6165 A>G/ ESR1 rs9340799 A>G AA+AG/AA+AG 259 (81.7) 116 (87.2) 1.000 (reference) AA+AG/GG 8 (2.5) 7 (5.3) 1.979 (0.700–5.599) 0.198 GG/AA+AG 47 (14.8) 8 (6.0) 0.373 (0.171–0.816) 0.014 GG/GG 3 (0.9) 2 (1.5) 1.450 (0.239–8.817) 0.687 FSHR rs6165 A>G/ ESR1 rs2234693 T>C AA+AG/TT+TC 224 (70.7) 101 (75.9) 1.000 (reference) AA+AG/CC 43 (13.6) 22 (16.5) 0.604 (0.123–2.966) 0.686 GG/TT+TC 43 (13.6) 8 (6.0) 0.401 (0.181–0.886) 0.024 GG/CC 7 (2.2) 2 (1.5) 0.604 (0.123–2.966) 0.534 FSHR rs6165 A>G/ BMP15 rs17003221 C>T AA+AG/CC+CT 267 (84.2) 123 (92.5) 1.000 (reference) GG/CC+CT 50 (15.8) 10 (7.5) 0.430 (0.210–0.877) 0.020 FSHR rs6165 A>G/ BMP15 rs3810682 C>G AA+AG/CC+CG 267 (84.2) 123 (92.5) 1.000 (reference) GG/CC+CG 50 (15.8) 10 (7.5) 0.430 (0.210–0.877) 0.020 ESR1 rs9340799 A>G/ ESR1 rs2234693 T>C AA+AG/TT+TC 266 (83.9) 104 (78.2) 1.000 (reference) AA+AG/CC 40 (12.6) 20 (15.0) 1.270 (0.709–2.276) 0.422 GG/TT+TC 1 (0.3) 5 (3.8) 12.930 (1.492–112.052) 0.020 GG/CC 10 (3.2) 4 (3.0) 1.024 (0.314–3.340) 0.968 Note: AOR was adjusted by age. RIF, recurrent implantation failure; 95% CI, 95% confidence interval; AOR, adjusted odds ratio; NA, not applicable. Table 5. Clinical variables in RIF patients and controls stratified by FSHR , INHA , ESR1 , and BMP15 polymorphisms status by ANOVA and Kruskal–Wallis test. Genotypes BMI (kg/m 2 ) Homocysteine (μmol/L) Folate (ng/mL) BUN (mg/dL) Creatinine (mg/dL) Uric Acid (mg/dL) Total Cholesterol (mg/dL) TSH (mU/L) Mean ± SD Mean ± SD Mean ± SD Mean ± SD Mean ± SD Mean ± SD Mean ± SD Mean ± SD FSHR rs6165 A>G AA 21.4 ± 3.6 7.0 ± 2.3 16.8 ± 11.8 9.2 ± 3.0 0.7 ± 0.1 3.9 ± 1.0 199.1 ± 53.0 1.9 ± 1.4 AG 21.5 ± 3.2 6.3 ± 1.8 13.3 ± 6.2 9.5 ± 2.9 0.7 ± 0.2 3.9 ± 1.0 209.0 ± 55.6 1.9 ± 1.2 GG 21.5 ± 2.4 7.0 ± 3.8 9.0 ± 4.0 9.5 ± 2.9 0.7 ± 0.2 4.0 ± 1.2 202.0 ± 52.2 2.3 ± 1.2 p b 0.676 0.273 0.175 0.533 0.576 0.987 0.366 0.209 INHA rs11893842 A>G AA 21.4 ± 3.2 6.4 ± 1.8 13.9 ± 8.7 9.5 ± 3.0 0.7 ± 0.2 4.0 ± 1.0 205.2 ± 57.2 2.1 ± 1.6 AG 21.7 ± 3.6 6.9 ± 2.5 16.2 ± 12.2 9.3 ± 2.9 0.7 ± 0.1 3.8 ± 1.0 205.6 ± 53.3 1.7 ± 1.0 GG 20.9 ± 2.6 7.0 ± 2.4 13.9 ± 5.5 9.4 ± 2.8 0.7 ± 0.2 4.0 ± 1.1 198.7 ± 52.1 1.9 ± 1.3 p b 0.77 0.576 a 0.838 0.873 0.789 0.381 0.532 0.332 INHA rs35118453 C>T CC 21.3 ± 3.1 6.6 ± 1.9 14.7 ± 10.7 9.4 ± 3.1 0.7 ± 0.2 3.9 ± 1.1 203.1 ± 56.5 2.0 ± 1.4 CT 21.8 ± 3.9 7.1 ± 2.8 14.4 ± 7.8 9.2 ± 2.7 0.7 ± 0.2 4.0 ± 0.9 206.6 ± 51.4 1.8 ± 1.0 TT 19.8 ± 1.7 6.3 ± 1.3 18.3 ± 6.5 9.9 ± 2.2 0.7 ± 0.2 3.6 ± 1.0 196.8 ± 34.7 2.1 ± 1.3 p b 0.317 0.716 0.389 0.559 0.746 0.531 0.785 0.789 ESR1 rs9340799 A>G AA 21.6 ± 3.6 6.7 ± 1.9 14.3 ± 7.6 9.4 ± 3.0 0.7 ± 0.2 3.9 ± 1.0 200.7 ± 51.7 1.8 ± 1.2 AG 21.0 ± 2.6 7.3 ± 3.0 17.9 ± 14.6 9.3 ± 2.8 0.7 ± 0.2 3.9 ± 1.1 214.5 ± 59.5 2.1 ± 1.4 GG 22.3 ± 4.2 5.6 ± 2.7 11.1 ± 7.7 9.7 ± 2.9 0.7 ± 0.1 4.1 ± 0.8 184.5 ± 42.4 2.2 ± 1.3 p b 0.662 0.276 a 0.502 0.847 0.726 0.618 0.116 0.42 ESR1 rs2234693 T>C TT 21.8 ± 4.1 6.5 ± 1.6 12.9 ± 6.0 9.0 ± 3.0 0.7 ± 0.1 4.0 ± 1.2 204.8 ± 51.0 1.9 ± 1.4 TC 21.2 ± 2.7 6.8 ± 2.2 15.4 ± 11.4 9.5 ± 3.0 0.7 ± 0.2 3.8 ± 0.9 205.5 ± 58.8 1.9 ± 1.3 CC 21.5 ± 3.0 7.3 ± 3.4 18.3 ± 10.9 9.9 ± 2.5 0.7 ± 0.2 4.0 ± 1.0 197.6 ± 45.9 1.8 ± 1.0 p b 0.974 0.456 0.294 0.085 0.76 0.92 0.872 0.927 BMP15 rs17003221 C>T CC 21.4 ± 3.3 6.7 ± 2.3 15.1 ± 9.7 9.4 ± 3.0 0.7 ± 0.2 3.9 ± 1.1 204.5 ± 55.1 1.9 ± 1.3 CT 22.4 ± 4.0 7.5 ± 1.0 8.5 ± 1.0 9.1 ± 2.3 0.7 ± 0.2 3.9 ± 0.8 199.4 ± 44.7 1.8 ± 1.0 TT 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 p b 0.243 a 0.238 0.149 0.832 0.444 0.902 0.977 0.94 BMP15 rs3810682 C>G CC 21.5 ± 3.3 6.7 ± 2.3 15.0 ± 9.7 9.4 ± 3.0 0.7 ± 0.2 3.9 ± 1.0 203.8 ± 54.3 1.9 ± 1.3 CG 20.6 ± 3.5 7.1 ± 1.3 10.9 ± 3.9 9.2 ± 1.8 0.7 ± 0.2 4.0 ± 0.9 208.4 ± 51.4 1.6 ± 1.1 GG 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 p b 0.303 a 0.636 0.537 0.909 0.928 0.674 0.785 0.741 Genotypes E2 (pg/mL) FSH (U/L) LH (U/L) WBC (10 3 /μL) Hgb (g/dL) PLT (10 3 /μL) PT (sec) aPTT (sec) Mean ± SD Mean ± SD Mean ± SD Mean ± SD Mean ± SD Mean ± SD Mean ± SD Mean ± SD FSHR rs6165 A>G AA 29.0 ± 14.4 8.5 ± 4.4 4.2 ± 2.4 7.9 ± 2.6 12.4 ± 1.3 235.6 ± 62.3 11.1 ± 1.8 29.5 ± 3.5 AG 33.0 ± 16.8 8.2 ± 3.2 4.3 ± 2.9 7.9 ± 2.5 12.4 ± 1.4 231.1 ± 60.5 10.8 ± 0.9 29.3 ± 3.5 GG 25.9 ± 12.9 8.8 ± 2.2 4.0 ± 1.5 6.8 ± 2.1 12.0 ± 1.5 234.0 ± 89.9 10.7 ± 1.0 28.4 ± 2.8 p b 0.076 0.11 0.843 0.119 0.429 0.576 0.207 0.334 INHA rs11893842 A>G AA 27.2 ± 13.3 8.6 ± 3.5 3.9 ± 2.0 7.6 ± 2.3 12.4 ± 1.2 236.4 ± 73.8 10.8 ± 0.8 29.3 ± 3.9 AG 30.4 ± 16.1 8.1 ± 4.2 4.1 ± 2.9 7.9 ± 2.7 12.3 ± 1.4 233.5 ± 64.7 11.0 ± 1.8 29.1 ± 3.2 GG 34.0 ± 15.7 9.0 ± 1.7 4.7 ± 2.2 7.6 ± 2.2 12.3 ± 1.4 228.2 ± 47.5 10.8 ± 0.8 29.7 ± 3.4 p b 0.067 0.113 0.077 0.682 0.812 0.97 0.788 0.521 INHA rs35118453 C > T CC 29.0 ± 15.0 8.5 ± 4.0 4.1 ± 2.7 7.6 ± 2.4 12.3 ± 1.3 236.6 ± 67.6 10.9 ± 0.9 29.3 ± 3.5 CT 31.0 ± 15.4 8.4 ± 3.0 4.3 ± 1.9 7.9 ± 2.6 12.4 ± 1.4 229.0 ± 60.4 11.0 ± 2.0 29.3 ± 3.5 TT 44.9 ± 15.0 7.1 ± 2.8 6.1 ± 1.7 8.6 ± 2.5 12.6 ± 1.6 217.6 ± 50.1 10.7 ± 0.6 29.8 ± 2.8 p b 0.067 0.066 0.017 0.232 0.262 0.482 0.878 0.634 ESR1 rs9340799 A>G AA 30.5 ± 16.2 8.5 ± 4.0 4.3 ± 2.7 7.8 ± 2.5 12.4 ± 1.4 234.6 ± 69.6 10.9 ± 0.8 29.2 ± 3.4 AG 28.8 ± 13.4 8.4 ± 3.0 4.0 ± 2.1 7.8 ± 2.5 12.4 ± 1.3 233.3 ± 55.8 11.0 ± 2.2 29.4 ± 3.6 GG 30.2 ± 12.6 7.1 ± 2.8 3.3 ± 1.1 6.9 ± 1.9 12.0 ± 1.2 219.8 ± 48.4 11.1 ± 1.0 29.7 ± 3.7 p b 0.882 0.267 0.576 0.423 0.381 0.644 0.51 0.918 ESR1 rs2234693 T>C TT 28.7 ± 14.2 8.4 ± 3.6 4.1 ± 3.0 7.9 ± 2.6 12.4 ± 1.5 232.3 ± 59.5 10.9 ± 0.9 29.3 ± 3.7 TC 30.9 ± 15.5 8.1 ± 3.3 4.3 ± 2.3 7.7 ± 2.5 12.4 ± 1.2 237.7 ± 71.0 11.0 ± 1.8 29.4 ± 3.4 CC 30.0 ± 16.8 9.2 ± 4.7 4.0 ± 1.8 7.7 ± 2.3 12.2 ± 1.3 223.1 ± 55.8 10.8 ± 0.8 29.0 ± 3.1 p b 0.58 0.446 0.601 0.861 0.761 0.543 0.775 0.625 BMP15 rs17003221 C > T CC 30.1 ± 15.0 8.5 ± 3.7 4.2 ± 2.5 7.8 ± 2.6 12.4 ± 1.4 233.2 ± 64.2 10.9 ± 1.4 29.4 ± 3.5 CT 27.0 ± 19.8 7.8 ± 2.5 4.0 ± 0.9 7.7 ± 1.6 12.0 ± 1.0 234.9 ± 69.6 10.8 ± 0.8 28.5 ± 3.1 TT 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 p b 0.55 0.5 0.524 0.68 0.039 0.812 0.553 0.146 BMP15 rs3810682 C>G CC 29.7 ± 15.2 8.4 ± 3.7 4.2 ± 2.5 7.7 ± 2.5 12.3 ± 1.3 233.4 ± 65.1 10.9 ± 1.4 29.3 ± 3.4 CG 35.6 ± 15.6 8.4 ± 3.4 4.4 ± 1.5 8.7 ± 2.2 12.4 ± 1.8 231.9 ± 53.6 10.6 ± 0.6 30.0 ± 5.2 GG 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 P b 0.248 0.902 0.34 0.105 0.68 0.732 0.363 0.761 ANOVA, Analysis of variance; BMI, body mass index; BUN, blood urea nitrogen; TSH, thyroid stimulating hormone; E2, estradiol; FSH, follicle stimulating hormone; LH, luteinizing hormone; WBC, white blood cell; Hgb, hemoglobin; PLT, platelet; PT, prothrombin time; aPTT, activated partial thromboplastin time; SD, standard deviation. a p -values were calculated by ANOVA. b p -values were calculated by Kruskal–Wallis tests. 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Refworld | Hunan police delete Chinese filmmaker's documentary footage Refworld is the leading source of information necessary for taking quality decisions on refugee status. Refworld contains a vast collection of reports relating to situations in countries of origin, policy documents and positions, and documents relating to international and national legal frameworks. The information has been carefully selected and compiled from UNHCR's global network of field offices, Governments, international, regional and non-governmental organizations, academic institutions and judicial bodies. Hunan police delete Chinese filmmaker's documentary footage Publisher Radio Free Asia Publication Date 16 January 2014 Cite as Radio Free Asia, Hunan police delete Chinese filmmaker's documentary footage , 16 January 2014, available at: https://www.refworld.org/docid/532adeb014.html [accessed 31 May 2023] Disclaimer This is not a UNHCR publication. UNHCR is not responsible for, nor does it necessarily endorse, its content. Any views expressed are solely those of the author or publisher and do not necessarily reflect those of UNHCR, the United Nations or its Member States. 2014-01-16 Independent Chinese filmmaker Liu Yimu has demanded a public explanation from authorities after raw footage for his planned documentary was deleted by state security police in his hometown of Changsha, the capital of Hunan province. Liu, who had recently finished interviewing 23 local people, including democracy activists and petitioners, was about to begin editing the film, "These Changsha People," when police intervened, he told RFA's Mandarin Service this week. On Jan. 11, they ordered him to halt production, as their "leaders" were very concerned by the film, without specifying which leaders. Liu said all of his interviewees had "different political views," and that the film, while avoiding highly sensitive material, had been intended as a channel for communication between local people and officials. "Changes are taking place in every city, and someone should be doing something to document them," he said. State security police deleted the original footage as shot by Liu and his team since Jan. 3, however. Cheng Qiubo, who worked with Liu on the film, said documentary films in China usually only represent the view of the government. "In documentary, the state media ... uses only one model and one voice," he told New Tang Dynasty Television in a recent interview. "We wanted to record and to observe society as it truly is." Request launched Liu said state security police had initially approached him and questioned him about the project shortly after it began. Interviewees included independent labor leader Zhang Jingsheng, a student follower of late supreme leader Mao Zedong, petitioners, forced evictees, and veterans rights activists, including retired People's Liberation Army (PLA) soldiers. Liu also spoke to university professors, lawyers, fashion models, and entrepreneurs. Cheng told NTDTV: "The producers feared oppression by the regime, so they decided not to include anything that would be deemed too sensitive by the regime: they took a neutral stance." Liu said he has since lodged a formal request for public disclosure regarding the deletion of his raw footage, but has yet to receive a reply. Nationwide crackdown Last year, the ruling Chinese Communist Party stepped up a nationwide crackdown on independent filmmakers and festivals, rounding up film students who attended an independent film summer school in Beijing last July. Employees told RFA at the time that the Li Xianting Film Foundation and its sister project the Beijing Independent Film Festival (BIFF) are no stranger to harassment from the authorities. The BIFF and the school are hotbeds of fresh documentary talent intent on filming topics and stories that the authorities would rather didn't reach the public domain. Meanwhile, the Yunnan Multicultural Visual Festival, which showcases some of China's most influential and cutting-edge documentaries, was forced to cancel a public screening in March. Reported by Xin Yu for RFA's Mandarin Service. Translated and written in English by Luisetta Mudie.

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MitoKATP regulating HIF/miR210/ISCU signaling axis and formation of a positive feedback loop in chronic hypoxia-induced PAH rat model MitoKATP regulating HIF/miR210/ISCU signaling axis and formation of a positive feedback loop in chronic hypoxia-induced PAH rat model Authors: Yang Lu Jing Huang Shuang Geng Hao Chen Cheng Song Shan Zhu Su Zhao Mingli Yuan Xueying Li Hongling Hu Affiliations: Department of Respiratory Medicine, Key Laboratory for Molecular Diagnosis of Hubei Province, Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430017, P.R. China Published online on: February 23, 2017 https://doi.org/10.3892/etm.2017.4161 Pages: 1697-1701 Abstract In the present study, we studied the mechanism of mitochondrial ATP-sensitive potassium (mitoKATP) channels regulating hypoxia-inducible factor (HIF)-1α/microRNA (miR)-210/mitochondrial iron‑sulfur protein integrin (ISCU) signaling axis and forming a positive feedback loop in chronic hypoxia‑induced pulmonary arterial hypertension (PAH) by using in vivo animal model. Two hundred healthy adult SPF Sprague‑Dawley rats were randomly divided into five groups: Control, a mimic miR-210 agent (mimic‑210) intervention, a miR-210 inhibitor (anti‑210) intervention, a chronic PAH and an anti‑210 intervention PAH groups, with 40 rats in each group. After the chronic PAH rat model was successfully established, the rats were intervened with mimic‑210 and anti‑210. The pulmonary artery smooth muscle cells (PASMCs) of rats in each group were acutely isolated and the activity of mitoKATP and mitochondria‑derived oxygen free radicals reactive oxygen species (ROS) was detected. RT-qPCR was used to detect the gene of HIF-1α/miR-210/ISCU and western blot analysis was used to detect the protein of HIF-1α and ISCU. The gene and protein expression were detected again after mitoKATP-specific opener diazoxide and blocker 5-HD was given via tail vein and took effect on each group of rats, respectively. Additionally, the indicators were detected again after ISCU recombinant protein was given via tail vein and ISCU small interfering RNA (siRNA) via nasal feeding and took effect on each group of rats, respectively. It was found that the activity of mitoKATP and ROS and the gene and protein levels of HIF-1α/miR-210/ISCU of the mimic-210 group were significantly higher than those of the control group while that of the anti-210 group was significantly reduced (P<0.05). The indicators in the chronic PAH group were significantly higher than those of the control group while those of the anti-210 intervention PAH group were significantly reduced (P<0.05). The indicators of all the groups were increased after being given mitoKATP specific opener diazoxide. The indicators of all the groups were significantly reduced after receiving blocker 5-HD (P<0.05). The indicators of all the groups were significantly reduced after given ISCU recombinant protein. The indicators of all the groups increased following ISCU siRNA, and there was a statistically significant difference (P<0.05). In conclusion, the mechanism of mitoKATP regulating the HIF-1α/miR-210/ISCU signaling axis and formation of a positive feedback loop exists in the PAH rat model. Introduction Chronic hypoxia-induced pulmonary arterial hypertension (PAH) is an important pathogenesis mechanism in the progress of chronic obstructive pulmonary disease to chronic pulmonary heart diseases ( 1 ). PAH can be caused by chronic hypoxia-induced pulmonary vasoconstriction and reconstruction but the exact mechanism is unclear ( 1 ). There are not enough effective clinical interventions. Under hypoxia, mitochondrial electron transport chain (ETC) regulates the production of mitochondrial-derived oxygen-free radicals reactive oxygen species (ROS) ( 2 ), especially H 2 O 2 , which is permeable and can activate the transcription factor, low oxygen-induced factor hypoxia-inducible factor (HIF)-1α ( 3 ), causing the mitochondrial ATP-sensitive potassium (mitoKATP) channel of pulmonary artery smooth muscle cells (PASMCs) to be open and activated ( 4 ). It gives a positive feedback on ROS level and the expression of HIF-1α ( 5 ), promoting the proliferation of PASMCs and inhibiting their apoptosis ( 6 ) and taking part in the remodeling process of chronic hypoxic pulmonary blood vessels. In addition, ~6% of human microRNAs (miRNAs) including the miR-210 have HIF binding sites in the promoter region, with which HIF can combine to regulate the expression of miRNA ( 7 ). It is confirmed that the expression of HIF-1α and miR-210 increases and forms a positive feedback loop ( 8 ) when human mesenchymal stem cells are under hypoxia. The highly expressed miR-210 under hypoxia stress is able to make target regulation to the mitochondrial iron-sulfur protein integrin (ISCU) of tumor cells, vascular endothelial cells and many other cells, affecting the circulation of Krebs, electron transport, ion metabolism and the production of ROS, thus affecting the function of mitochondria ( 9 ). Previous studies confirmed that the ETC complex II inhibitor can activate mitoKATP and mitoKATP-specific openers can also inhibit the activity of complex II ( 10 ). Those two are closely related, suggesting that ETC complex II may be a component of mitoKATP, composition or an regulatory factor ( 10 ). Based on the above, we found in in vitro cell experiments that the mechanism of the mitoKATP regulating HIF-1α/miR-210/ISCU signaling axis forming a positive feedback loop in PAH is involved in the remodeling process of chronic hypoxic pulmonary blood vessels ( 11 ). However, to the best of our knowledge, there are few studies in vivo on animals. The current study examined the relationship between the mitoKATP and the signaling axis from the PAH rat model to provide strong evidence for clinical treatment. Materials and methods Animals and model building Two hundred healthy adult SPF SD rats, each weighing 150–200 g were fed normally. The rats were housed in a temperature controlled room (21 ± 2°C) on a 12:12-h light:dark cycle (lights on at 06:00) with free access to water and food. The rats were randomly divided into five groups: A normal control, a mimic miR-210 agent (mimic-210) intervention, a miR-210 inhibitor (anti-210) intervention, a chronic PAH group and an anti-210 intervention PAH groups, with 40 rats in each group. Establishment of the chronic PAH rat model The rats were placed in an open normobaric low-oxygen cabin in which the oxygen concentration was kept at 10.0±0.3% and the carbon dioxide concentration was kept <3%. The low-oxygen condition was kept for 8 h daily, 4 weeks continuously. The intervention in rats with mimic-210 and anti-210 (both from R&D Systems, Minneapolis, MN, USA) was made by the nasal topical drug delivery method (compared with systemic drug delivery, nasal topically drug delivery is lower in dosage, works better in targeting and miRNA positioning, and can reduce side effects on other organs). Several studies have shown that small interfering RNA (siRNA) delivered topically though nasal cavity or trachea can have a significant target gene effect in the lungs ( 12 ). Approval for the animal studies was received from Wuhan Central Hospital Affiliated to Huazhong University (Hubei, China). Research methods The rat PASMCs in each group were acutely isolated (each PASMC was obtained using the enzyme digestion method and its shrinkage was observed by phenylephrine to identify its physiological activity). The flow cytometry method was used to detect immunofluorescent activity of mitoKATP and ROS, the RT-qPCR assay was used to detect the gene of HIF-1α/miR-210/ISCU and western blot analysis was used to detect the protein of HIF-1α and ISCU. The gene and protein expressions were detected again after mitoKATP-specific opener diazoxide and blocker 5-HD (both from R&D Systems) was given via tail vein and took effect on each group of rats, respectively. The indicators above were detected again after ISCU recombinant protein was given via tail vein and ISCU siRNA (both from R&D Systems) via nasal feeding and took effect on each group of rats, respectively. Detection methods Flow cytometry FITC-labeled anti-human R-123, ROS monoclonal antibody, immunoglobulin G (IgG) isotype control with corresponding fluorescence tags and a BD FACSAria sorting flow cytometer (both from BD Biosciences, Frankin Lakes, NJ, USA). Separately 10 µl of different fluorescently labeled R-123, ROS antibodies and isotype control was added to the tubes of the PASMC suspension. Three sets of peripheral blood were added with anticoagulant (50 µl each set) and was incubated at 25°C, in the dark for 15 min. Hemolysin (1 ml) was added and reacted at room temperature in the dark for 15 min and washed with phosphate-buffered saline (PBS), followed by centrifugation at 1,200 × g for 5 min, and 500 µl of PBS was added for cell suspension. Flow cytometry and the CellQuest software (Version 3.1, BD Biosciences, San Jose, CA, USA) were used to detect and corresponding fluorescence-tagged IgG staining cells isotype control was used as a negative control. ROS is the oxygen sensing function indicator of the mitochondrial respiratory chain. R-123 fluorescence intensity is proportional to the mitochondrial membrane potential, indirectly detecting the activity of mitoKATP. RT-qPCR assay Primer sequences were synthesized by Shanghai Shenggong Biological Engineering Technology & Services Co., Ltd. (Shanghai, China). The fluorescence quantitative PCR instrument 7900HT was purchased from Applied Biosystems (Foster City, CA, USA). Conventional TRIzol reagent was used to extract total RNA, and the ultraviolet spectrophotometric method was used to detect the concentration and purity. The reverse transcription kit indicated the synthesis of cDNA, designed primer sequences and amplified PCR ( Table I ). The reaction system consisted of 2X Taq Master Mix (25 µl), forward and reverse primers (10 µM) of 2 µl, template DNA (4 µl) and ddH 2 O (17 µl) to make a total reaction volume of 50 µl. <table><tbody><tr><td> <here is a image 17abcfc219395351-f039551d52bc6801> </td><td> Table I. Primer sequences used in this study.</td></tr></tbody></table> Table I. Primer sequences used in this study. <table><tr><th rowspan=1 colspan=1> Genes</th><th rowspan=1 colspan=1> Forward primer</th><th rowspan=1 colspan=1> Reverse primer</th><th rowspan=1 colspan=1> Band length (bp)</th></tr><tbody><tr><td rowspan=1 colspan=1> HIF-1α</td><td rowspan=1 colspan=1> 5′-CGTTCCTTCGATCAGTTGTC-3′</td><td rowspan=1 colspan=1> 5′-TCAGTGGTGGCAGTGGTAGT-3′</td><td rowspan=1 colspan=1> 143</td></tr><tr><td rowspan=1 colspan=1> miR-210</td><td rowspan=1 colspan=1> 5′-CGCCTGTGCGTGTGACAGCG-3′</td><td rowspan=1 colspan=1> 5′-GTGCAGGGTCCGAGGT-3′</td><td rowspan=1 colspan=1> 71</td></tr><tr><td rowspan=1 colspan=1> ISCU</td><td rowspan=1 colspan=1> 5′-GGCAAACCAGGCAGAGCCAGAG-3′</td><td rowspan=1 colspan=1> 5′-GGATGGTACGGCCCGAGGTG-3′</td><td rowspan=1 colspan=1> 95</td></tr><tr><td rowspan=1 colspan=1> β-actin</td><td rowspan=1 colspan=1> 5′-CTGGAACGGTGAAGGTGACA-3′</td><td rowspan=1 colspan=1> 5′-AAGGGACTTCCTGTAACAATGCA-3′</td><td rowspan=1 colspan=1> 140</td></tr></tbody></table> [i]HIF, hypoxia-inducible factor; miR, microRNA. The amplification conditions were pre-denaturation at 94°C for 4 min, denaturation at 94°C for 30 sec, annealing at 56°C for 30 sec, extension at 72°C for 1 min for total of 35 cycles, with another extension at 72°C for 10 min. The agarose gel electrophoresis was performed by loading 5 µl of PCR product in each well, with an voltage of 110V for 40 min. The gel image was captured under ultraviolet analyzer and the results were expressed as a ratio of the target gene and reference gene, 2 −ΔΔCq . Western blot analysis Total protein was extracted according to protein extraction kit (BCA protein). The BCA protein assay kit was used for the determination of protein concentration to make up 5.0 mg/ml of concentration to store at −80°C. SDS-PAGE electrophoresis method (Beijing Liuyi Instrument Factory, Beijing, China) was used to separate HIF-1α and the ISCU proteins. The gel was stained and placed in fast dye Coomassie brilliant blue to analyze with the gel imaging system (Syngene, Frederick, MD, USA) for the grayscale value, while using β-actin as the internal control. Statistical analysis SPSS 19.0 statistical software (Chicago, IL, USA) was used for inputting and analysis, all the quantitative data were expressed as mean ± standard deviation. The single factor ANOVA analysis was used for group comparison, and LSD or Bonferroni test was used for pairwise comparison; qualitative data were expressed as a number or percentage (%), χ 2 test was used for group comparison; P<0.05 was considered to indicate a statistically significant difference. Results Comparison of the activity of mitoKATP and ROS The activity of mitoKATP and ROS of the mimic-210 group was significantly higher than that of the control group while that of the anti-210 group was significantly reduced (P<0.05). The activity of mitoKATP and ROS of the chronic PAH group was significantly higher than that of the control group while that of the anti-210 intervention PAH group was significantly reduced (P<0.05) ( Fig. 1 ). <table><tbody><tr><td> <here is a image 3a75d1db0ed0e6ec-2f405a6a16f6a5ad> </td><td> Figure 1.Activity of mitoKATP and ROS detected by flow cytometry method (control, mimic-210, anti-210, chronic PAH and anti-210 intervention groups). mitoKATP, mitochondrial ATP-sensitive potassium; ROS, reactive oxygen species.</td></tr></tbody></table> The activity of ROS in all the groups increased after given mitoKATP specific opener diazoxide via the tail vein. ROS activity in all the groups reduced significantly after given blocker 5-HD (P<0.05). The activity of mitoKATP and ROS in all the groups reduced after given ISCU recombinant protein via the tail vein (P<0.05). In addition, the activity of mitoKATP and ROS in all the groups significantly increased after given ISCU siRNA via nasal feeding (P<0.05). Gene level of HIF-1α/miR-210/ISCU The levels of HIF-1α/ miR-210/ISCU of the mimic-210 group were significantly higher than that of the control group while that of the anti-210 group was significantly reduced (P<0.05). The gene level of HIF-1α/miR-210/ISCU of the chronic PAH group was significantly higher than that of the control group while that of the anti-210 intervention PAH group was significantly reduced (P<0.05) ( Fig. 2 ). <table><tbody><tr><td> <here is a image 230c889de9d8bff3-a1ec6f774847f23a> </td><td> Figure 2.Levels of HIF-1α/miR-210/ISCU detected by RT-qPCR assay. HIF-1α, miR-210 and ISCU; lane 1, control (β-actin); lane 2, mimic; lane 3, anti-210; lane 4, chronic PAH; and lane 5, anti-210 intervention groups. HIF, hypoxia-inducible factor; PAH, pulmonary arterial hypertension.</td></tr></tbody></table> The gene level of HIF-1α/miR-210/ISCU in all the groups increased after diazoxide was given. The gene level of HIF-1α/miR-210/ISCU in all the groups decreased following 5-HD, with a statistically significant difference (P<0.05). The gene level of HIF-1α/miR-210/ISCU in all the groups decreased following ISCU recombinant protein, whereas the gene level of HIF-1α/miR-210/ISCU in all the groups increased following ISCU siRNA, with a statistically significant difference (P<0.05). Protein level of HIF-1α and ISCU The protein levels of HIF-1α and ISCU of the mimic-210 group were significantly higher than that of the control group while that of the ant-210 group was significantly reduced (P<0.05); the protein level of HIF-1α and ISCU of the chronic PAH group was significantly higher than that of the control group while that of the anti-210 intervention PAH group was significantly reduced (P<0.05) ( Fig. 3 ). <table><tbody><tr><td> <here is a image 540d0a569d5168c0-8193176ead56c5c9> </td><td> Figure 3.Protein level of HIF-1α and ISCU detected by western blot analysis. HIF-1α, and ISCU; lane 1, control (β-actin); lane 2, mimic-210; lane 3, anti-210; lane 4, chronic PAH; and lane 5, anti-210 intervention groups. HIF, hypoxia-inducible factor; PAH, pulmonary arterial hypertension.</td></tr></tbody></table> The protein level of HIF-1α and ISCU in all the groups significantly increased after diazoxide was given, whereas the protein levels of HIF-1α and ISCU in all the groups were significantly reduced after 5-HD was given (P<0.05). The protein level of HIF-1α in all the groups significantly reduced after ISCU recombinant protein was given (P<0.05). In addition, the protein level of HIF-1α in all the groups significantly increased after ISCU siRNA was given (P<0.05). Discussion Our previous experiments in cell lines has confirmed that activated mitoKATP under hypoxia can increase the production of ROS, which can activate the HIF ( 13 ). The mitoKATP of PASMCs opens and regulates the signaling pathway of ROS/HIF/miR-210/ISCU under hypoxia, forming a positive feedback loop that continually induce a high expression of miR-210, thus repetitively stimulate the proliferation of PASMCs. The opening and activating of mitoKATP can cause an influx of potassium ions and mitochondrial swelling, being an important factor that affects the mitochondrial membrane potential. It can protect a variety of cells such as nerve cells and heart muscle cells and promote cell survival and inhibit apoptosis ( 14 ). Previous findings have shown that miR-210 has an anti-apoptotic function of hypoxic PASMCs ( 15 ). We assume that the opening and activating of mitoKATP changes oxygen-free radical levels by affecting the oxygen sensing function of mitochondrial respiratory chain complex, thus activating increased HIF and regulating the HIF/miR-210/ISCU signaling pathway while ISCU affects the activity of mitoKATP by affecting the activity of ETC compound II. In this way, a positive feedback loop is formed, repeatedly stimulating the proliferation of PASMCs to promote the remodeling of pulmonary blood vessels. Through establishment of the PAH rat model and in vivo study, we found that the activity of mitoKATP and ROS and the gene, protein levels of HIF-1α/miR-210/ISCU of the mimic-210 group were significantly higher than that of the control group, while that of the anti-210 group was significantly reduced. These indicators of the chronic PAH group were all significantly higher than those of the control group while those of the anti-210 intervention PAH group was significantly reduced. The indicators of all the groups significantly increased after mitoKATP-specific opener diazoxide was given, and the indicators of all the groups significantly reduced after blocker 5-HD was given. The indicators of all the groups significantly reduced after ISCU recombinant protein was given, the indicators of all the groups significantly increased following ISCU siRNA. Thus, the mechanism of mitoKATP regulating HIF-1α/miR-210/ISCU signaling axis and formation of a positive feedback loop exists in the PAH rat model. We further explored, giving mitoKATP-specific inhibitors in vivo , pulmonary arterial pressure and pathological changes in reversible pulmonary hypertension rats ( 16 ). In this way, we completely explained the mechanism of mitoKATP regulating HIF-1α/miR-210/ISCU signaling axis and the formation of a positive feedback loop in chronic PAH, providing theoretical basis for filtering new chronic hypoxic PAH therapeutic targets. Currently there are no studies on chronic hypoxic PASMC mitoKATP regulating miR-210 and its target gene ISCU. This study explored that the chronic hypoxic PASMC mitoKATP regulating HIF/miR-210/ISCU signaling axis and forms a positive feedback loop, repeatedly stimulating the proliferation and migration of PASMCs. Since they horizontally regulate the expression of genes after transcription, miRNAs can regulate the function of mitochondria at the genetic level, providing experimental basis for chronic hypoxic pulmonary hypertension gene therapy. This in vivo study on the function of mitoKATP regulating HIF-1α/miR-210/ISCU signaling axis and formation of a positive feedback loop based on previous research has improved value and significance due to the complexity of in vivo experiments. Acknowledgements This study was supported by the Health and Family Planning Commission of Hubei Province (no. WJ2015MB140). References <table><tbody><tr><td rowspan=1 colspan=1> 1</td><td rowspan=1 colspan=1>Zhang S, Liu B, Fan Z, Wang D, Liu Y, Li J, Wang N, Liu Y and Zhang B: Targeted inhibition of survivin with YM155 promotes apoptosis of hypoxic human pulmonary arterial smooth muscle cells via the upregulation of voltage-dependent K +channels. Mol Med Rep. 4:3415–3422. 2016.</td></tr><tr><td rowspan=1 colspan=1> 2</td><td rowspan=1 colspan=1>Zhang B, Chu W, Wei P, Liu Y and Wei T: Xanthohumol induces generation of reactive oxygen species and triggers apoptosis through inhibition of mitochondrial electron transfer chain complex I. Free Radic Biol Med. 89:486–497. 2015.View Article:Google Scholar:PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 3</td><td rowspan=1 colspan=1>Comito G, Calvani M, Giannoni E, Bianchini F, Calorini L, Torre E, Migliore C, Giordano S and Chiarugi P: HIF-1α stabilization by mitochondrial ROS promotes Met-dependent invasive growth and vasculogenic mimicry in melanoma cells. Free Radic Biol Med. 51:893–904. 2011.View Article:Google Scholar:PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 4</td><td rowspan=1 colspan=1>Hu HL, Zhang ZX, Chen CS, Cai C, Zhao JP and Wang X: Effects of mitochondrial potassium channel and membrane potential on hypoxic human pulmonary artery smooth muscle cells. Am J Respir Cell Mol Biol. 42:661–666. 2010.View Article:Google Scholar:PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 5</td><td rowspan=1 colspan=1>Vriend J and Reiter RJ: Melatonin and the von Hippe-Lindau/HIF-1 oxygen sensing mechanism: a review. Biochim Biophys Acta. 1865:176–183. 2016.PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 6</td><td rowspan=1 colspan=1>Dong L, Li Y, Hu H, Shi L, Chen J, Wang B, Chen C, Zhu H, Li Y, Li Q, et al: Potential therapeutic targets for hypoxia-induced pulmonary artery hypertension. J Transl Med. 12:392014.View Article:Google Scholar:PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 7</td><td rowspan=1 colspan=1>Wang X, Ren H, Zhao T, Ma W, Dong J, Zhang S, Xin W, Yang S, Jia L and Hao J: Single nucleotide polymorphism in the microRNA-199a binding site of HIF1A gene is associated with pancreatic ductal adenocarcinoma risk and worse clinical outcomes. Oncotarget. Feb 8–2016.(Epub ahead of print).</td></tr><tr><td rowspan=1 colspan=1> 8</td><td rowspan=1 colspan=1>Guo S, Bai R, Liu W, Zhao A, Zhao Z, Wang Y, Wang Y, Zhao W and Wang W: MicroRNA-210 is upregulated by hypoxia-inducible factor-1α in the stromal cells of giant cell tumors of bone. Mol Med Rep. 12:6185–6192. 2015.PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 9</td><td rowspan=1 colspan=1>Cawley K, Logue SE, Gorman AM, Zeng Q, Patterson J, Gupta S and Samali A: Disruption of microRNA biogenesis confers resistance to ER stress-induced cell death upstream of the mitochondrion. PLoS One. 8:e738702013.View Article:Google Scholar:PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 10</td><td rowspan=1 colspan=1>Cuong DV, Kim N, Joo H, Youm JB, Chung JY, Lee Y, Park WS, Kim E, Park YS and Han J: Subunit composition of ATP-sensitive potassium channels in mitochondria of rat hearts. Mitochondrion. 5:121–133. 2005.View Article:Google Scholar:PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 11</td><td rowspan=1 colspan=1>Jin Y, Xie WP and Wang H: Hypoxic pulmonary hypertension and novel ATP-sensitive potassium channel opener: the new hope on the horizon. Zhongguo Ying Yong Sheng Li Xue Za Zhi. 28:510–523. 2012.(In Chinese).PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 12</td><td rowspan=1 colspan=1>Moschos SA, Spinks K, Williams AE and Lindsay MA: Targeting the lung using siRNA and antisense based oligonucleotides. Curr Pharm Des. 14:3620–3627. 2008.View Article:Google Scholar:PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 13</td><td rowspan=1 colspan=1>Guo Y, Yang T, Lu J, Li S, Wan L, Long D, Li Q, Feng L and Li Y: Rb1 postconditioning attenuates liver warm ischemia-reperfusion injury through ROS-NO-HIF pathway. Life Sci. 88:598–605. 2011.View Article:Google Scholar:PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 14</td><td rowspan=1 colspan=1>Zuo X, Zong F and Wang H, Wang Q, Xie W and Wang H: Iptakalim, a novel ATP-sensitive potassium channel opener, inhibits pulmonary arterial smooth muscle cell proliferation by downregulation of PKC-α. J Biomed Res. 25:392–401. 2011.View Article:Google Scholar:PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 15</td><td rowspan=1 colspan=1>Jin Y, Pang T, Nelin LD, Wang W, Wang Y, Yan J and Zhao C: MKP-1 is a target of miR-210 and mediate the negative regulation of miR-210 inhibitor on hypoxic hPASMC proliferation. Cell Biol Int. 39:113–120. 2015.View Article:Google Scholar:PubMed/NCBI</td></tr><tr><td rowspan=1 colspan=1> 16</td><td rowspan=1 colspan=1>Wang H, Tang Y and Zhang YL: Hypoxic pulmonary hypertension (HPH) and iptakalim, a novel ATP-sensitive potassium channel opener targeting smaller arteries in hypertension. Cardiovasc Drug Rev. 23:293–316. 2005.View Article:Google Scholar:PubMed/NCBI</td></tr></tbody></table>

