N = 55), particularly for more ecological images, such as photos (N = 14). Among the several dimensions in which the items were assessed, the most commonly referred in our sample were familiarity, visual complexity and name agreement, illustrating some consistency across the reported dimensions while also indicating the limited examination of other potentially relevant dimensions for image processing. The lack of normative studies simultaneously examining affective, perceptive and semantic dimensions was also documented. The number of such normative studies has been increasing in the last years and published in relevant peer-reviewed journals. Moreover, their datasets and norms have been complying with current open science practices. Nevertheless, they are still scarcely cited and replicated in different linguistic and cultural contexts. The current study brings important theoretical contributions by characterizing images of common objects stimuli and their culturally-based norms while highlighting several important features that are likely to be relevant for future stimuli selection and evaluative procedures. The systematic scrutiny of these normative studies is likely to stimulate the production of new, robust and contextually-relevant normative datasets and to provide tools for enhancing the quality of future research and intervention.Common objects comprise living and non-living things people interact with in their daily-lives. Images depicting common objects are extensively used in different fields of research and intervention, such as linguistics, psychology, and education. Nevertheless, their adequate use requires the consideration of several factors , and careful validation procedures. The current study presents a systematic review of the available published norms for images of common objects. A systematic search using PRISMA guidelines indicated that despite their extensive use, the production of norms for such stimuli with adult populations is quite limited ( Objects constitute a distinctive type of stimuli that entail specific visual processing as compared, for example, to faces or words , contextual occurrence and also the emotions they elicit  as well as cross-country evaluations that consider the influence of language variations and cultural specificities. Moreover, the norms already produced are not always available, and even when they are, the stimuli selection must often be adapted to the researchers' goals and to the specific cultural contexts. However, the current demands of scientific practice and the pressure for publication often conflict with the time-consuming steps prior to experimental studies. As a consequence, the control for stimuli diversity and related confounding factors established in previous normative procedures are often misinterpreted as an obstacle instead of a step toward increasing quality in research.In sum, the process of producing and selecting stimuli constitutes an important but also complex and costly task. Therefore, a systematic review of the standardization studies of interest might be highly relevant in assisting this first and essential phase of planning the research in order to identify, access and select adequate stimuli and potentially relevant dimensions. Given its particularities and its widespread use, it is crucial to systematically examine the available normative studies using image datasets of common objects to uncover their specificities, their standardization practices as well as the potential gaps in those studies. Finally, the systematic information about which normative studies have been produced for common objects constitutes a valuable resource for electing adequate procedures and databases as well as acknowledge common objects as a relevant general category.The systematized knowledge from these normative studies, constitutes an important resource regarding the stimuli characteristics but also a valuable asset to identify well-established practices.https://protocols.io/view/a-systematic-review-of-normative-studies-using-ima-bbysipwe [doi: 10.17504/protocols.io.bbysipwe]) Specifically, the present systematic review aimed to:A systematic review of the literature was conducted using the PRISMA  Identify and characterize the published normative studies of images of common objects with adults, in order to assist the selection and further production of such stimuli and new databases (2) Determine and critically examine the most reported evaluative dimensions and their parameters in common objects normative studies, to uncover the most relevant properties to examine in normalizing images of common objects (3) Critically appraise the reliability of the norms produced, without losing their cultural specificity, by inspecting the coherence of ratings and their correlations reported across normative studies of images of common objects [How are images of common objects rated across studies? How are the main evaluative dimensions correlated across studies?](4) Uncover the application potential of each elected normative study by an exploration of the availability of the databases  and their