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For Freedmen's Sake: The Story of the Native Blacks of the Muscogee Nation and Their Fight for Citizenship Post-Mcgirt Excerpted From: Trevion Freeman, For Freedmen's Sake: the Story of the Native Blacks of the Muscogee Nation and Their Fight for Citizenship Post-mcgirt, 57 Tulsa Law Review 513 (Winter, 2022) (425 Footnotes) (Full Document) For Freedmen's Sake: The Story of the Native Blacks of the Muscogee Nation and Their Fight for Citizenship Post-Mcgirt Parent Category: Rights of Indigenous Peoples Category: Tribal Sovereignty Abstract Excerpted From: Trevion Freeman, For Freedmen's Sake: The Story of the Native Blacks of the Muscogee Nation and Their Fight for Citizenship Post-Mcgirt, 57 Tulsa Law Review 513 (Winter, 2022) (425 Footnotes) ( Full Document ) On August 23, 2020, Vanessa Hall-Harper, a councilwoman for the City of Tulsa representing District 1 in Oklahoma, hosted a discussion with Eli Grayson on the Creek Freedmen, “The Forgotten Black People.” Like many events and conversations in 2020, the Zoom discussion was devoted to the subject of McGirt v. Oklahoma, the recent Supreme Court decision, and how its ruling affects Black people in Oklahoma. Grayson, a tribal citizen of the Muscogee (“Creek”) Nation, began his discussion on how he became involved with the Creek Freedmen issue as the former president of the California Muscogee Creek Association. A Racoon Clan and Eufaula Canadian tribal town descendant, Grayson described receiving a phone call from the Creek Chief who explained that he had received a message from a fellow Creek citizen stating “Eli Grayson opened up the California Creek meetings to niggers.” This illuminating scenario describes a vital issue that is a reality for the Freedmen of the great five Sovereign Nations of Eastern Oklahoma: “Oklahoma tribes are exclusive if the hyphenated Indian is Black and inclusive if the hyphenated Indian is white.” In Article 2 of the Treaty of 1866, the treaty rights relevant to the McGirt decision, the Muscogee Nation and the United States Government affirmed that freed Slaves formerly in bondage by Creek Indians were entitled to full citizenship rights as tribal members of the Muscogee Nation. Today, over 155 years have passed since the signing of the Treaty, and the Black descendants of AfricanSlaves of the Muscogee Nation (commonly referred to as Creek Freedmen) should be enjoying the benefits and privileges of their rightful tribal citizenship. However, when a nation (e.g., the Muscogee Nation) “commit[s], in the words of W.E.B. DuBois, 'hideous mistakes' and 'frightful wrongs”’ under the guise of sovereignty without any proper repercussions or redress for the victims, there lies the tragedy of the Creek Freedmen. By extinguishing tribal citizenship of the Creek Freedmen in the decades since the agreement, the Muscogee Nation has failed to honor the Treaty by turning a blind eye to the “Native Black[s]” of the Muscogee Nation. Recently, the Supreme Court in McGirt affirmed that treaty rights granted in the Treaty of 1866 are still intact when it held that Congress never disestablished the Creek reservation under Article III. For the Freedmen of the Muscogee Nation, the reasoning of the McGirt decision was simple. Since the highest court in the land provided a newly illuminated path for discussions of the Treaty of 1866, courts should find that the Freedmen are rightfully entitled to full citizenship under the same Treaty. However, the truth is much more perplexing. Because of Sovereign Immunity and the lack of clarity regarding if tribes can abrogate their own treaties, there may not be a clear legal path for the Creek Freedmen to legally ensure their citizenship rights. This Comment confronts the past and ongoing tragedy of the Creek Freedmen and argues that in light of McGirt (which is premised on the importance of honoring treaty promises) Congress, courts, and Tribal Nation's should therefore honor their solemn promise to the “African Creeks” of the Muscogee Nation under the Treaty of 1866. Part II of this Comment first provides a historical foundation of Federal Indian Law, chiefly highlighting areas relevant to the Freedmen issue. This is principle in understanding the rights and governance of a federally recognized tribe such as the Muscogee Nation. First, this section explains tribal sovereignty by examining federal recognition of Indian Self-Government. It will then define what an “Indian” is by analyzing the varying definitions of the term to understand that an individual can effectively be defined as an “Indian” under a specific tribe but not under federal law. Next, this section defines the scope and limitations of the “Contours of Federal Plenary Power,” including the government's power over Indian Affairs. This part will also analyze and interpret tribal membership. Part II concludes by inspecting the relationship of tribes and the federal government in their tribal treaty rights and assessing the power of tribal sovereignty in membership acceptance, with particular focus on courts' use of Sovereign Immunity to bar suits against the tribe. The courts and Congress have given Creek Freedmen, and others seeking tribal membership, an uphill battle that will always place them on the losing side if they are barred from filing suit. If the Freedmen are barred from filing suit via the federal court system and can only use the tribal courts to seek a remedy, then will they truly get justice? Part III provides a brief history of each of the five major Oklahoma tribes relationships with AfricanSlaves and their descendants, the Freedmen. Chiefly, it assesses the legal controversies between the Freedmen and the respective tribe in their fight for citizenship rights. This provides the opportunity to understand that following the Civil War, the Cherokee, Chickasaw, Choctaw, Seminole, and Muscogee “identit[ies] [were] socially and politically constructed around hegemonic notions of blood, color, race, and culture that permeate[ed] discourses of social belonging” within the Tribes and the United States. An understanding of previous Freedmen litigation lays the foundation for what the Creek Freedmen have to overcome to reach full citizenship within the Tribe. Part IV presents a deeper focus on the Muscogee Creek Freedmen's most recent legal controversies in their fight for inclusion and citizenship. Representative of this fight is Muscogee Creek Indian Freedmen Band, Inc. v. Bernhardt, in which the Supreme Court of the Creek Muscogee held that the Creek Freedmen “failed to produce sufficient evidence that applying for citizenship through the [Muscogee Nation] ... Board of Citizenship, and appealing any adverse decisions through the tribal courts, would be futile.” Federal and tribal court suits in cases such as Muscogee Creek Indian Freedmen Band, Inc. and Ron Graham v. Muscogee (Creek) Nation Citizenship Board present evidence that the Creek Freedmen have to form a clearer stratagem in their uphill battle for citizenship, one void of simple justiciability errors. Part V critiques the Muscogee Nation's use of Sovereign Immunity as a sword to damage the Creek Freedmen's ability to gain citizenship. If tribes such as the Muscogee Nation have the ability to argue Sovereign Immunity under Santa Clara Pueblo v. Martinez, then the Muscogee Nation will always have the upper hand against the Freedmen within the courts. Fundamentally, “racism is fueled toward Freedmen descendants every time [the Creek] dismisses concerns about the nation's action based on sovereignty.” Further, the Muscogee Nation's Constitution of 1979, which mandates that citizenship is determined by blood and excludes the Freedmen, is in violation of Article 2 of the Treaty of 1866. Article 2 provides that former freed Slaves and their descendants are subject to the same rights as “native citizen[s].” Since the enactment of the Constitution in 1979, the question remains: can tribes abrogate their own treaties? Lastly, McGirt is analyzed as an aid to the Creek Freedmen in their fight. [. . .] Declarative cries of “I can't breathe” and “Black Lives Matter” spattered across the days, weeks, and months of the summer of 2020. This international protest for social justice and equality did not just erupt as the nation gripped with horror and anger as Breonna Taylor and George Floyd's Black lives were overtaken by White supremacy. No, it erupted because in every city, there is a proverbial George Floyd or Breonna Taylor, individuals and families who feel in one way or another that they have been let down by the entity which swore to protect them. The idea of broken promises for the AfricanAmerican within the United States is not a new issue. As the Civil War reached its end, Union General William T. Sherman issued Special Field Order No. 15 on January 16, 1865. The order awarded recently freed Slaves a right to forty acres and a mule each. However, this promise was overturned in the fall of 1865, by Lincoln's successor and southern sympathizer Andrew Johnson. Like the Creek Freedmen, it is the idea of broken promises that laid the foundation for the demand of justice, equality, and equity in the summer of 2020. Therefore, it should not be considered ironic that the Supreme Court decision of McGirt fell right upon the heartbeat of this social and civil rights movement. Although laid in legal reasoning, for its holding of the non-disestablishment of the Muscogee Nation, at its core, Justice Gorsuch's opinion is about keeping promises. Speaking for every AfricanAmerican living under the JimCrow rules of the 1960s, Civil Right Activists Fannie Lou Hamer stated in a now-famous speech, “'[i]'ve been tired so long, now I am sick and tired of being sick and tired, and we want a change. We want a change in this society in America because you see, we can no longer ignore the facts.” On February 23, 2021, the Cherokee Nation could no longer ignore the facts. In a unanimous decision, the Tribe's Supreme Court ruled in favor of removing the word “by blood” from all tribal laws, including the Cherokee Nation's Constitution. Like the Creek Freedmen, this decision uplifted the burden which denied descendants of the Cherokee Freedmen the rights and obligations of a Cherokee citizen. According to the Cherokee Nation's Attorney General, Sara Hill, “Cherokee citizens of Freedmen descents are simply this: Cherokee citizens.” U.S. Representative Maxine Waters, in her capacity as chairwoman over the powerful House Committee for Financial Services has also decided to not ignore the facts. On September 7, 2021, Representative Waters introduced H.R. 5195 which provided for the reauthorization of the Native American Housing Assistance and Self Determination Act (“NAHASDA”). A law that grants the “Five Civilized Tribes” of Eastern Oklahoma millions in yearly housing and community development funds could now be withheld if tribes such as the Muscogee Nation fail to comply with their promise to their Freedmen. As a senior member of the Congressional Black Caucus, Representative Waters understands that the liberation of AfricanAmericans in their fight for equity and justice is tied up with the Creek Freedmen issue. Nevertheless, introduction of a house bill such as H.R. 5195 is meaningless if the truth of Representative Waters words are not felt by the entirety of Congress with the passage of such a legislation. After approving the new Constitution for the Cherokee Nation, Secretary of the Interior Deb Haaland stated that the Cherokee Nations “actions demonstrate that Tribal self-governance is the best path forward to resolving internal Tribal conflicts. We encourage other Tribes to take similar steps to meet their moral and legal obligations to the Freedmen.” However, what if the racism is far too great of burden to overcome for the Creek Freedmen? The Muscogee Nation has continued to ignore the facts of their promise. The “sick and tired” Freedmen are still within that proverbial burning building, with the only exits being blocked by Sovereign Immunity and the Creek's unlawful abrogation of its promise of 1866. Ultimately, there is not one clear solution for the Creek Freedmen to receive their rightful citizenship rights. However, the idea of keeping promises which was foundational in McGirt must be the key to any solution for the Creek Freedmen. Thus, for the sake of keeping promises; for the sake of doing what is right; for Freedmen's sake; the Muscogee Nation, Congress, and the courts must uphold the promise of Article II of the Treaty of 1866, giving those who are “Creek to the bone” their citizenship. Trevion Freeman is a Juris Doctor candidate at the University of Tulsa College of law and currently serves as an Articles Editor of Tulsa Law Review.

https://racism.org/articles/citizenship-rights/rights-of/223-tribal-sovereignty/10431-for-freedmen

Falcetta v. State, 06-98-00035-CR - Texas - Case Law - VLEX 892097958 0: [object Object]. 1: [object Object]. 2: [object Object]. 3: [object Object]. 4: [object Object]. 5: [object Object] Falcetta v. State, 06-98-00035-CR <table><tbody><tr><td> Court</td><td> Court of Appeals of Texas</td></tr><tr><td> Writing for the Court</td><td> Before CORNELIUS</td></tr><tr><td> Citation</td><td> 991 S.W.2d 295</td></tr><tr><td> Parties</td><td> Joseph James FALCETTA, Jr., Appellant, v. The STATE of Texas, Appellee.</td></tr><tr><td> Docket Number</td><td> No. 06-98-00035-CR,06-98-00035-CR</td></tr><tr><td> Decision Date</td><td> 06 January 1999</td></tr></tbody></table> Page 295 991 S.W.2d 295 Joseph James FALCETTA, Jr., Appellant, v. The STATE of Texas, Appellee. No. 06-98-00035-CR. Court of Appeals of Texas, Texarkana. Submitted Dec. 9, 1998. Decided Jan. 6, 1999. Page 296 William Brandon Baade, Tyler, for appellant. Edward J. Marty, District Attorney's Office, Tyler, for appellee. Before CORNELIUS, C.J., GRANT and ROSS, JJ. OPINION Opinion by Justice GRANT. Joseph James Falcetta, Jr. appeals his conviction and sentence for aggravated robbery. Falcetta contends he was denied due process and a fair trial under the Fourteenth Amendment to the United States Constitution and Article I, Section 19 of the Texas Constitution by (1) the admission of an audio-videotape that was not properly authenticated and (2) the denial of a mistrial after a juror improperly communicated with a witness. 1 On November 7, 1996, Falcetta boarded a bus traveling from Dallas to the Isle of Capri Casino in Bossier City, Louisiana. While the bus was traveling through Smith County, Texas, a robbery occurred. Falcetta was accused of robbing one of the passengers, Melody Ann Dailey, and was indicted for the offense of aggravated robbery. Trial was to a jury. The jury found him guilty, fined him $5,280.82, and assessed his punishment at forty-four years in the Texas Department of Criminal Justice, Institutional Division. Falcetta contends he was denied due process and a fair trial under the Fourteenth Amendment to the United States Constitution and Article I, Section 19 of the Texas Constitution by admission of a audio-videotape that was not properly authenticated. Falcetta asserts the necessary predicate for the admission of his statement was set forth in Edwards v. State. 2Edwards was overruled by the Court of Criminal Appeals in Angleton v. Page 297 State. 3 The court held that to determine if an audio tape is authenticated, Rule 901 of the Rules of Criminal Evidence is applied instead of Edwards. 4 Both Edwards and Angletoninvolved the proper predicate necessary for the admission of a sound recording which did not involve an oral statement by the accused made as a result of custodial interrogation. While Falcetta contends his rights to due process and a fair trial were violated, the court in Edwards did not address any constitutional (federal or state) requirements for due process or a fair trial when dealing with the admissibility of a tape recording. Due process applies under both the United States and Texas Constitutions when the statement was not freely and voluntarily given. 5Falcetta does not contend his statement was not freely and voluntarily given, he only challenges the predicate necessary for the admission of the audio-videotape. Furthermore, the standard in Edwards has been superseded by the Rules of Evidence. 6 In the present case, Article 38.22 of the Code of Criminal Procedure is controlling. 7The audio-videotape contained the defendant's oral statement confessing to the robbery, which was elicited by the officers during custodial interrogation. In Swann v. State, 8the defendant complained his statement was not admissible without a proper predicate complying with the seven-pronged test set forth in Edwards. 9The court held Article 38.22 provided the procedure for the introduction of statements made by an accused. 10Article 38.22's recording provision is specific to Texas and provides greater protection than the federal constitutional confession provisions. 11Article 38.22 precludes the use of statements from custodial interrogation without compliance with its procedural safeguards. 12For these reasons, we will examine whether the audio-videotape met the requirements for admissibility under Article 38.22. Article 38.22 mandates that an oral statement is not admissible unless (1) an electronic recording, which may include motion picture, videotape, or other visual recording, is made of the statement; (2) prior to the statement but during the recording, the accused is given the warning required in Section 2(a) of the article and the accused knowingly, intelligently, and voluntarily waives any rights set out in the warning; (3) the recording device was capable of making an accurate recording, the operator was competent, and the recording is accurate and has not been altered; (4) all voices on the recording are identified; and (5) not later than the twentieth day before the date of the proceeding, the attorney representing the defendant is provided with a true, complete, and accurate copy of all recordings of the defendant made under this article. 13These requirements are strictly construed with the exceptions Page 298 that only material voices need be identified and the warnings required by Section 2(a) or their fully effective equivalent are admissible. 14 First, Falcetta argues the State did not show that the operator of the machine was competent because Special Agent Dennis Murphy did not know who turned on the audio-video machine (although Murphy turned it off) and he could not testify from personal knowledge about that person's training. Second, Falcetta argues there was no showing that changes, additions, or deletions had not been made because Murphy did not view the original and did not compare the original with the copy he viewed. Third, Falcetta contends the State did not establish the authenticity and correctness of the tape because Murphy viewed a copy of the original, did not compare the copy with the original, and testified that the copy was a fair and accurate representation from his memory rather than from any written notes he had taken on the day Falcetta gave the statement. Contrary to Falcetta's contentions, the record reflects that Murphy did view the original audio-videotape and the copy in their entirety. Last, Falcetta argues the State did not present any evidence on the manner in which the copy or the original was preserved. Murphy testified that the audio-videotape shows Falcetta was given the warnings contained in Section 2(a) and that he knowingly, intelligently, and voluntarily waived his rights prior to giving his statement. Falcetta also signed a written waiver... 7 practice notes Fluellen v. State, 06-01-00132-CR. United States Court of Appeals of Texas March 25, 2003 ...that the final product accurately portrays the interview, then it may be inferred that the operator was competent. Falcetta v. State, 991 S.W.2d 295, 298-99 (Tex.App.-Texarkana 1999, pet. ref'd). Fluellen's contention regarding the authentication of the tape is without Fluellen next contend...... Pina v. State, 06-99-00082-CR United States Court of Appeals of Texas January 17, 2001 ...due process protection, he was not provided with the required greater protection afforded by Article 38.22. See Falcetta v. State, 991 S.W.2d 295, 297 (Tex. App. Texarkana 1999, no pet.). We find that the admission of Detective Warren's testimony about the contents of these statements was T...... Sanchez v. State, No. 03-03-00139-CR (TX 6/30/2005), 03-03-00139-CR United States Supreme Court of Texas June 30, 2005 ...If the final product accurately portrays the interview, then it can be inferred that the operator was competent. Falcetta v. State, 991 S.W.2d 295, 298-99 (Tex. App.-Texarkana 1999, no Sanchez challenged the competency of the operator by pointing to the incorrect time and date shown on the ...... Baez v. State, No. 14-07-00426-CR (Tex. App. 11/18/2008), 14-07-00426-CR. United States Court of Appeals of Texas November 18, 2008 ...is an accurate portrayal of the interview. See Maldonado v. State, 998 S.W.2d 239, 246 n.9 (Tex. Crim. App. 1999); Falcetta v. State, 991 S.W.2d 295, 298 (Tex. App.-Texarkana 1999, pet. ref'd). A person with knowledge of the interview need only testify that the video is an accurate portraya...... 7 cases Fluellen v. State, 06-01-00132-CR. United States Court of Appeals of Texas March 25, 2003 ...that the final product accurately portrays the interview, then it may be inferred that the operator was competent. Falcetta v. State, 991 S.W.2d 295, 298-99 (Tex.App.-Texarkana 1999, pet. ref'd). Fluellen's contention regarding the authentication of the tape is without Fluellen next contend...... Pina v. State, 06-99-00082-CR United States Court of Appeals of Texas January 17, 2001 ...due process protection, he was not provided with the required greater protection afforded by Article 38.22. See Falcetta v. State, 991 S.W.2d 295, 297 (Tex. App. Texarkana 1999, no pet.). We find that the admission of Detective Warren's testimony about the contents of these statements was T...... Sanchez v. State, No. 03-03-00139-CR (TX 6/30/2005), 03-03-00139-CR United States Supreme Court of Texas June 30, 2005 ...If the final product accurately portrays the interview, then it can be inferred that the operator was competent. Falcetta v. State, 991 S.W.2d 295, 298-99 (Tex. App.-Texarkana 1999, no Sanchez challenged the competency of the operator by pointing to the incorrect time and date shown on the ...... Baez v. State, No. 14-07-00426-CR (Tex. App. 11/18/2008), 14-07-00426-CR. United States Court of Appeals of Texas November 18, 2008 ...is an accurate portrayal of the interview. See Maldonado v. State, 998 S.W.2d 239, 246 n.9 (Tex. Crim. App. 1999); Falcetta v. State, 991 S.W.2d 295, 298 (Tex. App.-Texarkana 1999, pet. ref'd). A person with knowledge of the interview need only testify that the video is an accurate portraya......

https://case-law.vlex.com/vid/falcetta-v-state-no-892097958

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https://lasp.colorado.edu/maven/sdc/public/data/sites/site-20180507T025005/anc/eng/usm6/sci_anc_usm617_214_217.drf

'Kobalos' Linux Malware Targets Supercomputers Worldwide A newly identified Linux malware variant dubbed & quot;Kobalos& quot; is targeting high-performance computing clusters and supercomputers running multiple operating Cybercrime,Fraud Management & Cybercrime,Malware as-a-Service 'Kobalos' Linux Malware Targets Supercomputers Worldwide Report: Malware Steals SSH Credentials From High-Performance Computers How 'Kobalos' malware compromises supercomputers and other high-performance systems (Source: ESET) A newly identified Linux malware variant dubbed "Kobalos" is targeting high-performance computing clusters and supercomputers running multiple operating systems, a new report by security firmESETfinds. Kobalos functions primarily as a backdoor and is designed to steal SSH credentials - aka Secure Shell or Secure Socket Shell - a cryptographic network protocol for providing secure remote login, even over unsecured networks. ESET notes Kobalos is "generic malware," but a highly versatile Linux strain and is equipped with a unique infrastructure - most notably, its command-and-control structure. "The fact that any system compromised with Kobalos can be turned into a C&C server of other compromised hosts is quite unique," says Marc-Etienne M. Léveillé, a senior malware researcher with ESET and a co-author of the report. "The code for running such a server is in the malware itself. They are also using other Kobalos-infected hosts to control the backdoor." As part of the investigation, the ESET researchers worked with the security team from the European Organization for Nuclear Research, or CERN. Targeting Academia ESET says that since the campaign began in 2019 and continued through 2020, the developers of Kobalos malware have compromised supercomputers and high-performance clusters belonging to academic researchers as well as private businesses and organizations. These targets include an Asian ISP, a U.S. endpoint security vendor and several privately held servers, among other victimized organizations. The malware targets a wide range of operating systems, including Linux, BSD and Solaris, and Kobalos might also have the ability to compromise supercomputers that run AIX and Windows, the report notes. Analysis According to the ESET report, since the malware contains a broad set of commands that are defined using a single function, it is difficult to detect. "Analyzing Kobalos isn't as trivial as most Linux malware because all of its code is held in a single function that recursively calls itself to perform subtasks," the report notes. "This makes it more challenging to analyze. Additionally, all strings are encrypted so it's more difficult to find the malicious code than when looking at the samples statically." While it's not clear how the initial attack begins, the researchers say that once the malware compromises a supercomputer or high-performance cluster, it will embed itself within the system's OpenSSH server executable. From there, the backdoor code will activate if Kobalos receives a command over a specific TCP port. The researchers also found variants of the Kobalos malware that can act as "middlemen" for other types of command-and-control connections. The malicious code will also turn an infected server into a command-and-control server. "Any server compromised by Kobalos can be turned into a command-and-control server by the operators sending a single command," the report says. "As the command-and-control server IP addresses and ports are hardcoded into the executable, the operators can then generate new Kobalos samples that use this new command-and-control server." In most systems compromised by Kobalos, the SSH client is altered to steal credentials, which could be one way the malware propagates, according to the report. Kobalos will also grant remote access to the file system of a compromised supercomputer, which then gives the attackers the ability to spawn terminal sessions and also allows proxy connections to other Kobalos-infected servers. Other HPC Attacks In May 2020, the University of Edinburgh in the U.K. was forced to cut off research access to its supercomputer called ARCHER after it was infected with Linux malware as part of a wider cryptomining botnet campaign. Following the incidents, victims across Canada, China, the U.S. and parts of Europe reported their high-performance computing labs had been affected by malware (see: Supercomputer Intrusions Trace to Cryptocurrency Miners). In the same month, the security team atEuropean Grid Infrastructure, which coordinates supercomputer research across Europe, said that its members had witnessed similar attacks. The ESET report notes the Kobalos attacks predate these other incidents involving cryptominers and since the Linux malware functions differently, it's likely these malicious campaigns are not connected. API Security Cybercrime Fraud Management & Cybercrime Malware as-a-Service Asokan is a U.K.-based senior correspondent for Information Security Media Group's global news desk. She previously worked with IDG and other publications, reporting on developments in technology, minority rights and education.

https://www.bankinfosecurity.com/kobalos-linux-malware-targets-supercomputers-worldwide-a-15916

(PDF) Temporal Analysis and Forecast of Surface Air Temperature: case study in Colombia PDF | In this work, we study the short-term dynamics of the Surface Air Temperature (SAT) using data obtained from a meteorological station in Bogotá... | Find, read and cite all the research you need on ResearchGate Temporal Analysis and Forecast of Surface Air Temperature: case study in Colombia DOI: 10.22059/POLL.2021.330761.1187 Authors: <here is a image a0696da2915cb0b5-70f25f8b035f540f> Jhoana Romero University of Manitoba <here is a image f2f62089ad6ff61c-d19572ff1996375a> Diego Torres Diego Torres <here is a image f2f62089ad6ff61c-d19572ff1996375a> Manuel Romero Manuel Romero Abstract and Figures In this work, we study the short-term dynamics of the Surface Air Temperature (SAT) using data obtained from a meteorological station in Bogotá from 2009 to 2019 and using time series. The data that we used correspond to the monthly mean of the historical registers of SAT and three pollutants. A descriptive analysis of the data follows. Then, some predictions are obtained from two different approaches: (i) a univariate analysis of SAT through a SARIMA model, which shows a good fit; and (ii) a multivariate analysis of SAT and pollutants using a SVAR model. Suitable transformations were first applied on the original dataset to work with stationary time series. Subsequently, A SARIMA model and a VAR(2) with its associated SVAR model are estimated. Furthermore, we obtain one-year forecasts for the logarithm of SAT in both models. Our forecasts simulate the natural fluctuation of SAT, presenting peaks and valleys in months when SAT is high and low, respectively. The SVAR model allows us to identify certain shocks that affect the instant relationships among variables. These relations were studied by the impulse-response function and the VAR model variance decomposition. Although the statistical methods used in this study are classical, they continue being widely used in the environmental field, presenting god fits, and the results obtained in this study are consistent with environmental theories. <here is a image f6004b4fed4f1e9f-f030d849aca79aff> A Google Maps view of Colombia (left-hand side) and close-up of Bogota (right-hand side) showing the location of the monitoring station Puente Aranda. … <here is a image 5e2fcb26e4a7a31d-022d246ac40b7023> Box-plots for SAT and pollutants. … <here is a image 87a3be07b21aa1af-6fd5ba21b4b559a2> Individual impulse-response diagrams obtained from the SVAR model that show the periodicity of impulses due to the presence of seasons during the year, and the response variables. … <here is a image 47c7e7d2917cb289-c9337c30ab0827f3> FEVD diagrams: the instantaneous influence between variables is observed. … <here is a image d70c666c15331d9f-74b8a42b0cb6ec64> Descriptive information of the variables. … Figures - uploaded by Jhoana Romero Pollution 2022, 8(1): 269- 279 DOI: 10.22059/POLL.2021.330761.1187 RESEARCH PAPER Temporal Analysis and Forecast of Surface Air Temperature: case study in Colombia Jhoana P. Romero – Leiton 1*, Diego Torres 2and Manuel Romero 3 1. Facultad de Ingeniería, Universidad Cesmag, Pasto, Colombia 2 , 3. Fundación Universitaria los Libertadores, Bogotá, Colombia Received: 19.09.2022, Revised: 25.10.2022, Accepted:24.11.2021 ABSTRACT Inthiswork,westudytheshort-termdynamics oftheSurfaceAirTemperature(SAT)usingdata obtained from a meteorological station in Bogotá from 2009 to 2019and using timeseries. The data that weusedcorrespond tothemonthly meanof thehistoricalregisters ofSATand threepollutants. Adescriptive analysisofthe datafollows.Then, somepredictions areobtainedfromtwodifferent approaches: (i) aunivariate analysis ofSAT through aSARIMAmodel, which showsa good fit;and (ii)a multivariateanalysisofSATandpollutantsusingaSVARmodel.Suitabletransformations were first applied on theoriginal dataset to work withstationary time series. Subsequently, A SARIMA model anda VAR(2)with itsassociated SVARmodel areestimated. Furthermore, we obtain one-year forecasts for the logarithm ofSATin bothmodels. Ourforecasts simulate thenatural fluctuation ofSAT,presenting peaksandvalleys inmonthswhen SATishigh andlow,respectively. The SVAR modelallows ustoidentify certainshocks thataffect theinstant relationshipsamong variables. These relations were studied by the impulse-response function and the VAR model variance decomposition.Although thestatisticalmethodsusedinthisstudyareclassical,theycontinuebeing widelyused intheenvironmentalfield,presentinggodfits,andtheresultsobtainedinthisstudyare consistent with environmental theories. Keywords: Time series, Pollutants, SARIMA, SVAR. INTRODUCTION SAT isanimportant variable thathas been studiedina widerangeofenvironmental applicationsincludingvector-bornediseases,bionomics,terrestrialhydrology,biosphere processes, climatechange,among others(Benalietal.,2012). Even,thetemporalchanges of SAThavebeen usedas aprominent indicatorofglobal climatechange (Aghelpouret al., 2019). The spatio-temporalSAT patterns can often be highly variable and complex dueto the heterogeneity of theenvironmental factorsthat controltheenergy balanceof theland- atmosphere system(Benalietal.,2012).Thesefactorsincludethepresenceofcertain atmosphericpollutants.Theatmosphericpollutantsthatarenormallymeasuredintheurban atmosphere basically come fromvehicles usedfor transport, fromstationary sources of combustion (industries)andfromwastedisposalprocesses.Someofthemostrelevant atmospheric pollutantsaresuspendedparticles,sulfur compounds,nitrogen compounds, carbon oxidesandphotochemicaloxidants.However,thepresenceofpollutants inthe atmosphere notonly affectthebehaviourofSATbuttheyalso affect publichealth.Mostair * CorrespondingauthorEmail: [email protected] 270 J. Romero-Leiton et al. pollutantshaveeffects onhuman health,although theireffectsare different.Indeed, some studieshaverevealedthatparticulate matter(PM)canpenetrate therespiratorysystemvia inhalation,causingrespiratoryandcardiovasculardiseases,reproductiveandcentralnervous systemdysfunctions,and cancer(Grivas etal., 2008).Therefore,many researchershave focused on studyingSAT andthe behaviourof atmosphericpollutants toobtain useful informationtomakeforecastsandtakecontrolmeasures.Infact,information onSATand atmosphericpollutant concentrationsusuallycomes fromautomatic continuousmonitoring stations.Thesestationscapturerecordsatdifferentfrequenciesofthepollutant’s concentrationsandtheyalsorecordtheintensityofmeteorologicalvariables.Many mathematical and statistical studies have used these data; for instance, through multiple regression andinterpolationmethods,timeseries methods,geographicalinformationsystems (GIS)techniques,machineanddeeplearningmethods(seee.g.,(AlonsoandRenard,2019, Agbazo et al., 2019, Shen et al., 2020)), among others. In this work, we will focus on time series techniques. Time series models have been widely usedinabroadrangeofscientificapplicationstoforecastvariables,includingclimatology. Among thetimeseriesmodelsfrequentlyusedonenvironmentalstudies,we havethe SeasonalAutoregressiveIntegratedMovingAverage(SARIMA)andtheVector Autoregressive (VAR) models (Aghelpour et al.,2019, Wang & Niu,2009). Thus, the present studyaims toanalysethe individualSAT andthe SATatmosphericpollutants short –term dynamicsusingdatareportedforonemonitoringstationoftheBogot áAirQuality Monitoring Network (RMCAB, by its acronym in Spanish). This data isassessed by means of adescriptiveanalysisandthen itis processedusingan univariateand amultivariate time series models. For the univariatecase, the SARIMA model (Section 2.3.1) isproposed according totheseasonaltrendofSAT.Thismodelshowsagood fitintheerrorsbehaviour anditisbeabletoforecastaccuratemonthlymeanvaluesforlogarithmofSATduringall 2020.Forthemultivariatecase(Section2.3.2),aVARmodelanditsstructuraland unrestrictedformSVARareestimated.Thus,SATisrelatedwithatmosphericpollutantsby meansofarestrictedVAR(2)modelanditsassociatedSVARmodeltodeterminepotential instantaneous relations among thevariables. The unrestrictedform of aVAR model identifies thestructuralimpactsandstorestheminamatrix,whichisobtainedviadata-driven identificationtechniques,suchasChangesinVolatility (CV)(Rigobon,2003).Diagnosisof therestrictedVARmodelhassomefitproblems,butinstantrelationsbetweenendogenous variables identifiedbythe SVARmodelaredetermined. Finally,aforecastfrom VAR(2)for all 2020 is obtained. MATERIALS AND METHODS BogotáisthecapitalofColombiaanditisoneofthelargestcitiesofLatinAmerica(see Figure1). Itis located inthe center ofthe countryat analtitude ofapproximately2600 meters.The populationin Bogotáfor2020 isnow estimatedat10,978,360 andit hasan annual growthrate of2.08 (Lozano,2004). Bogotáalsohas oneof thehighest environmental deterioration rates of the country. Air pollutionhas increased dramatically recently, due mainly tothe uncontrolled increase inthe number ofvehicles in thecity. Althoughair pollution hasbeenmonitored inBogot ásince 1967, itwasnotuntil 1990that themonitoring stations werespread widelythroughout thecity. Ithas sincebeen reportedthat themost important sourcesof pollutionin Bogotá areautomobiles followedby bricksand battery plants,amongothers.TheareaswiththehighestlevelsofatmosphericpollutionarePuente Aranda,Carvajal,andKennedy,whicharemostlyaffectedbyPM 10andPM 2.5. Pollution 2022, 8(1): 269-279 271 Furthermore,in 1990the Secretaryof Healthofthe Districtwiththe collaborationof the JapaneseInternationalCooperation Agency (JICA) identifiedfor thefirsttimethefollowing components ofairpollutioninBogotá:Sulfur Dioxide(SO 2),Nitrogen Oxides(NO x),Total SuspendedParticles(TSP), CarbonMonoxide(CO),Hydrocarbons(HC),andOzone(O 3). Localauthoritiesnowfacethechallengeofsupportingthegrowthanddevelopmentofthe city, while atthe same time minimizing theadverse effects ofthe associatedair pollution and itsconsequencesonhealth.Permanentsurveillanceisanimportanttoolinthisprocess.The RMCABcurrentlyoperates13fixedand1mobilestationthroughoutthecity.TheRMCAB measuresPMusingDASIBI7001(MetOneInstruments,Inc.,UnitedStates)and METONE –BAM1020(MetOne Inc.,USA) βattenuationparticlemonitors forPM 10 and PM 2.5 , respectively. Fig 1: A Google Maps view of Colombia (left –hand side) and close –up of Bogota (right –hand side) showing the location of the monitoring station Puente Aranda. The data analysedin this workcorresponds toPuente Aranda station(see Figure1), which is oneofthe 15stationsthatare part oftheRMCAB thatarespatially distributed throughout thecity. PuenteArandaisoneofthemostrepresentativestationsbecauseitislocatedinthe heartoftheindustrialzoneofthecityanditisofmixedmonitoring(i.e.itcoversbothair qualitydataandmeteorologicalvariables).Hourlyrecordsofseveralpollutantsand meteorologicalparameterswereobtainedfromPuenteArandastationfromJanuary2009to December2019.However,duetothenotableabsenceofrecordsinmanyoftheirvariables, onlythemeasurementofSATandthreeairpollutants:PM 10,O 3andNO 2withcomplete recordsgreaterthan60%weretaken. Abriefdescriptionofthose pollutantsfollows:(a) : PM 10 ,isparticulatematter10micrometersorlessindiameter.Particlesinthissizerange make upalarge proportionofdust thatcanbe drawndeepinto thelungs.The specificeffect of particlesdepends ontheircomposition, concentrationand the presenceofother pollutants, suchasacidforminggases.(b)O 3isahighlyreactivepale-bluegasthatisformedinthe Earth'sloweratmosphere, neargroundlevel. Ozoneis formedwhenpollutants emittedby cars,powerplants,industrialboilers,refineries,chemicalplants,andothersources(usually NO x andNO x ) react chemically inthe presence of daylight UV rays and is usually measured in ppb. (c) NO 2isformed asasub-productfromburningfossilfuelsanditisaprecursorof ozone.Breathing airwith ahighconcentration of NO 2canirritate airwaysin thehuman respiratorysystem. Theseexposures overshort periodscan aggravaterespiratorydiseases, 272 J. Romero-Leiton et al. particularlyasthma, leadingto delicaterespiratory symptoms.Likeozone, NO 2isusually 278 J. Romero-Leiton et al. taking intoaccount thegeographical distribution ofthe informationcollected,which is usuallyhowatmosphericdynamicsarestudied.Thiscouldbeachievedthroughthe implementationofgeostatisticalmethods(GISsystems)withmorethanonemeteorological station data, also considering geomorphology and relief, or the implementationof other meteorological variables or other atmospheric pollutants to the model. ACKNOWLEDGEMENT Theauthors thankto RMCABfor thefree dataavailability. JhoanaP. Romerothanks to Fundación Ceiba,Colombia.D.TorresandM.Romerothank toFundaciónUniversitarialos Libertadores, Colombia. GRANT SUPPORT DETAILS The present research did not receive any financial support. CONFLICT OF INTEREST The authorsdeclare thatthereisnotany conflictofinterests regardingthepublication ofthis manuscript.Inaddition,theethicalissues,includingplagiarism,informedconsent, misconduct,datafabricationand/orfalsification,doublepublicationand/orsubmission,and redundancy has been completely observed by the authors. LIFE SCIENCE REPORTING No life science threat was practiced in this research. REFERENCES Agbazo, M., Koto N’gobi, G., Alamou, E.,Kounouhewa, B., Afouda, A .and Kounkonnou, N. (2019). Multifractalbehaviorsofdailytemperaturetimeseriesobservedover beninsynoptic stations (west africa). Earth Sciences ResearchJournal, 23(4):365 –370. Aghelpour,P.,Mohammadi,B.andBiazar,S.M.(2019).Long-termmonthlyaveragetemperature forecasting in some climate types ofiran, using themodels sarima,SVR, andSVR- FA . Theoretical and Applied Climatology, 138(3-4):1471 –1480. Alonso,L.andRenard,F.(2019).Integratingsatellite –deriveddataasspatialpredictorsinmultiple regression modelsto enhancethe knowledge ofairtemperature patterns. UrbanScience, 3(4):101. Benali, A.,Carvalho, A.,Nunes, J.,Carvalhais, N.and Santos,A. (2012).Estimatingair surface temperature in portugal using modis lst data. Remote Sensing of Environment, 124:108 –121. Box, G.E.and Jenkins,G. M.(1976).Time seriesanalysis: forecastingand controlrevised.Holden – Day. Grivas, G., Chaloulakou, A. andKassomenos, P. (2008). An overview ofthe pm10 pollution problem, in the metropolitan area of athens, greece: assessment of controlling factors and potential impact of long range transport. Science of the Total Environment, 389(1):165 –177. Lozano,N.(2004).AirpollutioninBogotá,colombia:aconcentration –responseapproach.Revista Desarrollo y Sociedad, (54):133 –177. Ninyerola, M., Pons, X. and Roure,J. M. (2000). A methodological approach of climatological modellingofairtemperatureandprecipitationthroughgistechniques.InternationalJournalof Climatology: A Journal of the Royal Meteorological Society, 20(14):1823 –1841. Pollution 2022, 8(1): 269-279 279 Rigobon,R.(2003).Identificationthroughheteroskedasticity.ReviewofEconomicsandStatistics, 85(4):777 – 792. Shen, H.,Jiang, Y., Li,T., Cheng, Q.,Zeng, C. andZhang, L. (2020).Deep learning-based air temperaturemapping byfusing remotesensing,station, simulationand socioeconomicdata. Remote Sensing of Environment, 240:111692. Singh, K.P., Gupta,S. andRai, P.(2013). Identifying pollutionsources andpredicting urbanair quality using ensemble learning methods. Atmospheric Environ- ment, 80:426 –437. Wang, W. and Niu, Z. (2009). Var model of pm2.5, weather and traffic in los angeles –long beach area. In 2009InternationalConferenceon EnvironmentalScienceand InformationApplication Technology, volume 3, pages 66 –69. IEEE. Integrating Satellite-Derived Data as Spatial Predictors in Multiple Regression Models to Enhance the Knowledge of Air Temperature Patterns Sep 2019 <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Alonso Lucille Renard With the phenomenon of urban heat island and thermal discomfort felt in urban areas, exacerbated by climate change, it is necessary to best estimate the air temperature in every part of an area, especially in the context of the on-going rationalization weather stations network. In addition, the comprehension of air temperature patterns is essential for multiple applications in the fields of agriculture, hydrology, land development or public health. Thus, this study proposes to estimate the air temperature from 28 explanatory variables, using multiple linear regressions. The innovation of this study is to integrate variables from remote sensing into the model in addition to the variables traditionally used like the ones from the Land Use Land Cover. The contribution of spectral indices is significant and makes it possible to improve the quality of the prediction model. However, modeling errors are still present. Their locations and magnitudes are analyzed. However, although the results provided by modelling are of good quality in most cases, particularly thanks to the introduction of explanatory variables from remote sensing, this can never replace dense networks of ground-based measurements. Nevertheless, the methodology presented, applicable to any territory and not requiring specific computer resources, can be highly useful in many fields, particularly for urban planners. Long-term monthly average temperature forecasting in some climate types of Iran, using the models SARIMA, SVR, and SVR-FA Nov 2019 THEOR APPL CLIMATOL <here is a image 33d4158ffdcaca86-17686088c2807b10> Pouya Aghelpour Babak Mohammadi <here is a image e1ca6aacd46cb7aa-72b7ea8e10baef22> Seyed Mostafa Biazar Temporal changes of the global surface temperature have been used as a prominent indicator of global climate change; therefore,making dependable forecasts underlies the foundation of sound environmental policies. In this research, the accuracy of theSeasonal Autoregressive Integrated Moving Average (SARIMA) Stochastic model has been compared with the Support VectorRegression (SVR) and its merged type with Firefly optimization algorithm (SVR-FA) as a meta-innovative model, in long-termforecasting of average monthly temperature. For this, 5 stations from different climates of Iran (according to the Extended DeMartonne method) were selected, including Abadan, Anzali, Isfahan, Mashhad, and Tabriz. The data were collected during1951–2011, for training (75%) and testing (25%). After selecting the best models, the average monthly temperature has beenforecasted for the period 2012–2017. The results showed that the models had better performances in Extra-Arid and Warm(Abadan) and after that Extra-Arid and Cold (Isfahan) climate, in long-term forecasting. The weakest performances of the modelswere reported in Semi-Arid and Cold climate, including Mashhad and Tabriz. Also, despite the use of the non-linear SVR modeland its meta-innovative type, SVR-FA, the results showed that, in the climates of Iran, the linear and classical SARIMA modelstill offers a more appropriate performance in temperature long-term forecasting. So that it could forecast the average monthlytemperature of Abadan with root mean square error (RMSE) = 1.027 °C, and Isfahan with RMSE = 1.197 °C for the 6 yearsahead. The SVR and SVR-FA models also had good performances. The results of this checking also report the effectiveness ofthe merging SVR model with the Firefly optimization algorithm in temperature forecasting in Iran’s climates, so, compared withthe SVR model, it is suggested to use SVR-FA for temperature forecasting.

https://www.researchgate.net/publication/357238114_Temporal_Analysis_and_Forecast_of_Surface_Air_Temperature_case_study_in_Colombia

(PDF) 12288 2014 Article 384 PDF | On Sep 11, 2015, Arjun Datt Law and others published 12288 2014 Article 384 | Find, read and cite all the research you need on ResearchGate September 2015 Authors: Arjun Datt Law The Princess Margaret Hospital Subhash Varma Postgraduate Institute of Medical Education and Research Neelam Varma Postgraduate Institute of Medical Education and Research Alka Khadwal Postgraduate Institute of Medical Education and Research Show all 8 authors Hide Download file PDF Read file Download citation Copy link Link copied Read file Download citation Copy link Link copied References (26) Figures (2) Figures Characteristics of patients with eosinophilic vasculitis and thrombotic complications … Healing ulcers on lower extremity (after steroid therapy) … Figures - uploaded by Arjun Datt Law Author content All figure content in this area was uploaded by Arjun Datt Law Content may be subject to copyright. Discover the world's research 25+ million members 160+ million publication pages 2.3+ billion citations Join for free Public Full-text 1 Content uploaded by Arjun Datt Law Author content All content in this area was uploaded by Arjun Datt Law on Sep 11, 2015 Content may be subject to copyright. A preview of the PDF is not available Citations (0) References (26) ResearchGate has not been able to resolve any citations for this publication. Long-term safety of mepolizumab for the treatment of hypereosinophilic syndromes Article Full-text available Oct 2012 Florence Roufosse Jean-Emmanuel Kahn Gerald Joseph Gleich Amy D Klion Background: Hypereosinophilic syndromes (HESs) are chronic disorders that require long-term therapy to suppress eosinophilia and clinical manifestations. Corticosteroids are usually effective, yet many patients become corticosteroid refractory or develop corticosteroid toxicity. Mepolizumab, a humanized monoclonal anti-IL-5 antibody, showed corticosteroid-sparing effects in a double-blind, placebo-controlled study of FIP1L1/PDGFRA-negative, corticosteroid-responsive subjects with HESs. Objective: We evaluated long-term safety and efficacy of mepolizumab (750 mg) in HES. Methods: MHE100901 is an open-label extension study. The primary end point was the frequency of adverse events (AEs). Optimal dosing frequency, corticosteroid-sparing effect of mepolizumab, and development of antimepolizumab antibodies were also explored. Results: Seventy-eight subjects received 1 to 66 mepolizumab infusions each (including mepolizumab infusions received in the placebo-controlled trial). Mean exposure was 251 weeks (range, 4-302 weeks). The most common dosing interval was 9 to 12 weeks. The incidence of AEs was 932 events per 100 subject-years in the first year, declining to 461 events per 100 subject-years after 48 months. Serious AEs, including 1 death, were reported by the investigator as possibly due to mepolizumab in 3 subjects. The median daily prednisone dose decreased from 20.0 to 0 mg in the first 24 weeks. The median average daily dose for all subjects over the course of the study was 1.8 mg. Sixty-two percent of subjects were prednisone free without other HES medications for ≥ 12 consecutive weeks. No neutralizing antibodies were detected. Twenty-four subjects withdrew before study completion for death (n = 4), lack of efficacy (n = 6), or other reasons. Conclusion: Mepolizumab was well tolerated and effective as a long-term corticosteroid-sparing agent in PDGFRA-negative HES. View Show abstract The hypereosinophilic syndrome. Analysis of fourteen cases with review of the literature Article Feb 1975 Michael Joseph Chusid David C. Dale Burton C. West Sheldon M. Wolff Several closely related disease entities make up the idiopathic hypereosinophilic syndrome (HES). The syndrome is manifest by persistent and prolonged eosinophilia with organ damage. A group of 14 patients had hematologic, cardiac, and neurologic abnormalities attributable to this disease. Patient survival and response to chemotherapy was significantly better in this group than in previously reported patients. The etiology of HES remains unknown, as does the mechanism of tissue damage. View Show abstract Major basic protein binding to thrombomodulin potentially contributes to the thrombosis in patients with eosinophilia Article Aug 1995 BRIT J HAEMATOL Harumi Y Mukai Haruhiko Ninomiya Koji Ohtani Tsukasa Abe The contribution of an eosinophil granule protein, major basic protein (MBP), to the pathogenesis of thrombosis seen in patients with eosinophilia was investigated. The sera from eosinophilic patients containing elevated levels of MBP inhibited thrombomodulin (TM) function as a cofactor for the thrombin-catalysed activation of protein C more significantly than those from normal individuals (means 48.5%v 17.4%, respectively). It was suggested that the binding of mature MBP in the sera to TM was electrostatic, because mature MBP (pi 10.9) bound to TM, whereas pro-MBP (pi 6.2) did not. The inhibition of TM cofactor activity by eosinophil granule proteins was mainly attributed to the mature MBP, because MBP-depleted eosinophil granule proteins did not inhibit TM cofactor activity significantly. This inhibition seemed to be due to the specific thrombin-binding to TM being blocked. We concluded that eosinophil granule proteins, particularly MBP, potentially contribute to the hypercoagulation seen in some conditions of eosinophilia, at least because of the inhibition of TM function as a cofactor of the anticoagulation system. View Show abstract Acute pulmonary embolism in a patient with hypereosinophilia and psoriasis Article Nov 2011 Jui-Hung Ko Jheng-Wei Lin Rosaline Hui Peripheral blood hypereosinophilia reflects various underlying disorders. However, its thromboembolic consequences are not often highlighted. We report a case of acute pulmonary embolism in a 42 year-old male prisoner hospitalized for erythrodermic psoriasis who presented with generalized edema and shortness of breath. Severe eosinophilia (absolute eosinophil count 6232/ΜL) was also noted. Although severe psoriasis can be associated with eosinophilia and metabolic syndrome, acute pulmonary embolism is very rare in these patients. Immobilization secondary to severe psoriatic arthritis and prolonged use of leg cuffs can exacerbate the formation of thromboembolism. Although investigating the underlying causes of eosinophilia is important, we would like to highlight the importance of being aware of thromboembolic events, especially in patients with other thrombotic risk factors. View Show abstract Contemporary consensus proposal on criteria and classification of eosinophilic disorders and related syndromes Article Mar 2012 Peter Valent Amy D Klion Hans-Peter Horny Gerald Joseph Gleich Eosinophilia is an important indicator of various neoplastic and nonneoplastic conditions. Depending on the underlying disease and mechanisms, eosinophil infiltration can lead to organ dysfunction, clinical symptoms, or both. During the past 2 decades, several different classifications of eosinophilic disorders and related syndromes have been proposed in various fields of medicine. Although criteria and definitions are, in part, overlapping, no global consensus has been presented to date. The Year 2011 Working Conference on Eosinophil Disorders and Syndromes was organized to update and refine the criteria and definitions for eosinophilic disorders and to merge prior classifications in a contemporary multidisciplinary schema. A panel of experts from the fields of immunology, allergy, hematology, and pathology contributed to this project. The expert group agreed on unifying terminologies and criteria and a classification that delineates various forms of hypereosinophilia, including primary and secondary variants based on specific hematologic and immunologic conditions, and various forms of the hypereosinophilic syndrome. For patients in whom no underlying disease or hypereosinophilic syndrome is found, the term hypereosinophilia of undetermined significance is introduced. The proposed novel criteria, definitions, and terminologies should assist in daily practice, as well as in the preparation and conduct of clinical trials. View Show abstract Treatment of Hypereosinophilic Syndromes-The First 100 Years Article Apr 2012 Joseph H. Butterfield Catherine R Weiler Treatment of the hypereosinophilic syndrome (HES) has advanced rapidly and prevention of end organ damage previously associated with the disorders is now possible in most patients who have had a timely diagnosis. Tried and true medications such as prednisone, hydroxyurea, and interferon-alpha (IFN-aα) continue to play a valuable role in treating HES and their cost is modest. Newer medications included pegylated forms of IFN-aα and IFN-α2b, first- and second-generation tyrosine kinase inhibitors (imatinib mesylate, nilotinib), and monoclonal antibodies to interleukin (IL)-5 and CD52. The combination of better understanding of HES and better medications now provide the clinician with an improved ability to control unregulated proliferation of eosinophils. View Show abstract Eosinophilic vasculitis: An inhabitual and resistant manifestation of a vasculitis Article Nov 2010 Vasa Peter Jandus Nicola Bianda Mario Alerci Claudio Marone A 55-year-old woman was referred because of diffuse pruritic erythematous lesions and an ischemic process of the third finger of her right hand. She was known to have anaemia secondary to hypermenorrhea. She presented six months before admission with a cutaneous infiltration on the left cubital cavity after a paravenous leakage of intravenous iron substitution. She then reported a progressive pruritic erythematous swelling of her left arm and lower extremities and trunk. Skin biopsy of a lesion on the right leg revealed a fibrillar, small-vessel vasculitis containing many eosinophils.Two months later she reported Raynaud symptoms in both hands, with a persistent violaceous coloration of the skin and cold sensation of her third digit of the right hand. A round 1.5 cm well-delimited swelling on the medial site of the left elbow was noted. The third digit of her right hand was cold and of violet colour. Eosinophilia (19 % of total leucocytes) was present. Doppler-duplex arterial examination of the upper extremities showed an occlusion of the cubital artery down to the palmar arcade on the right arm. Selective angiography of the right subclavian and brachial arteries showed diffuse alteration of the blood flow in the cubital artery and hand, with fine collateral circulation in the carpal region. Neither secondary causes of hypereosinophilia nor a myeloproliferative process was found. Considering the skin biopsy results and having excluded other causes of eosinophilia, we assumed the diagnosis of an eosinophilic vasculitis. Treatment with tacrolimus and high dose steroids was started, the latter tapered within 12 months and then stopped, but a dramatic flare-up of the vasculitis with Raynaud phenomenon occurred. A new immunosuppressive approach with steroids and methotrexate was then introduced. This case of aggressive eosinophilic vasculitis is difficult to classify into the usual forms of vasculitis and constitutes a therapeutic challenge given the resistance to current immunosuppressive regimens. View Show abstract Thrombotic eosinophilic vasculitis Article May 2009 CLIN EXP DERMATOL H Nakajima T Nakamura Eosinophilic vasculitis is characterized by pruritic erythema, papules and angio-oedema. We report a case of thrombotic eosinophilic vasculitis with leg oedema and widespread purpura, which resulted in extensive gangrene. Corticosteroids and anticoagulants relieved the symptoms; however, deep ulcers remained after the removal of gangrene. An attempt to decrease corticosteroid dose caused a recurrence of purpura. This case strongly suggests that eosinophilis evoke serious cutaneous vasculitis in the absence of hypereosinophilia. View Show abstract Enhancement of F XII-dependent reactions by eosinophil cationic protein Article Feb 1979 THROMB RES Per Venge Ronald Dahl R Hällgren The cationic protein derived from the granules of human eosinophil granulocytes (ECP) shortened the coagulation time of normal plasma in a time and dose dependent manner. The stimulating effect of ECP could not be brought about in plasma deficient of the factor XII. ECP also enhanced the formation of plasma-kallikrein activity upon glass activation of normal plasma. The lowest effective concentrations of ECP for both reactions were approximately 100 μg/l plasma. The results suggest a role of the eosinophil granulocyte in factor XII-dependent reactions. View Show abstract Cytotoxic Properties of the Eosinophil Major Basic Protein Article Jan 1980 Gerald Joseph Gleich Evangelo Frigas David A. Loegering David Steinmuller The eosinophil major basic protein (MBP) damages cells including those from organs whose dysfunction is associated with eosinophil infiltration. Concentrations of MBP in the toxic range were found in sputa from asthmatics, in pleural fluid and in sera. These findings suggest the hypothesis that the eosinophil produces damage to cells in disease. View Show abstract Show more Linked Research Eosinophilic Vasculitis: Time for Recognition of a New Entity? Article September 2014 Arjun Datt Law · Subhash Varma · Neelam Varma · Alka Khadwal · Pankaj Malhotra Download Recommendations Discover more Article Shame, the Affective Side of Secrets: Commentary on Barth’s Hidden Eating Disorders December 2008 · Clinical Social Work Journal Cathy Siebold This paper expands on the ideas presented by Barth’s article on hidden eating disorders. The topic of eating disorders continues to be an important focus of clinical work. Applying constructs related to the experience of shame, I explore the way that feelings of shame may impact an exploration of eating disorders by the patient and clinician. Read more Chapter Homological Perturbation Theory and Homological Mirror Symmetry Hiroshige Kajiura In this article, we discuss an application of homological perturbation theory (HPT) to homological mirror symmetry (HMS) based on Kontsevich and Soibelman’s proposal [Kontsevich, M., Soibelman, Y. (2001) Homological mirror symmetry and torus fibrations]. After a brief review of Morse theory, Morse homotopy and the corresponding Fukaya categories, we explain the idea of deriving a Fukaya category ... [Show full abstract] from a DG category via HPT, which is expected to give a solution to HMS, and apply it to the cases of \({\mathbb{R}}^{2}\) discussed in [Kajiura, H. (2007) An A ∞ -structure for lines in a plane] and then T 2. A finite dimensional A ∞ -algebra obtained from the Fukaya category on T 2 is also presented. Mathematics Subject Classification: 18G55, 53D37 Read more Article Functional equations with a restricted domain December 1977 · Milan Journal of Mathematics Roman Ger This is a survey of some results on Cauchy type functional equations whose domain of validity is supposed to be restricted in various ways. The behaviour of solutions is discussed, certain methods of solving such functional equations are presented jointly with a number of examples. Applications to the classical functional equation theory are given. Questo articolo è una rassegna di alcuni ... [Show full abstract] risultati riguardanti equazioni funzionali del tipo di Cauchy nelle quali le variabili sono soggette a vari tipi di restrizione. In esso viene discusso il comportamento delle soluzioni e vengono presentati, insieme ad un certo numero di esempi, alcuni metodi di soluzione di tali equazioni; da ultimo sono illustrate delle applicazioni alla teoria delle equazioni funzionali. Read more Article Evidence-based decisions for the treatment of osteoporosis January 2001 E. Field-Munves The author discusses the epidemiology and prevalence of osteoporosis, including male osteoporosis, and presents the evidence-based medicine approch to evaluating therapies. The article also reviews the literature on several pharmacologic and hormone replacement therapies, and discuss the importance of bone mineral density testing and the option of nonpharmacologic therapy. Read more Discover the world's research Join ResearchGate to find the people and research you need to help your work. Join for free ResearchGate iOS App Get it from the App Store now. 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Fruits in the wilderness: A potential of local food resource Download Citation | Fruits in the wilderness: A potential of local food resource | The present study deals with the documentation and future potential of 29 wild edible fruit plants consumed by tribal communities from... | Find, read and cite all the research you need on ResearchGate Article Fruits in the wilderness: A potential of local food resource January 2010 International Journal of Pharma and Bio Sciences1(2) Authors: <here is a image f2f62089ad6ff61c-d19572ff1996375a> B.S. Deshmukh B.S. Deshmukh This person is not on ResearchGate, or hasn't claimed this research yet. <here is a image f2f62089ad6ff61c-d19572ff1996375a> V. Shinde V. Shinde Abstract The present study deals with the documentation and future potential of 29 wild edible fruit plants consumed by tribal communities from Kalsubai-Harishchandragad wild-life Sanctuary fall in Maharashtra state-India. Several plant species have been collected from the study areas which are of high economic food potential. Many plants reported are used in different formulation of 'Ayurveda' in Indian folk-medicine. They provide fibers which prevent constipation. Some examples are, Cordia dichotoma Frost.f.Prodr., Diospyros peregrina (Gaertn.)Gurke, Carrisa congesta Weight., Grewia abutilifolia Vent, Grewia villosa Willd, Rhus sinuata Thunb., Meyna laxiflora Robyns, Securinega leucopyrus (willd.)Muell., Elaeagnus conferta Roxb., Feronia elephuntum Corr., Cucumis setosus Cogn., Capparis zeylanica L. etc. Fruits of these species are edible and having nutritious food value which provides us the minerals like Sodium, Potassium, Iron, Calcium, Magnesium, phosphorus etc. <here is a image 175d1bd4b7439d95-754927124da715c1> ... The unripe fruits act as vegetables and can be eaten pickled as well as cooked. While Ripe fruits can be eaten raw [12] The botanical/taxonomic description and vernacular names of M. spinose are presented in Table 1 and 2, while the images of different parts of M. spinose are presented in Fig. 1. ... ... Fruiting season: August to December Shedding season: The leaves start to shed at the end of autumn and undergo a brief period of hibernation for winter where by January, new shoots start appearing Ecology and cultivation: It grow in very humid as well as hotter climate of Hamito-Semitic region which have acidic to neutral (pH 6.3-7.3) soil [7,[10][11] [12] . ... ... The MIC values of different extracts against multiple gram positive and gram-negative bacteria ranges from 10 to 26 mg/ml and 9-22 mg/ml, respectively. From the Table 10, it can be stated that the stem part of M. spinosa has the least antibacterial effect as compared to the fruit and leaf [12, 23,37]. ... A review on phytochemistry and pharmacology of an unexplored ethnomedicinal plant: Meyna spinosa Roxb. Ex Article Oct 2021 Aheibam Denish Singh <here is a image ab82b6021aeac042-f723c8c7fac7cac1> Ajay Sharma <here is a image fc40cb19c068f733-18939735e6afb37c> Vishal Mutreja <here is a image 82ca7e7e224eac0c-da7083e4b100d852> Garima Bhardwaj The significance of medicinal plants to modern allotropic healthcare is increasingly recognized in every part of the world. A global resurgence in the exploration and identification of plants of restorative significance is the outcome. Since ancient times, people are using the variety of ethnomedicinal plants for their primary health care and Meyna spinose Roxb. Ex. is one of them. It belongs to the genus Meyna and the family Rubiaceae. The plant is spread over a vast area of tropic and sub-tropical zones of the world. It is mainly found in north-east and southern part of India. It is used by the local people in various ethnomedicinal practices ranging from a simple cure for the common cold to diabetic conditions, skin infection, painful urination, hepatic disorder, intestinal worm, dysentery, indigestion, and abortifacient activity. Apart from variety of ethnomedicinal uses, it is also known for its wide range of pharmacological uses such as antimicrobial, antioxidant, cytotoxic, antidiabetic, nephroprotective and hepatoprotective potential. M. spinose also act as a stockroom for innumerable bioactive secondary metabolites (such as triterpenes, phytosterols, flavonoids, fatty alcohol, hydroxy acid etc.) and minerals (like Fe, Cu, Zn, Mo, Mn Cr. Etc.). The aim of present article is to summarize the key findings regarding the folk medicinal uses, phytochemical composition and pharmacological potential of different parts of M. spinosa along with their future prospective. ... Wild fruits can provide an open-access source of food and income, especially to vulnerable groups such as the poor, malnourished children (McGarry and Shackleton, 2009). Some botanical exploration and publications have emphasized the diversity and food value of wild edible fruit plants (Vartak, 1959;Billore & Hemadri, 1969;Maikhuri et al., 2000;Sundriyal, 2001;Bhattacharjee et al., 2008; Deshmukh and Shinde, 2010) . So, from the above-mentioned data, we can say that Purulia is very rich in wild edible fruit-yielding plants and their fruits and byproducts play the important role in the socio-economic upliftment of tribal communities. ... Wild Edible Fruits Used by Tribal and Rural Communities of Purulia District, West Bengal Chapter Full-text available Jun 2023 <here is a image 3d04c5a6f5f5074b-427d0779bf92d442> Asit Mahato <here is a image 430e0d40711bdaaa-d72dbcae79f86e35> Subrata Raha <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Kashinath Katary <here is a image 2675cb16748311ab-e411197a35f2125a> Manoranjan Paramanik ... Conventionally, fruits of E. conferta have shown antitumor, anti-microbial, anti-viral, anti-fungal, antioxidant, antiinflammatory, and anti-diabetic activities. [41,[103][104][105][106] [107] [108][109][110] ... Medicinal and nutritional aspects of wild edible fruits from Western Ghat of India Article Full-text available Jan 2023 Int J Green Pharm <here is a image 78a580a2674fa4a7-9a272026f145f91c> Deepdarshan Urs <here is a image 5e78bd88b0072d76-83bea43a351f4a9c> Sophiya P. <here is a image 59c7fbb74a451790-a2cfb3d94efe55be> M. Narayanappa <here is a image 042c845c07b18cec-1a351878d96f92fa> Dharmappa Kattepura Krishnappa The Western Ghats is a mountain range western coast of the Indian peninsula that covers an area of 160,000 km 2. It is one of the eight biodiversity hotspots in the world. It contains a very large proportion of the country's flora and fauna, many of which are endemic to this region. According to a survey, the Western Ghats are older than the Himalayas and home to many wild edible fruits that are rarely eaten and are traditionally being used as medicine for several ailments. These wild edible fruits provide a variety of nutrients, vitamins and secondary metabolites. Hence, this review, we documented 28 important wild edible fruiting plants from Western Ghat. The most species of wild edible fruit plants belong to the families of Anacardiaceae, Clusiaceae, Malvaceae, Myrtaceae, Phyllanthaceae, Moraceae, Rutaceae, etc. Conventionally, tribal and rural people often rely on wild edible fruits for their food, which could provide primary dietary constituents and natural bioactive compounds. In this study, we have precisely recorded the medicinal importance, nutritional constituents, occurrence, and the biological activities of wild edible fruits prevalent to Western Ghat. ... Elaeagnus fruits contains carotenoids, anthocyanin, carbohydrates, lipid, phytic acid, proteins, ascorbic acid, amino acids, flavonoids, minerals, plant sterols, oxalate, catalase, peroxidase, superoxide dismutase which have been used as anti-tumour, anti-microbial, anti -viral, anti-fungal, antioxidant, anti-inflammatory, anti-diabetic and cytoprotective. Bastard oleaster fruits are known for its antirheumatic, haematinic, analgesic properties [8][9] [10] [11] .Kaempferol is a strong antioxidant which helps in preventing oxidative damage of cells, lipids, and DNA. It is also known to be a chemopreventive agent [12]. ... Prospective reserves of bioactive compounds and their effect on human health: A review on Elaeagnus conferta roxburgh, an edible wild fruit Article Full-text available Dec 2022 S.V.K. Selvakumar P. Rajesh M. Ezhil Inban J. Manikandan Fruits are largely consumed throughout the world; they are not only the sources of energy, but also rich in the sources of nutritive compounds, carotenoids, polyphenols, organic acids etc. Fruits provide an optimal mixture of antioxidants such as vitamins, polyphenols, carotenoids along with complex carbohydrate, protein, fat and fiber. Bioactive compounds are molecules with therapeutic potential and have a positive effect on human health and could reduce the risk of numerous chronic diseases. Bioactive compounds are capable of modulating metabolic processes and possess antioxidant, anti-inflammatory, antifungal, antiviral, antibacterial, anticancer, analgesic and hepatoprotective properties both in vivo and In vitro. Epidemiological information substantiates that higher the consumption of fruits, lowers the risk of chronic diseases and fosters good health in humans. The following is a review on Elaeagnus conferta roxburgh fruits prospective reserves of bioactive compounds and their effect on human health. ... [8] Ripe fruits are used in the preparation of sharbat. [9] The decoction of the whole plant is consumed by the Malay community in Malaysia as a remedy for cancer. [10] The mashed whole plant is added to the palm wine and taken to cure fever. ... ANTI-INFLAMMATORY AND ANTIOXIDANT POTENTIALS OF PHYSALIS MICRANTHA LINK Article Full-text available Jul 2022 Mohammad Iftahaz Bin Habib <here is a image 5615ea826b9d27d4-cd2cfa18699fbc88> Md. Shakhuat Hossain Tusher Mohammed Mashrur Chowdhury <here is a image e1fa5b8132f59c26-a571ce983f14ae60> Muhammad Torequl Islam Many species of Physalis have been used ethnobotanically for treating various ailments in humans worldwide. Ethnobotanically, P. micrantha is used to treat skin infections, diabetes, abdominal pain, inflammations, gonorrhoea, spleen enlargement, ear problems, fever, and cancer. In the present study, the methanol extract of P. micrantha (MEPM) has been evaluated for its in vitro anti-inflammatory and antioxidant activities. The phytoconstituents analysis revealed that the methanol extract of P. micrantha contains reducing sugar, glycosides, tannins, alkaloids, and saponins. The extract showed moderate anti-inflammatory and antioxidant activity in the in vitro test models. Further research is necessary on this hopeful medicinal plant. ... and red-veined, in small groups along with the leafless shoots, in the axils of the spines. Fruits are small, many-seeded, ovoid or sub-globulous (Padhan et. al., 2010). It shows some medicinal properties like hypercholesterolemic, anti-inflammatory, analgesic, antidiabetic, antimicrobial, antihypertensive, antihelmintic, and purgative activities (Deshmukh et. al., 2010 and Bokhad et. al., 2013). Due to these medicinal properties, it is necessary to investigate and compare phytochemical analysis of these plants. ... Comparative Preliminary Phytochemical Analysis Of Capparis Decidua (Forsk.) And Capparis Zeylanica Linn Conference Paper Apr 2022 <here is a image 3eb5e8483b8ccf7d-399faa66a4cc45cd> Ajay B. Jadhao <here is a image e0e019626be00917-2ace2ee2b432e201> Manoj R. Kakpure The present study was aimed to evaluate the phytoconstituents of the leaf extracts of Capparis decidua Forsk. and Capparis zeylanica Linn. plants of an ethno-botanical importance belonging to the family Cappariceae using ethanolic extracts. The result reveals the presence of bioactive compounds (phytoconstituents) such as anthraquinones, terpenoids, flavonoids, saponins, tannins, alkaloids, and cardiac glycosides. Both the Capparis species have an ethno-medicinal importance such as being anti-inflammatory, analgesic, antioxidant, antipyretic and antimicrobial properties. The results showed the dominantly presence of alkaloids, flavonoids, phenols and saponins in both plants. So, further research is necessary to isolate and determine the identity of the bioactive compounds present in this plants species. ... In general information on edibility and therapeutic properties of wild fruits is scanty and data on their nutritional composition is negligible. A number of recent studies have indicated that the dietary use of wild fruits appears in numerous records especially in underdeveloped countries and some botanical studies and publications have emphasized on the diversity and food value of wild edible fruit plants (Reddy et al., 2006, Mishra et al., 2007 Deshmukh and Shinde, 2010) . ... Nutritional and Quality Evaluation of Selected Wild Edible Plants as a Source of Functional Food - PROJECT COMPLETION REPORT Technical Report Full-text available Dec 2020 <here is a image 219899fb56885f6f-535cf439b49fdf07> YOGESH CHANDRA TRIPATHI <here is a image 074652eef36a5aa2-0f48308dfab63091> Rakesh Kumar <here is a image 182ae8a3389a540d-6cfbbf975b32c74b> Praveen Kumar Verma The project was aimed to explore the nutritional status of five wild edible fruits of Garhwal Himalaya by profiling their nutritional and phytochemical characteristic as well as their multifunctional antioxidant activity. Fruits of selected wild plant species namely Rhus parviflora, Rubus ellipticus, R. niveus, Ficus palmata, and Coriaria nepalensis were investigated for their physicochemical attributes (extractable juice content, moisture content, ash values, TSS, and OM), nutritional profile (crude fat, crude protein, total carbohydrate, and crude fibre), ualitative and quantitative phytochemical composition (phenoilic, flavonoid, anthocyanin and ascorbic acid), and multifunctional antioxidant property. The study led to the conclusion that fruits of all the investigated wild fruits contain varying but considerable amount of nutritional constituents and diverse of bioactive phytochemicals. Further, high phenolic content and remarkable antioxidant activity of all the fruits ascertain these wild fruits as better candidates for potential use in functional foods and nutraceuticals. The physicochemical values like moisture content, ash values, etc. and phytochemical profile of all the five fruits have been established to substantiate the standardization data on the plants. The knowledge of the nutritional properties of studied wild fruits may be useful as dietary information. The result of proximate analysis of fruits revealed that these wild edible fruits may be good substitute to cultivated fruits for their substantial nutrient content. The extractive values of fruits in solvents of different polarity have shown that extract obtained with solvents of higher polarity have notable antioxidant capacity which might be associated with the high value of TPC, TFC, TAC and AA recorded in these extracts. In the present study methanol and aqueous methanol has been found as best extractant for most advantageous extraction of phytochemical having immense biological and protective functions. The relationship between total phenolics, flavonoids, anthocyanin and ascorbic acid contents with the antioxidant activities of all the five fruits was established by correlation analysis. Further, higher correlation with values of free radical scavenging assays suggested most likely hydrophilic antioxidant constituents of extracts/fraction contribute to the antioxidant capacity. The edible fruits of Rhus parviflora, Rubus ellipticus, R. niveus, Ficus palmata, and Coriaria nepalensis can be considered a source of important phytochemicals with bioactive properties, thus may be explored for nutraceutical and pharmaceutical applications. Utilization of these wild fruits for production of bioactive extracts/fractions/compounds and their applications in food, cosmetic, pharmaceutical, and chemical industries, could be a practical proposition that could also benefit rural and forest-dependent communities. ... We have also observed that some of the wild fruits are abundantly produced in the forest and are available in urban and rural markets ( (Patkar et al., 2012), Capparis zeylanica (Mishra, 2006), Careya arborea (Ambardar and Aeri, 2013), Phyllanthus emblica (Paria, 2005;Sarkar et al., 2016), Solanum torvum (Paria, 2005), Tamilnadia uliginosa (Sudhakar et al., 2012), Terminalia bellirica (Paria, 2005), Terminalia chebula (Paria, 2005) Some botanical exploration and publications have emphasized the diversity and food value of wild edible fruit plants (Vartak, 1959;Billore & Hemadri, 1969;Maikhuri et al., 2000;Sundriyal, 2001;Bhattacharjee et al., 2008; Deshmukh and Shinde, 2010) . So, from the above-mentioned data, we can say that Purulia is very rich in wild edible fruit-yielding plants and their fruits and byproducts play the important role in the socio-economic upliftment of tribal communities. ... Plant-A valuable resource of Sustainable Agriculture, Food and Medicine Book Nov 2021 <here is a image 5b5c7c12aaab0967-f5a9913c8c2d06af> Dwaipayan Sinha Plants are a remarkable collection of organisms that have ruled the planet for 400 million years. They’ve gone through a series of evolutionary changes to fit in with their surroundings during evolution. These evolutionary changes included not only morphological changes to adapt to different climates, but also sophisticated physiological changes to synchronize with the former and strengthen adaptability. These physiological changes in the plant subsequently proven to be highly beneficial to humans, who appeared between 6 million and 2 million years ago. Plants’ physiological and biochemical evolution, which coincided with the emergence of various taxa, led to the formation of innumerable biochemical pathways that produced a large number of secondary metabolites, the primary goal of which was to protect the plants from herbivores and insects. Secondary metabolites, on the other hand, have shown to be extremely useful to humans since antiquity, who have unintentionally relied on plants for food and medicine since prehistoric times. Humans became aware of the chemical constituents of plants in the last hundred years or so and began researching their varied useful effects. Agricultural activities evolved in tandem with human civilization, and as the population grew, improved yields and crop protection from disease attacks became a requirement. Protection of plants from pathogens rapidly became a priority in the last fifty years and people and initially chemical methods were adopted to protect the plants from pathogen attack which later proved to be disadvantageous from an environmental point of view. Thus the scientific community delved further in search of biological agents which can prevent infection of plant pathogens. This paved the way for biocontrol agents and consequent development of bio fungicides, biopesticides, and bioinsecticides with fewer side effects on humans and animals but with a more green approach towards fertility enhancement. In this book, an attempt is made to emphasize the usage pattern of plants as a source of food and medicine. The book also covers the role of various microbes which can act as pathogen control and also disease management. Thus ‘Plant - A valuable resource of sustainable agriculture, food and medicine’ provides information related to the mentioned theme. The main purpose of the book is to provide relevant information to the readers on aspects largely cantered on plants. The book is divided into five sections namely Phytodiversity, agriculture and sustainable development, natural resource, and usage pattern, microbes and role in agriculture and disease management, plants as sources of medicine, exploration, and conservation of the genetic resource. Selected chapters in relevance to the sections have been accommodated to provide an overview. The first section deals with plant diversity, and agricultural practices in rural Bengal. This section culminates in the impact of urbanization on agriculture which is of high relevance at present. The second section illustrates the usage of indigenous patterns of plants as food and also the conservation of plants from religious aspects through the concept of sacred groves. The third section contains selected chapters on bioprospection of microbes for agricultural sustainability, disease management, and mitigation of stress. The fourth section contains selected chapters on the uses of plants as medicines and herbal excipients and highlights the usage of herbal remedies for the cure of physical ailments. The book concludes with the fifth section containing a chapter on the importance of field gee banks for the conservation of genetic resources of agricultural importance. Within each chapter of the book, various tables provide a clear and methodical description of the issue being explored. The book is an academic endeavor that will assist the scientific community as well as readers with an interest in plant sciences. Publisher link: http://www.absbooksindia.com/shop/art-humanities/agriculture/plant-a-valuable-resource-of-sustainable-agriculture-food-and-medicine ... Ever since ancient times, their fruits, leaves, seeds, even roots and branches have been used to meet personal and social needs such as severing food and beautifying the planet [7,8]. Some of them may also possess medicinal properties, and are therefore used in treatment of ailments as a part of herbal medicine [9, 10]. Crataegus preparations are among the most commonly used herbal medicines worldwide for the treatment of declining cardiac performance according to Stage II of the New York Heart Association (NYHA) [11]. ... The effect of hawthornflower and leaf extract (CrataegusSpp.) on cardiachemostasis and oxidative parameters in Sprague Dawley rats Article Full-text available Jul 2020 <here is a image 308f1305ad861ecd-efb894d73f8a6337> Abeer Rababa'h <here is a image e9d2ea0d2adf7d28-d71703677d75d22d> Omar Al Yacoub Tamam El-Eliamt <here is a image 111d350a58c08f6c-ac0446c741fb2d47> Karem Alzoubi Cardiovascular diseases are described as disorders of heart and vessels that involve stroke and coronary heart diseases. People in the Middle East converged to complementary medicine as an economic alternative to expensive healthcare services. Crataegus monogyna Jacq. (Lindm.) Rosacea is among the most commonly used herb for the treatment of declining cardiac performance, hypertension, and arrhythmias. Previously, we had shown that Crataegus Spp. (Hawthorn) extract increased the tendency of bleeding among patients undergoing coronary artery bypass grafting. Herein, the effects of Crataegus Spp. extract on oxidative stress, cardiac and hematological parameters were evaluated in Sprague Dawley rats. Male rats were randomly assigned into four groups. Group 1 served as control while groups 2–4 served as the experimental groups and were administered extract at doses of 100, 200, and 500 mg/kg. All the doses were given orally once/day and the treatment was continued for three weeks. Hawthorn treatment resulted in a significant decrease in the liver thiobarbituric acid reactive substances level in a dose-dependent manner compared to the control (1.258 (3, 24); P < 0.0001). We found a significant increase in the cardiac antithrombin III among hawthorn treated group compared to the control (4.18 (3, 24); P < 0.0001). On the other hand, hawthorn treatment decreased significantly the liver factor-X level (0.1341 (3, 22); P < 0.0001), while no significant changes were seen in soluble-platelet endothelial cell adhesion molecule-1 (P-value = 0.0599). In conclusions, hawthorn extract possesses an antioxidant effect and blood-thinning properties. Hence, we recommend attention when using this herbal extract with other anticoagulation and/or antiplatelet drugs or undergoing major cardiac surgery. ... Arora and Pandey (1996), reported that there are 647 species of wild edible fruit species under 112 families in India. There are plenty literature available on ethnobotany of wild edible products in different parts of Western Ghats (Vartak 1980(Vartak , 1982Gunjatkar 1985;Bhandary et al. 1995;Alagesaboopathi et al. 1996;Nair and Jayakumar 1999;Patil and Patil 2000;Hebber et al. 2003Hebber et al. , 2010Joseph et al. 2003;Nadanakunjidam 2000Nadanakunjidam , 2003Rajasab and Isaq 2004;Sivakumar and Murugesan 2005;Ignacimuthu et al. 2006;Arinathan et al. 2007;Narayanan and Anilkumar 2007;Narayanan et al. 2011a;Yesodharan and Sujana 2007;Ramachandran et al. 2009;Avdabhai 2009;Anuradha et al. 2010;Bhogaonkar et al. 2010;Hebber et al. 2010;Binu 2010;Bhogaonkar et al. 2010; Deshmukh and Shinde 2010; Paulsamy et al. 2010;Arinathan et al. 2011;Deshmukh and Waghmode 2011;Jadhav et al. 2011;Sasi et al. 2011;Ravikrishna 2011;Valvi et al. 2011;Mallesh Reddy 2012;Dhore et al. 2012;Sarvalingam et al. 2014;Sathyavathi and Janardhanan 2014;Singh and Kumar 2014;Nandini and Shiddamallayya 2014;Satpute 2015: Vijigiri andBembrekar 2015;Patale et al. 2015;Setiya et al. 2016;Prashanth and Shiddamallayya 2016;Niveditha 2017 etc.,) and from these literatures, we compiled 237 wild edible fruits consumed by the different tribal groups in different regions. Fruits are important source of food for the tribal communities and tribals have good knowledge about the sites and seasonal availability of forest products. ... An overview of wild edible fruits of Western Ghats, India Article Full-text available Oct 2020 GENET RESOUR CROP EV Sreekumar VB <here is a image d7d7383c7130a86f-e0d739a69c793769> Sreejith Ashtamoorthy <here is a image a4d079d81f3949f8-58333ce57a5df866> Hareesh V. S. <here is a image 347f966b18e810b7-64c0adb38d31aff7> Sanil M S Fruits are the nature’s gift to mankind, they are not only delicious and refreshing, but also nutritionally rich and supplementing the daily diet of the people. This paper reviews the ethnobotanical knowledge on wild edible fruits traditionally used by the tribal and rural peoples of the Western Ghats, India. There are 237 wild fruiting plants belonging to 136 genera, comes under 65 families used by tribal community for edible purpose, which includes 37 endemic and 11 red listed plants. Among 237 species majority were trees (107), followed by shrubs (75), climbers (42) and herbs (13). The highest numbers of species in the families are Malvaceae, Myrtaceae, Phyllanthaceae, Moraceae, Rutaceae etc. The wild edible plants are integral part of food sources of the tribals and rural peoples and are considered as the oldest food stuff of peoples and provide primary dietary constituents of their daily life. Moreover, it plays a vital role in combating food security also. Most of the wild edible plants have additional usages, some of them have medicinal properties while some others having anti-nutritional factors and are poisonous too. The uses of wild edibles are now mainly confined to tribal or allied community. Scientific research on suitable species is suggested to identify the competent species to replace/improve/substitute the common cultivated species in chemical constitutent, nutritional value and also to explore the possibilities of wild edible plants to provide food security for the growing population. ... Several workers viz. Balkundi (1998) [3] ; Sundriyal and Sundriyal (2004) [37] ; Kulkarni (2006) [22] ; Bhogaonkar et al. (2010) [4] ; Deshmukh and Shinde (2010) [8] ; Dhore et al. (2012) [10] ; Rasingam (2012) [31] ; Deshpande and Kulkarni (2013) [9] ; Rekha and Kumar (2014); Vijigiri et al. (2015) [41] ; Satpute (2015) [34] worked on the wild edibles from various tribal areas in Maharashtra. Earlier work on wild edible plants from Maharashtra like Nasik, Amravari, Buldhana, Kolhapur, Jawhar were carried out by Vartak (1959); Vartak and Kulkarni (1987); Kulkarni and Kumbhoikar (1992), Patil and Patil (2000) [28] , Bhogaonkar et al. (2010) [4] , Kshirsagar et al. (2012) [18] , Mahadkar and Jadhav (2013) [23] , Joshi et al.(2013) [14] . ... Exploration and documentation of wild food plants from Satara district, Maharashtra (India) Research Full-text available Jan 2019 <here is a image d555758366b741d4-b05d96d758eb5e1e> Swapnaja Deshpande <here is a image 8b89978915320fa4-e2c44028f4fd61df> Uday Pawar Rajendra Kumbhar Traditional knowledge throughout the world includes health, nutrition, agriculture, management and utilization of natural resources. In past, tribals consume main staple diet along with wild food supplements which helps to balance their diet. The 21 century is coming with the food scarcity as the global population is increasing day by day. But the available food sources are limited which are not sufficient. The present paper deals with the survey and documentation of food potential of edible plants consumed by people in rural areas from the Satara district. Total ninety plants were recorded belonging to 46 families and 73 genera. The edible part of these species include rhizomes, corms, stem, leaves, petiole, inflorescence, flowers, petal, thalamus, fruits, seeds and aril. ... 2008, Khyade et.al. 2008 , Deshmukh and Shinde 2010 . However there is no concrete report about the nutritional potential of wild edible fruits from the study area. ... INTERNATIONAL JOURNAL OF PHARMACY & LIFE SCIENCES Role of wild edible fruits as a food resource: Traditional knowledge Jul 2011 <here is a image 2555f89bc35462e4-c51ad4aac25ffae6> Babasaheb Deshmukh <here is a image d15615c763527e47-2e012ae93325ba47> Ahilya Vitthal Waghmode The present study deals with the documentation and study of food potential of some traditional wild edible fruits, consumed by tribal communities in Western Ghats region of Maharashtra. Numbers of plant species were documented as wild plants used for food purposes. Among those 11 fruit plant species were investigated for their nutritional food value and medicine from the study area. It has been observed that the traditional knowledge is sharp and valuable. The fruits are rich in minerals and carbohydrates. They provide the minerals like sodium, potassium, magnesium, iron, calcium, phosphorus etc. They can be used as remedy for various diseases. This type of study could contribute to educate the younger generation about the importance of wild fruits. These wild fruit plants can be incorporated in commercial crop plants. This will improve food scarcity, economy in tribal areas and helps in regeneration of barren lands. ... The analysis reveals that out of 187 species, 41.18 % species harvested are consumable fruits while 22.99 % species harvested are leafy shoots. Similar results were also reported from other regions of the Himalaya [13][14] [15] [16] . This indicates the rich gene pool diversity of wild fruits and leaves in the region that confers scopes for further agro-horticultural and biotechnology research in the region by targeting commercially viable wild edible fruit and leaf plants for ensuring sustainable economic and livelihood security of the local residents. ... Ethnobotanical appraisal on wild edible plants used by the Monpa community of Arunachal Pradesh Oct 2017 <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Jambey Tsering Baikuntha Jyoti Gogoi <here is a image 5a84949fbe0a4055-20df52d45507b433> Pallabi Kalita Hui <here is a image 4e22ab93589ca9b0-3a5322ce5d6697f1> Dr HUI Tag The present study based on ethnobotanical exploration deals with such aspects as documentation of traditional uses, diversity, distribution, threat status and marketing potentialities of wild edible plants consumed by the Monpa community of Arunachal Pradesh. A total of 187 wild edible species belonging to 137 genera of 76 families were documented, of which almost 82.88 % were recorded from subtropical to temperate region, 11.23 % from tropical areas and 5.88 % of the alpine meadows. Rosaceae (19 species) was the most dominant family distributed in high Montane slopes and temperate to subtropical forests of Tawang and West Kameng districts. Among genera, Berberis and Rubus were the most widely consumed genera with 06 species each. Fruits (77 species) happen to be the most common edible plant organ getting consumed mostly raw (81.81 %) whereas leaves and shoots are mostly cooked. As many as 48 wild edible species were found to be extensively exploited by the local residents from wild habitat for food, medicines, rituals, traditional paper making, firewood and construction work. Thirty-two species were found to have local market value and 08 species were identified to be facing high degree of threat in the study area. Twenty two plants are reporting for the first time as edible and many other plant uses are novel. ... WEFs are among the most commonly used NTFPs [28,29], and some may also possess medicinal properties, and are therefore used in treatment of ailments [30] [31][32]. WEFs are used for a range of other purposes, such as cosmetics [33], crafts [34,35], fiber [36,37], and fuel [38][39][40]. In the nutrition and pharmaceutical literature, WEFs have been widely studied and recommended as rich sources of antioxidants, minerals, and vitamins [41][42][43]. ... Wild Edible Fruits: A Systematic Review of an Under-Researched Multifunctional NTFP Full-text available May 2019 <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Mallika Sardeshpande <here is a image 0739c6d98b64a574-6155aca5c40ddda4> Charlie Shackleton Wild edible fruits (WEFs) are among the most widely used non-timber forest products (NTFPs), and important sources of nutrition, medicine, and income for their users. In addition to their use as food, WEF species may also yield fiber, fuel, and a range of processed products. Besides forests, WEF species also thrive in diverse environments, such as agroforestry and urban landscapes, deserts, fallows, natural lands, and plantations. Given the multifunctional, ubiquitous nature of WEFs, we conducted a systematic review on the literature specific to WEFs and highlighted links between different domains of the wider knowledge on NTFPs. We found that literature specific to WEFs was limited, and a majority of it reported ethnobotanical and taxonomic descriptions, with relatively few studies on landscape ecology, economics, and conservation of WEFs. Our review identifies priorities and emerging avenues for research and policymaking to promote sustainable WEF management and use, and subsequent biodiversity and habitat conservation. In particular, we recommend that ecosystem services, economic incentives, market innovations, and stakeholder synergies are incorporated into WEF conservation strategies. ... Studies by Ndoye et al. (2004), Shackleton and Shackleton (2005), and Schreckenberg et al. (2006) revealed the important contribution that indigenous fruit trees make towards poverty reduction. In India, the indigenous fruits collected from the wild, play an important role in meeting the food and cash income requirement of the rural poor and the tribals (Deshmukh and Shinde 2010; Mahapatra and Panda 2012). They are good substitutes for exotic fruits which are not easily accessible to a majority of the poor population. ... Marketing of Indigenous Fruits: A Source of Income among Khasi Women of Meghalaya, North East India Jul 2014 <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Ksanbok Makdoh <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Marvellous B. Lynser Kevin Howard M. Pala The present research attempted to document the usage of important marketable indigenous fruits of Meghalaya, India and evaluated their role in addressing livelihood and food needs of the people of the state. Information on common marketable indigenous fruits were collected through informal interviews and market surveys conducted for a period of one year. Ten indigenous fruits were found to be widely sold in Iewduh, one of the largest open markets in the state. These fruits are available during different seasons of the year. Women are mostly involved in marketing and selling of these fruits, enabling them to generate additional cash income. The status of 5 out of 10 indigenous fruit trees were found to be wild, 3 semi-domesticated and 2 domesticated. Five fruits were found to have value addition, which is a way out to reduce the perishability rate of fruits. The study contributed towards knowledge based on indigenous plant resources and recommends measures for conservation and livelihood enhancement through value addition of this hitherto less researched bioresource of north-east India. ... The analysis reveals that out of 187 species, 41.18 % species harvested are consumable fruits while 22.99 % species harvested are leafy shoots. Similar results were also reported from other regions of the Himalaya [13][14] [15] [16] . This indicates the rich gene pool diversity of wild fruits and leaves in the region that confers scopes for further agro-horticultural and biotechnology research in the region by targeting commercially viable wild edible fruit and leaf plants for ensuring sustainable economic and livelihood security of the local residents. ... Ethnobotanical appraisal on wild edible plants used by the Monpa community of Arunachal Pradesh Oct 2017 INDIAN J TRADIT KNOW <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Jambey Tsering Baikuntha Jyoti Gogoi <here is a image 5a84949fbe0a4055-20df52d45507b433> Pallabi Kalita Hui <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Hui Tag The present study based on ethnobotanical exploration deals with such aspects as documentation of traditional uses, diversity, distribution, threat status and marketing potentialities of wild edible plants consumed by the Monpa community of Arunachal Pradesh. A total of 187 wild edible species belonging to 137 genera of 76 families were documented, of which almost 82.88 % were recorded from subtropical to temperate region, 11.23 % from tropical areas and 5.88 % of the alpine meadows. Rosaceae (19 species) was the most dominant family distributed in high Montane slopes and temperate to sub-tropical forests of Tawang and West Kameng districts. Among genera, Berberis and Rubus were the most widely consumed genera with 06 species each. Fruits (77 species) happen to be the most common edible plant organ getting consumed mostly raw (81.81 %) whereas leaves and shoots are mostly cooked. As many as 48 wild edible species were found to be extensively exploited by the local residents from wild habitat for food, medicines, rituals, traditional paper making, firewood and construction work. Thirty-two species were found to have local market value and 08 species were identified to be facing high degree of threat in the study area. Twenty two plants are reporting for the first time as edible and many other plant uses are novel. © 2017, National Institute of Science Communication and Information Resources (NISCAIR). All rights reserved. ... Being a part of a balanced diet, fruits play a vital role in human nutrition by supplying the necessary growth regulating factors such as vitamins, minerals, antioxidants and fiber that are essential for maintaining normal health. Cultivated as well as wild edible fruits have nutritious food value which provides us the minerals like sodium, potassium, iron, calcium, magnesium, phosphorus 1 . ... Role of expansin genes in softening of fruits during fruit ripening Jul 2017 IJPBS AS HA ... Almost all the parts i.e. root, bark, fruits, leaves and seeds are used for different purposes. The root and bark of the plant are bitter and useful as tonic, expectorant, anthelmintic, emmenagogue, analgesic and also used in rheumatism, paralysis, tooth ache, enlarged spleen [8, 9] . In Unani medicine, the decoction of the root bark is prescribed as deobstruent to liver and spleen and as an anthelmintic and anti-inflammatory agent [10] . ... Phytochemical screening and antimicrobial activity of Capparis zeylanica stem extract Oct 2016 Vidyadhara Suryadevara Triveni Jasti Likhitha Sai Krothapalli <here is a image b25ae48d96b4100b-f73ad0b8e6d3a2ee> Vijetha Pendyala Medicinal plants are alternative rich source of antibacterial agents. The present study was an attempt to isolate the phytoconstituents present in the ethyl acetate fraction of Capparis zeylanica and to assess in vitro antimicrobial activity of different fractions of Capparis zeylanica. Capparis zeylanica belongs to Capparidaceae, found useful in various system of medicine. It possesses numerous pharmacological and phytochemical properties. The stems are shade dried powdered and subjected to extraction and fractionation. The fractions were subjected to column chromatography followed by isolation and characterisation of active components using TLC and spectrophotometric methods. The present research lead to the separation of β-sitosterol, α-amyrin and kaempferol a flavonoid derivative were identified and characterized by physical methods. The methanol, ethyl acetate, hexane and hydroalcoholic extracts of stems of Capparis zeylanica were evaluated for antimicrobial activity against gram+ve, gram-ve bacteria and fungi. Zone of inhibition diameters were observed. This activity was evaluated using disc diffusion method with a standard antibiotic and anti-fungal drug, penicillin and fluconazole. The antibacterial activity was not present in hexane fraction. Methanolic extract showed moderately good activity. However, highest antibacterial activity was found in Ethyl acetate fraction. The anti-fungal activity of the extract and fractions was moderately active compared to standard. ... Capparis zeylanica Linn. is commonly known as Indian caper; a climbing Scandant shrub and found throughout India 1, 2 . Capparis zeylanica Linn is belonging to the family Capparidaceae 3 . ... PHYTOCHEMISTRY AND PHARMACOLOGICAL ACTIVITIES OF CAPPARIS ZEYLANICA: AN OVERVIEW Lather Amit <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Amrendra Kumar Chaudhary Kumar M Pharm Student Capparis zeylanica Linn. (Capparidaceae) has been used as a 'Rasayana' drug in the traditional Ayuevedic system of medicines. Capparis zeylanica Linn. is reported to posses antioxidant, antipyretic, analgesic, anti-inflammatory, antimicrobial and immunostimulant activity. Phytochemical screening of the plant has shown the presence of fatty acids, flavonoids, tannins, alkaloids, E-octadec-7-en-5-ynoic acid, saponins glycosides, terpenoids, saponin, p-hydroxybenzoic, syringic, vanillic, ferulic and p-coumanic acid. This review explains the evidence based information regarding the phytoconstituents and pharmacological activities of Capparis zeylanica plant which helps the researcher to investigate more about this important rasayana. ... This is a substantial number and comparable to those documented from other Indian states, e.g. Tripura (Sankaran et al. 2006), Maharashtra (Deshmukh and Shinde 2010) , Western Himalayas (Parmar and Kaushal 1982), Western Ghat of Tamil Nadu (Arinathan et al. 2007), Andhra Pradesh (Reddy et al. 2007a,b) and the Andaman & Nicobar Islands (Singh et al. 2001). The fact that such varieties of woodland fruits are used for consumption and sale indicates the extent of traditional ecological knowledge of the tribal communities living in the region. ... Wild edible fruit diversity and its significance in the livelihood of indigenous tribals: Evidence from eastern India. Food Security 4 (2): 219-234. <here is a image 2ca7adbd4021baa8-e9e32746fca94027> Ajay Mahapatra <here is a image a3f475b005fde003-445f4cb37ba0c5f6> Pratap Chandra Panda A number of wild plants, used by rural and tribal populations and contributing significantly to their livelihood and food security have escaped recognition and scientific inquiry. Their distribution, conservation, mode of harvest by locals and optimal use require region-specific assessment in order to integrate them into developmental interventions. This study analyzed the collection, consumption, sale and income from edible forest fruits in 49 tribal villages spread over five districts of Orissa State in eastern India. Density, dominance and diversity of species yielding wild fruit were measured by studying ecological parameters in the sample plots. We estimated an average of 48 fruit plants per hectare of deciduous forests. Fifty-six wild edible fruit species belonging to 40 genera in 26 families were recorded in the study region, many of which have multiple uses. Indigenous fruits formed part of the family diet with average annual consumption of 73 kg per household. Sale of wild fruits contributed 15 % of income for tribal households. Despite their good knowledge of indigenous fruits, the tribal populations have not adopted fruit tree farming which would enhance their nutrition and income. Traditional herbal plant useful for treatment of inflammatory bowel disease Anju Balkrishna Bhandole Harish N. Kakrani As per WHO Traditional medicine is the sum total of the knowledge, skill, and practices based on the theories, beliefs, and experiences, used in the maintenance of health as well as in the prevention, diagnosis, improvement or treatment of illness. World Health Organization define Traditional herbal medicines as naturally occurring, plant-derived substances with minimal or no industrial processing that have been used to treat illness within local or regional healing practices. Meyna is a genus of shrub and the plant is Small or medium size tree, Bark light black, smooth. Leaves opposite or whorled distributed in tropical and subtropical region. Earlier Vangueria spinosa covers a group of plants from Meyna genus recently the plants has been separated and classified into eleven different species of Meyna with the help of molecular phylogenetics The plant was reported to be used for treatment of diseases of digestive system, anti- inflammatory ,antioxidant, anti-dysentery and constipation .It also exhibits anti-fungal and anti-bacterial activities which indicate its usefulness as a promising drug in treatment of IBD. Bioautography and liquid chromatography‒mass spectrometry studies of Meyna spinosa Roxb. ex Link leaf methanolic extracts <here is a image 445795231a938424-3520d79fdbd48215> Sharon Kadirvelu Sagarika Damle A. Pillai Meyna spinosa Roxb. ex Link, locally known as “Haloo”, was sampled from Surgana, Nasik, and is reported to have many ethnobotanical uses. This study aims at analyzing and isolating the bioactive components responsible for the antimicrobial activity of the plant’s leaf methanolic extracts. Antimicrobial activity was studied using ditch-plate technique and bioautography. Ditch-plate method showed antimicrobial activity against Staphylococcus aureus. Bioautography in which thin-layer chromatography (TLC) is coupled to bioassay technique showed antimicrobial activity at the application spot against Pseudomonas aeruginosa. The compound/s were collected and concentrated by preparative TLC. The concentrated bioautography isolate (BI) and the leaf methanolic extract (ML) of M. spinosa Roxb. ex Link were subjected to liquid chromatography (LC) followed by electrospray ionization‒quadrupole-time-of-flight‒mass spectrometry (ESI‒Q-TOF‒MS), to compare the bioactive components. It was found that BI showed 10-deoxymethymycin, dihydrodeoxystreptomycin, 5E,8E,11E-hexadecatrienoic acid, deferoxime that have antibacterial properties. ML in addition shows quercetin, kaempferol, 3,4,5-trihydroxystilbene, swietenine, which have antioxidant and antimicrobial activities. We conclude that the antimicrobial activity exhibited by the ML was due to the synergistic action of all these compounds. The minimum inhibitory concentration (MIC) of ML by microdilution revealed to be 375 mg/mL for S. aureus and 125 mg/mL for P. aeruginosa. Documentation of wild edible fruits (WEFs) from mon District of Nagaland, India <here is a image 6a96049a4fe4e467-eebe4c2c262e366f> Zenwang Konyak E Phongle Konyak Mon District of Nagaland, India has tremendous potential in providing valuable Wild edible Fruits (WEFs) and very few works has been done on it. This paper deals with documentation of 65 species of WEFs belonging to 36 families and 51 genera. An attempt has been made to record the use of fruits and its treatment in various ailments. Besides medicinal values it also has high market values. Grewia Species: Diversity, Distribution, Traditional Knowledge and Utilization Dec 2019 Rahul Dev <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Venkatesan Kannan M. Suresh Kumar <here is a image 65ef56df7a1ae88e-080de77e74b0a71f> Rohitkumar Patel Grewia species (Tiliaceae) are medium-size shrub to small tree species and have multipurpose uses. Their fresh and dry fruits are utilized as food, leaves as forage for livestock and plant parts like fruit pulp and root bark are used to cure various ethnomedicinal and ethnoveterinary purposes. Due to their drought and salt tolerance ability, these species are abundantly distributed in arid and semiarid regions of world including India. Though they have multi-purpose utilization including medicinal values, only limited/scanty information are available on public domain on the aspects of systematic utilization and domestication. In this Chapter, the detailed informations about different Grewia species on the aspects of taxonomy, botanical description, distribution and diversity, traditional knowledge and its utilization potential were provided as ready reference in future use to different stakeholders like researchers, farmers and others. Exploration and documentation of some wild edible plants used by the aboriginals from Gadchiroli District (M.S.) India <here is a image 6783beca45914e2f-e968979550548876> Amit Setiya <here is a image 26744038a01c47ff-ab90d4c2150344d5> Nitin Dongarwar <here is a image 34030c1b6acfbcaa-151ba5af397650e1> Sushama Narkhede The present study deals with the exploration, identification, documentation and ethnobotanical aspects with respect to food value of wild edible plants consumed by aboriginals from Gadchiroli District of Maharashtra StateIndia. The Forest adjoin is of dry deciduous to moist deciduous type. The prominent tribes in Gadchiroli District are Raj Gond, Gond, Madia, Pardhan and Kanwar, etc. and other than these tribes the large population is scheduled caste and other backward cast and nomadic tribes which are dwelling on this land since ancient time and no such investigation has been undertaken in this District. Investigation results in 61 plant species of 40 families consumed as food during various situations. Forest Foods of northern region of Western Ghats <here is a image cb766f71b021d5f3-09b86e6a174dd805> Mandar N. Datar <here is a image c4fa3fbd022b31a1-d13add17d1e02a3e> Upadhye Anuradha The book gives information about 142 wild edible species consumed by local inhabitants of northern region of Western Ghats A Comprehensive Review on Meyna laxiflora Robyns (Rubiaceae) <here is a image 1937ee36a50dd27a-fa33d150fae83bfe> Molvi Khurshid Quazi Majaz A Meyna laxiflora is a small or medium size tree, which having traditional importance for its medical uses, for treatment of Inflammation, Gastrointestinal disorder etc. It is distributed in tropical and subtropical region all over the world. In India it is available in Assam, Bengal, Konkan, Deccan etc and wildly available throughout Satpuda region mainly in Sawarimal, Umarpata, Morkaranja and kondaibari villages of Nandurbar district. This review presents detailed survey of literature on phytochemical and medicinal properties of the plant which will be beneficial for exploring its phytochemical and pharmacological properties. The chemicals reported from the plant belong to different classes such as carbohydrates, starch, proteins, tannins, saponins and alkaloids. Thus, indicating the potential of the plant in pharmaceuticals and in indigenous systems of medicine. The notable pharmacological properties include anti-diabetic, antioxidant, antibacterial, antifungal and many more activities which are yet to be explored. Wild edible plants of Chandrapur district, Maharashtra, India This paper is a synthesis of local traditional knowledge of plants used as medicine and western biological science. In many areas women are responsible for curing their family members and others for various illness. Ethnomedical uses of 51 plants by women in hamlets/villages of Jawhar and Mokhada talukas in Thane district, Maharashtra, have been recorded. Information from scientific literature for each plant collected has been incorporated with traditional knowledge to explain or substantiate traditional medicinal uses.