impact  as potentially relevant indicators for selecting, producing or replicating normative studies [How accessible and widespread these databases are (availability and impact)?].https://protocols.io/view/a-systematic-review-of-normative-studies-using-ima-bbysipwe [10.17504/protocols.io.bbysipwe]) The PRISMA checklist is included as The systematic review conducted followed the PRISMA guidelines  were based on dimensions commonly reported in the literature. The (d) search entries were included to filter an extensive list of articles that refer to the words “image,” “picture,” and “norms.” The search was conducted without year restrictions or entry boundaries . An additional search was conducted on Scopus and on Web of Science databases with the same Boolean criteria and without year parameter, defining as search criteria: type of document “article” and “English” language. On Scopus, the search was conducted in title, abstract, and keywords. On WoS, the search was limited to the title. The analysis of overlapping articles and the management of the selected articles was made using EndNote X8 software. A complementary hand search phase was also conducted based on known authors/papers including pertinent normative studies using images of common objects not captured by the automatic search. The search procedures and the collection of the articles were completed by June, 2019.The search strategy included a first stage of systematic electronic search in online sources to identify the relevant normative studies published in English in academic peer-reviewed journals. Four databases were explored in the EBSCOhost platform to find potentially relevant studies: Academic Search Complete (1976–2019), PsycINFO (1948–2019), Psychology and Behavioral Science (1950–2019), and PsycARTICLES (1948–2019). The search terms entered in a Boolean phrase search mode using all possible combinations were the following: (a) validation OR norms; AND (b) pictures OR images; AND (c) typicality OR familiarity OR name-agreement OR valence OR arousal OR aesthetic OR “visual complexity” OR categories; (d) NOT social OR body parts OR face OR emotionThe inclusion criteria for electing potential studies involved three cumulative conditions: (1) the inclusion of healthy adult participants (minimum of 18 years-old); (2) the standardization of images of common objects into categories of the living and/or non-living domains ; (3) at least one the following dimensions as independent variable: semantic dimensions , affective dimensions  and perceptive dimensions .The second stage of the search (see PRISMA guidelines) involved the screening of the data by title, abstract, and full text, by three independent judges using the Rayyan QCRI  web application. The selection by title and abstract intended to control for subjective bias in the selection of the articles as well as to efficiently filter the relevant articles, confirming the inclusion criteria. With this procedure, articles from the previously comprehensive search that did not include image validation studies  or studies that were not pertinent to the current review  were excluded. Subsequently, the full-text examination ensured the eligibility of the selected studies to be retained based on all inclusion criteria. Disagreements on retaining or excluding on each screening phase were discussed until a consensus was reached.To our knowledge there is no specific standardized tool available for quality assessment in normative studies. In the present research the Cochrane Collaboration Risk of Bias tool . The extracted information included: (a) bibliometric information and indicators ; (b) general characteristics and standardization practices of images of common objects ; (c) dimensions reported in the standardization of images of common objects ; (d) assessment of images of common objects ; (e) accessibility and application potential of the normative databases (availability and citation impact).t-test, ANOVA). In (c), a qualitative appraisal of item norms was provided by examining the dimensions reported . Main findings for the most reported dimensions were also included in (d), namely assessment of images  of all studies reporting overall results for images of common objects. Correlational results for each dimension between studies were also considered for the main dimensions. Finally, information about the impact  and availability  of the database was also collected (e). Such indicators are predictors of the scientific impact of the articles in their respective areas  and also reflect their potential for replication. The overall findings were summarized in qualitative  and quantitative  tables and figures.For the general characteristics of the studies, sample characteristics comprised N of participants, mean age, age range, schooling, schooling range; language and cultural variations included language/cultural context and cross-cultural comparisons; procedures entailed data collection procedure; and stimulus characteristics discriminated if it was S&V stimuli replication/adaptation/extension, the stimuli description, stimuli type, image resolution, number of stimuli, number of stimuli/participant, categories of the items—number and types. Moreover, the cross-cultural comparisons described the presence of comparisons, the type of comparison (if between or within studies), the sample source (if they compared the same database or not) and the statistical methods used for cross-cultural analysis , 69 from Scopus and 21 from WoS. Four additional relevant studies were inserted on the data  presenting norms for images of common objects grouped in two distinct types of visual representations: line-drawings , photographs , or both . The analysis of the publications examining the different stimuli type  across the years  revealed an earlier trend for publishing datasets using drawings  and an increased interest in ecological stimuli or its comparison with drawings in recent years . The following subsections present the systematization of the information available in the studies sample considering the previously reported categories of data treatment. For each of these categories, the results of normative studies using photographs are emphasized because of their relevance in introducing ecological validity and the documented increased interest in this type of stimuli. A descriptive summary of the results is presented in The qualitative appraisal of the retained studies focused on the identification and categorization of their characteristics that were relevant to our aims. Overall, the final sample included a reasonable number of papers (Behavior Research Methods (n = 27), which is not surprising given the scope of this publication. However, several recent normative studies with photos have been published in open access journals, such as PLoS ONE, Frontiers in Psychology and Frontiers in Human Neuroscience. To assess the impact of the journals in which the reviewed articles were published, the h-indexes were obtained using the SCImago platform (www.scimagojr.com) , but mostly in the last 10 years  reviewed used samples of University students , with fewer studies recruiting participants outside the academic environment . This review also indicated that most of the studies included young adults (with ages between 18 and 35 years old) and only 14 studies (24%) included broader age samples . Studies using photographs were mostly conducted with undergraduate student samples, with a narrow age range  were the most recruited samples. Other languages referred across the study sample were: Indian, Greek, Persian, Icelandic, and Thai. Only, five studies (9.1%) examined more than one language/culture and three did not specify the native language of the sample. The 14 studies using photographs of common objects were mostly conducted in English, Spanish, and Portuguese , and the remaining in other language communities  for validity and reliability purposes, studies with a specific purpose of cross-cultural comparisons  were rare. In addition to the scarce examination of cross-linguistic/cultural reported in the entire sample of studies  to avoid fatigue and bias in the ratings across dimensions , even when response times were not a variable of interest  or photographs  or both . From the line-drawing's studies sample, the majority  included the Snodgrass and Vanderwart . The number of categories varied across studies ranging from one broad concept  across dimensions was not consistently examined, particularly in those using real-world photographs , perceptual , and affective  domains  or in some cases presented inconsistencies between the instruction focus  and scale  , in both articles norming photographs and line-drawings. Only a few exceptions did not examine this dimension  and Image Agreement  were the most reported dimensions. Moreover, Imageability  and Picture-name agreement  were also examined. Additionally, a few studies  addressed Manipulability, particularly for objects and tools  and Valence  being examined but only in real-word photographs of common objects  and perceptive dimensions , but affective variables were rarely examined. Additionally, none of the reviewed studies simultaneously explored the relations between dimensions of these three domains. The most frequent combinations were the perceptive and semantic domains. The semantic and affective or perceptive and affective combinations were also found, albeit scarce. Interestingly, the simultaneous examination of affective and other domains was only present in photographs of common objects norms. The results of the association between dimensions in the reviewed studies and the comparisons between stimuli type (line-drawings vs. photographs) are presented in Overall, the correlations scores indicate consistency in the direction of the correlations across studies, with a few exceptions . Correlations between semantic dimensions were overrepresented. Name agreement measures were negatively correlated for both type of items. The correlation between name agreement (%) and familiarity was positive and from moderate to strong, independently of the stimuli type. Moreover, visual complexity was negatively correlated with familiarity . Overall, the