https://www.researchgate.net/publication/289714516_Fruits_in_the_wilderness_A_potential_of_local_food_resource

Should I Exercise During the Coronavirus Pandemic? Experts Explain the Just Right Exercise Curve - EcoWatch So here we are, perfecting our social distancing skills while schools, sports and other forms of social engagement are on indefinite hold, by a dangerous virus named after a (regal) crown. The coronavirus is named because the center envelope is surrounded by small protein spikes called peplomers. These little protein spikes... Should I Exercise During the Coronavirus Pandemic? Experts Explain the Just Right Exercise Curve Health + Wellness Pexels By Tamara Hew-Butler and Mariane Fahlman So here we are, perfecting our social distancing skills while schools, sports and other forms of social engagement are on indefinite hold, by a dangerous virus named after a (regal) crown. The coronavirus is named because the center envelope is surrounded by small protein spikes called peplomers. These little protein spikes wreak havoc when they attach to lung tissue and hijack otherwise healthy tissue into building a potentially lethal coronavirus army of invaders. Because the virus settles primarily with the respiratory tract — the nose, mouth and lungs — it is highly contagious when people sneeze, cough or exchange respiratory droplets with others. Despite its importance,social distancinghas been a social disappointment for many weekend warriors, team sport athletes, fitness fanatics and sports fans who find camaraderie, biochemical joy from dopamine rushes or stress reduction through regular exercise and sport. Wearebothsports scientists who study athlete health and safety. We’re also proud exercise addicts who find the prospect of not exercising almost as disturbing as the prospect of the disease itself. Here’s how exercise affects the immune system in response to the flu and some practical tips on how much people should (and should not) exercise. Look for the ‘Just Right’ Amount Both too much and too little are bad while somewhere in the middle is just right. Scientists commonly refer to this statistical phenomenon as a “J-shaped” curve. Research has shown exercise caninfluence the body’s immune system. Exercise immunity refers to both the systemic (whole body cellular response) and mucosal (mucous lining of the respiratory tract) response to an infectious agent, which follows this J-shaped curve. A largestudyshowed that mild to moderate exercise — performed about three times a week — reduced the risk of dying during the Hong Kong flu outbreak in 1998. The Hong Kongstudywas performed on 24,656 Chinese adults who died during this outbreak. This study showed that people who did no exercise at all or too much exercise — over five days of exercise per week — were at greatest risk of dying compared with people who exercised moderately. Additionally, studies performed on mice demonstrated that regular exercise performed two to three months prior to an infection reducedillness severityand viral load inobese and non-obese mice. Thus, limited animal and human data cautiously suggest that exercise up to three days per week, two to three months prior, better prepares the immune system to fight a viral infection. What if we have not exercised regularly? Will restarting an exercise routine be good or bad? Limited data, also obtained from mice, suggests thatmoderate exercisefor 20 to 30 minutes a day after being infected with the influenza virus improves the chances of surviving. In fact, 82 percent of the mice who exercised 20-30 minutes a day during the incubation period, or the time between getting infected with flu and showing symptoms, survived. In contrast, only 43 percent of the sedentary mice and 30 percent of the mice who performed strenuous exercise — or 2.5 hours of exercise a day — survived. Therefore, at least in laboratory mice, mild to moderate exercise may also be protective after we get infected with the flu virus, whereas a little exercise is good while no exercise — or even too much exercise — is bad. For those who are “committed exercisers,” how much exercise is probably too much during a flu pandemic? It is clear that bothtoo much exerciseandexercising while sickincreases the risk of medical complications and dying. We conducted studies on both collegiatefootball playersandcross-countryrunners, which showed a decrease in secretory immunoglobulin A, or “sIgA” when athletes competed and trained hard. SIgA is an antibody protein used by the immune system to neutralize pathogens, including viruses. SIgA is also closely associated with upper respiratory tract infections (URTI). When sIgA levels go down, URTI’s usually go up. We saw this relationship in football players, whereas the players showed the most URTI symptoms when their sIgA levels were lowest. This indirectly suggests that over-exercise without adequate recovery may make our body more vulnerable to attack, especially by respiratory viruses. So, when it comes to immunity, our studies show that more exercise is not necessarily better. Here are some guidelines based on just the right amount — for most people. Do perform mild to moderate exercise (20-45 minutes), up to three times per week. Strive to maintain (not gain) strength or fitness during the quarantine period. Do avoid physical contact during exercise, such as playing team sports, that is likely to expose you to mucosal fluids or hand-to-face contact. Wash and disinfect equipment after use. If you use a gym, find one that is adequately ventilated and exercise away from others to avoid droplets. Remain engaged with teammates through social media, rather than social gatherings or contact. Eat and sleep well to boost your immune system. Remain optimistic that this too shall pass. How Much Exercise May Be Too Risky? Here are somethings not to do: Do not exercise past exhaustion, which increases the risk of infection. An example would include marathon running, which increases the risk of illness from 2.2 percent to 13 percent after the race. Do not exercise if you have any flu-like symptoms. Do not exercise more than five days a week. Do not exercise in crowded, enclosed spaces. Do not share drinks or eating utensils. Do not overdrink fluids, especially when sick, to try and “flush out” the toxins or preventdehydration. It is not true that you can “flush out” toxins. The J-shaped (“just right”) curve suggests that exercise, like most things, is best in moderation. Stay safe out there and be creative — our game is not over, just temporarily suspended. coronavirus covid-19 health public health wellness

https://www.ecowatch.com/exercise-coronavirus-pandemic-2645566737.html

(PDF) BIOLOGICAL ACTIVITY OF EXTRACTS AND ESSENTIAL OILS OF TWO ERYNGIUM (APIACEAE) SPECIES FROM THE BALKAN PENINSULA PDF | The present study describes antioxidant and antimicrobial activity of water, methanol, acetone and ethyl acetate extracts from Eryngium maritimum... | Find, read and cite all the research you need on ResearchGate BIOLOGICAL ACTIVITY OF EXTRACTS AND ESSENTIAL OILS OF TWO ERYNGIUM (APIACEAE) SPECIES FROM THE BALKAN PENINSULA November 2019 Acta Medica Medianae 58(3):24-31 DOI: 10.5633/amm.2019.0304 Authors: Jelena S. Matejić University of Niš Zorica Stojanović-Radić University of Niš Zoran Dj. Krivošej Zoran Dj. Krivošej Bojan Zlatković University of Niš Abstract and Figures The present study describes antioxidant and antimicrobial activity of water, methanol, acetone and ethyl acetate extracts from Eryngium maritimum L. from Greece and Eryngium serbicum Pančić, growing wild in Serbia. Also, antimicrobial activity of essential oils from aerial parts was analysed. Spectrophotometric methods were used for measuring of total phenols, total flavonoids, as well as for antioxidant potential, using DPPH and ABTS methods. The total phenolic content in the extracts was determined using Folin-Ciocalteu reagent and their amounts ranged between 7.47 and 121.35 mg GAE/g. The concentrations of flavonoids in the extracts varied from 8.98 to 48.68 mg QU/g. Antioxidant activity ranged from 1.247 to 31.19 IC50 (mg/ml) and from 0.109 to 3.36 mg AA/g when tested with the DPPH and ABTS reagents, respectively. The antimicrobial activity of the extracts and essential oils was investigated using a micro well-dilution assay against the most common human gastrointestinal pathogenic bacterial strains. The most resistant bacterium was Srteptococcus pyogenes, while Staphylococcus aureus showed high sensitivity in presence of all tested extracts exept on water extract of E. maritimum. Essential oil of E. serbicum showed better antimicrobial activity than E. maritimum oil. This finding suggests that investigated Eryngium species may be considered as a natural source of antioxidant and antimicrobial agents. Acta Medica Medianae 2019;58(3):24-31. The extracts yield for investigated Eryngium species … Comparative antimicrobial activity of extracts and essential oils from two Eryngium species … Figures - uploaded by Jelena S. Matejić Author content All figure content in this area was uploaded by Jelena S. Matejić Discover the world's research Content uploaded by Jelena S. Matejić 24 www.medfak.ni.ac.rs/amm Original article UDC: 615.322:582.794.1 doi:10.5633/amm.2019 .0 304 BIOL OGIC AL AC TIVI TY OF EXT RACTS AND ESSEN TIAL OILS OF TWO ERYN GIUM ( API ACEAE ) SP ECIES FRO M THE BALK AN P ENIN SULA Jelena S. Matejić 1 , Zorica Z. Stojanović - Radić 2 , Zoran Dj. Krivošej 3 , B ojan K. Zlatković 2 , Petar D. Marin 4 , Ana M. Džamić 4 Th e pres en t stu dy de scr ibe s ant iox ida nt an d ant imi cr obi al ac tivi ty of wat er, meth ano l, ace to ne and ethy l aceta te ext rac ts from Ery ngi um mari ti mum L. from Gre ece an d Ery ngi um ser bi cum Pan či ć, gr owi ng w il d in Se rbia . Al so, ant imi cro bia l ac tiv ity of es sen tia l oi ls fr om aeri al par ts was anal yse d. Spec tro pho to met ric meth od s wer e used for measu ri ng of tot al phen ols , tot al flav ono ids , as we ll as for an ti oxi dan t pot ent ial , usi ng DP PH an d AB TS me tho ds. The tota l phe no lic con ten t i n t he ext rac ts was de te rmi ned usi ng Fol in- Cio cal teu re ag ent an d t hei r am oun ts ra nge d bet we en 7. 47 an d 12 1.3 5 mg GA E/ g. Th e con cen tr ati ons of fla von oid s in the ext ra cts vari ed fr om 8. 98 to 48.6 8 mg QU /g. Anti oxi da nt ac tiv ity rang ed fr om 1.247 to 31. 19 IC 50 (m g/m l) a nd fro m 0. 10 9 to 3.3 6 m g AA /g w hen te ste d wi th the DPP H an d A BTS rea ge nts , res pe cti vel y. Th e an tim icr obi al acti vit y of the ex tra ct s and ess ent ial oils was inve st igat ed usin g a mic ro we ll- dilu ti on ass ay again st th e most comm on hu man gast roi nte sti nal path oge nic bac- te rial st rain s. The mos t re sis tan t ba ct eri um w as S rte pto coc cu s pyo ge nes , wh ile Sta phy loc oc cus au reu s show ed hig h sens iti vi ty in pre sen ce of all teste d ext rac ts exe pt on water extra ct of E. ma riti mu m. E sse nti al o il o f E. ser bic um s how ed b et ter anti mi cro bia l ac tiv ity tha n E. mar iti mu m oil . This fin din g sugg est s that inv est ig ate d Eryng iu m speci es may be con sid ere d as a nat ura l sou rc e o f a nti oxi dan t and an tim icr obi al ag ent s. Acta M edica Me dianae 201 9 ;5 8(3 ): 24 - 31 . Key wor ds: Eryn gium ma ritimum , E. se rbicum, extract s, essen tial oil s, antio xidant, antimi crobial activit y 1 University of Niš, Faculty of Medicine, Departme nt of Physics and Biology with Human G enetics, Ni š, Serbia 2 University of Niš, Faculty of Science and Mathe mati cs, Department of Biology and Ecology, Niš, Serb ia 3 University of Priština, Faculty of Natural Science s, Department of Biology, Kosovska Mitrovica, Serbia 4 University of Belgrade, Faculty of Biology, Inst itute of Botany and Botanical Garden “Jevremovac”, B e lgrade, Serbia Contact: Jelena S. Matejić Blvd dr Zorana Đinđića 81 , 18000 N iš , Serbia E-mail: [email protected] Introduc tion The gen us Eryng ium , be lon gin g to the sub - fam ily Sa nicu lo idea e o f th e A pia cea e fa mi ly, is r epr e- sen ted by 317 accep ted taxa worl dwi de, known by thi er hig h cont ent of ace tyl ene s, flav on oids , coum a- rin s and tri ter pen e sapon ins (1). Amon g them , sev- era l Eryn giu m spec ies have bee n used as ornam en- tal pla nts , condi men ts (2) or in tra diti ona l medic ine (3, 4) . Ery ng ium m ar iti mum L. ( se a ho lly ) is a p ere n- nia l pl an t (3 0-6 0 cm hig h) with ma uve flo wer s gr ow- ing wil d o n th e s and y b each es of W es t Eu rop e, Med i- ter ran ean basi n and Bl ack Sea (5) . You ng ro ots and sho ots of E. ma rit imum ar e e at en a s a ve get abl e a nd you ng leave s are consu med as a sal ad in north ern Eur ope and Gre ece. Cand ied ro ots of E. marit imu m , hav e been val ued as an aphr odis iac tonic esp eci ally in E ngl and (6) . E. m ari tim um h ave als o bee n re por t- ed to ex hib it dif fer ent thera peu tic uses in folk me di- cin e as diure tic or hypo gly cem ic (7) . E. mar it imum was in the past a wid ely used medic ina l her b and in mod ern phyto ther apy it is con si der ed a re med y in ren al dis ord ers (3, 8). The main seco nda ry met abo - lit es isola ted from E. mar itim um extra cts were gly- cos ide s of kae mpf ero l, iso que rce tin a nd ast rag ali n (9) . Ery ngi um s erb icum Pan čić is a per enn ial plan t gro win g to a hei ght of 40 -75 cm. T his spec ies is di s- tri but ed i n Se rbi a an d con si der ed a reg iona l en dem ic (10 , 1 1). In this work the ant iox ida nt activ ity of fo ur dif fer ent extra cts for E. mari ti mum and E. ser bicu m is rep ort ed. Al so, we co mpa red an tim icr obi al eff ect of ess ent ial oils and ext rac ts from aeria l part s of this spe cie s. To our knowl edg e, the comp arat ive biolog - ica l effec t of thi s two sp ecie s has not been previ- ous ly rep ort ed. Biological activity of extracts and es sential oils of two Eryngium... Jelena S. Mateji ć et al. 25 Mat eri al an d m eth ods Che mic als Org ani c sol ven ts we re pur cha se d from "Zor ka pha rma " Ša bac , Se rbi a. Galli c aci d, 3 -ter t-bu ty l-4 - hyd rox yan iso le (BHA ) an d 2, 2-d yph eny l-1 -pi cry lhy - dra zyl (DPPH ) were obtai ned from Sig ma Che mic als Co. , St Louis , MO, USA. Foli n-C ioc alt eu ph eno l rea g- ent was pur cha sed from Mer ck, Dar mst adl , Germa- ny. So diu m carbon at e an hyd rou s ( Na 2 CO 3 ), potas - siu m acet ate (C 2 H 3 KO 2 ), pot ass ium perox idi sulp ha te (K 2 O 8 S 2 ) and L(+ ) - Asco rbic acid (Vit ami n C) wer e pur cha sed fr om Anala R Norma pur , VW R, Geld ena- aks eba an, Leuv en Be lgiu m. Alumi niu m nitr at e non- ahy dra te (Al (NO 3 ) 3 x 9H 2 O) was pur cha se d fr om F luk a Che mie AG , B uch s, Swi tze rla nd. AB TS (2, 2'- azi no- bis (3 -e thy lb en zo thi az ol ine -6 -s ul pho ni c acid )) an d qu er - cet in hydra te were obta in ed fr om TCI Eu rop e NV , Boe ren vel dsw eg, Belg ium . All ot her solv ent s and che mic als we re of ana lyt ica l g rad e. Pla nt ma teri al Aer ial pa rts fro m wild grow ing spe cie s of E. mar iti mum (Gly fad a, Corfu - Gree ce) and E. Serb i- cum (coa st of th e Iba r riv er – Serb ia) wer e col lec ted in Jul y 2015. A vou che r spec ime n (108 76, 10 877 ) was dep osi ted in th e "He rba riu m Mo esi acu m Niš " , Un iver sit y o f N iš. Pre par at ion of pla nt ext rac ts Pla nt mat eri al was ai r dried in the dark and gro und to pow der . The aeri al pl ants par ts (1 0 g) wer e extra cte d wit h 100 ml wat er (H 2 O), meth ano l (M eOH) , a ce tone (A cet ) o r e thy l a cet ate (E tOA c). Tab le 1. Th e e xt ract s yie ld for in ve sti gat ed Er yng ium sp eci es Yield of extract (%, mg per g) H 2 O MeOH Acet EtoAc E. maritimum 0.945 0.958 0.336 0.382 E. serbicum 0.993 1.043 0.275 0.186 The mix tur e was expo sed to ultra sou nd bath for 30 min and after 24 h stand ing in the dar k was fil ter ed. MeOH , EtO Ac and Ac et solven ts w er e re- H 2 O ext ra ct o f ae ria l pa rts fro m E. mar it imum and E. ser bic um poss ess ed the stro ng est ant iox idan t activ - ity compa red to other s. For both speci es, EtOAc ex- tra cts sh owe d t he low est ac tiv ity . ABT S s cav eng ing ac tiv ity The res ult s fr om t he A BTS as say are show n i n Ta ble 2. The am oun t ran ged from 0.10 9 to 3.36 m g AA/ g. The high er ABTS valu es pres ent the stro nge r ant iox ida nt act ivit y. The hig hes t cont ent wa s iden- tif ied in H 2 O ex tra ct and the low est in EtOA c ext rac t for bo th sp ecie s. Tab le 2. Co mpa rat iv e a nal ysi s o f T PC , T FC, AB TS an d D PPH t est s b etw een e xtra ct s f rom tw o E ryn gi um spe ci es Antioxidant activity TPC (mg GAE/g) TF C (mg Qu/g) ABTS (mg AA/g) DPPH IC 50 (mg/ml) E. maritimum (H 2 O) 1.84±0.023 c 2.82±0.033 e E. maritimum (MeOH) 49.01±0.016 d 8.98±0.007 e 1.058±0.026 b 4.82±0.012 d E. maritimum (Acet) 77.08±0.138 bc 48.68±0.026 a 0.769±0.0 45 b 12.52±0.033 b E. maritimum (EtOAc) 7.47±0.005 f 17.55±0.014 d 0.109±0.003 a 31.19±0.051 a E. serbicum (H 2 O) 90.1±0.004 b 17.49±0.004 d E. serbicum (Acet) / 0.054±0.002 Ea ch valu e in the tab le was obt ai ned by cal cul ati ng the avera ge of thr ee ana lys es ± stan dar d devi ati on. Diff ere nt lett er s abo ve ba rs i ndi ca te s tat ist ic all y si gni fic ant d iff ere nce s o nly am ong th e t rea tm ent s p erf orm ed for ea ch a ss ay a cc ord ing to th e Tu key te st (p ≤ 0.0 5) Ant im icr obia l a cti vit y Res ult s o bta ine d fo r th e ex tr act ’s a nt imic rob ial act ivi ty are prese nte d in Table 3. Two antim icr obi al com mer cia l ag ent s, namel y Ch lor amp hen ico l (a nti- bac ter ial ) an d N yst at in ( ant ifu nga l) w er e us ed as ref - ere nce s fo r comp ari son of the inv est iga ted ext rac t’s act ivi tie s. Ess ent ial oils of the two Eryn gi um spe cies sho wed a cti vit y at conce ntr ati ons rang ing from <0. 00 1- 15 mg/m l, wh ile th e sol ven t ext rac t’s activ i - tie s rang ed fro m 0.15 -30 mg/ml . In the cas e of the two esse nti al oil s, the one isol ate d fro m E. ser bic um pla nt ma ter ial showe d sign ifi can tly high er ant imi cro- bia l pot ent ial (inh ibi tio n from < 0.00 1-> 2.5 mg /m l) in com par iso n to the oil iso lat ed fro m E. mari timu m (3. 75- 15 mg/ml) . The menti- one d ess ent ial oil sho wed ver y in ten siv e inh ibi tor y ac tiv ity , ex hib ite d aga ins t a ll t est ed mic robi al str ain s wi th the exc ept ion sum pt ion of s econ dar y m etab ol ite s is ola ted fro m th is two spec ies may pro vid e posi tiv e con sequ en ces for hum an hea lth . Ac kno wle dge men ts The a uth ors a re gra tef ul to the Min ist ry of Edu cat ion , Scienc e and Techn olo gic al Devel opm ent of th e Rep ub lic of Serb ia fo r fin anc ial supp ort (Gra nt No. 17 302 9). Referenc es 1. Erd elm ei er CAJ , Sti che O. A cycl ohe xen on e and a cy- clo he xad ien one glyco sid e from E ryn giu m camp est re . Ph yto che mis try 1 986; 2 5:7 41- 3. [ Cro ss Ref ] 2. Ram a C PP, Sud ha kar RC, Ra za S H, D utt CB S. Fol klo re med ic ina l pl ant s o f N ort h A nda ma n I sla nds , I ndi a. Fit ot era pia 20 08; 7 9:4 58- 64. [C ros sRe f] [P ubM ed] 3. Kü peli E, Kart al M, Aslan S, Yesi lad a E. Com par ati ve eva lua ti on of the ant i-in fla mm ato ry and anti no cic ep- tiv e a cti vit y of Tu rki sh E ryn giu m s pec ies . J Eth nop ha r- ma col 20 06; 10 7:3 2- 7. [Cr oss Ref ] [ Pu bMe d] 4. Yan iv Z , Da fni A, Fr ied man J, Pa lev it ch D. Pla nts use d for the trea tme nt of dia bet es in Isr ael . J Eth nop ha r- ma col 19 87; 19 :14 5- 51. [C ros sRe f] [P ubM ed] la Cor se et des cont ré es limi tr oph es. Paris : Libr ai rie sci ent if iqu e e t t ech ni que Al ber t B la nch art ; 1 980 . 6. Kh osh bak ht K, Hamm er K, Pistr ic k K. Ery ngi um cau- cas icu m T rau tv . c ult iva ted as a v eg eta ble in th e El bu rz Mou nt ain s (Nort he rn Iran ). Gen et Re sou r Crop Ev ol 200 7; 54 :44 5- 8. [C ros sR ef] 7. Lis cia ni R, Fatt oru sso E, Su ran o V, Co zzo lin o S, Gia nn att asi o M, Sor ren tin o L. Ant i-in fl amm ato ry acti - vit y of Ery ngi um m ari tim um L . rh izo me ex tr act s in in- tac t rat s. J E thn oph ar mac ol 198 4; 12 :26 3- 70. [Cr os sRe f] [Pu bM ed] 8. Suc iu S, Bod oki E. Study of tri te rpe ne sap oni ns fro m Ery ngi um mar iti mum L. ( Api ace ae ). TM J 20 05; 55( 5): 348 - 50. 9. Hil ler K, Poh l B , Fr ank e P . Fl avo no id s pec tru m o f Ery ngi um mari tin um L. part 35. Com po nen ts of Sani - cul oid ae . P har maz ie 19 81; 36 (6) :45 1- 2. 10. Ch ate r AO . G enu s Er yn giu m L . In: Tu tin TG, He ywo od VH , Bu rg es N A, M oor e DM , Valen ti ne D H , Walt er s SM , Web b DA, e dit ors . Flo ra Eu rop aea 2. L ond on , UK: Cam bri dg e U niv er sit y P res s; 1968 p. 320 - 4. 11. St eva no vić V, T an K, I atr ou G. Di st rib uti on of t he en dem ic Bal kan flo ra on ser pen tin e I. – obli ga te ser - pen tin e end emi cs. P l S yst Ev ol 200 3; 242 :1 49- 70. [Cr os sRe f] 12. Si ngl eto n VL , Or th ofe r R, Lam uel a-R ave nt os R M. An a- lys is of total pheno ls and othe r oxid ati on subst rat es an d an tio xid ant s by me ans of F oli n – C ioc alt eu rea gen t. Met h Enz ymo l 1 99 9; 299 :15 2- 78. [C ros sRe f] 13. Wo is ky R, S al at in o A. An al ys is of p ro po li s: so me p ar am - et ers an d proce du res fo r chem ica l quali ty co ntr ol. J Api c Res 19 98; 37 :9 9- 105. [Cr oss Ref ] 14. Bl ois MS. Antio xid ant det er mina ti ons by the use of a sta ble f ree ra dic al. N atu re 195 8; 181 :1 199 - 200. [Cr os sRe f] 15. Mi lle r N, Rice -E van s C. Fact ors inf lu enci ng the anti oxi - dan t acti vit y dete rm ine d by the ABT S radi cal catio n ass ay . F ree Ra dic R es 199 7; 26: 195 -9 . [Cr os sRe f] [Pu bM ed] 16. NC CLS – Na tion al Commi tte e for Cli nic al Lab ora tor y Sta nda rd s, Perf orm anc e stand ard s for anti- mic rob ial su sce pti bili ty t est in g: eleve nth i nf orm ati ona l su ppl e- men t, Docum ent M10 0-S 11, Natio nal Comm itt ee for 17. Mo lyn eux P. The use of the stabl e fre e radi cal di phe- nyl pic ry lhy dra zyl (DPP H) fo r est ima tin g ant iox ida nt act i- vit y. Son gkl ana ka rin J Sci Tech nol 2004; 26 (2) :2 11- 9. 18. Rj eib i I, Saa d AB, Ncib S, Soui d S. Ph eno lic comp o si - ti on an d ant i ox id a nt pr op e rt ie s o f Ery n gi um mar iti mu m- (s ea hol ly) . J Co ast Li fe Me d 2 017 ; 5 (5) :2 12- 5. [Cr os sRe f] 19. Mo et- Du ros L, Le Fl och G, Ma gne C. Rad ica l sca ve ng- ing , a nt iox ida nt and an tim ic rob ial ac tiv iti es of hal oph y- tic spec ies . J Ethn oph arm aco l 2008 ; 116 (2) :2 58- 62. [Cr os sRe f] [Pu bM ed] 20. Da rri et F, Ben dah ou M, Des job ert JM, C osta J, Mu sel li A. Bic ycl o[ 4.4 .0] dec an e oxyg ena ted sesqu ite rp ene s fro m Eryn giu m mari tim um esse nt ial oi l. Plan ta me d 201 2; 78 :38 6- 9. [ Cr oss Ref ] [ Pub Me d] 21. Ca pet an o s C, Sar ogl ou V, Mar in PD, Simić A, Skalt sa HD. E sse nti al oil anal ysi s of two endem ic Ery ngi um spe cie s f rom Ser bia . J S erb Che m So c 20 07; 72 (1 0) : 961 - 5. [ Cr oss Ref ] 22. Me ot- Du ros L, Floc h G, Ma gné C. Radi cal scav eng ing , ant io xid ant and ant im icr obi al acti vit ies of halo phy tic [Cr os sRe f] [Pu bM ed] 23. T h iem B, Gośl iń ska O, Kiko wsk a M, Bud zi ano wsk i J. Ant im icr obi al a cti vit y o f th ree Ery ngi um L. s pe cie s (Ap ia cea e). He rba p olo nic a 2 010 ; 5 6( 4): 52- 8. 24. Kh olk hal W, Il ias F, Be kh ech i C , Be kk ara FA . E ryn giu m ma rit imu m : a ri ch med ici na l plan t of po lyp hen ol s and fla von oi ds comp oun ds wit h antio xid ant , anti bac ter ial an d a nti fu nga l act ivi tie s. Cu rr Res J Bi o S ci 20 12 ; 4(4 ): 437 - 43. Biological activity of extracts and es sential oils of two Eryngium... Jelena S. Mateji ć et al. 31 Originalni rad UDC: 615.322:582.794.1 doi:10.5633/amm.2019 .0 304 BIOL OŠKA AKT IVNOS T E KS TRAKA TA I ETAR SKIH ULJA DVE VRST E RODA ERY NGIUM ( AP IACEA E ) SA BALK ANSKO G PO LUOST RVA Jelena S. Matejić 1 , Zorica Z. Stojanović - Radić 2 , Zoran Dj. Krivošej 3 , Bojan K. Zlatković 2 , Petar D. Marin 4 , Ana M. Džamić 4 1 Univerzitet u Nišu, Medicinski fakultet, Katedra za fiziku i biolog iju sa humanom genetikom, Niš, Srbija 2 Univerzitet u Nišu, Prirodno - matemat ički fakultet, Odsek za biologiju i eko logiju, Niš, Srbija 3 Univerzitet u Prištini, Prirodno - mate matički fakultet, Odsek za b iologiju, Koso vska Mitrovica, Srbija 4 Univerzitet u Beogradu, Biološki fakultet, Institut za botaniku i Botanička bašta “Jevre movac”, Beograd, Srbija Bulevar dr Zorana Đinđića 81 , 18000 Niš , Srb ija E-mail: [email protected] U ov oj st udi ji o pis ana je an tio ksi dat ivn a i an tim ikr ob na ak ti vno st vo den ih , met an olni h, ace to nsk ih i etil- ace tat nih eks tra kat a do bij enih od vrs ta Eryn giu m mar iti mu m L. iz Grč ke i Ery ngi um serbi cu m Pa nči ć, samon ikl e u Srb iji . Tak ođe, anal izi ran a je an tim ikr obn a akt ivn ost eta rs kih ulj a iz olo van ih iz n adz emn ih d el ova ovih vr sta . Sp ekt rof ot ome tri js ke m eto de k ori š će - ne su za me ren je kon cen tr aci je uku pni h fen ola, flav ono ida , kao i za odr eđi van je ant iok sid a - tiv no g pot en cija la uzo rak a up otr ebo m DP PH i ABT S me tod a. U kup na koli čin a fe nol a u e kst ra -

https://www.researchgate.net/publication/337063280_BIOLOGICAL_ACTIVITY_OF_EXTRACTS_AND_ESSENTIAL_OILS_OF_TWO_ERYNGIUM_APIACEAE_SPECIES_FROM_THE_BALKAN_PENINSULA