most reliable dimensions across studies comparisons were age-of-acquisition and image agreement. In such dimensions, the correlations found (between moderate to strong) represented a comparison between very distinct cultural and linguistic contexts . The high variability on Naming agreement scores indicates their sensibility to changes in cultural/linguistic variations  and present strong correlations in similar linguistic backgrounds . The results extracted from the articles reporting correlations between the original (reference) and previous studies using the same database are presented in In order to evaluate the potential use of the databases, we collected information about their availability and their application.n = 43, 78%) and only eight (14%) did not refer how to access the database or presented an unavailable link  and presented free access to the database . The most cited articles from the overall sample refer to normalization of line-drawing images , Scopus , and Google Scholar  is relatively high. However, a closer inspection to these numbers revealed that there are very few articles with more than 100 citations  bibliometric information and indicators ; (b) general characteristics and standardization practices of images of common objects ; (c) dimensions reported in the standardization of images of common objects ; (d) assessment of images of common objects ; (e) accessibility and application potential of the normative databases (availability and citation impact).Overall, the results indicated 55 published normative studies using images of common objects. The bibliometric indicators examined revealed that normative studies of images of common objects have been increasing in the last 10 years and published in quality peer-review journals. These indicators document the recent efforts that have been made in the field to provide stimuli and to produce valid norms that support adequate manipulations and enhance quality and replicability in experimental research , and the sample size constitutes an important criterion for statistical purposes Cohen, . AlthougAnother feature of the samples in the reviewed studies is their limited age distribution and consequent scarcity of aging and development effect analysis. In conducting norms for adult populations, it is relevant to fully consider their developmental process once with increasing age some abilities  are known to decrease  are known to influence the processing of meaningful stimuli, such as common objects  and others may present conceptual similarities with well-reported dimensions .The inclusion of innovative dimensions, such as measures of color, manipulability, and ambiguity in image normative studies has been increasing in recent years inspired by developments in related fields. Therefore, it is important to contextualize the emergence of such dimensions in the broader scientific context. Particularly, visual cognition and picture processing research fields have been examining surface features and cognitive processing involved in object perception. For instance, the influence of surface features, such as color, amount of details or size, in the way visual items are perceived and retrieved  regarding their modal name and category, which vary across studies. Some of the correlations between dimensions were strong, suggesting the need for examining some potential confounds, namely between typicality and other semantic dimensions . Moreover, unexplored dimensions, such as aesthetic appeal and arousal/valence, should be examined for their impact on the processing of images of common objects. The correlation scores provided indicators of consistency in the direction of the correlations across studies, with a few exceptions . These exceptions seem to reflect the selective influence of context on meaningfulness-based dimensions in each stimuli type. Of special interest and despite their consistency in direction, it was observed that the range of the strength of the reported correlations presents some variation across studies, which is expected once the geographical context and language vary across studies. The results suggest that norms are sensible to both cultural and linguistic variations and their use should be restricted to the populations in which they were produced and also that they might depend on methodological differences . However, the present study only examined the ratings across studies in a qualitative manner which limits the possibility of drawing substantial conclusions and generalizations on this issue.The availability and application potential of the normative databases showed that most studies, their materials and data, have been made increasingly accessible in the last years, which favors reproducibility and application potential in the upcoming years. However, this accessibility could be further boosted, namely by increasing open access practices that facilitate the access to the stimuli, allowing replication studies across different cultural contexts and languages. Open practices are likely to stimulate the enhancement/extension of visual normative databases and the examination of new dimensions. The past use of these norms indicates a tendency to use well-established databases  as well as help preventing the impact of undesirable confounding variables. Finally, normalizing materials with the purpose of safeguarding the quality of procedures—being they experimental or interventional—is an important research field on its own. Therefore, the current review emphasizes the normalization of visual stimulus as more than a procedure related to the researchers' everyday practices, but as a wide and rich research topic that should be acknowledged as such.CS, MVG, and JCC contributed equally to design the procedures, to select the articles, and to draft the manuscript. CS executed the comprehensive search identification and selected all the relevant information for this review. MVG and JCC supervised and coordinated the whole process of producing this work and provided theoretical comments on the subject based on their expertise. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Stage III disease affects approximately one-third of patients diagnosed with NSCLC each year and prognosis is poor, with a low cure rate. The optimal treatment regimen for patients with NSCLC is most frequently multi-modal, with systemic and local therapies for distant and local disease control, respectively. The exact sequence and modality used is debated and case-specific. The first and most important question in the management of stage III NSCLC, whether the disease is potentially resectable or not, can often be difficult to answer. If the disease is resectable, surgery after preoperative treatment (chemotherapy or chemoradiotherapy (CRT)) is an option, particularly in patients with optimal pulmonary function, mediastinal tumour clearance following upfront treatment and no requirements for pneumonectomy. However, for the remaining patients, or those who have been diagnosed with unresectable stage III NSCLC, platinum-based concurrent chemoradiotherapy (cCRT) is the standard of care versus sequential CRT.2 The supplement concludes with a review of the practical implications, such as prehabilitation and rehabilitation, that could mitigate the potential adverse events of radical therapy.This supplement reviews the clinical evidence for the current European and UK treatment strategies for stage III NSCLC, whilst also providing an overview of the PACIFIC clinical trial (NCT02125461) as the first immunotherapy for unresectable stage III NSCLC whose tumours express PD-L1 on ≥1% of tumour cells following completion of cCRT. This is the first significant treatment advance for these patients in many years.3 and cCRT is not frequently applied. The shared decision-making expertise of the treating multidisciplinary team (MDT) is critical in determining the most appropriate treatment regimen for individual cases, which is cCRT for those with unresectable stage III NSCLC. The scientific rationale for this treatment is based on the results of two large Phase 3 randomised studies and two meta-analyses (predominantly in patients with stage III NSCLC), which demonstrated the superiority of cCRT over sequential chemotherapy and radiotherapy treatment.6 The second article elaborates on how to determine optimal treatable areas for radiotherapy, outlined in the Advisory Committee on Radiation Oncology Practice (ACROP) guidelines,7 as well as optimising dose fractionation and chemotherapy regimens to balance the benefits of treatment versus toxicity. In addition, the article discusses the rationale for the benefits that immunotherapies, such as durvalumab , could provide following cCRT.The first of these articles reviews the key surgical, chemotherapy and radiotherapy trials and guidelines that currently outline radical therapy options for patients with stage III NSCLC. This paper highlights that only one in five of these patients has radical intent multi-modality therapy in the United Kingdom,®; Bristol-Myers Squibb Company, USA) and pembrolizumab  for the treatment of stage IV metastatic disease. The role of immunotherapy in advanced NSCLC and the rationale for its use following cCRT, in light of the pivotal Phase 3 PACIFIC clinical trial of durvalumab in patients with stage III, locally advanced, unresectable NSCLC whose disease had not advanced following cCRT, is described in this supplement. The PACIFIC study reported significant improvements in the co-primary endpoints of progression-free survival (PFS) and overall survival (OS) following treatment with durvalumab versus placebo in the overall population, as well as in most pre-specified subgroups.9 Furthermore, the recently reported 3-year overall survival analysis of the PACIFIC study demonstrates long-term clinical benefit with durvalumab following cCRT, further establishing the PACIFIC regimen as the standard of care in this population.10 The authors of this supplement conclude that the PACIFIC study, along with ongoing studies of other treatment modalities, are providing significant progress in the treatment of patients with stage III unresectable NSCLC, offering a positive outlook through the