USDOS – US Department of State (Author): “2022 Report on International Religious Freedom: Bahrain”, Document #2092183 - ecoi.net Annual report on religious freedom (covering 2022) Document #2092183 USDOS – US Department of State (Author) 2022 Report on International Religious Freedom: Bahrain Executive Summary The constitution declares Islam to be the official religion and sharia the principal source for legislation. It provides for freedom of conscience, the inviolability of places of worship, and freedom to perform religious rites. The constitution guarantees the right to express and publish opinions, provided these do not infringe on the “fundamental beliefs of Islamic doctrine.” The law prohibits anti-Islamic publications and mandates imprisonment for “exposing the state’s official religion to offense and criticism.” There is no legal prohibition against apostasy. The penal code punishes any individual who mocks or disdains another religious group. In public schools, Islamic studies are mandatory for Muslim students in grades one through 12. The constitution guarantees women’s equality with men, “without breaching the provisions” of sharia. In March, the UN Economic and Social Council’s Committee on Economic, Social, and Cultural Rights (ECOSOC) expressed concern about reports of harassment, intimidation, arrests, and detention of Shia religious figures. In November, the UN Human Rights Council (UNHRC) expressed concerns about reports that authorities restricted the rights of the Shia community to worship and profess their religious beliefs. Reflecting the country’s demographics, the majority of citizen detainees and prisoners were Shia. Human rights groups reported security officials mistreated prisoners. In March, a prominent Shia prisoner accused guards of attempting to assault him. The ombudsman’s office and the government-affiliated National Institution for Human Rights (NIHR) said no physical abuse occurred. Nongovernmental organizations (NGOs), media, and opposition outlets reported that the government continued to summon Shia rights activists for questioning. NGOs and human rights activists continued to state Shia prisoners received inadequate medical treatment. The government reportedly investigated 12 individuals for unspecified “crimes affecting religion,” charged nine with publicly violating Ramadan restrictions, and imprisoned three for insulting religious symbols. In June, the Ministry of Social Development (MOSD) allowed the Islamic Enlightenment Shia Society, the charity wing of the banned al-Wifaq opposition Shia political party, to reopen; the government had ordered it shut down in 2016. The government continued to regulate the content of religious sermons of both Sunni and Shia religious leaders. Shia sources said that unlike in prior years, the government did not summon Shia clerics for questioning related to their sermons. According to procession organizers, the government permitted large groups to gather in Manama and in Shia-majority villages to observe Ashura – among the most significant days of the Shia religious calendar. Some Shia prisoners reported that during Ashura, prison authorities gave them additional time and resources to practice Ashura rituals in common areas, in contrast to previous years, but a media source said this was not true for all prisoners. Some prisoners stated authorities punished them for observing other Shia holidays. The ombudsman’s office reported that based on its prison visits, it found Shia prisoners received adequate religious accommodation. In July, authorities prohibited individuals from gathering to sing a song praising a ninth-century Shia cleric; authorities said the song was Iranian propaganda and summoned and fined individuals who disobeyed. During the year, the government funded the construction and renovation of several Sunni and Shia mosques in multiple parts of the country. Shia sources expressed dissatisfaction that Shia doctrine was not included in Islamic studies in public schools, which they said focused on Sunni Islam. According to Shia leaders and community activists, the government continued to discriminate against Shia citizens and to give Sunni citizens preferential treatment for Ministry of Education (MOE) scholarships and positions in the Ministry of Interior (MOI) and military, and government policy effectively gave preference to Sunni applicants for citizenship. In June, the King appointed a new cabinet of 22 ministers that included nine professed Shia. One critic said Shia representation should be “at least” half to reflect the country’s religious demographics. In July, the government issued the National Human Rights Plan for 2022-2026, which it developed in consultation with the Office of the UN High Commissioner for Human Rights, setting forth government goals for advancing religious freedom and freedom of expression, among other priorities. On April 18, the King met with senior Shia cleric Sayed Abdulla al-Ghuraifi, and they discussed the value of religious discourse, according to reports. Social media commentators described the visit as a trust-building exercise between the government and the Shia community. In November, Pope Francis, the Grand Imam of al-Azhar, and other religious leaders from around the world participated in a conference in Manama on East-West peaceful coexistence, and Pope Francis conducted an open-air Mass attended by 30,000 persons. According to non-Muslim religious groups, there was a high degree of government support for their worship activities. Anti-Shia and anti-Sunni commentary appeared in social media. NGOs reported on the adverse economic effect of Sunni-Shia tensions and local political divisions. Shia human rights and political activists reported persistently higher unemployment rates, limited prospects for upward social mobility, and lower socioeconomic status for that community compared with the Sunni population. According to minority religious groups, there was a high degree of tolerance within society for minority religious beliefs and traditions, although not for conversion from Islam or for atheistic or secularist views. Non-Muslim religious community leaders again reported converts from Islam were unwilling to speak publicly or privately to family or associates about their conversions for fear of harassment or discrimination. In September, there was a brief upsurge in antisemitic rhetoric online following an erroneous report that the government intended to establish a Jewish neighborhood in Manama. U.S. government officials, the Ambassador, and other U.S. embassy representatives met with senior government officials, including the Prime Minister, Foreign Minister, and the Minister of Justice, Islamic Affairs, and Endowments, and national human rights monitoring institutions to urge respect for freedom of religion or belief and expression, including the right of religious leaders to speak and write freely, and to advocate the full and equal participation of all citizens, irrespective of religious or political affiliation, in political and social activities and economic opportunities. The Ambassador and other embassy officials met regularly with religious leaders and community members from a broad spectrum of faiths, representatives of NGOs, and political groups to discuss freedom of religion and freedom of expression as they related to religious practice. In October, the visiting Deputy Special Envoy to Monitor and Combat Antisemitism met with senior government officials, civil society leaders, youth, students, and members of the local Jewish community. Section I. Religious Demography The U.S. government estimates the total population at 1.5 million (midyear 2022).  According to the national government, there are approximately 712,000 citizens, constituting less than half of the total population.  According to 2020 national government estimates, Muslims make up approximately 74 percent of the country’s total population.  Boston University’s 2020 World Religions Database states 82 percent of the total (citizen and noncitizen) population is Muslim and 18 percent adheres to other religions and beliefs, such as Christianity (12 percent), and Hinduism (6 percent).  According to the database, other groups comprising less than 1 percent total include Baha’is, Buddhists, Jews, agnostics, and atheists.  According to Jewish community members, there are between 20-40 Jewish citizens encompassing six families. The government does not publish statistics regarding the breakdown between the Shia and Sunni Muslim populations.  Most estimates from NGOs and the Shia community state Shia Muslims represent a majority (55 to 70 percent) of the citizen population. Most foreign residents are migrant workers from South Asia, Southeast Asia, Africa, and other Arab countries.  According to national government 2020 census data, approximately 401,500 foreign residents are Muslim; 387,800 are Hindu, Buddhist, Baha’i, Sikh, or Christian (primarily Roman Catholic, Protestant, Syrian Orthodox, and Mar Thoma Syrian from South India). Section II. Status of Government Respect for Religious Freedom Legal Framework According to the constitution, Islam is the official religion, and the state safeguards the country’s Islamic heritage. The constitution provides for freedom of conscience, the inviolability of places of worship, freedom to perform religious rites, and freedom to hold religious parades and religious gatherings “in accordance with the customs observed in the country.” The constitution provides for the freedom to form associations as long as they do not infringe on the official religion or public order, and it prohibits discrimination based on religion or creed. All citizens have equal rights by law. According to the constitution, all persons are equal without discrimination on the basis of gender, origin, language, or faith. The constitution states that sharia forms the principal basis for legislation, although civil and criminal matters are governed by a civil code. The labor law pertaining to the private sector prohibits discrimination against workers on grounds of gender, origin, language, religion, or belief. The labor law deems religion-based dismissal to be arbitrary and illegal but does not provide an automatic right to reinstatement. The law also prohibits wage discrimination based on religion, among other factors. The Ministry of Labor (MOL) investigates claims of discrimination where there is an existing labor relationship; it may escalate violations to the Public Prosecution Office. The MOL does not have the authority to receive or manage complaints of religion-based discrimination in hiring. There is no explicit law on discrimination in public sector employment, although such discrimination is governed by the constitution’s broader provisions. The constitution guarantees the right to express and publish opinions, provided these do not infringe on the “fundamental beliefs of Islamic doctrine” and do not prejudice the “unity of the people” or arouse discord or sectarianism. The law prohibits anti-Islamic publications and broadcast media programs but does not define “anti-Islamic.” It mandates imprisonment of no less than six months for “exposing the state’s official religion to offense and criticism.” The law states, “Any publication that prejudices the ruling system of the country and its official religion can be banned from publication by a ministerial order.” The Ministry of Information Affairs determines which materials are governed by this law. Islamic religious groups must register with the Ministry of Justice, Islamic Affairs, and Endowments (MOJIA) to operate. Sunni religious groups register with the ministry through the Sunni Waqf (endowment), while Shia religious groups register through the Jaafari (Shia) Waqf. MOJIA endowment boards supervise, fund, and perform a variety of activities related to mosques and prayer halls. Non-Islamic religious groups must register with and receive a license from the MOSD as a civil society organization in order to operate. Civil society organizations that are Islamic in nature must also register with the MOSD. To register, a group must submit an official letter requesting a license to operate; copies of minutes from the founders’ committee meeting; and a detailed list of founders and board members, including names, ages, nationalities, occupations, and addresses. It must also submit other information, such as the group’s bylaws, candidates who seek election to the organization’s governing board, a physical address, and evidence of a bank account in a bank registered with the Central Bank of Bahrain. Religious groups also may need approval from the MOE if the public education curriculum is involved; the Ministry of Information Affairs if the group intends to issue religious publications; or the MOI, in coordination with the Central Bank of Bahrain, if the organization receives funding from foreign sources. Individuals affiliated with Islamic organizations seeking to collect money for religious purposes must be Bahraini citizens and must receive approval from the MOJIA. The law prohibits civil society organizations from engaging in politics. The law prohibits activities falling outside an organization’s charter. The penal code does not specifically address the activities of unregistered religious groups but provides for the closing of any unlicensed branch of an international organization plus imprisonment of up to six months and fines of up to 50 dinars ($130) for the individuals responsible for setting up the branch. The law prohibits any individual from being a member of a political society or becoming involved in political activities, including running in elections, while serving in a clerical role at a religious institution, including on a voluntary basis. According to the MOSD’s website, the following non-Islamic religious groups are registered with the ministry: the National Evangelical Church, Bahrain Malaylee Church of South India Parish, Word of Life International Church, St. Christopher’s Cathedral (Anglican), Church of Philadelphia, St. Mary and Anba Rewis Church (St. Mary’s Indian Orthodox Cathedral), Jacobite Syrian Christian Association and St. Peter’s Prayer Group (St. Peter’s Jacobite Syrian Orthodox Church), St. Mary’s Orthodox Syrian Church, Sacred Heart Catholic Church, The Church of Jesus Christ of Latter-day Saints, Church of Christ, Greek Orthodox Church, Pentecostal Church, Baps Shri Swaminarayan Mandir Bahrain (Hindu Temple), Indian Religious and Social Group (Hindu Temple), Spiritual Sikh Cultural and Social Group, St. Thomas Evangelical Church of Bahrain, Marthoma Parish, House of Ten Commandments Synagogue, Shri Krishna Hindu Temple, and Baha’i Social Society. Some religious groups, including Shia religious education centers ( hawzas) seek government licensing for their places of worship, while others choose to operate in private residences. There are 16 registered churches and numerous other Christian places of worship, mostly in Manama. There is no legal prohibition against apostasy. The penal code calls for punishment of up to one year’s imprisonment or a fine of up to 100 dinars ($260) for offending one of the recognized religious groups or their practices or openly defaming a religious figure considered sacred to members of a particular group. It punishes an individual for desecration of religious books with up to one year in prison and a fine of 100 dinars ($260). The law also prohibits any person from imitating in public a religious ritual or ceremony with the intention of ridiculing it. The law stipulates fines or imprisonment for insulting an institution, announcing false or malicious news, spreading rumors, encouraging others to show contempt for a different religious denomination or sect, illegally gathering, or advocating a change of government, among other offenses. The MOI’s Office of the Ombudsman, the Prisoners’ and Detainees’ Rights Commission, and the NIHR address the rights of prisoners, including the right to practice their religion while incarcerated. The law guarantees inmates of correctional facilities the right to attend burials and receive condolences outside prison. The law allows prisoners and criminal defendants at their sentencing to receive an “alternative non-custodial sentence” in lieu of incarceration, provided such a sentence would not endanger public security. The judge and the public prosecutor determine an individual’s eligibility and conditions for an alternative sentence and the MOI supervises individuals following their release on an alternative sentence. Alternative sentences may include community service, home detention, electronic surveillance, no-contact orders, or participation in rehabilitation programs. The MOJIA oversees the activities of both the Sunni Waqf and the Jaafari Waqf; the King appoints their governing boards with recommendations from the president of the government-run and funded Supreme Council for Islamic Affairs (SCIA). The respective waqf boards supervise the activities of mosques and prayer halls, review and approve clerical appointments for religious sites under their purview, and fund expenses for the building and maintenance of religious sites. The government allocates 2.7 million dinars ($7.1 million) annually to each endowment board. Zakat (Islamic tithes), income from property rentals, and other private sources fund the remainder of the waqf boards’ operations. The waqf boards may pay flat commissions and bonuses to preachers and other religious figures. By law, the government regulates and monitors the collection of money by religious and other organizations. Islamic organizations wishing to collect money must first obtain authorization from the MOJIA. Authorizations are valid for one year. Non-Islamic organizations must obtain authorization from the MOSD. A 2021 amendment to a royal decree requires the Sunni and Jaafari waqfs to submit annual reports to the MOJIA on funds they collect for religious purposes, including for the construction or renovation of places of worship. The amendment also bans the waqfs from receiving money from abroad without MOJIA approval. The SCIA oversees general Islamic religious activities taking place within the country as well as the publication of Islamic studies curricula and official religious texts. The council consists of a chair, a deputy chair, and 16 religious scholars – eight Sunni and eight Shia, most of them prominent preachers or sharia judges. The King appoints all council members to a four-year term. Independent of other government scholarship programs, the council offers university scholarships to low-income students for advanced Islamic studies. The SCIA reviews all legislation proposed by parliament to ensure draft laws comply with sharia. The council also consults with other government entities before issuing permits to new Islamic societies or centers. The council is responsible for reviewing the content of Islamic programs broadcast on official government media outlets, such as the official television and radio stations. The council also organizes interfaith conferences and workshops. The government licenses houses of worship. Permission for construction of a new mosque, whether Shia or Sunni, requires a government determination of the need for a new mosque in the area based on population. The government also determines the need for non-Islamic houses of worship. The King has sole legal authority to allocate public land, including for religious purposes, although he may delegate this authority to government officials. Citizens may also offer private land to build mosques. By law, construction of Islamic places of worship requires MOJIA approval. Non-Islamic groups must obtain MOSD approval. Municipal authorities provide final approval for construction. The law permits non-Islamic houses of worship to display crosses or other religious symbols outside their premises. The law regulates Islamic religious instruction at all levels of the education system. In public schools, Islamic studies are mandatory for all Muslim students in grades one through 12 and are optional for non-Muslims. Private schools must register with the government and, with a few exceptions (for example, a foreign-funded and foreign-operated school), are also required to provide Islamic religious education for Muslim students. Private schools wishing to provide non-Islamic religious education to non-Muslims must receive permission from the MOE. Outside of school hours, Muslim students may opt to enroll in Quran recitation courses and religious studies sponsored by the MOJIA, as their parents deem fit. According to the government, the SCIA provides financial assistance to the six registered hawzas(Shia seminaries); other hawzas choose to be privately funded. The government does not permit foreign donors to contribute to privately funded hawzas. There are no restrictions on religious studies abroad. Specialized MOE-run religious schools provide more thorough religious instruction – the Jaafari Institute for instruction in Shia Islam and the Religious Institute for instruction in Sunni Islam – for students from elementary through high school. The remainder of the curriculum is consistent with the nonreligious curriculum in other public schools. The constitution also guarantees the duties and status of women and their equality with men, “without breaching the provisions” of sharia. The personal status law states that either the Sunni or Shia interpretation of sharia, depending on the religious affiliation of the parties, shall govern family matters, including inheritance, child custody, marriage, and divorce. Mixed Sunni-Shia families may choose which court system will hear their case. The provisions of the law on personal status apply to both Shia and Sunni women, requiring a woman’s consent for marriage and permitting women to include conditions in the marriage contract. Under sharia, Muslim men may marry non-Muslim Christian or Jewish women, but not women of other religions; the law considers offspring of such a marriage to be Muslim. Muslim women may not marry non-Muslim men. Non-Muslims may marry in civil or religious ceremonies; however, all marriages must be registered with a civil court. Civil courts also adjudicate matters such as divorce and child custody for non-Muslims. A woman’s court testimony is given equal weight to that of a man. The constitution states inheritance is a guaranteed right governed by sharia, meaning women are entitled to half the share of men. The government does not designate religious affiliation on national identity documents, including birth certificates. Applications for birth certificates and national identity documents, however, record a child’s religion (either Muslim, Christian, Jewish, or other), but not denomination. Hospital admission forms and school registration forms may also request information on an individual’s religion. The constitution says the state shall strive to strengthen ties with Islamic countries. It specifies that succession to the position of king is hereditary, passing from eldest son to eldest son. The royal family is Sunni. The country is party to the International Covenant on Civil and Political Rights, with reservations stating it interprets the covenant’s provisions relating to freedom of religion, family rights, and equality between men and women before the law as “not affecting in any way” the prescriptions of sharia. Government Practices On March 4, the UN ECOSOC released a report that stated, “Despite the constitutional prohibition of discrimination based on religious affiliation, members of the Shia community reportedly face discrimination in education, employment, and in the exercise of their cultural rights.” The committee stated it was “also concerned about reports of harassment, intimidation, arrests, and detention of religious and cultural figures.” In November, the UNHRC released its fourth Universal Periodic Review (UPR) of the country. The UNHRC stated it was “concerned about the existence of practices that adversely affected the exercise of freedom of conscience, at reports that members of the Shia community had been subjected to restrictions on their rights to worship and profess their religious beliefs, and that liberty of conscience was not guaranteed effectively.” Among other recommendations, it called on the government to decriminalize blasphemy. In the government’s national report submitted to the UNHRC ahead of the UPR, it stated that the constitution “guarantee[d] the right to freedom of expression, association and peaceful assembly,” that labor laws “prohibit[ed] discrimination against employees on grounds of gender, origin, language, religion or creed,” and that “in order to promote social security and peaceful coexistence, the Ministry of Justice [took] great care to control, monitor, and analyze the content of religious sermons, and [had] a comprehensive national strategy to address social issues and negative phenomena resulting from extremist ideology by updating the language used in such sermons in such a way as to strengthen social unity, maintain cohesion in the national social fabric and put an end to deviant ideology which gives rise to extremism and bigotry.” Reflecting the country’s demographics, the majority of citizen detainees and prisoners were Shia. Human rights groups reported security officials mistreated prisoners. NGOs reported prison authorities routinely denied Shia prisoners needed medical treatment more often than Sunni prisoners. On February 13, NIHR said it investigated complaints submitted by detainees in prison, primarily about lack of access to medical services. At year’s end, NIHR had not yet published its annual report or final figures on the outcomes of its investigations. On June 2, Amnesty International Bahrain posted a statement on Twitter accusing the authorities of “acting with gross negligence in the face of confirmed tuberculosis infections” at Jaw Prison. Members of the Shia opposition held multiple small-scale demonstrations on the evening of Friday, June 3, alleging medical mistreatment of Shia political prisoners. Shia opposition figure Shaikh Abduljalil al-Meqdad said in an audio recording released September 29 from his cell in Jaw Prison that five prison guards attempted to assault him for refusing to attend his medical appointment. Al-Meqdad was serving a life sentence for forming a terrorist group with intent to overthrow the system of government and change the constitution and for collaboration with a foreign state. The NGO Americans for Democracy and Human Rights in Bahrain (ADHRB) stated in October that his conviction was politically motivated. The ombudsman and the NIHR investigated al-Meqdad’s allegations, reviewing CCTV footage and interviewing prison guards, and said they found no evidence of physical abuse. They stated al-Meqdad and the prison guards had a verbal altercation, but no physical assault occurred. Other sources, including ADHRB, the Gulf Institute for Democracy and Human Rights, and Middle East Eye, stated some prison officers attempted a physical assault but others stopped it and that al-Meqdad had numerous health conditions that were being neglected by the prison authorities. The online opposition media outlet Bahrain Mirrorreported that in March, authorities sentenced Shia scholar Mohammed al-Madi to one year in prison for insulting seventh-century Umayyad ruler Mu’awiya bin Abi Sufyan (Mu’awiya I), a companion of the Prophet Muhammad. Al-Madi was released early by royal decree after serving only 15 days of his sentence, according to opposition sources. Several Shia clerics arrested during the country’s 2011 antigovernment protests, including banned Shia political party al-Wifaq’s Secretary General Shaikh Ali Salman, remained in prison at year’s end, serving sentences ranging from 15 years to life in prison on charges related to terrorist activity, seeking to overthrow the monarchy, treason, or inciting hatred. Some human rights NGOs considered them to be political prisoners. According to sources, peaceful protesters on February 13 and 14 commemorated the anniversary of the 2011 protests, chanting slogans against King Hamad bin Isa Al Khalifa and displaying pictures of exiled and jailed opposition figures. The ombudsman stated the office received 631 complaints and assistance requests from inmates and detainees from May 1, 2021, to April 30, 2022, compared with 691 complaints and assistance requests filed in the preceding 12-month period. The ombudsman stated most prisoners’ mistreatment complaints were related to verbal abuse and excessive use of solitary confinement. During the year, several inmates received permission to leave the prison to attend funerals. The government continued to enforce Ramadan fasting restrictions in public spaces on non-Muslims. The MOI said images widely circulated on social media of foreign workers in Manama eating during Ramadan led the Capital Governorate Police Directorate to take legal action. During the year, the government charged nine individuals with violating Ramadan fasting restrictions. The government reported that during the year, it imprisoned three persons for insulting religious symbols. Six individuals were imprisoned, four received fines, one received probation, and one investigation was pending at year’s end for other unspecified “crimes affecting religion.” Zuhair Ebrahim Jassim and Hussain Abdulla Khalil Rashid, two Shia prisoners, remained on death row at year’s end. In 2020, they were convicted of involvement in targeting security forces and killing one police officer in a police bus explosion in November 2017 and killing another officer in a bomb explosion in Damistan village in 2014. NGOs said their confessions were obtained through torture and that the trial proceedings were unfair. King Hamad pardoned 160 prisoners by royal decree and the MOI released 69 prisoners on alternative sentences on April 28 for Eid al-Fitr. King Hamad pardoned 58 prisoners and the MOI released 49 others on alternative sentences on July 9 for Eid al-Adha. MOI’s Anticybercrime Directorate reported it continued to monitor social media for indecency, blasphemy, incitements to sectarianism, and offenses to the sanctity of religion. In August, the directorate arrested two citizens, one of them a 17-year-old, for posts on TikTok that insulted religious figures and for misuse of telecommunications equipment. The courts sentenced the 17-year-old to a one-month suspended prison sentence and the adult to a two-month suspended sentence. According to the government, during the year, the MOI did not investigate any individuals for defamation of religion, a charge usually stemming from statements made during sermons. This compared with 26 such investigations and six prosecutions in 2021. The government reported in November that it detained four persons on charges of witchcraft and sorcery but did not provide further details. On June 9, the MOSD permitted the reopening of the Islamic Enlightenment Shia Society, the charity wing of the banned al-Wifaq Shia political party, three years after the Supreme Civil Court reversed a 2016 MOSD order to dissolve the party, liquidate its assets, and place it under judicial receivership over charges related to misappropriation of funds. NGOs reported the government continued to closely monitor the collection of funds, including charitable donations, by religious organizations, with some NGOs describing this as government overreach. Religious groups stated the arduous bureaucratic process to receive funding and send donations abroad constituted a de facto ban on receiving grants, donating to, or transferring money from entities outside of the country. In July, the MOJIA launched a campaign urging people to check the ministry’s list of individuals authorized to collect funds for religious purposes before sending money to an organization. According to the MOJIA, the awareness campaign was part of the government’s efforts to combat money laundering and terrorism financing. Of 72 fundraiser requests received during the year, the MOJIA reported approving 51 individuals to accept funds from local sources. There were 60 licensed fund collectors on the MOJIA’s website at year end. The government continued to monitor and provide general guidance on the content of sermons by sending circulars to mosques. The MOJIA continued to monitor clerics’ adherence to a pledge of ethics it created for individuals engaged in religious discourse. According to the MOJIA, preachers who diverged from the pledge were subject to censure or removal by authorities on the grounds that their actions jeopardized national security. The MOJIA reported reviewing on a weekly basis sermons preachers submitted to the government. The MOJIA reported regularly visiting mosques on unannounced visits to ensure that preachers’ sermons were “moderate,” avoided discussing controversial topics, did not incite violence, and did not use religious discourse to serve political purposes. According to members of the Shia community, unlike in previous years, MOI did not summon clerics for questioning related to the content of their sermons. There were no reports of the government detaining or arresting Shia clerics for what the government deemed objectionable content in their sermons during Ashura. According to social media posts associated with the political opposition, the MOI summoned several Shia activists for questioning in early April. Shia political activist Ali Muhanna, one of those summoned, later posted a video stating the MOI summoned 13 additional persons for participating in regular peaceful protests in Sanabis, a suburb of Manama. Al-Wifaq reported that the MOI summoned several individuals in early May for participation in Jerusalem (Quds) Day protests on April 29, adding that several Shia opposition figures had been summoned on repeated occasions for questioning in connection with protest activities. According to the NGO Armed Conflict Location & Event Data Project, in March, authorities punished Shia prisoners for participating in commemoration events in prison marking the anniversary of the death of Imam Musa al-Kadhim, the seventh imam of Shia Islam, by prohibiting them from contacting their families for a week following the commemorations. On July 14, opposition social media accounts reported the MOI removed banners that were placed in the village of Barbar to commemorate Eid al-Qadr, which took place on July 16-17. According to government supporters, the MOI intervened whenever home or business owners filed a complaint because individuals had placed the banners or signs outside their premises without the owners’ consent. Sources reported that, unlike in prior years, authorities did not take down banners during the holy month of Muharram (July 31-August 28). Opposition social media reported that the government banned two Shia religious gatherings, both called Salam Ya Mahdi ,that were planned in Dair village on July 13 and in Sitra village on July 16. The gatherings were named after the song Salam Ya Mahdi(“Hello Oh Mahdi”) – the Arabic version of an Iranian song called Salam Farmandeh(“Hello Commander”). The original Farsi song expressed devotion to Iranian Supreme Leader Ali Khamenei and made references to former Islamic Revolutionary Guard Corps general Qassim Soleimani. The song was rewritten in Arabic with lyrics expressing devotion to Muhammad al-Mahdi (the Hidden Imam), a ninth-century Shia cleric and messianic figure, and sung by a Bahraini singer. Radio Free Europe/Radio Liberty reported (RFE) reported authorities viewed the song as Iranian propaganda aimed at children. During his July 13 sermon, Shia scholar Abdulla al-Ghuraifi criticized authorities for banning the Salam Ya Mahdievents and accused them of inciting hatred against Shia. On July 17, al-Wifaq reported authorities summoned the caretaker of the Imam al-Sadeq Mosque in Diraz and warned him against accommodating a Salam Ya Mahdigathering that was planned for July 18. In response, protestors organized a car rally in Manama during which they chanted Salam Ya Mahdiand played it on loudspeakers. The Traffic Directorate reportedly summoned several individuals, fined them, and seized their vehicles for impeding traffic. In most villages, organizers incorporated the Salam Ya Mahdisong in Eid al-Qadr celebrations, including in Sitra, Markh, and Buri maatams(Shia prayer houses, sometimes called husseiniyasin other countries) and near al-Ajam al-Kabeer maatam in Manama. Speaking at a maatam, Shia scholar al-Ghuraifi stated he disapproved of the MOI summoning individuals for “chanting peacefully” and said that “people have the right to express their beliefs.” The government in February lifted COVID-19 restrictions on Ashura gatherings, which authorities had imposed in 2020. According to procession organizers, more than 100,000 Shia Muslims gathered in central Manama to mark Ashura on August 7-8. According to prisoners’ families, prisoners reported significant positive changes from previous years in conducting religious processions during Muharram, including for Ashura, such as authorities giving prisoners additional time to commemorate Ashura and gather after the religious processions. During site visits to the Dry Dock detention facility and Jaw Prison in August, the NIHR stated authorities gave Shia inmates additional time to practice Ashura rituals in common areas. They also reported the prison provided books, banners, and religious chanters to facilitate Ashura commemorations as well as TV screens on which to watch processions and allowed prisoners to gather in designated areas to commemorate Ashura en masse. Bahrain Mirror, however, reported authorities in Jaw Prison prevented some prisoners from practicing religious rituals during Muharram. Human rights activist Yousif al-Muhafda told the news outlet prison guards “mobilized” to prevent detainees from exercising their religious freedoms. He said prisoners feared guards would beat them if they did not comply. On August 2, Sayed Ahmed al-Wadaei of the London-based Bahrain Institute for Rights and Democracy posted on Twitter that prison authorities did not provide access to certain Shia religious books during this time. According to a July statement by the Minister of Interior, the government permitted individuals and families from outside of the country to participate in Ashura rituals, but, in response to commercial advertisements inviting tourist groups from Kuwait to participate in Ashura processions in Bahrain, for the first time barred large groups from abroad arranged by travel agencies. The government said it wanted to “protect the spirituality” of the event and not let Ashura devolve into a tourist spectacle. On October 12, ADHRB called on Pope Francis to reconsider his decision to visit the country on November 3-6, citing religious discrimination against Shia individuals and mistreatment of Shia political prisoners. Other media sources reported human rights activists said the pontiff should use his visit to speak out for Shia rights and meet with prisoners. On November 1, Amnesty International’s Bahrain researcher told the Associated Press, “The watchwords of this [papal] visit are coexistence and dialogue, and the Bahraini government suppresses civil and political freedoms, without which coexistence and dialogue cannot be sustained.” Vatican Newsreported that on November 5, Pope Francis presided at a Mass at Bahrain National Stadium in Riffa, a suburb of Manama, before a crowd of approximately 30,000 Catholics and others from across the region. Voice of America stated one of King Hamad’s sons and several government ministers also attended. The MOI stated its community policing program enlisted individuals directly from local communities, including Shia citizens, to act as informal community police, with the goals of maintaining local peace and security, resolving local issues at the community level, and avoiding escalating conflicts to law enforcement authorities. The MOI stated these informal community police monitored religious gatherings and funerals to prevent those gatherings from degenerating into protests or acts of violence. The government-affiliated Bahrain News Agency reported in August that police directorates “continued their field visits to inspect the needs of maatams” and that “community police provided facilities and services around community centers [maatams] and were involved in safety awareness campaigns.” In June, opposition social media stated that authorities required Shia citizens seeking to travel through Bahrain International Airport to visit religious shrines in Iran, Iraq, and Syria to register their travel and receive permission from the government, citing security issues. Some progovernment commentators said the government imposed the restrictions due to suspected links between Shia pilgrims and extremists based in these countries. Local sources said the government did not enforce the policy uniformly. According to reports, many travelers went through Kuwait, Saudi Arabia, and the UAE instead. Individuals posted videos to Twitter of protestors in the village of Sanabis chanting antiregime slogans in response to the reported travel restrictions. Other opposition figures condemned the government for operating direct flights to Israel while restricting Shia individuals from visiting religious shrines in Iran, Iraq, and Syria. Some travel agents and citizens said the registration and permission procedures were smooth and they were allowed to travel to Iran, Iraq, and Syria. There were conflicting reports of whether citizens needed government permission to travel to Iraq for al-Arbaeen ceremonies, a major religious ritual in which Shia Muslims commemorate the martyrdom of the Prophet Muhammad’s grandson, Hussein bin Ali. On September 9, Safa al-Khawaja, a member of the dissolved Bahrain Center for Human Rights, stated on social media that she was able to travel to Iraq. On September 9 and 10, however, the leaders of the dissolved Shia Scholars’ Council reported on their social media accounts that authorities prevented them from traveling to Iraq. Media reported that in December, thousands of Shia commemorated the martyrdom of Fatima, daughter of the Prophet Muhammad, in street processions and at maatams and mosques throughout the country. Alternative noncustodial sentencing beneficiaries stated authorities prohibited them from participating in social, cultural, and religious activities, including visiting mosques and maatams, or attending religious commemorations while serving their sentences. The MOJIA opened all places of worships at full capacity starting on February 15. No physical distancing or proof of vaccination was required. Masks remained mandatory in religious venues until May. On November 10, in advance of parliamentary elections on November 12, Amnesty International issued a statement calling on the government to lift its ban on the Shia-affiliated al-Wifaq and Amal opposition parties and the Wa’ad secular opposition party. Amnesty International said, “The banning of parties which have peacefully sought to change the system of government by legal means such as participating in elections is a flagrant violation of the right to freedom of association.” The law prohibiting political activities at worship centers remained in effect. The government did not allow individuals to use mosques, maatams, or other religious sites for political gatherings. Adherents of minority religious groups reported they were able to produce religious media and publications and distribute them in bookstores and churches, although the government did not permit publications it perceived as criticizing Islam. The Ministry of Information Affairs continued to review all books and publications prior to issuing printing licenses. The MOJIA also continued to review books that discussed religion. According to representatives from the Christian and Hindu communities, the government did not interfere with their religious observances. Government officials made public statements encouraging tolerance for minority religious beliefs and traditions. The government reported there were 575 licensed Sunni mosques, a decrease from 598 in 2021. Authorities decreased the number of licensed Shia mosques to 762 (from 763 in 2021) and increased the number of maatams to 634 (from 624 in 2021). No mosques or maatams were closed during the year for violating COVID-19 guidelines. The MOJIA’s website listed 100 Quran centers (where children memorize the Quran), 78 religious centers, 7 community awareness centers, and 5 sharia institutes. During the year, the government granted permits to build 14 new Sunni mosques and seven Shia mosques and maatams. In April, Prime Minister Salman bin Hamad Al Khalifa, who is also the Crown Prince, announced the restoration of 20 Sunni and Shia mosques, the construction of 12 new mosques (six Sunni and six Shia) in Salman City, and the launch of a development plan for mosques across the governorates. According to the Jaafari Waqf, 13 of the 20 to be restored would be Shia. On April 13, Minister of Justice, Islamic Affairs, and Endowments Nawaf bin Mohammed al-Maawda stated that six mosques would open during the year in Isa Town, Jurdab, Salhiya, Khalifa Town, Eastern Hidd, and Jaw. Fourteen other mosques in Manama, Juffair, Naeem, Eker, Salmaniya, Sitra, Sadad, Adhari, Jid el-Haj, and Hamad Town were under renovation at year’s end. On April 9, Minister al-Maawda said the Sunni Waqf allocated 800,000 dinars ($2.1 million) during the year for the maintenance of all existing Sunni mosques. Photographs posted on social media in April and May showed that worshippers gathered occasionally for prayer at the former sites of Shia mosques that the government demolished in the 2011 uprising, which appeared in the photographs to be empty plots or parking lots. On February 1, Indian Prime Minister Narendra Modi thanked Prime Minister Salman via Twitter for the government’s decision to allot land for the construction of an additional Hindu temple in Manama. The Hindu religious group Bochasanwasi Shri Akshar Purushottam Swaminarayan Sanstha was expected to oversee construction of the complex, with a planned prayer hall, community center, amphitheater, food court, and other facilities. Construction had not begun on the new Hindu temple as of year’s end. As of year’s end, the municipality of Awali had not approved requests from three congregations seeking since 2014 to construct new churches on land donated by the King for that purpose; municipal authorities cited unspecified security concerns. The municipality made no public statements about the delay in responding to the congregations’ requests. The Christian Cemeteries Committee began a campaign to raise $50,000 (133,000 dinars) to construct a new Christian cemetery in Salmabad. The new cemetery would be the third Christian cemetery in the country. The government officially allocated the land in late February, eight years after Christian community leaders notified the government in 2014 that they had filled the country’s second Christian cemetery’s final burial plot. The government reported it did not receive any new requests during the year from religious groups seeking formal registration. It continued to permit both registered and unregistered non-Muslim religious communities to maintain identifiable places of worship, hold religious gatherings, and display religious symbols, such as crosses outside churches. Shia sources expressed dissatisfaction that Shia doctrine was not included in Islamic studies in public schools, which they said focused on Sunni Islam. The independent but government-affiliated King Hamad Global Center for Peaceful Coexistence (King Hamad Center) continued to offer student exchanges and educational programs that it stated were focused on dispelling ignorance, discrimination, and intolerance, including religious intolerance. These included the King Hamad Faith in Leadership Program and the Cyber Peace Academy. The center’s board of trustees comprised representatives of the country’s Sunni, Shia, Christian, Baha’i, Hindu, and Buddhist communities. In April, the center signed a memorandum of understanding with the University of Haifa to promote coexistence between Arabs and Jews in the Middle East. The public University of Bahrain continued to offer degree programs in religious studies and Islamic jurisprudence for Shia and Sunni students. In January, Bahrain Mirrorreported that human rights activist al-Muhafda posted to Twitter that Shia students had complained to him that a required religious text in the Islamic Studies degree program “insults a symbol of the Shiite community.” He demanded an independent inquiry into the matter. He did not specify the nature of what he considered the objectionable material. All students, regardless of religious affiliation, were eligible to participate in the Crown Prince International Scholarship Program (CPISP); the government did not provide a statistical breakdown of participants by religious affiliation. CPISP published a list of scholarship recipients’ names, fields of study, and schools on its website. Human rights activists continued to state the MOE discriminated against Shia students in university scholarship distribution. There were reports that the MOE refused to recognize degrees granted by higher education institutions in foreign countries serving students who were unable to access scholarships in Bahrain or had limited financial means. Some activists said these refusals disproportionately affected Shia students, who were more likely to be in those circumstances. A member of parliament criticized the MOE for failing to accredit advanced degrees obtained from foreign universities. In response, then Minister of Education Majid bin Ali al-Nuaimi stated during the April 5 session of the Council of Representatives that the Diplomas Evaluation Committee (which assesses the qualifications of foreign advanced degrees in the humanities and social sciences) approved more than 6,000 masters and doctoral theses from foreign universities and rejected 12 on the grounds they harmed national unity or incited sectarianism, after requiring another 20 graduates to revise their theses. On March 16, the Ministry of Works, Municipalities Affairs, and Urban Planning announced the beginning of construction of the Sheikh Abdulla bin Khalid Faculty for Islamic Studies, to be located in the Southern Governorate. The project was expected to cost 1.1 million dinars ($2.9 million) and be finished in March 2023. The government-run television station Bahrain TV broadcast Friday sermons from the country’s official al-Fateh Mosque and other Sunni mosques, such as Sabeeka bint Ebrahim Mosque and Sabeeka al-Nusf Mosque, but not sermons from Shia mosques or clerics. Some Shia activists said this was discriminatory, while others said it was better not to be subject to government broadcasting restrictions. Many Shia mosques disseminated sermons via social media. A government-affiliated human rights monitoring organization and family members of prisoners said Shia prisoners could view Shia sermon videos on their mobile telephones. A November article in the Economistquoted a local businessman as saying that hunger and limited opportunities could ignite a wave of protests among the Shia population. The article said the Prime Minister “won praise for rolling out COVID-19 vaccines to all Bahrainis, regardless of sect,” but that “his failure to soothe Sunni-Shia tensions [was] setting Bahrain up for trouble.” One Shia businessman told the Economist, “The sectarian divide from a decade ago [during the 2011 antigovernment protests] hasn’t healed.” On November 3-4, the government, in partnership with the Muslim Council of Elders, SCIA, and King Hamad Center, hosted the “Bahrain Forum for Dialogue: East and West for Human Coexistence” in Manama. Pope Francis, Grand Imam of al-Azhar Ahmed al-Tayyeb (the most senior figure in Sunni Islam), Ecumenical Patriarch Bartholomew I, and 200 other religious leaders and prominent academic and media figures from around the world attended. During the forum, some European and Bahraini participants said the government did not adequately address the vulnerability of atheist youths, who risked ostracism, homelessness, and sometimes physical harm for their lack of belief. While by law Arab applicants with 15 years’ residence and non-Arab applicants with 25 years’ residence are eligible to apply for citizenship, arbitrary implementation of the law from the application stage to approval remained a common criticism of both Shia and Sunni citizens, as well as migrant rights activists. The government stated foreign residents applying for citizenship were not required to report their religious affiliation. Shia politicians and community activists, however, continued to say the government’s naturalization and citizenship process favored Sunni over Shia applicants. They said the government continued to recruit Sunnis from other countries to join the security forces, granted them expedited naturalization, and provided them with public housing, while excluding Shia citizens from those forces. According to Shia community activists, the continued recruitment and expedited naturalization of Sunnis represented an ongoing attempt to alter the demographic balance of the country’s citizens. The King appointed a new cabinet, selected by the Prime Minister, whose members were sworn in on June 15. The government did not maintain official statistics on the religious affiliation of public sector employees, members of parliament, or ministers, but nine of the 22 ministers publicly identified as Shia. A June 16 editorial in Bahrain Mirrorstated the Prime Minister should have selected “at least” an equal number of Sunni and Shia individuals, considering the country’s Shia majority. Following parliamentary elections in 2018, of 40 seats on the elected Council of Representatives, 25 were won by members identified as Sunnis and 15 by members identified as Shia. According to informal estimates, the Shura Council, parliament’s upper house whose members are appointed by the King, included 19 Shia members, one Jewish member, and one Christian member, while the remaining 19 members were Sunni. According to Shia leaders and community activists, the government continued to give Sunni citizens preference for government positions, especially in the managerial ranks of the civil service, military, and security services. They also said Sunnis received preference for other government-related employment, especially in the managerial ranks of state-owned businesses. Few Shia citizens served in leadership posts in the defense and internal security forces. According to Shia community members, senior civil service recruitment and promotion processes favored Sunni candidates. Shia community members said educational, social, and municipal services in most Shia neighborhoods remained inferior to those in Sunni communities. The government stated it made efforts to support public schools in Shia and Sunni neighborhoods equally. The MOSD organized expositions, job fairs, professional guidance, and assistance to needy families in predominately Shia neighborhoods. The MOSD, which has a supervisory role in implementing labor law in the private sector, again said there were no reported cases of religious or sectarian discrimination during the year. Shia community activists again responded that they lacked confidence in the effectiveness of government institutions to address discrimination and therefore did not utilize them. In July, the government issued the National Human Rights Plan for 2022-2026. The plan, developed in consultation with the Office of the UN High Commissioner for Human Rights, included discussion of religious freedom and freedom of expression, among other topics. It called for implementing education reform, a review of relevant laws, capacity building through training and seminars, and other projects to advance the country’s objectives of promoting religious freedom and “enabling a culture of tolerance and peaceful coexistence.” The plan outlined the government’s intent to develop television programs and films on tolerance and religious coexistence, periodically review controls on religious discourse, and review relevant laws and develop amendments “to guarantee the enjoyment of religious freedom for all.” During a July international Ministerial Conference on Freedom of Religion or Belief in London, Minister of Foreign Affairs Abdullatif bin Rashid al-Zayani commended what he said was the positive state of freedom of religion and tolerance in the country. Al-Zayani said that freedom of religion was the main objective of the 2022-2026 National Human Rights Plan. In February, then-Israeli Prime Minister Naftali Bennett met with King Hamad and Prime Minister Salman in Manama, becoming the first Israeli head of state to visit the country. During his visit, Prime Minister Bennett met also with members of the Jewish community. Bahrain’s most senior Shia cleric, Sheikh Isa Qassim (whose citizenship the government revoked in 2016 and who resides in exile in Iran), and al-Wifaq representatives issued statements condemning the visit. There were also reportedly small nonviolent protests in some Shia villages marking the anniversary of the 2011 antigovernment protests and criticizing the government’s normalization of relations with Israel. Opposition-linked social media accounts shared pictures of signs in the town of Abu Saiba on February 14 bearing the al-Wifaq slogan “United on the Path of Righteousness” and picturing Sheikh Qassim next to what the accounts described as martyrs from the 2011 unrest. In April during Ramadan, local media outlet a l -W atanreported the King, as he had done in prior years, met with senior Shia cleric al-Ghuraifi, a founding member of Hizb al-Da’awa in the late 1960’s, later rebranded as al-Wifaq. The two men discussed the value of religious discourse. The King said clerics were “role models in religion, morals, and enlightened discourse.” Social media commentators described the visit as a trust-building exercise for the government and Shia community. From May 31-June 1, the King Hamad Center hosted the EU-Bahrain conference entitled “Broadening the Tent: Freedom of Religion and Belief” in Manama. The conference featured religious community leaders, and politicians from Europe and neighboring countries. Bahrain News Agency reported that on September 6, Shaikh Khalid bin Khalifa Al Khalifa, chairperson of the King Hamad Center, met with Captain Doug Newman, regional commander of the Salvation Army for the Middle East Region. They discussed future cooperation on projects to promote religious tolerance. Newman expressed the Salvation Army’s desire to make the country the headquarters of its regional activities. He praised what he said were the country’s efforts, led by King Hamad, in promoting peace in the region as well as what he termed the center’s efforts in spreading tolerance and coexistence as part of its humanitarian and charity work. On November 3, the President of the SCIA, Sheikh Abdulrahman bin Mohammed Al Khalifa, announced the new King Hamad Prize for Dialogue and Coexistence to recognize the importance of fostering peaceful dialogue between different cultures and religions. The Muslim Council of Elders, an international independent organization founded in Abu Dhabi by the Grand Imam of al-Azhar with the expressed aim of promoting peace in Muslim-majority countries, held its 16th session in Manama on November 3-4, during the Bahrain Forum for Dialogue. Section III. Status of Societal Respect for Religious Freedom Non-Muslim religious community leaders again reported that there was ongoing societal pressure on individuals not to convert from Islam.  Those who did so were unwilling to speak publicly or privately to family or associates about their conversions out of fear of harassment or discrimination. Sources reported local society continued to reject any forms of atheism. Both anti-Shia and anti-Sunni commentary appeared in social media.  Anti-Shia posts described Shia opponents of the government as “traitors,” “agents of Iran,” “terrorists,” “killers,” “criminals,” plotters,” and, occasionally, “ rawafid” (a derogatory term describing Shia who refused to accept the early caliphs).  Anti-Sunni posts described the ruling Al Khalifa family and its supporters as “ nawasib” (a derogatory term describing Sunnis who are hostile to the family of the Prophet Muhammad). NGOs working on civil discourse and interfaith dialogue reported Sunni-Shia tensions and historical political divisions continued to have negative economic effects.  Shia representatives stated the persistently higher unemployment rate among members of their community, limited prospects for upward social mobility, and the lower socioeconomic status of Shia added to the tensions between the two communities. According to minority religious groups, there was a high degree of tolerance within society for minority religious beliefs and traditions, although not for conversion from Islam or for atheistic or secularist views.  Holiday foods, decorations, posters, and books were widely available during major Christian, Jewish, and Hindu holidays, and Christmas trees and elaborate decorations remained prominent features in malls, restaurants, coffee shops, and hotels.  The news media continued to print reports of non-Muslim religious holiday celebrations, including Christmas and Jewish celebrations such as Hanukkah and Hindu festivals such as Diwali and Holi. In May and June, the UAE research and consulting firm ASDA’A-BCW conducted a poll of youth between the ages of 18 and 24 in 17 Arab states and territories and reported 59 percent of Bahraini respondents said preserving their religious and cultural identity was more important than creating a more globalized society. Conspiracy theories that the government intended to establish a Jewish neighborhood in Manama were accompanied by a brief surge in online antisemitic rhetoric in September, which included one social media account posting a political cartoon with the presumed likeness of a well-known member of the local Jewish community.  Online sources, including some from outside the country, said Jews would take over culturally important parts of Manama’s downtown area.  The House of Ten Commandments Synagogue and the government refuted the claims, and the controversy dissipated. According to social media sources, government opponents organized approximately six small-scale unauthorized demonstrations from December 18 to 25, coinciding with Hanukkah, in Sitra, Diraz, Almarkh, and Muqasha, all with high percentages of Shia residents, in addition to a larger demonstration in the market in Manama.  One social media post read, “In rejection of the Jewish Hanukkah events … the youth of Sitra invite you to the central march under the slogan ‘Do not desecrate our land’.”  On December 25, protesters staged peaceful demonstrations in the Manama souq in rejection of Hanukkah celebrations and normalization of relations with Israel.  Videos circulating online showed persons chanting “Death to Israel” near the House of Ten Commandments Synagogue.  The region’s oldest synagogue, it hosted approximately 300 Hanukkah observers from Israel, Europe, and the United States, as well as visitors from other local religious communities, throughout the week.  Several government-affiliated newspapers issued columns defending the right of Bahrainis and visitors to carry out their religious practices. According to Jewish community representatives, the House of Ten Commandments Synagogue publicly observed Jewish traditions and held prayers in the downtown area of the capital.  The synagogue continued to host visitors to the country and provide services to Jewish families from neighboring countries.  The Association of Gulf Jewish Communities launched a campaign to restore the area’s only Jewish cemetery, located in Manama.  Kosher-certified food products were available in supermarkets, and meals prepared under kashrut supervision. Section IV. U.S. Government Policy and Engagement The Ambassador, other senior U.S. government officials, and embassy representatives met with senior government officials, including the Prime Minister, Foreign Minister and the Minister of Justice, Islamic Affairs, and Endowments, and other cabinet officials, and at the MOSD, Ministry of Legal Affairs, and national human rights monitoring institutions to urge respect for freedom of religion and expression, including the right of clerics and other religious leaders to speak and write freely.  Embassy officials encouraged continued government efforts to counter religious extremism and to ensure full inclusion of all citizens, including members of the Shia majority, in political, social, and economic opportunities.  U.S. officials publicly and in private meetings advocated that the government pursue political reforms that took into consideration the needs of all citizens regardless of religious or prior or current political affiliation by, for example, engaging civil society in reconciliation dialogue, encouraging an impartial application process for government positions, and allowing individuals previously connected to religiously based opposition political groups to run for public office. The Ambassador and other embassy officials met regularly with religious leaders from the Muslim, Christian, Hindu, Jewish, and other communities as well as representatives of NGOs and political groups to discuss freedom of religion and freedom of expression as it related to religious practices.  Embassy representatives and senior U.S. officials visited various houses of worship and attended religious events during the year, including observations of Ashura, Ramadan, Eid al-Fitr, Christmas, Hannukah, and Diwali.  At these events, they discussed issues related to religious tolerance with participants and emphasized the U.S. government’s commitment to religious freedom. On October 12 to 13, the U.S. Deputy Special Envoy to Monitor and Combat Antisemitism met with senior government officials to discuss joint efforts to combat antisemitism in all its forms.  Government officials made public statements in support of the visit and emphasizing the principles of peace and coexistence.  The Deputy Special Envoy and the chair of the King Hamad Center met to discuss potential opportunities for cooperation on combating antisemitism. The Deputy Special Envoy also met with civil society leaders and members of the Jewish community.  He gave an interview with Arabic daily al-Ayam in which he stressed the need to decouple geopolitics from religious issues and clarified that the Israeli-Palestinian conflict and antisemitism were separate questions.  During the Deputy Special Envoy’s visit, the Ambassador hosted an event with current and former government officials and others at which participants discussed the importance of educating young people about the country’s Jewish history as well as the Holocaust.  The embassy subsequently hosted an October discussion on antisemitism with local youth and university students; the students highlighted the presence of an Indigenous Jewish population and what they stated was the country’s tradition of inclusion and respect for various religions. The embassy continued to encourage the participation of religious leaders in exchange programs in the United States designed to promote religious tolerance and a better understanding of the right to practice one’s faith as a fundamental human right and source of stability.  The embassy regularly highlighted on social media high level engagements with religious leaders and the embassy’s participation in religious observances, for example, during Eid al-Fitr, Ashura, Diwali, and Ramadan, as part of its efforts to advance freedom of religion and belief. ecoi.net description: Annual report on religious freedom (covering 2022) Country: Bahrain Source: USDOS – US Department of State (Author) Original link: https://www.state.gov/reports/2022-report-on-international-religious-freedom/bahrain Document type: Periodical Report Language: English Published: 15 May 2023 Available on ecoi.net since: 22 May 2023 Document ID: 2092183