advancement of standards of care.In recent years, immunotherapy has gained wide popularity in NSCLC in light of the positive findings from trials of nivolumab .On the basis of the data from the PACIFIC clinical trial,However, it is unclear as to whether the set-up of the current standard-of-care procedures and management steps for stage III NSCLC are in place to enable the translation of these results to real-world practice. Increased adoption of cCRT across the United Kingdom will positively impact the proportion of eligible patients with unresectable stage III NSCLC who are able to gain benefit from durvalumab treatment. Patient selection prior to treatment with cCRT will be critical; however, there is currently a lack of comprehensive guidelines for the assessment of patient fitness prior to cCRT. Although often not formally evaluated within the context of clinical trials, there are a number of simple supportive measures (i.e. aggressive symptomatic interventions) that may improve outcomes to cCRT treatment, which is addressed in the final article of this supplement.In summary, within the heterogeneous treatment landscape for patients with locally advanced, unresectable stage III NSCLC, durvalumab, as a consolidation therapy following two or more cycles of platinum-based cCRT, is emerging as a viable option to treat patients whose tumours express PD-L1 on ≥1% of tumour cells in this setting with curative intent.


Aspergillus-produced mycotoxins can enter the feed and food chain at many points in both pre-harvest and post-harvest. Although current climate changes seem to speed up the world-wide spread of mycotoxigenic fungi including the Aspergilli and also facilitate the production of these harmful secondary metabolites the factors governing these disadvantageous global processes are only partly understood or even have remained completely hidden until now. This Research Topic summarizes our knowledge on Aspergillus-derived mycotoxins especially focusing on three major areas of on-going research: (i) toxicological, medical, veterinary aspects, prevalence, detection, risk assessment, control strategies, (ii) ecology and biological control of mycotoxigenic Aspergilli in the fields, and (iii) pre-harvest and post-harvest management of mycotoxigenic Aspergilli and their mycotoxin production. We hope that the wealth of information generously provided by the Aspergillus mycotoxin research community will help the hard work of all those experts, who are active in this important field, and the papers collected here will be instructive and illuminating readings for students and the public as well.Aspergillus-derived mycotoxins  represent a remarkably versatile group of fungal secondary metabolites considering both their chemical structures and adverse physiological effects in humans . Although current food safety measures are often adequate to prevent the accumulation of these mycotoxins in the food chain further interdisciplinary research is eagerly needed to elaborate more effective prevention strategies of mycotoxicoses, to reach a deeper understanding of the deleterious consequences of both sole and combined mycotoxin exposures at various stages of life, and to invent novel diagnostic tools and therapeutic procedures to mitigate both acute and chronic mycotoxin poisonings . To prevent primary and secondary  aflatoxicoses in humans is of paramount importance . Fortunately, the accurate physiological effects, the existent transformation and detoxification pathways and the mechanisms of channeling of harmful aflatoxins into food raw materials have been elucidated in important livestock like poultry, pigs and ruminants, and a wide spectrum of biocontrol and detoxification products are available to prevent harmful aflatoxins from entering the feed and food chain in animal husbandry .Aspergillus section Flavi and the occurrence of aflatoxins in raw peanuts and peanut-based products  and in food and feed . Since even a low aflatoxin concentration is hazardous for human and livestock, the identification and quantification of AFs is a major challenge to guarantee food safety. The demand for determination of aflatoxins triggered extensive research and method development, and, in the last decades, increasingly faster and more sensitive analytical techniques proved to be promising, but, only a few of them have gained applicability in routine analysis. The study of Miklós et al. provides guidance on the current performance characteristics of various analytical methods for determination of aflatoxins in different food and feed matrices, and highlights their limitations for practical use, i.e., the absence of processes applied for reduction of large laboratory samples to the few grams for extraction  or the fact that the repeatability or reproducibility, if reported, was based on a few spiked samples . This guide helps in the decision to choose the most appropriate method that meets the practical requirements of fast and sensitive control of their contamination. Special references are devoted to new methods developed for masked AFs that are unable to be identified by