https://www.ecoi.net/en/document/2092183.html

The Job Season Without In-Person Interviews How a virtual search process can lead to better, more equitable hiring. The Job Season Without In-Person Interviews How a virtual search process can lead to better, more equitable hiring. Getty Images On the first day of September, I welcomed a new member of my team in the dean’s office via Zoom. I was on the campus, but our new hire — let’s call her C., so as not to put her on the spot — was still more than 1,000 miles away. C. had asked to start work remotely for three weeks so she could have time to pack up, find a place to live, and move, all of which would take more time than usual given the restrictions of pandemic life. Just that morning, she had closed on a new house. Had she come to New Orleans to look at our housing market? No, she replied, a realtor had showed her houses via FaceTime. “I’m committed!” she said. On the first day of September, I welcomed a new member of my team in the dean’s office via Zoom. I was on the campus, but our new hire — let’s call her C., so as not to put her on the spot — was still more than 1,000 miles away. C. had asked to start work remotely for three weeks so she could have time to pack up, find a place to live, and move, all of which would take more time than usual given the restrictions of pandemic life. Just that morning, she had closed on a new house. Had she come to New Orleans to look at our housing market? No, she replied, a realtor had showed her houses via FaceTime. “I’m committed!” she said. Speaking with her that first day on the job, it struck me how unusual all of this would have seemed only six or eight months ago. Her success remains to be tested, of course. Still, I am confident that our hiring process not only led to the best choice, but that it screened out aspects of academic hiring that have long bothered me for how they replicate forms of privilege and implicit bias. As a dean and professor, I’ve done my share of hiring, and chaired or been involved with dozens of faculty, staff, and administrative searches. In this case, C. holds a senior administrative position. But what we learned from this virtual search seems relevant to both faculty and administrative hiring going forward. Driven out of necessity, the way we structured our search process helped us focus more on the talents of the candidates and the likelihood of their success in the job than on the superficial aspects of traditional interviews which tend to leave cultural self-replication unchallenged. If we are serious about transforming academic culture and leaving behind the pernicious category of “fit,” we need to reconsider how we go about hiring. I wasn’t thinking about behavioral economics when I started this job search, nor the deep tension between intuition and judgment that can plague in-person interviews. In fact, the job description itself had been crafted before the pandemic forced us to take a new approach, and I thought little of the means by which we’d go from approval to hiring. We were about to post the job ad when Covid-19 hit, which led to a hiring freeze for all but essential personnel. A significant portion of this position is essential to our educational mission, so we got approval to restart. But now we were in pandemic mode and had to improvise. First, I asked a larger number of colleagues than usual to play a role on the search — not just people in the dean’s office but also faculty and staff members — and divided them into teams of two or three. The first team screened applications based on CVs, focusing on qualifications. Then a second team conducted Round 1 interviews, with questions and interview duration standardized. Yet another team interviewed those who advanced to Round 2. And I recruited another dean to interview the final two candidates in Round 3. My executive assistant coordinated the process and asked everyone to write up their ratings of the candidates at each stage. I asked my executive assistant not to tell me what the teams were reporting until the end of each round. I also interviewed the candidates at each stage — briefly for Round 1, and more extensively as we went forward. I compared my notes against those of that round’s interview team, before deciding which candidates would move into the next round. This allowed for dissent throughout the process, and heightened the likelihood that judgment, rather than intuition, would take precedence in our decision. As it turned out, there was agreement at each stage. The final decision was mine, but in making it, I had maximized the amount of input I had to draw on and purposely slowed down the process. Why so much structure? Job interviews are one of those tasks in which the two competing aspects of decision making — intuition and judgment — are particularly likely to be thrown out of balance. In his 2011 book , Thinking, Fast and Slow, Daniel Kahneman , an emeritus professor of psychology at Princeton University and winner of the Nobel Prize in economics, argues that there are two aspects to how we make decisions. He calls them “System 1” — driven by intuition, speed, and emotions — and “System 2” —more deliberative and slow, associated with judgment. System 1 is highly valued in our society. We often celebrate leaders for their quick decisions and “expert intuition.” Intuitive decision making frequently carries the day, particularly in job interviews. Even those committed to working against implicit biases or the so-called halo effect (the influence of positive first impressions) have a hard time resisting the impulse to rely on intuition. Anyone who has served on a faculty search committee, working through dozens or hundreds of applications in order to derive a short list, has witnessed how conference interviews or campus visits can turn the tables. We argue against fetishizing the brilliant job talk but find ourselves unable to resist the candidate who “interviews well.” In his work on interviews, Kahneman tried to temper the powerful charisma of System 1 thinking by developing System 2 mechanisms. He asked colleagues to decide in advance which aspects of doing a particular job mattered most, and then to use interviews to rate candidates numerically on how well they fulfilled each characteristic. Such simple algorithms, he argues, are better than so-called “expert” judgment. “Because interviewers are overconfident in their intuitions,” he writes, “they will assign too much weight to their personal impressions and too little weight to other sources of information.” Even before Covid-19, many academic departments were moving away from using annual scholarly conferences to conduct first-round, in-person interviews for a faculty opening. One of the reasons for that shift is budget cuts. high expense for graduate students or underemployed Ph.D.s to travel to conventions, frequently for a single job interview; and the environmental impact of so much travel. Those are all good reasons to start using Zoom interviews — even after the pandemic is over — to assemble the short list of candidates you plan to bring to town for campus interviews. Now, with Covid-19 continuing to limit travel this year and to force academic conferences to go virtual or be canceled, the question is: What might we gain by moving allstages of the hiring process online? (Some may object that on-campus interviews allow departments to recruit a candidate and show off their campus, colleagues, or city, but this function can be separated out till after a hiring decision is made. Too often, in my experience, the so-called “wining-and-dining” stage can go awry and privilege social niceties over academic and professional credentials.) As I look back on our experience running this administrative search, virtual interviews democratized our procedures and minimized aspects of hiring built around intuition and discriminatory assumptions about class, gender, disability status, and race — assumptions that I believe fall into the category of System 1 thinking. By focusing attention on the candidate’s talents and experience, virtual interviews can screen out many if not most of what decision makers, despite their best intentions, may be distracted by during initial in-person encounters. Sure there are multiple social cues and codes that can come across in a Zoom interview and put implicit bias back into play. Room Rater , a popular Twitter account that evaluates (tongue firmly in cheek) Skype and Zoom backdrops, has taught all of us to look at cues in the background of a room. Being able to captivate during a Zoom interview allows some candidates to thrive. Putting your laptop on a stack of books, aiming a warm light source at your face, and taking care with your background are simple hacks for job candidates that can be surprisingly effective. We need to be on the lookout for the candidate who “interviews well” on Zoom, too, despite lacking the background and training we need for the position. But the tendency to be influenced by a candidate’s charisma can be countered on the hiring side by a rigorous System 2 process that builds algorithms and rates candidates accordingly — like the process that we put together on the fly. For faculty hires, no less than administrative ones, quick judgments are activated by prestigious letterhead or famous reference writers. Even simple algorithms that isolate the characteristics for success as a faculty member at your particular institution — rather than a generalized image of success — forces interviewers to focus on those traits. After C. had accepted the position, I told our new chief diversity officer my theory about the all-virtual search: “I feel I know all that I need to know. When C. gets to New Orleans, I’ll find out if she’s short or tall.” That was a joke, of course, but intended to make a serious point. Like any of the candidates, C. might be short or tall, in a wheelchair, or have no legs at all, but none of that would matter for the job we’d hired her for. And while height, weight, the kinds of shoes you wear, or the briefcase you carry are not at all relevant to the job, so many of those cues do affect decision making in person, however implicitly, which is where the devil lies. A version of this article appeared in the October 30, 2020, issue .

https://www.chronicle.com/article/the-job-season-without-in-person-interviews

Aubrey Williams: Abstraction in Diaspora by Kobena Mercer | Article index | Articles | British Art Studies An online, open-access and peer-reviewed academic journal for research on all aspects of British art, architecture and visual culture. Aubrey Williams: Abstraction in Diaspora ArticlebyKobena Mercer Aubrey Williams, Death and the Conquistador (detail), 1959, oil on canvas, 83.5 × 133.8 cm. Collection of Tate (T13341). Digital image courtesy of Estate of Aubrey Williams (All rights reserved, DACS 2018). DOI Abstract DOI Moving to London in 1952, Aubrey Williams gained valuable distance on the Amerindian petroglyphs that inspired his abstract painting. But as he deepened his engagement with the indigenous cultures of the precolonial Caribbean during the 1970s—working in studios in Jamaica and in Florida—Williams was edged out of late modernism’s narrative of abstraction. While retrospective exhibitions highlight the Olmec-Maya and Now series and the Shostakovich DOI The word “diaspora” comes to us from the Greek verb “to sow” and by extension “to disperse” combined with the preposition for “over” which gives us the “dia” in diameter. 1 2 While I agree with Brett wholeheartedly, my critical concern is that if we focus only on the neglect, and attention toward the paintings themselves is thereby postponed, the danger is that we may end up reinforcing the oversight that isolated Williams from the narrative of post-war abstraction in the first place. This is the dilemma that snags Eddie Chambers’s account in his survey of black artists in British art. 3 Tate Britain’s posthumous acquisition, in 1993, of Shostakovich Symphony No 3 Opus 20 (1981) and Olmec-Maya, Now and Coming Time (1985), along with the 2007 study day held when Tate Britain acquired letters and drawings from Williams’s archive, are indeed belated steps when, as Chambers points out, the institution had the opportunity to purchase one of Williams’s paintings in 1961, yet chose to decline ( Fig. 1 Aubrey Williams: Atlantic Fire at the Walker Art Gallery, Liverpool, and Aubrey Williams: Now and Coming Time at the October Gallery, London—attests to the deep interest Williams’s work commands among contemporary audiences eager to engage with insights generated from a diasporic practice of abstraction whose globe-spanning viewpoint has grown ever more relevant since the artist’s death nearly thirty years ago. DOI Figure 1.Aubrey Williams, Olmec Maya - Now and Coming Time, 1985, oil on canvas, 119 × 178 cm. Collection of Tate (T06675).Digital image courtesy of Estate of Aubrey Williams (All rights reserved, DACS 2018). New Commonwealth Moments and the 1960s Crisis of Institutional Modernism When Stuart Hall characterised Aubrey Williams and other abstract painters such as Anwar Shemza (1928–1985), from Pakistan, and Frank Bowling (b. 1934), from Guyana, as members of a generation who were among “the last colonials” to arrive into the post-1945 London artworld, he underlined their “universalist and cosmopolitan outlook,” emphasising that they felt “they belonged to the modern art movement and, in a way, it belonged to them.” 4 DOI But as the modernism institutionalised in the 1950s was thrown into crisis during the mid- to late-1960s, mounting pressures forced cracks in such “colour-blind” internationalism. Where the crisis of modernism erupted most vividly in the United States, with protests against the Museum of Modern Art in New York, and the Whitney Museum of American Art, led by African American organisations such as the Black Emergency Cultural Coalition, Susan Cahan’s detailed account reveals that institutional decision makers rarely gave much thought to the foundational assumptions on which their collections and exhibitions were built. Only as a result of external pressure did an institution such as MoMA reflect on its acquisitions criteria. When the Art Workers Coalition called for a public debate on MoMA’s policies in 1969, executive director Bates Lowry declined. On the day of the boycott, the museum issued a press release in a Q&A format: In selecting works works of art ... does the museum consider the sex, nationality, religion, politics or race of an artist? NO. What criteria does the museum apply? Quality, historical significance, significance of the moment. 5 In the monologue whereby we witness an institution talking to itself so as not Disavowal is when everyone agrees not to see what everyone knows is there. Whether it is the elephant in the room or the emperor’s new clothes, what is left out of the discourse, and goes unspoken, is not the result of accidental oversight but, in psychoanalytic terms, is the outcome of a process of repression. 6 Hence, coming back to Britain, Commonwealth artists were genuinely welcomed in their individuality, but the post-war reception was short lived. This was not primarily because, by the mid-1960s, abstraction was displaced by pop, minimalism, and performance, but because object-directed formalist criticism rendered all aspects of social context and social identity into a constitutive absence , something that had to be left out if formalism was to provide the epistemic authority on which institutional decision making relied. 7 8 Rather, the aim of revising previous narratives in light of what we now know about their exclusionary structures is to recast our understanding of the structural interaction among artists, artworks, and artworlds so as to better see the contingent conditions under which their conjunctures can be opened to reinterpretation. The diaspora concept, I argue, helps us to do just that. Border-Crossing Migrations 9 As a youngster, Williams studied in the Working People’s Art Class set up in Georgetown by painter E.R. Burrowes, while training as a civil servant as his parents wished ( Fig. 2 After touring Europe for a year—Albert Camus introduced him to Pablo Picasso in Paris—Williams enrolled at St Martin’s in 1954, but decided not to pursue the diploma: in his second year he chose merely to use the art school’s facilities instead. 10 His first solo exhibition, in 1954, at the Archer Gallery in Westbourne Grove, was followed by two in 1959, and in 1960, at the New Vision Centre Gallery, a venue in Marble Arch set up by Denis Bowen. Born in South Africa to British parents, Bowen was a painter, art critic, and curator who ran the New Vision Centre Gallery from 1955 to 1966 with Polish painter, Halima Nalecz, and British artist, Frank Avray Wilson ( Fig. 3 ). In 1963, in the same year Williams enjoyed a further solo exhibition at the Grabowski Gallery (which had previously exhibited Bowling), Bowen played a leading role in selecting entries to the first Commonwealth Biennale of Abstract Art, held at the Commonwealth Institute in London, at which Williams was awarded a prize for Roraima (1963). 11 informel and tachisme . Williams readily acknowledged the inspiration he took from the Tate exhibitions, Modern Art in the United States , in 1956, and New American Painting in 1959—“Pollock was our god!” he said in a 1987 interview, “Kline, Newman, Rothko, de Kooning … they were all great” 12 —but did that necessarily mean he sought to emulate the abstract expressionist paradigm? Figural Mark Making: Displacing Primitivist Paradigms “Despite Williams’s attempt to work in the manner of his American heroes,” writes Gavin Butt in A Companion to Contemporary Art Since 1945 , “his paintings were repeatedly viewed in primitivising terms by British critics.” 13 To the extent that Williams opted for fairly small-scale canvases, employing a variety of brushwork rather than one committed exclusively to gesturalism, it is misleading to align him directly with American abstractionists he admired. Moreover, rather than “primitivising,” would it not be more precise to say the source of the problem in the British reception was that primitivism was the only interpretive paradigm through which institutional modernism attempted to address questions of cultural difference? With coarse crimson, gold, and black impasto markings atop an ochre ground, early paintings such as Earth and Fire (1959) address elemental or even cosmological themes rather than primitive ones as such ( Fig. 4 ). Indeed, the question to put to Butt’s characterisation of the reasons why Williams came to be marginalised by the mid-1960s is whether “primitivism” is the best fit for describing Williams’s relationship to the Amerindian petrogyphs that was a distinctive feature of his abstraction? In the earth-toned palette and dry impasto of Bone Heap (1959), the shapes made by whitened figures laid out on a dark ground are suggestive not only of archaic petroglyphs inscribed in the soil, but also skeletal remains, although the work’s title does not indicate whether the bones are human or animal ( Fig. 5 ). Similarly, one might say of Sleeping Rocks Crystallised Landscape (1959) bears a title that solicits a figurative reading at the same time as the Tachist element of pigment forcefully thrown against the canvas acts as a counterpoint to the more undulating tones and gauzy colours we find in Williams’s paintings ( Fig. 6 ). Figure 5. Aubrey Williams, Bone Heap, 1959, oil on canvas. 50.5 × 60 cm. Private Collection. Digital image courtesy of Estate of Aubrey Williams (All rights reserved, DACS 2018). Figure 6. Denis Bowen, Crystallised Landscape, 1958, alkyd paint on canvas, 63.2 × 97cm Collection of Tate (T07840). Digital image courtesy of Estate of Denis A. Bowen. Whether evoking a gravesite in Bone Heap , or a dormant state prior to awakening in Sleeping Rocks ecological relationship far more so than a “primitivizing” relationship. “Figural” is a term I’ve drawn from philosopher Jean-François Lyotard who uses it in contrast to both “figurative” and “discursive.” 14 prior to Two Insects, Yellow (1950–55), playfully introduced an element of colour ( Fig. 7 ). Devon and Cornwall’s megaliths dating to the Neolithic Age were important for abstract sculptors such as William Turnbull (1912–2012) as well. Similarly grounded in a sense of place were abstract paintings by the Cornish artist Peter Lanyon (1918–1964), with a work as lush and lyrical as Lost Mine (1961), speaking eloquently to the sense of belonging Lanyon found in Cornwall as the place where he was born, where he lived and worked, and where he died ( Fig. 8 ). In terms of a comparative approach, Brett’s argument for aligning Lanyon and Davie with Williams holds good in stylistic terms, to a degree, for we are looking at British variants of post-war abstraction that took a vastly different direction from American abstract expressionism by investigating premodern and prehistorical sources in a lived relationship to an untamed rural landscape. 15 But whereas Lanyon and Davie were regionalists committed to their rootedness in the specificity of the English West Country, the core emphasis Carew drew out in 1959 was on the uprootedness Scientific American , Omni , and other science periodicals. 16 The twin poles of Williams’s interest in the science of rock formation, and the science of galaxy formation, alongside his interest in archaic Amerindian petroglyphs was beautifully synthesised by Carew in the following lines: 17 I would add that at the 2014 Cambridge University symposium on Aubrey Williams, convener Tim Cribb invited Robin Catchpole from the Institute of Astronomy, who helpfully pointed out something of which those of us in the humanities might not be aware, namely that light and colour are mediums that astronomers rely on to glean data regarding the distance between, and relations among, planetary phenomena. 18 Where Williams revealed his investment in astronomy directly in such late works as Nebula in Orion (1985), his painterly handling of atmospherics—with whole portions of his keynote works swathed in the light and colour of nuanced haziness—showed that Williams encompassed macro- and micro-cosmic dimensions in his ecological outlook from the outset. Interstitial Ambiences 19 The sheer scope, and philosophical gravitas, of Williams’s combined interests, or more to the point, the inability of formalist critics to fathom their implications for late modernist art making, are among the reasons why his work came to be pushed aside. Fig. 9 ). 20 In Untitled (1969) we see how Williams’s ability to make lines and shapes “jump” from one plane of semantic association to another meant that the canvas surface, whose flatness and rectangularity were of utmost concern to formalists, had become instead a receptacle for a heterogeneous array of painterly mark making ( Fig. 10 ). Figure 9. Aubrey Williams, Untitled, 1969, oil on hessian, 61 × 74cm. Private Collection. Digital image courtesy of Estate of Aubrey Williams (All rights reserved, DACS 2018). Aubrey Williams, Details from Sketchbooks, 1970s, reproduced in Aubrey Williamsby Andrew Dempsey, Gilane Tawadros, Maridowa Williams (London: Institute for International Visual Arts, 1998). Digital image courtesy of The Institute for International Visual Arts / the Estate of Aubrey Williams. Optically speaking, red advances while blue recedes, yet in the very instant we are tempted to see and read the U-shape in Untitled 21 Magma, Lava, Volcano, Rockface .” Insightfully concluding his 1966 article, Locke said “What is fascinating about these paintings … is that they were closely related to the subject of pregnancy and parturition which has always fascinated him. He portrays the very molten rock as if it were flesh, finding a unique equation between two diametrically opposing natures .” 22 precognitive and preverbal level of affect . 23 interstitial mode of abstraction Diasporic Ancestralism Tribal Mark II (1960) is a work asking just such a question ( Fig. 11 Tribal Mark II creates no longer implies a viewer who stands only perpendicular to the picture plane. The painting suggests a viewpoint from which its markings are to be seen from an aerial position, as though the viewer were hovering “above” the picture plane, looking down as if suspended over an archeological excavation. DOI Figure 11.Aubrey Williams, Tribal Mark, 1961, oil on canvas, 76 2 × 101 5cm. Collection of Tate (T13342).Digital image courtesy of Estate of Aubrey Williams (All rights reserved, DACS 2018). Tribal Mark II actually depicts a foetus or a corpse. But if the aim instead is to demonstrate how figural practice—poised on the borderline between abstraction and figuration— produces a plurality of potential connotations by virtue of affecting us at the precodified level before such markings are fully formed , then we begin to see how Williams put the element of formlessness—the not-yet-fully-formedness—associated with informel in post-war European abstraction into dialogue with ancestralism, a strand of African American modernism centrally preoccupied with questions of absence and presence, life and death, by virtue of addressing the legacy of uprooting and loss among “diasporised” peoples of African descent scattered and dispersed into the New World as a result of transatlantic slavery. Introduced in the 1920s by philosopher Alain Locke, “ancestralism” entailed depictions of African objets d’art among the Harlem Renaissance generation, but in the post-war period Hale Woodruff (1900–1980) turned to motifs in Akan and Yoruba art as a starting point for abstract works such as Afro Emblems (1950) ( Fig. 12 ). 24 DOI Figure 12.Hale Woodruff, Afro Emblems, 1950, oil on linen, 45.7 × 55.9 cm. Collection Smithsonian American Art Museum (1984.149.2).Digital image courtesy of Estate of Hale Woodruff / DACS, London / VAGA, New York 2018 Put another way, the critical project in diasporic ancestralism was never the recovery or redemption of the lost ancestor or an absent ancestry—as if the past could be restored to a state of plenitude—but an investigation into the afterlife of the rupture the nucleus of traumatic affect that made its presence felt—like a phantom limb—through its unsettling aftereffects. Hauntological Traces Looking at Tribal Mark II Every Atom Glows (1951) Lewis spoke to the science of nuclear fission that led to the atom bomb. Lewis’s Harlem Turns White (1954) is not a depiction of an atomic aftermath, but it nonetheless engages abstraction to address traumatic events of world-historical magnitude whose scale exceeds our human ability to grasp them in consciousness ( Fig. 13 ). DOI Figure 13.Norman Wilfred Lewis, Harlem Turns White, 1955, oil on canvas, 127 × 162.5 cm. Collection of the Estate of Norman W. Lewis.Digital image courtesy of Estate of Norman W. Lewis / Michael Rosenfeld Gallery LLC, New York. Insofar as any attempt to “represent” such events diminishes the momentousness of the trauma, reducing it to mere anthropomorphic proportions, it is not that abstract paintings by Lewis or Williams put us in the presence of the unrepresentable—for as Mark Godfrey argues in Abstraction and the Holocaust (2007), the “unrepresentable” is too easily misappropriated by those who claim the events never took place—but that the entire interpretive centre of gravity shifts with regard to understanding abstraction as an inquiry into aftermaths that have been among the defining conditions of modernity. 25 Instead of the progress narrative whereby many formalists saw abstraction as the culmination of modernity in art, the interstitial outlook in Williams’s ecological approach leads instead to the counterview that modernity was a history of successive aftermaths, with one disaster piled up upon others. However, unlike American abstract artists responding to traumatic events within their own lifetime, when we see a work such as Death and the Conquistador (1959) we need to understand that, at age 33, Williams was metaphorically leaping back in world-historical time, inviting us to imagine the cataclysmic advent of 1492—Europe’s arrival into the New World—from the point of view of those whose ancestral homes were about to be decimated by the incoming colonists ( Fig. 14 ). As with another painting of this moment, El Dorado all identities are implicated in the historical trauma and its aftermath. Figure 14.Aubrey Williams, Death and the Conquistador, 1959, oil on canvas, 83.5 × 133.8 cm. Collection of Tate (T13341). Digital image courtesy of Estate of Aubrey Williams (All rights reserved, DACS 2018). Even as figural lines in crimson, black, and white seem to emerge into anthropomorphic shape in Death and the Conquistador they withdraw from legibility at the last minute, as it were, leaving only a tumult of fugitive traces on the picture plane. That colonial history weighed heavily on Williams’s mind in this period is evident in Revolt (1960), a figurative painting that depicts the “decapitation of [a] planter’s wife by an unshackled slave in the 1763 revolt.” 26 Although Williams returned to figuration when he produced four paintings in the Guyana Myths (1971) series (also in Guyana’s National Collection alongside Revolt Death and the Conquistador plunges the beholder into an immersive space in which figural evocations of life-and-death entanglements of coloniser and colonised pulsate with affective intensity precisely by virtue of the way the not-yet-formedness of the painterly markings stimulates our quest, as viewers, to decipher the inscriptions and, in the process, deepen our engagement with the sensuous materiality of the painting itself. social relationship, that the value of an artwork lies not “in” the work, as if it were self-sufficiently autonomous, but in our relationship to it Williams and the Caribbean Artists Movement 27 For my part I want to isolate just one moment, a statement that was part of Williams’s contribution to the June 2 1967 Symposium on West Indian artists that featured sculptor Ronald Moody, painter Karl “Jerry” Craig, and textile designer Althea McNish. Commenting on the work exhibited, Williams said: It has haunted me all of my life and I don’t understand it . 28 29 That Williams located himself in an art historical context that conjoins Latin Americans, Matta from Chile, Tamayo from Mexico—and Caribbeans, Lam from Cuba—clearly shows that however much he admired the North Americans, he did not identify himself with either the formalism or the transcendentalism that framed the dominant narratives of post-war abstraction. In 1967 Williams was making a lateral or transversal move to reframe his practice within an interpretive paradigm grounded in what today would be called “the global South,” and which at the time would have been referred to as “the Third World.” Implicit in such a move that no longer regarded New York or London as the epicentre of artistic life was a further shift away from an exclusively anglophone context to embrace a Spanish-speaking one as well by virtue of the prominence Williams gave to Latin American artists. In any event, the common factor in the lineage of the “claw-like forms” through which Williams connects Lam, Matta, and Tamayo is colonial history. In Science, Conscience et Patience du Vitruer (1944) the angular biomorphic shapes surrounding the “glass being” of the painting’s title bear “claw-like” forms that Matta arrived at through his conception of living beings as made up not of solid substances but of oscillating waves of matter and energy ( Fig. 15 ). Claw-like forms are found in Lam’s paintings such as Zambezia, Zambezia (1950), where a limb with three jagged edges pointing left rises from a blue female torso that also bears a horse’s tail. In this and many other works, Lam was addressing the femme-cheval , the metaphorically hybrid “woman-horse” evoked in self-descriptions among Vodun participants who enter a trance possession state ( Fig. 16 ). Where language conventionally separates agent and patient, subject and object, the folklorist Lydia Cabrera, with whom Lam was in conversation when he returned to Cuba in 1940, explained the undoing of such either/or dualisms in her following gloss on the femme-cheval : 30 the multifarious figural , marking the potential of the inchoate, the not-yet-fully-formed, to produce an excess or surplus of meanings all at once. Figure 15. Roberto Matta, Science, Conscience et Patience du Vitreur, 1944, oil on canvas, 200 × 450 cm. Private Collection, Paris. Digital image courtesy of Estate of Gordon Matta-Clark / Artists Rights Society (ARS), New York, DACS London 2018. Figure 16. Wifredo Lam, Zambezia, Zambezia, 1950, oil on canvas, 125.4 × 110.8 cm. Collection of Solomon R. Guggenheim Museum, New York (74.2095). Digital image courtesy of ARS, New York / ADAGP, Paris / DACS, London, 2018. Figure 17. Denis Williams, Composition I, 1954, watercolour. 30 × 20.3 cm. Private Collection. DOI My third example of such “claw-like” forms, Composition I (1954) is by Guyanese artist Denis Williams, who was not a family relation but an artist-cum-archeologist who, as the first Director of Guyana’s Walter Roth Museum of Anthropology , was a significant interlocutor for Aubrey Williams after Williams’s return visits to Guyana became more frequent from 1970. The potent red and black ground in Composition I intensifies the menacing aura that radiates from the skeletal biomorph that forms the watercolour’s white-coloured figure ( Fig. 17 Abstraction as a Site of Decolonisation In an earlier essay I sought to explore these aesthetic effects generated in Williams’s work—disturbing and alluring in equal measure—by turning to the concept of “fossil identities” put forward by Wilson Harris. 31 trace as a phenomenon of the differencing activity of language. Although he is addressing the repeatability or iterability of marks that eventually come to function as signs, marks that are formalised at the point where temporary fixity brings the slide of signifiers to the closure that correlates signifier (form) with signified (meaning), he also draws attention to the ambivalent temporality whereby trace is both absent and present at once: 32 Simultaneously embracing the future that is yet to come and the past that has not completely gone, trace anticipates the later concept of “hauntology,” in which Derrida discusses the persistence of utopian desires for the future, despite the eradication of hope as a result of the devastations left behind in the aftermath of political revolutions. 33 In light of such theoretical precepts, the many-sided significance of Triptych (1976)—one of the most important works in Williams’s oeuvre—takes on further ramifications, politically as well as aesthetically ( Fig. 18 ). DOI Figure 18.Aubrey Williams, “Arawak”, “Carib”, and “Warrau” Triptych panels, 1976, oil on canvas, 243.8 × 731.5 cm. Location unknown. From left to right, the panels are titled Arawak, Carib, Warrau, all peoples indigenous to the Caribbean region. “Claw-like” forms abound in Triptych hauntological mode of abstraction in which interstitial trace structures punctured the nucleus of disavowal encircling all that was unsaid and unspoken in the aftermath of a new nation having been formed. DOI We could say that what “moves” us, what “touches” us, what “affects” us, as we behold a work such as Triptych is that we have entered into a strangely double-facing relationship to a past that is not completely gone and a future that is yet to come. As with Bone Heap and Tribal Mark II 34 So deeply was disavowal implanted into the colonial formation of Caribbean societies, as Stuart Hall explains in his posthumous memoir, that it was not until after independence that Guyana or Jamaica, both very different in their own right, came to speak of themselves as multiethnic societies or plural societies. 35 To say Williams’s “claw-like shape” pierced the nucleus of disavowal that locked vast areas of historical experience into the realm of the unspeakable, the unrepresentable, is to say that in 1976 Williams was one of the first to demarcate the ambivalence of the postcolonial condition. Likewise Wilson Harris, in his 1976 essay “Fossil and Psyche,” spoke to the predicament that literary scholars Kirsten Holst Petersen and Anna Rutherford parse in the following exegesis of the “fossil identities” concept: each living person is a fossil in so far as each man carries within himself remnants of deep seated antecedents … By entering into a fruitful dialogue with the past one becomes able to revive the fossils that are buried within oneself and are part of one’s ancestors. Petersen and Rutherford add the all-important qualifier that while fossil identities may open human and ahuman antecedents can either act as positive forces or can become prejudices, hideous biases. 36 Disavowal is not something people do self-consciously (which means it is a psychic defence similar to but not equivalent to the ego’s acts of denial), yet the key issue here is the sheer amount of affective labour—psychic energy—involved in keeping the unspoken unsaid. Hall repeatedly made the point that everyone “knew” Jamaica, for instance, was a black majority society, but that it was only in the 1970s that Jamaica began to think of itself as a black majority society. 37 and I don’t understand it El Dorado in 1958, a dark star from the age of empire continuing to absorb the living present into the void of its black hole. “In art things get said in ways in which they can’t get said in any other domain,” said Hall in a 2007 conversation with Bill Schwarz. 38 39 Paradoxical it may be to suggest it was an abstract painter who made colonial trauma speakable, but we can come at the complex relationship between words and abstract art in another way. DOI Williams often seemed discomfited by the verbal medium, even though in his articles of the late 1960s and early 1970s he spoke to the decolonial moment with penetrating insight. Williams also contributed to far-reaching debates on the British-born generation of diaspora artists in events such as the 1987 Creation for Liberation panel held in London. 40 Watching Imruh Bakari Caesar’s important film documentary, The Mark of the Hand: Aubrey Williams (1986), one senses Williams’s discomfort, at times, with the interviewer’s presumption that the artist’s words will somehow “explain” his abstract paintings, as if their meaning will be finally fixed once we hear the artist himself speak. There is a fraught moment when, in the hinterland where his relationship to soil and land first took shape, Williams says he would prefer listening to Shostakovich rather than carry on with the interview. 41 It would be entirely wrong to think the artist was nonverbal. Far from it, in his 1970 essay, “Caribbean Visual Art: A Framework for Further Inquiry” (valuably reprinted in Anne Walmsley’s indispensable Guyana Dreaming anthology), Aubrey Williams was among the first to use “postcolonial” as a key term. At CAM’s first conference, held at the University of Kent in September 1967, Williams had said “Art is always in the foreground; it is the true avant-garde … It always seems in the history of man that the arts give the direction for the technology, for the philosophy and the very life of the people.” 42 43 Cenote , also spelled zenote , is a phonetic translation of an indigenous Amerindian word ( Fig. 19 ). In the language of natural history, cenote are circular pools opening onto underground caves, all concentrated geographically in the north east Yucatán peninsula because of an asteroid that created the Chicxulub crater 66 million years ago ( Fig. 20 ). But Williams was also using cenote 44 Cenote cannot be an ending if this is the point from which a future visual identity emerges. This is also to say cenote cannot be fixed in the precolonial past as something belonging to the Maya or to the Amerindians alone, for the word has now become cross-culturally translated as a double-facing trace structure , imbued with the pastness of ancient petroglyphs while at the same time radiant with future pluripotentiality. Aubrey Williams’s work deals with an ecological materialism on a planetary scale. Like a fossil identity harbouring utopian and authoritarian possibilities, cenote —in Williams’s handling of the term—is cataclysm’s trace, an underwater archive storing archaic treasures for posterity and a place of futurity anticipating the rediscovery of an extinct civilisation. For me, some of the most memorable scenes in Imruh Bakari Caesar’s invaluable film portrait are those in which we see Aubrey listening to Shostakovitch in the jungle, an image that is always called to mind for me when I see Quartet no. 15 opus 144 (1981) from the Shostakovich series ( Fig. 21 ). In a setting worlds away from the one inhabited by the Russian composer, we glimpse a pluriverse in which particles assemble, decay, and reassemble in the time-space continuum of cenote , which is not a void or a hole but a passageway from one realm into others. DOI Figure 21.Aubrey Williams, Quartet No. 15 Opus 144, 1981, oil on canvas, 132 × 208 cm. Private Collection.Digital image courtesy of Estate of Aubrey Williams (All rights reserved, DACS 2018). DOIFor generously sharing their insights, and for responses to the lecture on which this article is based, I would like to thank Tim Cribb, Mel Gooding, Anne Walmsley, and Maridowa Williams. About the author Kobena Mercer is Professor of History of Art and African American Studies at Yale University, where he teaches modern and contemporary art in the Black Atlantic, examining  African American, Caribbean, and Black British artists with critical methods from cultural  studies. He is the author of monographic studies of James VanDer Zee, Romare Bearden and Adrian Piper,  Isaac Julien and Rotimi Fani-Kayode, and he edited the Annotating Art’s Historiesseries whose titles are Cosmopolitan Modernisms[2005], Discrepant Abstraction[2006], Pop Art and Vernacular Cultures[2007], and Exiles, Diasporas & Strangers[2008]. Mercer previously taught at New York University and the University of California–Santa Cruz and was an inaugural recipient of the 2006 Clark Prize for Excellence in Art Writing awarded by the Sterling and Francine Clark Art Institute in Massachusetts. His most recent book is his essay collection Travel & See: Black Diaspora Art Practices since the 1980s[2016]. Footnotes A useful point of entry to the field is provided by Robin Cohen, Global Diasporas: An Introduction, London: University College Press, 1997. 1 Guy Brett, “A World Aesthetic” [1988] in Guyana Dreaming: The Art of Aubrey Williams, ed. Anne Walmsley, Sydney: Dangeroo Press, 1990, 97. 2 Eddie Chambers, Black Artists in British Art: A History Since the 1950s, London: I.B. Tauris, 2014, 10–24. Artist Sonia Boyce and art historian Leon Wainwright were among invited speakers at the Aubrey Williams: In ProfileStudy Day held at Tate Britain, 21 September 2007; the 2010 exhibitions were accompanied by the catalogue Aubrey Williams: Atlantic Fire, ed. Reyahn King, Liverpool: National Museums Liverpool and October Gallery, 2010. 3 Stuart Hall, “Black Diaspora Artists in Britain: Three ‘Moments’ in Post-War History,” History Workshop JournalVol 61 n 1, March 2006, 10. 4 The Museum of Modern Art visitor handout, March 30 1969, cited in Susan Cahan, Mounting Frustration: Art Museums in the Age of Black Power, Durham NC: Duke University Press, 2015, 207. 5 See, “Disavowal,” Jean Laplanche and Jean-Bertrand Pontalis, The Language of Psycho-analysis, 1968 trans. Donald Nicholson Smith, New York: W.W. Norton & Co., 1974, 118–121. 6 Barnor Hesse, “Symptomatically Black: A Creolization of the Political,” in Creolizing the Political, eds. Françoise Lionnet and Shu-mei Shih, Durham NC: Duke University Press, 2011, 37–61. 7 See Fredric Jameson, The Political Unconscious: Narrative as a Socially Symbolic Act,Ithaca: Cornell University Press, 1981. 8 Jan Carew, “Portrait of the Artist: Aubrey Williams” Art News and Review[1959] in Guyana Dreaming, ed. Walmsley, 1990, 66–67. 9 Rasheed Araeen, “Conversation with Aubrey Williams,” Third TextVol 1 no 2, 1987, 25–52. 10 According to one source, the “Assistant Curator of the Commonwealth Institute Art Gallery made contact with Denis Bowen and agreed … [to] … leave most of the organization up to Denis Bowen provided that … the final selection was to be decided in consultation with the Institute’s art advisor, Eric Newton, art critic of The Guardian,” Simon Pierse, Australian Art and Artists in London, 1950–1965: An Antipodean Summer, Farnham: Ashgate, 2012, 176. 11 Araeen, “Conversation with Aubrey Williams,” 1987, 36. 12 Gavin Butt, “America and its Discontents: Art and Politics, 1945–60,” in Companion to Art Since 1945, ed. Amelia Jones, Malden MA: Blackwell, 2006, 24. 13 Jean-François Lyotard, Discourse, Figure, 1971: trans. Antony Hudek and Mary Lydon, Minneapolis: University of Minnesota, 2011. 14 See The Dark Monarch: Magic and Modernity in British Art, eds. Michael Bracewell, Martin Clark, and Alun Rowlands, London: Tate Publishing, 2009. 15 Maridowa Williams, interview with the author, London, 10 October 2017. 16 Jan Carew, “Portrait of the Artist,” in Walmsley, Guyana Dreaming, 1990, 66–67. 17 Now & Coming Time: A Symposium on Aubrey Williams and the Textual and Visual Arts of Guyana and the Caribbean, 26 April 2017, University of Cambridge. 18 Clement Greenberg, “American-Type Painting,” [1955] in Art and Culture: Selected Essays, Boston MA: Beacon Press, 1961, 208–229. 19 Guy Brett, “A Tragic Excitement,” in Aubrey Williams, eds. Andrew Dempsey, Gilane Tawadros and Maridowa Williams, London: Institute of International Visual Arts, 1997, 30–31. 20 Mark Cheetham, The Rhetoric of Purity: Essentialist Theory and the Advent of Abstract Painting, Cambridge: Cambridge University Press, 1994. 21 Donald Locke, “Contemporary Guyanese Painters,” Kaie, National History and Arts Council, Guyana, no 2, May 1966 in Walmsley, Guyana Dreaming, 1990, 72, emphasis added. 22 Michael Fried, “Three American Painters: Kenneth Noland, Jules Olitski, Frank Stella,” [1965] in Art and Objecthood: Essays and Reviews, Chicago: University of Chicago Press, 1998, 213–266. 23 Alain Locke, “The Legacy of the Ancestral Arts,” in The New Negro: An Interpretation,ed. Alain Locke, [1925] New York: Simon & Schuster, 1992, 254–267. 24 Mark Godfrey, Abstraction and the Holocaust, New Haven CT: Yale University Press, 2007. 25 Denis Williams, “Aubrey Williams in Guyana,” in Aubrey Williams, eds. Dempsey, Tawadros and Williams, 1997, 40. 26 Anne Walmsley, The Caribbean Artists Movement, 1966–1972: A Literary and Cultural History, London and Port of Spain: New Beacon Books, 1992. 27 Aubrey Williams, “A Glimpse of Caribbean Art,” 1967 cited in Walmsley, The Caribbean Artists Movement, 1992, 13–14, emphasis added. 28 Cathy Caruth, Unclaimed Experience: Trauma, Narrative, and History, Baltimore MD: John Hopkins University Press, 1996. 29 Lydia Cabrera, El Monte, Havana: Editorial Letras Cubanas [1954] cited in Edna M. Rodriguez-Mangual, Lydia Cabrera and the Construction of Afro-Cuban Cultural Identity, Chapel Hill NC: University of North Carolina Press, 2004, 78–9 30 Kobena Mercer, “Black Atlantic Abstraction” in Discrepant Abstraction, ed. Kobena Mercer, London and Cambridge MA: Institute of International Visual Arts and MIT Press, 2006, 182–205. 31 Jacques Derrida, “Différance” in Margins of Philosophytrans. Alan Bass, Chicago: University of Chicago Press, 1984, 13. 32 See Jacques Derrida, Spectres of Marx: The State of the Debt, the Work of Mourning, and the New International, 1993; trans. Peggy Kamuf, New York: Routledge, 1994. 33 See Evelyn A. Williams, The Art of Denis Williams, London: Peepal Press, 2012. 34 Stuart Hall with Bill Schwarz, “Race and Its Disavowal,” in Familiar Stranger: A Life Between Two Islands, Durham NC: Duke University Press, 2017, 95–106. 35 Kirsten Holst and Anna Rutherford, “Fossils” from The Enigma of Values: An Introduction to Wilson Harris[1976], in The Postcolonial Studies Reader, eds. Bill Ashcroft, Gareth Griffiths and Helen Tiffin, New York: Routledge, 1995, 185–188, emphasis added. 36 See Stuart Hall, “Race, Class, and Pluralism in Caribbean Societies” in Race and Class in Post-Colonial Society, Paris: UNESCO 1977, 150–182, and Stuart Hall, “Cultural Identity and Diaspora” in Identity: Community, Culture and Difference, ed. Jonathan Rutherford, London: Lawrence & Wishart, 1990, 222–239. 37 Stuart Hall cited in Bill Schwarz, “Living with Difference: Stuart Hall in Conversation with Bill Schwarz,” Soundings27, 2007, 150–151. 38 Bill Schwarz, “Introduction,” West Indian Intellectuals in Britain, Manchester: Manchester University Press, 2003, 3. 39 Williams spoke at the seminar on “Seeking a Black Aesthetic,” chaired by artist Errol Lloyd and organised by Creation for Liberation at Brixton Village, Brixton Hill, following the Open Exhibition, 17 October–7 November 1987, selected by a panel that included black British artists, Eddie Chambers, Chila Kumari Burman and Eugene Palmer. See http: new. diaspora-artists.net/index.php. 40 Imruh Bakari Caesar, director, The Mark of the Hand: Aubrey Williams(1986), 52 min, produced by Arts Council of Great Britain, distributed on DVD by Concord Media. 41 Aubrey Williams, “The Predicament of the Artist in the Caribbean,” Savacouno 2, September 1970, in Walmsley, Guyana Dreaming, 1990, 15. 42 Aubrey Williams, “Caribbean Visual Art: A Framework for Further Inquiry,” The Literary Half-Yearly, Mysore, Vol XI no 2 July 1970, in Walmsley, Guyana Dreaming, 1990, 21. Following his return visits to Guyana, Williams developed a close relationship with the Jamaican art scene through his friendship with Karl Parboosingh, which is touched on by Claudia Hucke, Picturing the Postcolonial Nation: (Inter)Nationalism in the Art of Jamaica, 1962–1975, Kingston: Ian Randle Publishers, 2013. 43 Williams, “Caribbean Visual Art”, 1990, 28. 44 Bibliography Rasheed Araeen, “Conversation with Aubrey Williams,” Third TextVol 1 no 2, 1987, 25–52. Eddie Chambers, Black Artists in British Art: A History Since the 1950s, London: I.B. Tauris, 2014. Andrew Dempsey, Gilane Tawadros and Maridowa Williams eds. Aubrey Williams,London: Institute of International Visual Arts/Whitechapel Gallery, 1997. Stuart Hall, “Black Diaspora Artists in Britain: Three ‘Moments’ in Post-War History,” History Workshop JournalVol 61 n 1, March 2006, 1–24. Reyahn King ed., Aubrey Williams: Atlantic Fire, Liverpool: National Museums Liverpool and October Gallery, 2010. Kobena Mercer, “Black Atlantic Abstraction” in Discrepant Abstraction, ed. Kobena Mercer, London and Cambridge MA: Institute of International Visual Arts and MIT Press, 2006, 182–205. Leon Wainwright, “A Painter in the Aftermath of Painting,” Timed Out: Art and the Transnational Caribbean, Manchester: Manchester University Press, 2011, 22–53. Anne Walmsley ed., Guyana Dreaming: The Art of Aubrey Williams, Sydney: Dangeroo Press, 1990. Anne Walmsley, The Caribbean Artists Movement, 1966-1972: A Literary and Cultural History, London and Port of Spain: New Beacon Books, 1992. Imprint Author Kobena Mercer Date 08 June 2018 Category Article Review status Peer Reviewed (Double Blind) Licence CC BY-NC International 4.0 Downloads PDF format Article DOI https://doi.org/10.17658/issn.2058-5462/issue-08/kmercer Cite as Kobena Mercer, "Aubrey Williams: Abstraction in Diaspora", British Art Studies, Issue 8, https://doi.org/10.17658/issn.2058-5462/issue-08/kmercer Next“The Art Game”:Television, Monitor, and British Art at the turn of the 1960s

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Micromachines | Free Full-Text | Hybrid Optimization Algorithm Based on Double Particle Swarm in 3D NoC Mapping Increasing the number of cores on a chip is one way to solve the bottleneck of exponential growth but an excessive number of cores can lead to problems such as communication blockage and overheating of the chip. Currently, networks-on-chip (NoC) can offer an effective solution to the problem of the communication bottleneck within the chip. With current advancements in IC manufacturing technology, chips can now be 3D-stacked in order to increase chip throughput as well as reduce power consumption while reducing the area of the chip. Automating the mapping of applications into 3D NoC topologies is an important new direction for research in the field of 3D NoC. In this paper, a 3D NoC partitioning algorithm is proposed, which can delineate the 3D NoC region to be mapped. Additionally, a double particle swarm optimization (DPSO) based heuristic algorithm is proposed, which can integrate the characteristics of neighborhood search and genetic algorithms, and thus solve the problem of a particle swarm algorithm falling into local optimal solutions. It is experimentally demonstrated that this DPSO-based hybrid optimization algorithm has a higher throughput rate and lower energy loss than the traditional heuristic algorithm. Hybrid Optimization Algorithm Based on Double Particle Swarm in 3D NoC Mapping by Juan Fang * , Huayi Cai and Xin Lv Faculty of Information Technology, Beijing University of Technology, Beijing 100124, China Author to whom correspondence should be addressed. Micromachines 2023 , 14 (3), 628; https://doi.org/10.3390/mi14030628 Received: 6 February 2023 / Revised: 1 March 2023 / Accepted: 6 March 2023 / Published: 9 March 2023 Abstract : Increasing the number of cores on a chip is one way to solve the bottleneck of exponential growth but an excessive number of cores can lead to problems such as communication blockage and overheating of the chip. Currently, networks-on-chip (NoC) can offer an effective solution to the problem of the communication bottleneck within the chip. With current advancements in IC manufacturing technology, chips can now be 3D-stacked in order to increase chip throughput as well as reduce power consumption while reducing the area of the chip. Automating the mapping of applications into 3D NoC topologies is an important new direction for research in the field of 3D NoC. In this paper, a 3D NoC partitioning algorithm is proposed, which can delineate the 3D NoC region to be mapped. Additionally, a double particle swarm optimization (DPSO) based heuristic algorithm is proposed, which can integrate the characteristics of neighborhood search and genetic algorithms, and thus solve the problem of a particle swarm algorithm falling into local optimal solutions. It is experimentally demonstrated that this DPSO-based hybrid optimization algorithm has a higher throughput rate and lower energy loss than the traditional heuristic algorithm. Keywords: 3D NoC ; particle swarm optimization ; neighborhood search ; gene cross-mutation ; high performance computing Graphical Abstract 1. Introduction With the advent of the digital era, the improvement of arithmetic power has become an increasingly critical issue in the development of computers today. Integrated circuits have evolved rapidly in just a few decades. Hundreds of millions of transistors can now be integrated into a single chip, allowing for a great increase in computer processing power. However, the external I/O of the chip has become the bottleneck of the chip’s computing power, and so to solve this problem, system-on-chip (SoC) technology was born. With an increase in the number of cores in a single chip, inter-core communication becomes a key issue to be considered in SoC design, which uses a bus-based architecture wherein multiple cores and internal storage devices are connected to the same bus and exchange data through the bus. However, with the increase in the number of IP cores, the data exchange load on the bus increases. Therefore, network-on-chip (NoC) architecture is proposed in order to relieve the bus load. However, as the number of IP cores continues to increase, there are some issues to be faced with 2-Dimensional (2D) NoCs, such as chip area, performance, bandwidth, and power consumption. The 3-Dimensional (3D) NoC can effectively reduce the size of system interconnects for large-scale SoCs, eliminating the limitations of 2D NoCs in terms of chip area and performance. A 3D NoC can vertically accumulate multiple 2D NoCs and package them into a single chip, increasing the data transfer in the vertical dimension. The longitudinal interconnection of chips relies on through-silicon-via (TSV) technology. The technique of stacking 3D chips using TSV technology is a key to overcoming the problem of the chip performance bottleneck [ 1 ]. When the number of nodes is the same, 3D NoC has a smaller area, lower delay, and lower power consumption than 2D NoC, and can disperse data transmission in more links, therefore improving overall system performance and power consumption. Hybrid optimization algorithms are usually applied in complex nonlinear systems. Zhang et al. [ 16 ] proposed an orientation function-based prescribed performance control (PPC). Ding et al. [ 17 ] indicated that the conventional linear and nonlinear models would fail for long-term prediction and an evolutionary algorithm is utilized to obtain the optimal model for long-term prediction. Zhang et al. [ 18 ] proposed an image enhancement algorithm; by combining the rough set theory with a Gaussian mixture model, an algorithm with higher immunity is proposed. The task mapping is divided into two stages: the mapping of task nodes to IP cores and the mapping of IP cores to NoC maps. In this paper, based on the analysis of the network scale of 3D NoC and the existing NoC benchmark, we propose an IP core to NoC mapping by region, in order to achieve an even distribution of task load while taking into account the heat dissipation efficiency, in order to reduce the chip temperature as much as possible and improve the processing capacity of the chip. The problem of IP core to NoC node mapping is an NP-hard problem. In this paper, we propose a particle swarm-based thermal-aware mapping algorithm in order to find the optimum in the solution space. In this work, in order to prevent the PSO algorithm from prematurely falling into the local optimal solution, this paper proposes a double particle swarm optimization algorithm. One of the particle populations introduces cross-mutation to improve global search capabilities. Another particle population incorporates neighborhood search to improve local search capabilities. When the two populations update their vectors of positions to the global optimal solution and the local optimal solution, they also update to the optimal solution position of the other population. A dynamic inertia factor is proposed to control the convergence speed. 2. System and Computation Model In the field of on-chip networking, routing, scheduling, and task mapping are all hot topics. Each application can be divided into several independent subroutines that can be assigned in different IP cores at runtime to exchange data over links and thus coordinate the overall output of the final result. Thus, each application can be represented by a directed acyclic graph (DAG). Each node of the graph represents a subroutine, the edges represent the data transfer relationships of the subroutines, and the weights of the edges represent the amount of data communication of the connected subroutines. Each resource node can execute any subroutine (a node in the DAG) of the application. Determining how to assign the sub-nodes of the DAG graph to the resource node in the NoC is the problem addressed by task mapping. In this paper, the mapping problem is solved by the optimization of power consumption, latency and the three aspects of thermal energy. 2.1. Task Graph The task graph used is usually a weighted DAG. In a task graph T G V , K the vertex v i ∈ V , which represents a subtask in the application. The directed edge k m , n ∈ K , which contains both direction and weight information (i.e., the data transmission direction is from the e m node to the e n node). The weight indicates the amount of data communicated in these two subtasks. If no communication relationship is formed between the e m node and the e n node, then k m , n = 0 . Equation (1) represents the communication relatiohip matrix. K = k i , j = 0 ⋯ k 0 , n − 1 ⋮ ⋱ ⋮ k n − 1 , 0 ⋯ 0 (1) 2.2. 3D NoC Topology In this paper, the NoC mapping algorithm under a 3D mesh structure is discussed. The 3D NoC mapping system consists of resource nodes, routings, physical links and TSV links, as shown in Figure 1 . The NoC topology graph is defined as N o C ( U , L ) . Where the vertex u i ∈ U denotes the resource node in NoC, l i ∈ L represents the pathway of the communication link in NoC. N U = X × Y × Z and where N U is the number of resource nodes in the U set, X × Y represents the number of resource nodes in each layer; Z represents the number of layers in this 3D NoC system. The size of different layers in 3D NoC is the same. The vertical connection of nodes through TSV technology. For example, the size of 3D NoC shown in Figure 1 is 3 × 3 × 3 ; through the coordinates, from (1,1,1) to (3,3,3), the unique resource node can be identified. 2.3. Communication Power Consumption Model Communication power consumption is an important metric used for describing the performance of on-chip networks. It refers to the communication power consumption of each node in the NoC system and the power consumption generated by the data in the link. Its size is related to the distance the data slices are transmitted between nodes, the type of transmission path (Link or TSV), and the amount of communication transmitted. The model used in this paper is a generic power calculation model for a 3D-mesh structured NoC, i.e., the energy consumption required for 1-bit data transmission over the NoC, as shown in Equation (2). E b i t = E S b i t + E B b i t + E L b i t + E W b i t (2) E B b i t and E W b i t represent the energy consumption of 1-bit data on the cache and internal link lines, respectively. This energy consumption is relatively small and usually neglected; E S b i t and E L b i t represent the energy consumed by 1-bit data on the router and adjacent routing node links, respectively. Thus, Equation (1) can be simplified to Equation (3). E b i t = E S b i t + E L b i t (3) Therefore, the energy consumed by 1-bit data from the e m node to the e n , E b i t m n node is shown in Equation (4). E b i t m n = E S b i t m n + E L b i t m n E S b i t m n = N d i m x × E S b i t ′ + N d i m y × E S b i t ′ + N d i m z × E S b i t ′ + E L b i t m n = N d i m x × E L b i t ′ d i m y × E L b i t ′ + N d i m z × μ E L t ′ E S b i t ′ E S b i t ′ and E L b i t ′ N d i m x , N d i m y and N d i m z represent the number of links that pass through the X-dimension, Y-dimension, and Z-dimension from node e m to node , provided that the shortest possible routing algorithm is adopted. μ represents the energy consumption of data in transmission in TSV and the ratio of energy consumption in horizontal links (TSV consumes only 7.5% of the communication energy compared to normal horizontal links [ 4 ]). Assuming that the size of the data volume from the e m node to the e n node is B m n , the total power consumption E t o t a l m n from the e m node to the e n node can be derived according to Equation (4), as shown in Equation (5). E t o t a l m n = B m n × E b i t m n (5) The total communication energy consumption of the NoC chip is obtained by adding up the communication consumption of all resource nodes and transmission links in the NoC. This is shown in Equation (6). E t o t a l = ∑ m , n ∈ N o C E t o t a l m n (6) 2.4. Delay Model The delay of the NoC network is an important metric for describing the on-chip network performance. It refers to the time taken to perform the task of each node in the NoC system, the delay generated by the data in the link and the delay of data transmission in the router. The Delay in the NoC system is shown in Equation (7). D b i t = ∑ D l i n k + ∑ D c o r e + ∑ D r o u t e r (7) where D l i n k represents the delay in the data on the transmission link, D c o r e represents the delay in the data in the resource node, and D r o u t e r represents the delay in the data in the router, as shown in Equation (8). D l i n k m , n = β l i n k × d h o r i z o n m , n D c o r e n = β c o r e D r o u t e r m , n = β r o u t e r × d m , n + 1 β l i n k , β c o r e and β r o u t e r are constants that represent the transmission delay of 1-bit data in the link, resource node and router, and d h o r i z o n m , n is the shortest path length from the e m node to the e n node in the horizontal direction, because the data transmission path in the vertical direction is achieved by TSV technology and its delay is negligible compared to the normal technology. d m , n is the shortest path length from the e m node to the e n node. Assuming that the size of the data volume from the e m node to the e n node is B m n , the total communication delay D t o t a l m n from the e m node to the e n node can be derived according to Equations (7) and (8), as shown in Equation (9). D t o t a l m , n = B m n × D b i t (9) The total communication delay on the NoC chip is obtained by calculating the total communication consumption of all resource nodes and transmission links in the NoC. This is shown in Equation (10). D t o t a l = ∑ m , n ∈ N o C D t o t a l m n (10) 2.5. Thermal-Aware Model In this subsection, we formulate the computation offloading problem in order to optimize the overhead of applications generated by U E i . In Section 1 , the hazards of high chip temperatures are mentioned. Therefore, this paper introduces the processor’s thermal sensing model into consideration for algorithm optimization. The temperature of a single resource node depends on the energy consumption of the resource node, the temperature of neighboring nodes, and the distance between the processor and the heatsink [ 19 ]. Hotspot [ 20 ] is a model that can calculate the thermal energy efficiency of the resource node in a chip. It takes into account the heat dissipation within the resource node and the heat transfer between adjacent processing units, from which the temperature of the resource node is calculated. For adjacent nodes, Hotspot calculates the thermal resistance R m , n from node e m to node e n by calculating the length and ratio of the adjacent edges between two resource nodes. This is shown in Equation (11). R m , n = Δ T m , n / E n (11) where Δ T m , n represents the temperature at which the node e m rises, and E n represents the power consumption of the (previously mentioned) e n node. It is assumed that all resource nodes in a unified NoC system interact with each other, so the temperature of each resource node can be calculated by Equation (12). T i = ∑ j ∈ N o C R i , j × E j , i ∈ N o C (12) Assuming that the number of resource nodes in NoC is N , the peak temperature T p e a k in the model is shown in Equation (13). T p e a k = max T i , i ∈ N o C (13) The above calculation process is entirely integrated into the Hotspot thermal model. This is the most commonly used thermal sensing model in NoC computing systems. In this paper, Hotspot is used to quantify energy consumption. 3. Proposed Algorithm In this paper, the mapping algorithm is divided into two phases: division of the 3D NoC operation region, and the mapping of tasks to NoC resource nodes. In 2D NoC, because the chip area is limited, the number of integrated resource nodes is usually less than the number of subtasks of the application, so there can be multiple subtasks mapped to the same resource node. In contrast, 3D NoC uses chip stacking technology to increase the layers of the chip through TSV, and this increases the number of resource nodes so that the number of resource nodes is larger than the number of application subtasks. Part of the 3D NoC area is delineated to map subtasks, allowing subtasks to transfer data with the shortest possible communication distance, which usually results in the lowest power consumption and latency. 3.1. Regional Division In 3D NoC, the expansion of the network dimension makes the number of network computing nodes grow exponentially, and the performance of 3D NoC cannot be compared with traditional NoC, so concentrating the mapped nodes when running a single application can effectively avoid the delay and power consumption caused by long pathways during data transmission, and thus avoid the heat buildup generated by power consumption. In order to address these characteristics, we seek to improve the convergence speed of the particle swarm algorithm, by performing region partitioning on 3D NoC in advance based on the thermal resistance theorem and communication delay. The sub-nodes of the application are run in the partitioned region in order to achieve the lowest possible communication delay and heat buildup. Considering that the pathway distance directly affects the communication delay, we prioritize the node with the shortest pathway length in our NoC subgraph. If there are multiple nodes with the same pathway distance to the NoC subgraph, we calculate the impact of each of these nodes on the heat propagation of other nodes in the NoC subgraph according to the thermal resistance formula, and select the node corresponding to the smallest of them, then add it to the NoC subgraph. Because the TSV technique has a low delay on the vertical link, it is preferable to select the computational nodes in the vertical link. Figure 2 shows an example of regional division. First, we define the number of C V nodes of the T G graph to determine the minimum number of computational nodes for the partitioned NoC region. We also define a hypothetical required energy consumption value ϵt in each node to calculate the hypothetical temperature increase value of the node based on the thermal resistance formula in order to predict the performance of the partitioned NoC region in terms of communication delay and temperature when executing the application. The process of NoC regional division algorithm is shown in Algorithm 1. The temperature metrics of each node are calculated based on the energy consumption value ϵ and added to the temperature queue T L (lines 1–3), then arranged in ascending order. The smallest temperature node is found as the initial division node and added to the resource node group V G (lines 4–5). 2. Iterate through all nodes connected by TSV technology in the V G group and add them to the queue L (lines 7–9). If there are no more vertically adjacent (connected by TSV) nodes, the horizontally adjacent (connected by common link) nodes of all nodes in the V G group are added to the queue L . 3. The sum of the horizontal Manhattan distances from the nodes in the queue L to the nodes in the V G are calculated (lines 11–12). The values are then added to the queue and sorted, and the minimum value is selected (lines 23–29). If multiple nodes have the same horizontal distance to the V G , their effect on the thermal resistance value in the V G is calculated (lines 13–15). The minimum operator node is then selected and added to the V G group (lines 16–22). 4. If the number of nodes in the V G is less than the number of nodes C V in the T G graph, steps 2 to 4 are repeated. Algorithm 1 : NoC Regional Division Algorithm Inputs: N o C U , L : NoC topology graph X , Y , Z : The number of NoC resource nodes in the three dimensions u x , y , z : Resource node with coordinates x, y, z T G V , K : Application task graph of the application, where the number of the task graph is C V є t : Assumed power consumption of each alternative node Ensure: V G : Group of resource node Optimal offloading policy A , the total overhead C 1: For u є 𝑈 do 2:   temperature list: T L ← ∑ u ∈ N o C R i , j × ϵ t 3: End for 4: Get the coordinate of the v x t , y t , z t ← min T L 5: V G ← v x t , y t , z t . 6: While n u m V G < C V − 1 do 7: If Z t + 1 ≤ Z then 8: V G ← v x t , y t , z t 9: z t ← z t + 1 10: Else 11: d i s t x ← ∑ v ∈ V G x t + 1 − X v + y t − Y v 12: d i s t y ← ∑ v ∈ V G X t − X v + y t + 1 − Y v 13: If d i s t x = = d i s t y then 14: T x ← ∑ v ∈ V G R v x t + 1 , y t , v × ϵ _ t 15: T y ← ∑ v ∈ V G R v x t , y t + 1 , v × ϵ _ t 16: If T x ≤ T y then 17: V G ← v x t + 1 , y t 18: x t ← x t + 1 . 19: Else 20: V G ← v x t , y t + 1 21: y t ← y t + 1 22: End if 23: Else if d i s t x < d i s t y then 24: V G ← v ( x t + 1 , y t ) 25: x t ← x t + 1 26: Else 27: V G ← v x t , y t + 1 28: y t ← y t + 1 29: End if 30: End if 31: End while 3.2. Double Particle Swarm Based Thermal-Aware Mapping Algorithm 3D on-chip network task mapping is the process of finally mapping a DAG task graph to an NoC resource node. Based on the previous division of NoC resource nodes, the task nodes that are to be mapped and the NoC resource nodes waiting to be mapped have been identified at this point. This section will describe the use of the particle swarm algorithm to map tasks to a NoC. The particle swarm algorithm is a type of swarm intelligence algorithm, which has the characteristics of simple structure and fast convergence for both linear and discrete solution spaces. Each particle in the swarm algorithm represents a solution, which is influenced by the self-confidence value of the particle itself, the optimal solution in each round, and the global optimal solution during the iterative process. However, studies have shown that the particle swarm algorithm is prone to fall into local optimal solutions. Therefore, an improved discrete particle swarm algorithm is proposed in this paper. For the common discrete PSO algorithm, if the task graph has N nodes, we define the structure of particles as a tuple with N elements p k i = a k , 1 i , a k , 2 i , … , a k , N i , where p k i denotes the i -th particle in the k -th iteration, and a denotes the n -th particle in task graph node corresponding to the n-th resource node. We define as the process of particle update, which denotes the exchange sequence of the minimum number of exchanges required to go from particle to particle. For example, if and . We define as the local optimal solution, i.e., the solution with the minimum value of the loss function obtained immediately by the particle swarm algorithm at the k-th round; we define as the global optimal solution, i.e., the optimal solution of the particle swarm algorithm during the iteration. According to the traditional PSO algorithm, the iterative formula for the particle swarm is obtained as follows. where is the inertia factor, defining In this paper, we proposed a double particle swarm algorithm, the aim of which is to obtain an optimal solution with the fastest running time. In order to prevent the particle swarm algorithm from easily falling into the local optimal solution, the number of particle populations increased to 2. One of the particle populations introduces the property of cross-variation in the genetic algorithm. Another particle population introduces the feature of neighborhood search to increase the local optimization ability. When the two populations converge to the global optimal solution, they also converge to the optimal solution of the other population to avoid falling into the local optimal solution. Therefore, the number of particles that undergo crossover variation and the number of rounds in which the value of the loss function remains constant determine the convergence time of the algorithm. Thus, it achieves controllability in the algorithm execution time and accuracy. When converging, the inertia factor of the PSO is constant, which leads to premature falling into the local optimal solution. The dynamic transformation of the inertia factor is introduced for both particle populations in DPSO to control the speed of convergence to the global optimal solution. 3.2.1. Dynamic Transformation of the Inertia Factor The inertia index is an important parameter in the particle swarm algorithm, which is itself an important parameter for the regulation of the global search ability and convergence speed. A larger inertia index can strengthen the global search ability of t algorithm, but can result in the convergence speed becoming slower; conversely, with a smaller inertia index , the algorithm has poor global search ability, but stronger local search ability and faster convergence speed. Therefore, our goal is for the particle swarm algorithm to have a stronger global search capability in the early stage, and thus be able to cover a larger range in the solution space; in the later stage a rapid reduction in the inertia index is required in order to strengthen the local search capability. A more common way to adjust the weights dynamically is to apply a linear weight decreasing strategy, in which the weight is gradually decreased to a smaller value from the early stage of the algorithm. In this paper, an adaptive exponential dynamic transformation weight optimization formulation as in Equation (15) is incorporated. The algorithm ensures that the particle swarm algorithm has a high inertia index in the early stage, which improves the global search ability of the algorithm in this stage; while in the later stage, the inertia index decreases rapidly, which accelerates the convergence speed of the algorithm and the local search ability. The inertia weights are set to the maximum value in the initial stage of the algorithm. During the iteration of the algorithm, the states of all particles are used as input to get the return information, and the rate of inertia index decline is calculated according to the exponential formula. The algorithm uses the evolutionary step of the particles as a feedback parameter and dynamically adjusts the inertia weight strategy according to the particle states, as shown in Equation (15). where denotes the decreasing value of the inertia weight, denote the preset maximum inertia weight and minimum inertia weight, respectively. represent the maximum number of iterative rounds and the current number of iterative rounds. is the environmental feedback factor, and the interval of the value of is usually around [0, 0.5], which is used to describe the degree of evolution of the particle population in the last iteration. It indicates the feedback property of the particle population to the inertia weight. When the value of is larger, it means that the evolution of the particles in the last iteration is better, and when the value of is 0, it means that the algorithm has converged. 3.2.2. Neighborhood Search Based on the adaptive exponential dynamic transformation inertia weight optimization formulation proposed in Section 3.2.1 , the convergence ability and local search ability of the algorithm are enhanced significantly in the later stage. However, as a swarm intelligence algorithm, the particles with lower adaptability are still attractive to the particles with higher adaptability, and the algorithm rapidly increases the convergence speed in the late stage, which thus increases the probability of avoiding falling into the local optimal solution while improving the local search ability. In this study, to the particle swarm algorithm based on the introduction of adaptive exponential dynamic transformation weight optimization, the concept of neighborhood search is added to search for more optimal solutions in its neighborhood when the particle diversity is less than a set threshold, and the concept of the taboo table is introduced in order to prevent repeated searches. The local search capability of the particle swarm algorithm is increased by masking the particles that have fallen into local optimal solutions and searching for more optimal solutions in their neighborhoods. 3.2.3. Gene Cross-Mutation Drawing on the idea of a genetic algorithm for avoiding falling into local optimal solutions, in this paper the genetic variation property is introduced into the PSO algorithm. Because the PSO algorithm requires the exploration of a wider solution space upfront, particles must be able to migrate from the solution space more flexibly. We define the structure of particles as a tuple p k i = a k , 1 i , a k , 2 i , a k , N i with N elements, where p k i denotes the i -th particle in the k -th iteration, and a k , n i denotes the task graph node corresponding to the n -th resource node in k i . We define p k i c = c r o s s o v e r p k i , n d 1 , n , r a n d 1 , n as the mutation process of particle p k i , indicating that the generation of p k i c occurs by the swapping of particles in two random dimensions. The mutation process occurs randomly, which avoids the problem of slowing down the convergence of the particle swarm algorithm after the occurrence of cross mutations and better extends the optimal solution in the early stage of the algorithm. This section uses the environmental feedback factor , as previously mentioned, as an input to calculate the probability, as shown in Equation (16). The algorithm requires a larger search range of solution space in the early stage, while in the later stage, it must avoid affecting the convergence effect and local search effect, and the probability decreases with the increase of rounds. However, when the evolution of particles becomes slower, a larger variation probability is needed in order to extend the exploration of the solution space, so the addition of environmental feedback factors can appropriately adjust the search range of the algorithm for the solution space, in order to obtain the optimal solution. 3.2.4. Double Particle Swarm Optimization To combine the properties proposed above, a double particle swarm optimization algorithm based on cross-variance and neighborhood search is proposed in this section, which combines the strong global search capability of cross-variance and the strong local search capability of neighborhood search as well as the previously mentioned adaptive exponential dynamic inertia factor in order to enhance the integrated capability of the algorithm. Due to the parallel computation of the double particle swarm, a mutual disturbance term is added to Equation (14), as shown in Equation (17). b e s t is the optimal solution in the double particle swarm, which enhances the linkage of the double particle swarms and further improves the global search capability and local search capability of the algorithm. The algorithm flow is as follows. The process of DPSO is shown in Algorithm 2. Initialization: the velocity and position of the particles, based on the initialized position information, are substituted into Equations (9) and (12) to calculate the initial fitness value (lines 1–2). The forbidden table is initialized for the neighborhood search (line 3). The particle population that joins the neighborhood search feature is defined as p o p 1 , the list of fitness values as f i t n e s s 1 , the particle population that joins the genetic variation feature as p o p 2 , and the list of fitness values as f i t n e s s 2 (lines 5–7). Particle evolution: the inertia factor of particles is updated according to Equation (15) and the particle position is updated according to Equation (17) (lines 8–11). The cross-variation state of particles in p o p 2 is evaluated according to Equation (16) (lines 12–17). The diversity of particles in p o p 1 is evaluated, and if the diversity value is less than the predetermined threshold, g 1 , k b e s t is added to the forbidden table and g 1 , k b e s t is reset to the suboptimal solution of fitness in p o p 1 (lines 18–23). Step 2 is repeated. Algorithm 2 : DPSO Inputs: N o C U , L : NoC topology graph VG: Group of resource node X , Y , Z : The number of NoC resource nodes in the three dimensions v x , y , z : Resource node with coordinates x, y, and z Population: Maximum particle number i t e r m , a x : m a x i m u m n u m b e r o f i t e r a t i o n s Output: M A P P I N G T G V G : Mapping relation of TG to VG 1: Initializes particle positions and velocities in a random range: p o p 1 0 , p o p 2 0 2: Calculates the fitness of each particle: f i t n e s s 1 0 , f i t n e s s 2 0 3: Initializes forbidden table: F o r b i d d e n T a b l e 4: For i t e r < i t e r m a x do 5:   updates pop fitness with Equation (9) and Equation (12): 6: f i t n e s s 1 i t e r ← f i t n e s s 1 i t e r − 1 7: f i t n e s s 2 i t e r ← f i t n e s s 2 i t e r − 1 8:   updates inertia factor according to Equation (15) 9:      updates particle position according to Equation (17) 10: p o p 1 i t e r ← p o p 1 i t e r − 1 11: p o p 2 i t e r ← p o p 2 i t e r − 1 12:      updates mutation probability 13: For p o p ∈ p o p 2 i t e r do 14: I f Equation 16 t h e n 15: p o p ← c r o s s o v e r ( p o p , r a n d o m ( ) , r a n d o m ( ) ) 16: End if 17: End for 18:    sorts p o p 1 i t e r according to f i t n e s s 1 i t e r 19: If p a r t i c l e s d i v e r s i t y < t h r e s h o l d then 20: F o r b i d d e n L i s t ← p o p 1 i t e r 0 21: p o p 2 i t e r 0 searches neighborhood, updates if there is a better solution 22: p b e s t ← p o p 1 i t e r 1 23: End if 24: End of 4. Simulation and Results 4.1. Experience Environment This study used the Hotspot-integrated, event-driven C++ 3D NoC simulator. The simulator models the packet communication delays at the clock cycle level. The bandwidth ratio between the horizontal and vertical directions is 100:10. The environment configuration is shown in Table 1 . In this paper, we consider the four baseline algorithms to evaluate the performance of each one, which are simulated annealing algorithm (SA), genetic algorithm (GA), particle swarm optimization algorithm (PSO) and KBMA [ 15 ]. and make a comparative analysis of their experimental results. 4.2. Experimental Task Graph In this experiment we used the classical task graphs MWD, VOPD, and DVOPD of NoC as shown in Figure 3 , and random task graphs generated by Task Graphs For Free (TGFF) [ 21 ]. 4.3. Performance Comparison 4.3.1. Experimental Results of the Traditional NoC Task Map Shown in Figure 4 , Figure 5 and Figure 6 , the performance with different mapping algorithms at different classical task graphs is undertaken with 3D NoC scale set to . Communication delay is reduced by 23.5% on average, the throughput is improved by 8.08% and the energy consumption is reduced by 18.0% in DVOPD, VOPD and MWD task graphs. From the point of view of data transmission, the area division algorithm calculates the thermal resistance and the Manhattan distance between nodes. Due to the small number of nodes in the traditional DAG graph, the same application is divided into a relatively concentrated area. The data transmission passes through a smaller number of nodes and links. The DPSO algorithm can improve the overall performance. 4.3.2. Results of Randomized Task Graphs Because the number of task graph nodes and NoC are not in one-to-one correspondence, this paper uses an adaptive region partitioning algorithm to partition the 3D NoC and then maps the task graph to task nodes in the 3D NoC according to the partitioning result. As shown in Figure 7 , Figure 8 and Figure 9 , G64, G48, G32, G24, G16, and G10 are randomly generated task graphs with the number of child nodes of 64, 48, 32, 24, 16 and 10, respectively. It is evident that for the larger task graph G64, the maximum improvement in the average communication delay, throughput rate, and energy loss is 19.48%, 12.48%, and 26.99, respectively, while the context of smaller task graph sizes, such as the task graph with task size 10, has a worse effect. It is evident that the performance of the DPSO algorithm proposed in this paper improves with the increase of the task graph size in the scenario of 3D NoC scale of 4 × 4 × 4. This indicates that the proposed double particle swarm algorithm has better global search capability in a larger solution space, and that the larger task graph size also allows the region division strategy to make better use of TSV channels and reduce the information communication between layers. When the size of the task graph is smaller than 24 task nodes, the solution space is small, the global search ability and local search ability of the DPSO algorithm cannot be fully reflected, and the effect is close to that of the traditional heuristic algorithm; when the size of the task graph is larger than 24 task nodes, the local search ability and global search ability can be reflected, and the mapping effect is greatly improved. The traditional PSO algorithm is the earliest to fall into the optimal solution, so the overall effect is the worst. From the perspective of data transmission, applications are divided into a relatively concentrated area. When the number of nodes in the task graph is less than 48, there are less NoC nodes and links that data packets pass through. Therefore, this algorithm can achieve better results (lower delay, higher throughput, and lower heat loss) than other algorithms. When the number of nodes in the task graph is greater than 48, the effect of the region division algorithm is weakened. Because the number of nodes in the divided NoC area is close to the number of nodes in the NoC, the best result at this time depends more on the best convergence of the DPSO algorithm. 5. Conclusions In this paper, we first propose an adaptive 3D NoC module partitioning algorithm. The node with the lowest thermal resistance, the node with the lowest delay, and node with the lowest throughput in the neighborhood are searched, starting with the node with the lowest thermal resistance. The DPSO algorithm improves search capability in the early stage based on the PSO algorithm, slows down the convergence of particles in the later stage in order to reduce the probability of falling into local optimal solutions, and adds a neighborhood search function to search for more optimal solutions in the solution space near the optimal and suboptimal solutions. The experiments show that the average delay is reduced by up to 32.4%, the throughput is improved by up to 35.5%, and energy loss is reduced by up to 31.3% compared with the existing algorithm based on a 3D NoC scale of . Author Contributions Conceptualization: J.F. and H.C.; methodology: J.F. and H.C.; validation: J.F., H.C.; formal analysis: J.F.; investigation: J.F. and X.L.; resources: J.F. and H.C.; writing—original draft preparation: H.C.; writing—review and editing: J.F., H.C. and X.L.; supervision: J.F., H.C. and X.L.; project administration: J.F.; funding acquisition: J.F. All authors have read and agreed to the published version of the manuscript. Funding This work is supported by Beijing Natural Science Foundation (4192007), and by the National Natural Science Foundation of China (61202076), along with other government sponsors. Informed Consent Statement Informed consent was obtained from all subjects involved in the study. Data Availability Statement Not applicable. Acknowledgments The authors would like to thank the reviewers for their efforts and for providing helpful suggestions that have led to several important improvements in our work. We would also like to thank all teachers and students in our laboratory for helpful discussions. Conflicts of Interest The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results. References Skadron, K.; Stan, M.R.; Huang, W.; Velusamy, S.; Sankaranarayanan, K.; Tarjan, D. Temperature-aware microarchitecture. In Proceedings of the 30th Annual International Symposium on Computer Architecture, San Diego, CA, USA, 9–11 June 2003; pp. 2–13. [ Google Scholar ] [ CrossRef ][ Green Version ] Wang, F.; Xie, Y.; Vijaykrishnan, N.; Irwin, M.J. On-chip Bus Thermal Analysis and Optimization. In Proceedings of the Design Automation & Test in Europe Conference, Munich, Germany, 6–10 March 2006; pp. 1–6. [ Google Scholar ] [ CrossRef ][ Green Version ] Hertl, M.; Weidmann, D.; Ngai, A. An advanced reliability improvement and failure analysis approach to thermal stress issues in IC packages. In Proceedings of the Physical and Failure Analysis of Integrated Circuits, IPFA 2009, 16th IEEE International Symposium on the IEEE, Suzhou, China, 30 June–4 July 2009. [ Google Scholar ] Liu, G.; Ming, F.; Gang, Q. Neighbor-aware dynamic thermal management for multi-core platform. In Proceedings of the Conference on Design, Automation and Test in Europe, Dresden, Germany, 12–16 March 2012. [ Google Scholar ] Chakraborty, A.; Duraisami, K.; Sathanur, A.; Sithambaram, P.; Macii, A.; Macii, E.; Poncino, M. Implementation of a thermal management unit for canceling temperature-dependent clock skew variations. Integr. VLSI J. 2008 , 4 , 2–8. [ Google Scholar ] [ CrossRef ] Shafifique, M.; Garg, S.; Henkel, J.; Marculescu, D. The EDA challenges in the dark silicon era. In Proceedings of the Design Automation Conference, San Francisco, CA, USA, 1–5 June 2014; pp. 1–6. [ Google Scholar ] Shafifique, M.; Garg, S.; Henkel, J.; Marculescu, D.J.; Henkel, H.; Khdr, S.; Pagani, A.; Shafifique, M. New trends in dark silicon. In Proceedings of the Design Automation Conference, San Francisco, CA, USA, 7–11 June 2015; pp. 1–6. [ Google Scholar ] Fang, J.; Yu, L.; Liu, S.; Lu, J.; Chen, T. KL_GA: An application mapping algorithm for mesh-of-tree (MoT) architecture in network-on-chip design. J. Supercomput. 2015 , 71 , 4056–4071. [ Google Scholar ] [ CrossRef ][ Green Version ] Jayshree, D.; Seetharaman, G.; Pati, D. Energy consumption and performance comparison of DE optimization and PSO-based IP-core mapping technique for 2D and 3D network-on-chip. Semicond. Sci. Technol. 2021 , 36 , 085015. [ Google Scholar ] [ CrossRef ] De Barros, J.B.; Sampaio, R.C.; Llanos, C.H. 2019 An adaptive discrete particle swarm optimization for mapping real-time applications onto network-on-a-chip based MPSoCs Proc. In Proceedings of the 32nd Symposium on Integrated Circuits and Systems Design ser. SBCCI 19, Sao Paulo, Brazil, 26–30 August 2019; ACM: New York, NY, USA, 2019; pp. 5–6. [ Google Scholar ] Keley, M.; Khademzadeh, A.; Hosseinzadeh, M.; Center, I. Efficient mapping algorithm on mesh-based NoCs in terms of cellular learning automata. Int. Arab J. Inf. Technol. 2019 , 16 , 312–322. [ Google Scholar ] Li, B.; Wang, X.; Singh, A.K.; Mak, T. On runtime communication- and thermal-aware application mapping in 3D NoC. In Proceedings of the 2017 Eleventh IEEE/ACM International Symposium on Networks-on-Chip (NOCS), Seoul, Republic of Korea, 19–20 October 2017; pp. 1–8. [ Google Scholar ] Gan, Y.; Guo, H.; Zhou, Z. 3D NoC Low-Power Mapping Optimization Based on Improved Genetic Algorithm. Micromachines 2021 , 12 , 1217. [ Google Scholar ] [ CrossRef ] [ PubMed ] Aravindhan, A.; Gopalakrishnan, L.; Ko, S. KBMA: A knowledge-based multi-objective application mapping approach for 3D NoC. IET Comput. Digit. Tech. 2019 , 13 , 324–334. [ Google Scholar ] Nalci, Y.; Kullu, P.; Tosun, S.; Ozturk, O. ILP formulation and heuristic method for energy-aware application mapping on 3D-NoCs. J. Supercomput. 2021 , 77 , 2667–2680. [ Google Scholar ] [ CrossRef ] Zhang, J.X.; Yang, G.H. Low-Complexity Tracking Control of Strict-Feedback Systems with Unknown Control Directions. in IEEE Trans. Autom. Control 2019 , 64 , 5175–5182. [ Google Scholar ] [ CrossRef ] Ding, W.; Wang, Q.G.; Zhang, J.X. Analysis and prediction of COVID-19 epidemic in South Africa. ISA Trans. 2022 , 124 , 182–190. [ Google Scholar ] [ CrossRef ] [ PubMed ] Zhang, X.; Dai, L. Image enhancement based on rough set and fractional order differentiator. Fractal Fract. 2022 , 6 , 214. [ Google Scholar ] [ CrossRef ] Manna, K.; Swami, S.; Chattopadhyay, S.; Sengupta, I. Integrated through-silicon via placement and application mapping for 3D mesh-based NoC design. ACM Trans. Embed. Comput. Syst. 2016 , 16 , 24. [ Google Scholar ] [ CrossRef ] Cheng, Y.; Zhang, L.; Han, Y.; Li, X. Thermal-Constrained Task Allocation for Interconnect Energy Reduction in 3-D Homogeneous MPSoCs. IEEE Trans. Very Large Scale Integr. (VLSI) Syst. 2013 , 21 , 239–249. [ Google Scholar ] [ CrossRef ] Dick, R.P.; Rhodes, D.L.; Wolf, W. TGFF: Task graphs for free. In Proceedings of the Sixth International Workshop on Hardware/Software Codesign. (CODES/CASHE’98), Seattle, WA, USA, 18 March 1998; pp. 97–101. [ Google Scholar ] [ CrossRef ] Figure 1. 3D NoC architecture graph. Figure 1. 3D NoC architecture graph. Figure 2. Example map of region division. Figure 2. Example map of region division. Figure 3. Classic Task Graph. Figure 3. Classic Task Graph. Figure 4. Communication delay at DVOPD, VOPD and MWD on 4 × 4 × 4 3D mesh structure. Figure 4. Communication delay at DVOPD, VOPD and MWD on 4 × 4 × 4 3D mesh structure. Figure 5. Throughput rate at DVOPD, VOPD and MWD on 4 × 4 × 4 3D mesh structure. Figure 5. Throughput rate at DVOPD, VOPD and MWD on 4 × 4 × 4 3D mesh structure. Figure 6. Heat loss at DVOPD, VOPD and MWD on 4 × 4 × 4 3D mesh structure. Figure 6. Heat loss at DVOPD, VOPD and MWD on 4 × 4 × 4 3D mesh structure. Figure 7. Communication delay at different sizes of random task graphs on 4 × 4 × 4 3D mesh structure. Figure 7. Communication delay at different sizes of random task graphs on 4 × 4 × 4 3D mesh structure. Figure 8. Throughput at different sizes of random task graphs on 4 × 4 × 4 3D mesh structure. Figure 8. Throughput at different sizes of random task graphs on 4 × 4 × 4 3D mesh structure. Figure 9. Energy loss at different sizes of random task graphs on 4 × 4 × 4 3D mesh structure. Table 1. The simulation parameters. Table 1. The simulation parameters. Network parameters Flit size 128 bits Latency Router 2 cycles, link 1 cycle Buffer depth 4 flits Routing algorithm XYZ routing Baseline topology 4 × 4 × 4 Configuration of Many-core Simulator Core Architecture Intel Pentium 4 Baseline Frequency 1.9 GHz Hotspot Parameters Die size [mm] 0.5 × 0.5 Specific heat capacity [J/(m 3 × K)] 1.75 × 10 6 Resistivity [(m − K)/W] 0.01

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COURT OF APPEALS COURT OF APPEALS DECISION DATED AND FILED March 28, 2012 Diane M. Fremgen Clerk of Court of Appeals NOTICE This opinion is subject to further editing. �If published, the official version will appear in the bound volume of the Official Reports. � A party may file with the Supreme Court a petition to review an adverse decision by the Court of Appeals. � See Wis. Stat. � 808.10 and Rule809.62. � Appeal No. � 2011AP1327 Cir. Ct. No. � 2009FA315 STATE OF WISCONSIN � IN COURT OF APPEALS DISTRICT II In re the marriage of: Joyce A. Hiller, ���������Petitioner-Respondent, ����v. George C. Hiller, ���������Respondent-Appellant. ����������� APPEAL from a judgment of the circuit court for Washington County : � James G. pouros , Judge. � Affirmed . � �����������Before Brown, C.J., Neubauer, P.J., and Reilly, J. �1 �������PER CURIAM.   George C. (�Chuck�) Hiller appeals from the maintenance and property division portions of the judgment divorcing him and Joyce A. Hiller. �We interpret the trial court�s decision, issued after a three-day trial and consideration of the parties� posttrial memoranda, as an effort to balance the equities in light of the parties� particular circumstances and as an appropriate exercise of the court�s discretion. �We affirm. �2 �������Chuck and Joyce married in 2002 after cohabiting for approximately twenty years. �It was a second marriage for both. �They each have children from their prior marriages. �Chuck is retired from the carpentry business he owned. �His long-time avocation has been collecting and restoring classic cars. �Throughout their years together, Joyce cared for the home. �She also worked as a retail clerk until retiring in 2007. �The parties maintained separate checking accounts but generally used the funds for marital purposes. �3 �������The trial court ordered Chuck to pay Joyce maintenance of $2000 per month for an indefinite period, reflecting a 55/45 income split in Chuck�s favor. �The court valued the marital estate at $1,003,383. �It deviated from the presumptive fifty-fifty property division under Wis. Stat. � 767.61(3) (2009-10), [1]awarding Joyce approximately thirty-seven percent of the estate and Chuck approximately sixty-three percent. �We will supply additional facts as necessary. Property Division � 4 ������� Property division lies within the sound discretion of the trial court. � LeMere v. LeMere , 2003 WI 67, �13, 262 Wis. 2d 426, 663 N.W.2d 789. � We sustain discretionary decisions where the trial court examines the relevant facts, applies a proper standard of law and, using a demonstrated rational process, reaches a conclusion a reasonable judge could reach. � Id. � Findings of fact are affirmed unless clearly erroneous. � Wis. Stat. � 805.17(2). � The weight and credibility to be given to testimony is uniquely within the province of the trial court, Siker v. Siker , 225 Wis. 2d 522, 528, 593 N.W.2d 830 (Ct. App. 1999), �because of the court�s superior opportunity to judge such matters,� Gardner v. Gardner , 190 Wis. 2d 216, 230, 527 N.W.2d 701 (Ct. App. 1994). � 5 ������� Due to its underlying philosophy that �marriage is akin to an economic partnership between the spouses,� Rodak v. Rodak , 150 Wis. 2d 624, 630, 442 N.W.2d 489 (Ct. App. 1989), Wis. Stat . � 767.61(3) creates a rebuttable presumption that the marital estate is to be divided equally at the time of divorce. � �[S]pouses are presumed to be equal contributors to the accumulated wealth of a marriage, regardless of whether their contributions are principally made inside or outside the home.� � LeMere , 262 Wis. 2d 426, �19 (citation omitted). � Courts may deviate from an equal property division, however, upon considering numerous statutory factors. � Sec. 767.61(3). 1.Attorney fee �add-backs� �6 �������When Joyce filed for divorce in June 2009, Chuck�s checking account had a balance of $63,575. �From June 2009 through January 24, 2011, just before trial commenced, Chuck deposited $92,093 in social security, pension and annuity income into the account, so that, had there been no expenditures, a balance of $155,668 should have existed at the time of divorce. �Instead, the balance was $3,325. �The trial court found that some of Chuck�s expenditures�$20,000 in attorney fees paid from joint monies, $39,000 in gifts to his adult children, and a $20,000 �loan� to a friend�constituted �marital waste or depletion of assets� and therefore should be added back into the marital estate. � �7 �������On appeal, Chuck counts as error only the adding back of the $20,000 he paid toward attorney fees during the pendency of the action. �He argues that paying for one�s own attorney fees is not �waste� under Wis. Stat.� 767.63 and there is no other authority for subjecting that sum to an �add-back.� � � 8 ������� The �waste� phraseology notwithstanding, we disagree in the final analysis that the court looked at Chuck�s legal expenses in the framework of the �waste� statute. [2] � Rather, the court expressly noted that the so-called �add-back� line of cases comes under the �contribution to the marriage� factor of Wis. Stat. � 767.61(3)(d). � This factor allows the court to consider each party�s efforts to preserve the marital assets. � See Anstutz v. Anstutz , 112 Wis. 2d 10, 12, 331 N.W.2d 844 (Ct. App. 1983). � � 9 ������� With that premise, we turn to the �add-backs.� � Whether an asset is subject to division under Wis. Stat. � 767.61 is a question of law. � See Weiss v. Weiss , 122 Wis. 2d 688, 692, 365 N.W.2d 608 (Ct. App. 1985). � While paying one�s own attorney fees may not be �waste,� nonmarital liabilities incurred after the petition for divorce is filed should not be subtracted from the marital estate. � See id. at 699. � Personal expenditures may bear on the property division and a party�s ability to pay maintenance. � Id. at 699-700. � Furthermore, just as assets acquired during the pendency of a divorce action are included when determining the size of the marital estate at the time of the divorce, Overson v. Overson , 125 Wis. 2d 13, 21, 370 N.W.2d 796 (Ct. App. 1985), by the same logic, the trial court should be able to add back the value of assets spent or disposed of during the pendency of the action if they were acquired during the marriage and if the court deems their expenditure to be an unjustified depletion of the estate. � 10 ����� Here, the court found that Joyce paid her attorney fees from an inheritance and �contributed mightily to the homemaking� and to hostessing and catering large events related to Chuck�s car hobby, and that Chuck negatively impacted the marital estate by depleting assets. � �[R]ecompense [is] available when one spouse has mismanaged or dissipated assets.� � Covelli v. Covelli , 2006 WI App 121, �30, 293 Wis. 2d 707, 718 N.W.2d 260. � The court�s analysis under Wis. Stat. � 767.61(3)(d) was proper. � We construe the attorney fee add-back as an appropriate effort to make the marital estate whole. ���� 2.Asset value add-back �11 �����The court also ordered an add-back of the value of cars and car parts Joyce claimed were missing at the time of divorce. �Suspecting that Chuck might conceal assets, Joyce and her son secretly photographed Chuck�s vehicle and parts inventory and photocopied his car records. �When court-appointed appraiser Larry Vanne later evaluated the collection, he could not locate five cars, eleven chrome bumpers and two performance carburetors (�tri-carbs�). �Chuck testified that he had dismantled one of the cars and sold another one; that Vanne overlooked some of the bumpers; and that the other three cars, the tri-carbs and the remaining bumpers actually were owned by third parties who had retrieved them. �Vanne appraised the value of the missing cars and parts between $77,300 and $101,400. �The court placed the value at $80,000 and ordered that amount added back. � �12 �����Chuck challenges only the add-back for the three cars and thetri-carbs. �He claims there is no evidence he ever owned those three cars and that the evidence is uncontradicted that he was working on them for the real owners. �13 �����A court may not disregard uncontradicted testimony absent something that discredits it or renders it against reasonable probabilities. � See Ashraf v. Ashraf, 134 Wis. 2d 336, 345, 397 N.W.2d 128 (Ct. App. 1986). �Here, the court had �something else.� �It had Joyce�s photographic documentation and Vanne�s appraisal, which the court deemed more credible than that of Chuck�s appraiser. �Both served to discredit Chuck�s self-serving explanations about the items� whereabouts and ownership. �14 �����Next, citing Dean v. Dean, 87 Wis. 2d 854, 865, 275 N.W.2d 902 (1979), Chuck asserts that the burden of proof was on Joyce as the party seeking inclusion in the marital estate of assets held in the name of a third party. �Third-party ownership never was established, however, and Chuck was better situated to prove that claim. [3] �The failure to produce relevant evidence in his control and that would have been in his interest to produce permits an inference that the evidence would have been unfavorable to him. � See Convey v. Milwaukee & Suburban Transp. Corp., 8 Wis. 2d 520, 526, 99 N.W.2d 713 (1959). �15 �����We conclude that these assets are subject to division under Wis. Stat.� 767.61. �We uphold the trial court�s finding that their value was $80,000 because that valuation is not clearly erroneous. � See Liddle v. Liddle, 140 Wis. 2d 132, 136, 410 N.W.2d 196 (Ct. App. 1987). �It was within the court�s discretion to add back their value. 3.Property division methodology �16 �����Chuck next takes issue with the court�s exercise of discretion in making the property division, through which the trial court awarded him approximately two-thirds of the property. �He asserts that, while the trial court �was conscientious in mentioning� the relevant statutory factors, its decision failed to reveal a reasoning process. �He contends the court�s lack of analysis stems from the court�s reluctance to divide the property, about ninety percent of which he claims he brought to the marriage, after only an eight-year marriage. �We disagree. �17 �����A court begins with the presumption that property should be divided equally. � Wis. Stat.�767.61(3). �It may deviate from that presumption after considering twelve listed factors. � See id.; see also LeMere, 262 Wis. 2d 426, �22. �The court may give varying weight to the factors it deems applicable. � See LeMere, 262 Wis. 2d 426, �25. �The court�s explanation �need not be a lengthy process. �While reasons must be stated, they need not be exhaustive.� � Burkes v. Hales, 165 Wis. 2d 585, 590, 478 N.W.2d 37 (Ct. App. 1991). � �18 �����Here, the trial court�s decision fully satisfied the standard we have set forth. �The court first observed that the case was �fact intensive and fact specific� making witness credibility �critical.� �It explained why it found Chuck�s testimony less credible than Joyce�s and Chuck�s appraiser�s testimony less credible and reliable than Vanne�s. �The court also observed that the parties did not have either a cohabitation agreement or a premarital agreement, either of which might have defined assets as not being subject to division. �19 �����The court then (1) determined what assets comprised the divisible estate, including those that should have been there but no longer were;(2) presumed an equal division; (3) assessed whether to deviate from the presumed fifty-fifty split by examining each statutory factor; and (4) determined how to accomplish the division in practical terms. �Several factors weighed heavily in the property division the court fashioned: �the substantial assets Chuck brought to the marriage, Joyce�s significant homemaking contributions, Chuck�s asset depletion, and Joyce�s lesser earning capacity. � See Wis. Stat.� 767.61(3)(b), (d) and (g). �It expressly noted that it was not considering the parties� two-decade cohabitation as an �other factor� relevant to property division. � See� 767.61(3)(m). �The weight given the various considerations is within the trial court�s discretion. � See Herlitzke v. Herlitzke, 102 Wis. 2d 490, 495, 307 N.W.2d 307 (Ct. App. 1981). � 4.Gifted and inherited assets �20 �����Chuck next criticizes the trial court for acknowledging gifted and inherited funds but not explaining the impact, if any, that they had on the property division. �He complains that the court simply �stated that it had taken the Schwartz v. Linderscase into account in making the property division.� �21 �����Chuck does not go even that far. �He does not provide a citation to Schwartz, [4]let alone discuss its application. �We could search the record for support for the court�s discretionary decision. � See Randall v. Randall, 2000 WI App 98, �7, 235 Wis. 2d 1, 612 N.W.2d 737. �But as Chuck�s undeveloped,four-sentence argument comes nowhere near persuading us that the court erroneously exercised its discretion, we decline to address it further. � See Barakat v. DHSS, 191 Wis. 2d 769, 786, 530 N.W.2d 392 (Ct. App. 1995). Maintenance �22 �����Like property division, the determination of the amount and duration of a maintenance award also is entrusted to the trial court�s discretion. � See Grace v. Grace, 195 Wis. 2d 153, 157, 536 N.W.2d 109 (Ct. App. 1995). � �23 �����In setting maintenance here, the trial court found authority in Meyer v. Meyer, 2000 WI 132, 239 Wis. 2d 731, 620 N.W.2d 382, to consider the parties� twenty years of premarital cohabitation. � Wisconsin Stat.� 767.56(9) permits a court to consider as a factor in setting maintenance �[t]he contribution by one party to the education, training or increased earning power of the other.� �In Meyer, where the issue was the wife�s support of the husband�s educational pursuits, the supreme court held that � 767.56(9) is broad enough to embrace premarital contributions. � Meyer, 239 Wis. 2d 731, ��1, 43. � �24 �����In view of Meyer�s interpretation of Wis. Stat.� 767.56(9), the court concluded that Joyce contributed to Chuck�s earning power throughout the parties� entire time together by performing services that allowed him to pursue his business and his avocation. �Given the plainly stated scope of the statute, we must reject Chuck�s claim that Meyeris limited to spousal contributions only to the other�s education. � �25 �����In addition, the trial court found that the twenty years the parties cohabited also was an �other factor� relevant to setting maintenance in this particular case. � See Wis. Stat.� 767.56(10). �We reject Chuck�s contention based on the Meyerdissent that this was error because �[i]f parties want the benefits of marriage, the Family Code makes it clear that the way to obtain them is to get married.� �First, a dissent �is what the law is not.� � State v. Perry, 181Wis. 2d 43, 49, 510 N.W.2d 722 (Ct. App. 1993). �More to the point, though, we disagree that the cohabitation drove the court�s decision. �Rather, it considered the parties� mutual contributions to their �economic partnership.� � See Rodak, 150 Wis. 2d at 630. �The court deemed the long-term nature of the contributions to be an �other factor� relevant to this particular case, a consideration that falls under the court�s general discretion and � 767.56(10). � �26 �����Chuck also complains that the court wrongly factored into his available income sums it had �deprived� him of in the property division and an amount he receives monthly from an inherited annuity. �Chuck is mistaken. �The court set maintenance based on income figures he provided which, according to his own posttrial brief, did not include the amounts he complains about. �27 �����In sum, the trial court carefully ticked through each of the statutory factors, explaining why it did or did not find them applicable to the maintenance award. �It awarded Joyce $2000 a month, one of Chuck�s alternate proposals on the Mac Davis calculation he submitted to the court. �The court�s findings are not clearly erroneous, and we are bound by them. �Those findings support the court�s exercise of discretion in awarding maintenance. �We see no error. ����������� By the Court.�Judgment affirmed. �����������This opinion will not be published. � See Wis. Stat. Rule809.23(1)(b)5. [1]All references to the Wisconsin Statutes are to the 2009-10 version unless noted. [2]Even if it did, we could affirm on different grounds. � Vanstone v. Town of Delafield, 191 Wis. 2d 586, 595, 530 N.W.2d 16 (Ct. App. 1995). � [3] Chuck supplies in his appendix a certificate of title indicating third-party ownership of one of the three cars. � The court did not admit it as an exhibit at trial because Chuck did not produce the alleged owner as a witness. � As the document is not part of the record, it will not be considered. � See Balele v. Wisconsin Pers. Comm�n , 223 Wis. 2d 739, 752, 589 N.W.2d 418 (Ct. App. 1998). � [4] Schwartz v. Linders , 145 Wis. 2d 258, 426 N.W.2d 97 (Ct. App. 1988).

https://www.wicourts.gov/ca/opinion/DisplayDocument.html?content=html&seqNo=79891

Smith v. Minster Mach. Co. - New York - Case Law - VLEX 884461524 0: [object Object]. 1: [object Object]. 2: [object Object]. 3: [object Object]. 4: [object Object] Smith v. Minster Mach. Co. <table><tbody><tr><td> Court</td><td> New York Supreme Court Appellate Division</td></tr><tr><td> Writing for the Court</td><td> All concur except BALIO; BALIO</td></tr><tr><td> Citation</td><td> 233 A.D.2d 892,649 N.Y.S.2d 257</td></tr><tr><td> Parties</td><td> Sheila SMITH, Appellant, v. MINSTER MACHINE COMPANY, Respondent.</td></tr><tr><td> Decision Date</td><td> 08 November 1996</td></tr></tbody></table> Page 257 649 N.Y.S.2d 257 233 A.D.2d 892 Sheila SMITH, Appellant, v. MINSTER MACHINE COMPANY, Respondent. Supreme Court, Appellate Division, Fourth Department. Nov. 8, 1996. Page 258 The Schiano Law Office by Charles Schiano, Jr., Rochester, for appellant. Connors and Corcoran by Gregg Redmond (Joseph Fritsch, of counsel), Rochester, for respondent. Before DENMAN, P.J., and PINE, FALLON, DOERR and BALIO, JJ. MEMORANDUM: Plaintiff commenced this action against defendant, Minster Machine Company (Minster), the manufacturer of a power-press, for personal injuries she sustained while operating the press at her work place. When the press was purchased by the original buyer, it was equipped with a standard two-hand control system, designed to engage both hands of the operator while the press was being cycled. The only warning placed on the machine pertained to lubrication instructions and press specifications. At the time of the accident, the two-hand control had been replaced by a movable barrier guard, which was installed on the front of the press frame and operated by a pneumatic foot-pedal valve. Neither the guard nor the foot pedal was manufactured or provided by Minster. An examination of the machine after the accident revealed that the operator's hand was exposed to the operation of the press because a guard had not been properly adjusted. Minster moved for summary judgment dismissing the complaint upon the grounds that post-sale modifications to the press had altered a key safety feature, and there was no showing that a warning would have given plaintiff any more appreciation of the dangers involved, which were obvious and apparent. Supreme Court agreed and dismissed the complaint in its entirety.That was error. It is now well established that " [m]aterial alterations at the hands of a third party which work a substantial change in the condition in which the product was sold by destroying the functional utility of a key safety feature, however foreseeable that modification may have been, are not within the ambit of a manufacturer's responsibility" (Robinson v. Reed-Prentice Div.of Package Mach. Co.,49 N.Y.2d 471, 481,426 N.Y.S.2d 717,403 N.E.2d 440). Nevertheless, post-sale modifications to a product do not defeat a products liability claim unless those modifications rendered a safe product defective and caused the injuries (Lamey v. Foley,188 A.D.2d 157, 168,594 N.Y.S.2d 490). Here, there is a question of fact whether the post-sale modifications to the press constituted material alterations that rendered the press defective. It cannot be determined on this record whether, as a matter of law, the modifications rendered an otherwise safe product defective. The issues whether the modifications were contemplated by Minster at the time of sale, and whether the modifications constituted material alterations sufficient to exculpate Minster from liability were within the province of the jury. There is also a question of fact whether the press was defectively designed and manufactured. Although the press was manufactured with a dual hand... Ganter v. Makita U.S.a., 01-01829 United States New York Supreme Court Appellate Division February 1, 2002 ...alternative designs at the time of manufacture (see, Eiss v Sears, Roebuck & Co., 275 A.D.2d 919, 919-920; Smith v Minster Mach. Co., 233 A.D.2d 892, 893; see also, Sanchez v Otto Martin Maschinenbau GmbH & Co., 281 A.D.2d 284, 285; see generally, Lopez v Precision Papers, 67 N.Y.2d 871, We...... Santangelo v. Fluor Constructors Int'l, 01-02354 United States New York Supreme Court Appellate Division May 3, 2002 ...decedent's injuries, i.e., that the subsequent modification rendered an originally safe product defective (see Smith v Minster Mach. Co., 233 A.D.2d 892, 893; see generally Robinson v Reed-Prentice Div. of Package Mach. Co., 49 N.Y.2d 471, 475). Lill alleges that there may be two contributi...... Hoang v. ICM Corp., PLAINTIFF-RESPONDENT United States New York Supreme Court Appellate Division July 3, 2001 ...with the foot switch creates a triable issue of fact with respect to the failure to warn claim (see, Smith v Minster Mach. Co., 233 A.D.2d 892, The court also properly denied those parts of defendants' motions with respect to the design defect claim, which is based on the alleged lack of pr...... Schneider v. Verson Allsteel Press Co. a Div. of Allied Products Corp. United States New York Supreme Court — Appellate Division February 7, 1997 ...Corp., rather than the lack of protective guards, were the proximate cause of plaintiff's injuries (see, Smith v. Minster Mach. Co., 233 A.D.2d 892, 649 N.Y.S.2d 257; Lamey v. Foley, supra, at 168, 594 N.Y.S.2d 490). Finally, whether the action of plaintiff in placing his hand inside the di...... 4 cases Ganter v. Makita U.S.a., 01-01829 United States New York Supreme Court Appellate Division February 1, 2002 ...alternative designs at the time of manufacture (see, Eiss v Sears, Roebuck & Co., 275 A.D.2d 919, 919-920; Smith v Minster Mach. Co., 233 A.D.2d 892, 893; see also, Sanchez v Otto Martin Maschinenbau GmbH & Co., 281 A.D.2d 284, 285; see generally, Lopez v Precision Papers, 67 N.Y.2d 871, We...... Santangelo v. Fluor Constructors Int'l, 01-02354 United States New York Supreme Court Appellate Division May 3, 2002 ...decedent's injuries, i.e., that the subsequent modification rendered an originally safe product defective (see Smith v Minster Mach. Co., 233 A.D.2d 892, 893; see generally Robinson v Reed-Prentice Div. of Package Mach. Co., 49 N.Y.2d 471, 475). Lill alleges that there may be two contributi...... Hoang v. ICM Corp., PLAINTIFF-RESPONDENT United States New York Supreme Court Appellate Division July 3, 2001 ...with the foot switch creates a triable issue of fact with respect to the failure to warn claim (see, Smith v Minster Mach. Co., 233 A.D.2d 892, The court also properly denied those parts of defendants' motions with respect to the design defect claim, which is based on the alleged lack of pr...... Schneider v. Verson Allsteel Press Co. a Div. of Allied Products Corp. United States New York Supreme Court — Appellate Division February 7, 1997 ...Corp., rather than the lack of protective guards, were the proximate cause of plaintiff's injuries (see, Smith v. Minster Mach. Co., 233 A.D.2d 892, 649 N.Y.S.2d 257; Lamey v. Foley, supra, at 168, 594 N.Y.S.2d 490). Finally, whether the action of plaintiff in placing his hand inside the di......

https://case-law.vlex.com/vid/smith-v-minster-mach-884461524

Strategic Dialogue on North Korea Global Policy Forum is a policy watchdog that follows the work of the United Nations. We promote accountability and citizen participation in decisions on peace and security, social justice and international law. Strategic Dialogue on North Korea Foreign Policy in Focus Brent Choi and Joowon Jung John Feffer is a North Korea specialist who is well known for his pro-engagement policy toward North Korea. In his article North Korea and Malign Neglect, it's not surprising that Feffer argues for an engagement approach as a solution to the North Korean nuclear issue. Such a point of view, however, results in unrealistic and unbalanced claims in several respects. First of all, Feffer fails to recognize the fact that it is North Korea that should be held accountable for the current standoff between the United States and the North. His assertion that the Obama administration has been using a "malign neglect" policy in dealing with the North over the past 9 months is not true. U.S. Secretary of State Hillary Clinton said in February that the Obama administration is willing to establish diplomatic relations with the North, replace the armistice treaty with a peace treaty, and provide it with economic aid. Stephen Bosworth, a special envoy for North Korea, even stated that the Unites States could make a visit to the North for a bilateral negotiation. Despite such continued U.S. efforts, however, North Korea launched a long-range rocket in April and rejected Bosworth's visit. Secondly, without reflecting on the current political situation in North Korea, Feffer maintains that the United States must take an engagement approach in dealing with the North. However, considering that the North threatening to go ahead with its second nuclear test, he should have been able to realize that the center of the power in the North Korean regime is moving in favor of the harder-liners. It also appears that North Korean leader Kim Jong-Il has started the selecting process for his successor, who is likely to be one of his sons. Therefore, Pyongyang's recent hard-line stance toward the United States is based on its effort to hide these political changes. Finally, when suggesting "an audacious and hopeful plan of action for North Korea" in the article, Feffer must lay out in detail the sorts of things that need to done by the Obama administration. For instance, he should discuss in a clear and specific manner what kind of economic and political aid, and how much, is needed to bring the North back to the negotiation table. John Feffer North Korea is the longest running U.S. policy failure. We've tried just about everything: war, containment, threats, isolation, an agreed framework, Six Party Talks, and bilateral discussions. The State Department has accumulated over 60 years of frustrations. The Pentagon is largely out of the equation given the devastating consequences of a military attack and subsequent war on the Korean peninsula. Now, the Obama administration comes into the White House under the general banner of "change." But with its default policy of malign neglect, it has only offered more of the same. Brent Choi and Joowon Jung, who are both very seasoned observers of North Korea, aren't optimistic about a breakthrough any time soon. They argue that North Korea has decided once and for all to be a nuclear power. Pyongyang isn't interested any more in the Six Party Talks. It wants bilateral negotiations with the United States on an equal basis - between two nuclear powers. The United States, they argue, should respond with a bigger stick (the threat of redeployment of nuclear weapons in South Korea) and a bigger carrot (high-level bilateral talks). This, Choi and Jung argue, is a new approach. But it isn't. The challenges of engaging North Korea have led many to walk away from the negotiating table in favor of a policy of brinkmanship. North Korea uses aggressive rhetoric, has flouted international norms by pursuing a nuclear program, and has ignored even the pressure of its putative allies in testing long-range rockets. At a certain level, North Korea is the most geopolitical of countries: it conducts its international relations almost exclusively on the basis of calculations of power and advantage. It wants a deal from the United States because it judges, correctly, that the United States is the most powerful country in the world and thus holds the key to North Korea's future. Despite Pyongyang's focus on power and Washington's role in resolving its myriad problems, we haven't had much luck using brinksmanship to force North Korea to change its pattern of behavior. We practically bombed the country out of existence during the Korean War and failed to dislodge its leadership. The numerous threats against the country during the Cold War likewise proved unsuccessful. The Clinton administration attempted to push North Korea to the brink in 1994 during the first nuclear crisis and Pyongyang didn't blink. During its first term, George W. Bush's administration did its best to destabilize the country, and again North Korea simply pushed back. The threat of placing nuclear weapons back in South Korea will have as much chance of success as these earlier attempts at brinksmanship. North Korea thrives on such threats. The leadership uses them to prove to the population that the country is surrounded and emergency measures are required in response. I, too, am worried that South Korea and Japan will decide to pursue nuclear programs of their own as a way to counter North Korea's capabilities. But bringing U.S. nuclear weapons back to the region in this way is no solution, for it doesn't address the underlying security threats. Even with a nuclear program, North Korea is in the same corner it's been in for years. The economy is in lousy shape. Relations with Russia and China are strained. Kim Jong Il isn't exactly a much-beloved leader. He's getting on in years, and there's no clearly popular successor in the wings. North Korea will only change internally when its external relations change dramatically, and that will require a new U.S. approach. We've tried brinksmanship and containment for over 60 years and the only change has been North Korea going nuclear. By following the example of détente with Vietnam and China, we can minimize the risk that North Korea poses to the international community and also encourage positive changes within the country. Only a fundamentally altered relationship with North Korea - economic engagement, a peace treaty to finally end the Korean War, diplomatic normalization - will change the dynamic by removing the threat that sustains the Kim Jong-Il regime and keeps the North Korean garrison mentality intact.

https://archive.globalpolicy.org/security-council/index-of-countries-on-the-security-council-agenda/north-korea/47847-strategic-dialogue-on-north-korea.html

Extending the straight leg raise test for improved clinical evaluation of sciatica: validity and diagnostic performance with reference to the magnetic resonance imaging-ICH GCP Pesonen J, Shacklock M, Suomalainen JS, Karttunen L, Maki J, Airaksinen O, Rade M. Extending the straight leg raise test for improved clinical evaluation of sciatica: validity and diagnostic performance with reference to the magnetic resonance imaging. BMC Musculoskelet Disord. 2021 Sep 21;22(1):808. doi: 10.1186/s12891-021-04649-z. Erratum In: BMC Musculoskelet Disord. 2021 Nov 10;22(1):937. BMC Musculoskelet Disord. 2022 May 31;23(1):518.。 ICH GCP。 Janne Pesonen, Michael Shacklock, Juha-Sampo Suomalainen, Lauri Karttunen, Jussi Mäki, Olavi Airaksinen, Marinko Rade, Janne Pesonen, Michael Shacklock, Juha-Sampo Suomalainen, Lauri Karttunen, Jussi Mäki, Olavi Airaksinen, Marinko Rade Abstract Background:The straight leg raise test (SLR) is one of the most utilized and studied physical tests in patients with low back pain (LBP) for the detection of lumbar disc herniation (LDH), showing high sensitivity and heterogeneous or low specificity. The high incidence of asymptomatic 'pathologic' findings in the magnetic resonance imaging (MRI) scans may cause verification bias to these results. We studied an extended SLR (ESLR) by adding location-specific structural differentiation movements (hip internal rotation or ankle dorsiflexion) to the traditional SLR for it to better differentiate neural symptoms from musculoskeletal. Previously, the ESLR has shown almost perfect interrater reliability between examiners and ability to detect sciatic patients. In this study, we investigated whether a 'positive' ESLR finding is associated with pathology seen on MRI. Methods:Forty subjects comprised the study population, 20 in sciatic group and 20 in control group. The ESLR was performed 'blinded' to the subjects. After the ESLR, each subject's lumbar MRI was evaluated. The MRIs were analyzed independently by 2 senior radiologists and a spine specialist clinician. The ESLR and MRI results were cross-tabulated. To obtain the odds ratio (OR) with positive ESLR or SLR results for LDH or nerve root compression (NC), a binary logistic regression analysis with subjects' age, gender, height and weight was performed. ESLR's validity was assessed by combination of interrater agreement and percentage prevalence of both LDH and NC. Results:Of sciatic (ESLR+) patients, 85 % had LDH and 75 % NC in the MRI. Not surprisingly, MRI showed a very high incidence of 'false-positive' findings with the ESLR negative group. The ESLR showed 0.85 sensitivity and 0.45 specificity for LDH and 0.75 sensitivity and 0.50 specificity for NC. A positive result in the ESLR was found to be strongly associated with for both LDH and NC: the OR was 8.0 (p = 0.028) and 5.6 (p = 0.041), respectively. Conclusions:The ESLR shows high validity in detecting neural symptoms and is strongly associated with pathology seen in the MRI when judged positive. We suggest the use of ESLR in clinical practice as a part of clinical examination, where it may prove to be a valuable tool in detecting patients with sciatic symptoms. Conflict of interest statement The authors declare they have no competing or conflict of interest. © 2021. The Author(s). Figures Fig. 1 Proximal structural differentiation for distal symptoms with hip internal rotation. Published earlier by Pesonen et al., BMC Musculoskeletal Disorders 2021 [14] Fig. 2 Distal structural differentiation for proximal symptoms with ankle dorsiflexion (also known as Bragard test). Published earlier by Pesonen et al., BMC Musculoskeletal Disorders 2021 [14] Fig. 3 MRI showing lumbar disc herniation on L5-S1 disc with neural compression on right S1 nerve root (T2-weighted sagittal and axial views) Fig. 4 Fig. 5 Crosstabulations between ESLR, traditional SLR and MRI findings for neural compression. ESLR = Extended straight leg raise test; MRI = Magnetic resonance imaging; Trad. SLR = Traditional straight leg raise test Fig. 6 Crosstabulations between ESLR and traditional SLR results. ESLR = Extended straight leg raise test, Trad. SLR = Traditional straight leg raise test References James SLG, Abate D, Abate KH, Abay SM, Abbafati C, Abbasi N, et al. Global, regional, and national incidence, prevalence, and years lived with disability for 354 diseases and injuries for 195 countries and territories, 1990–2017: a systematic analysis for the Global Burden of Disease Study 2017. Lancet. 2018; 10.1016/S0140-6736(18)32279-7. Hartvigsen J, Hancock MJ, Kongsted A, Louw Q, Ferreira ML, Genevay S, et al. What low back pain is and why we need to pay attention. Lancet. 2018 doi: 10.1016/S0140-6736(18)30480-X. Buchbinder R, van Tulder M, Öberg B, Costa LM, Woolf A, Schoene M, et al. Low back pain: a call for action. Lancet. 2018 doi: 10.1016/S0140-6736(18)30488-4. Maher C, Underwood M, Buchbinder R. Non-specific low back pain. Lancet. 2017 doi: 10.1016/S0140-6736(16)30970-9. Brinjikji W, Luetmer PH, Comstock B, Bresnahan BW, Chen LE, Deyo RA, et al. Systematic literature review of imaging features of spinal degeneration in asymptomatic populations. AJNR Am J Neuroradiol. 2015;36:811–816. doi: 10.3174/ajnr.A4173. Webster BS, Choi Y, Bauer AZ, Cifuentes M, Pransky G. The cascade of medical services and associated longitudinal costs due to nonadherent magnetic resonance imaging for low back pain. Spine (Phila Pa 1976) 2014 doi: 10.1097/BRS.0000000000000408. Foster NE, Anema JR, Cherkin D, Chou R, Cohen SP, Gross DP, et al. Prevention and treatment of low back pain: evidence, challenges, and promising directions. Lancet. 2018 doi: 10.1016/S0140-6736(18)30489-6. Chou R, Fu R, Carrino JA, Deyo RA. Imaging strategies for low-back pain: systematic review and meta-analysis. Lancet. 2009 doi: 10.1016/S0140-6736(09)60172-0. Ropper AH, Zafonte RD. Sciatica. N Engl J Med. 2015;372:1240–1248. doi: 10.1056/NEJMra1410151. Airaksinen O, Brox JI, Cedraschi C, Hildebrandt J, Klaber-Moffett J, Kovacs F, et al. Chapter 4. European guidelines for the management of chronic nonspecific low back pain. Eur Spine J. 2006;15(Suppl 2):192. doi: 10.1007/s00586-006-1072-1. Ekedahl H, Jönsson B, Annertz M, Frobell RB. Accuracy of clinical tests in detecting disk herniation and nerve root compression in subjects with lumbar radicular symptoms. Arch Phys Med Rehabil. 2018 doi: 10.1016/j.apmr.2017.11.006. van der Windt, D A, Simons E, Riphagen II, Ammendolia C, Verhagen AP, Laslett M, et al. Physical examination for lumbar radiculopathy due to disc herniation in patients with low-back pain. Cochrane Database Syst Rev. 2010;(2):CD007431. 10.1002/14651858.CD007431.pub2. Tawa N, Rhoda A, Diener I. Accuracy of clinical neurological examination in diagnosing lumbo-sacral radiculopathy: a systematic literature review. BMC Musculoskelet Disord. 2017 doi: 10.1186/s12891-016-1383-2. Pesonen J, Shacklock M, Rantanen P, Mäki J, Karttunen L, Kankaanpää M, et al. Extending the straight leg raise test for improved clinical evaluation of sciatica: reliability of hip internal rotation or ankle dorsiflexion. BMC Musculoskelet Disord. 2021 doi: 10.1186/s12891-021-04159-y. Stynes S, Konstantinou K, Ogollah R, Hay EM, Dunn KM. Clinical diagnostic model for sciatica developed in primary care patients with low back-related leg pain. PLoS ONE. 2018 doi: 10.1371/journal.pone.0191852. Sim J, Wright CC. The kappa statistic in reliability studies: use, interpretation, and sample size requirements. Phys Ther. 2005;85:257–68. Buckup K. Bragard Test. In: Buckup K, editor. Clinical Tests for the Musculoskeletal System. Stuttgart: Thieme; 2008. pp. 59–60. Boyd BS, Topp KS, Coppieters MW. Impact of movement sequencing on sciatic and tibial nerve strain and excursion during the straight leg raise test in embalmed cadavers. J Orthop Sports Phys Ther. 2013 doi: 10.2519/jospt.2013.4413. Bueno-Gracia E, Pérez-Bellmunt A, Estébanez-de-Miguel E, López-de-Celis C, Shacklock M, Caudevilla-Polo S, et al. Differential movement of the sciatic nerve and hamstrings during the straight leg raise with ankle dorsiflexion: Implications for diagnosis of neural aspect to hamstring disorders. Musculoskelet Sci Pract. 2019 doi: 10.1016/j.msksp.2019.07.011. Breig A, Troup JD. Biomechanical considerations in the straight-leg-raising test. Cadaveric and clinical studies of the effects of medial hip rotation. Spine (Phila Pa 1976) 1979 doi: 10.1097/00007632-197905000-00011. Li Y, Fredrickson V, Resnick D. How should we grade lumbar disc herniation and nerve root compression? A systematic review. Clin Orthop Relat Res. 2015 doi: 10.1007/s11999-014-3674-y. van Rijn JC, Klemetsö N, Reitsma JB, Majoie Charles BLM, Hulsmans FJ, Peul WC, et al. Observer variation in MRI evaluation of patients suspected of lumbar disk herniation. AJR Am J Roentgenol. 2005 doi: 10.2214/ajr.184.1.01840299. Fardon DF, Williams AL, Dohring EJ, Murtagh FR, Gabriel Rothman SL, Sze GK. Lumbar disc nomenclature: version 2.0: Recommendations of the combined task forces of the North American Spine Society, the American Society of Spine Radiology and the American Society of Neuroradiology. Spine J. 2014 doi: 10.1016/j.spinee.2014.04.022. Pesonen J, Rade M, Könönen M, Marttila J, Shacklock M, Vanninen R, et al. Normalization of spinal cord displacement with the straight leg raise and resolution of sciatica in patients with lumbar intervertebral disc herniation: a 1.5-year follow-up study. Spine. 2019 doi: 10.1097/BRS.0000000000003047. Sharma S, Traeger AC, Reed B, Hamilton M, O’Connor DA, Hoffmann TC, et al. Clinician and patient beliefs about diagnostic imaging for low back pain: a systematic qualitative evidence synthesis. BMJ Open. 2020 doi: 10.1136/bmjopen-2020-037820. Carragee E, Alamin T, Cheng I, Franklin T, van den Haak E, Hurwitz E. Are first-time episodes of serious LBP associated with new MRI findings? Spine J. 2006 doi: 10.1016/j.spinee.2006.03.005. 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