routine analysis processes  and for concomitant detection of aflatoxins and their major fungi precursors Aspergillus flavus and Aspergillus parasiticus in stored maize by a simultaneous run of a Display Mediated Immuno-polymerase Chain Reaction (PD-IPCR) for aflatoxins and a conventional real-time PCR (RT-PCR) for aflatoxin producers .Food and feed contamination by aflatoxins create food insecurity around the world. Two overviews detail the prevalence of Aspergillus-derived mycotoxins are still present in the feed and food chain, as well as the mycotoxicoses . Quantitative exposure assessment is a methodology developed to evaluate the probable intake of chemical substances via food. The study of Serraino et al. calculated the Estimated Daily Intake, the Hazard Index, and the fraction of hepatocarcinoma cases (HCC) due to AFM1 exposure in different population groups in Italy. A low risk of HCC was predicted but the variability of climatic conditions throughout years justifies a continuous monitoring of aflatoxins and an update of the risk assessment. To implement appropriate control measures, a special focus is devoted to the aflatoxin management and the impact of climate change on AFs production, and of control strategies of AFs in terms of innovative processing technologies applied for pre-and post-harvest aflatoxins management in combination with either biological, physical, chemical or genetic engineering methods .Despite of prevention methods and strict regulations, Aspergilli can enter and fill in is still waiting to be described and deciphered in details . Ecological factors influencing the production and fate of fungal secondary metabolites including mycotoxins like sterigmatocystin/aflatoxins, gliotoxin, ochratoxins, patulin, and cyclopiazonic acid as well as the versatile interactions of these molds with plants , other microorganisms  and animals  need to be clarified to reach a deeper understanding of these ecosystems and also to develop novel biocontrol and mycotoxin biodegradation technologies for plant and food protection . In the last decades, various and powerful omics techniques helped us to shed light on the fine details of the molecular mechanisms of host-pathogen interactions especially in A. flavus-maize and A. flavus-groundnut relations . Not surprisingly, a plethora of proteins and genes have been identified with definite or hypothesized functions in the resistance of these agricultural crops to aflatoxin contaminations, which has opened the way to the development of novel molecular breeding technologies in this important field .The remarkably complex and dynamic network of soil microbiota and macrobiota determining the ecological niches the mycotoxigenic A. flavus strains to prevent aflatoxin contamination under field conditions is one of the leading pre-harvest strategies to control this harmful, carcinogenic mycotoxin in the feed and food chain. Interactions of these biocontrol agents with the indigenous soil populations of aflatoxigenic fungi has been the subject of extensive research for a long time. Interestingly, the Afla-Guard strain originally isolated from naturally infected peanut in Georgia and belonging to lineage IB performed better in maize fields in the south-eastern United Stated  than the AF36 strain in lineage IC and isolated from cottonseed in Arizona as indicated by shifts in genetic diversities . In Ghana, 12 atoxigenic African A. flavus VCGs (AAVs) were identified and the biocontrol potential of a representative member of each AAV was tested under both laboratory (maize) and field  conditions . As a result, four-four well-preforming isolates were selected and incorporated into two biocontrol products, Aflasafe GH01 and Aflasafe GH02, for use in maize and groundnut cropping systems in Ghana . Importantly, each isolate has a unique simple sequence repeat (SSR) signature based on 17 SSR loci, which makes the tracking of each active ingredient possible under field conditions . Fast and reliable tracking and quantification of active ingredients of biocontrol products in crops are also required by farmers, the regulatory community and crop end-users . An array of quantitative pyrosequencing-based assays was developed and successfully applied in maize-associated fungal populations to monitor frequencies of SNPs characteristic of non-aflatoxigenic A. flavus isolates included in the Aflasafe product in Nigeria .Application of non-aflatoxigenic A. flavus is a prerequisite for creating an Integrated Pest Management (IPM) in order to protect plants and related products prone to aflatoxins contamination. Commercial biopesticides could offer an economically effective solution that may contribute to the exclusion of aflatoxigenic fungi from maize plants and minimize the mycotoxin production. The efficiency evaluation of these biopesticides in vitro assay is crucial for a future IPM system friendly and sustainable for the environment .The use of biological agents and biostimulants for the control of Peles et al.; Pfliegler et al.). For example, abiotic oxidative stress stimulates aflatoxin biosynthesis in A. flavus , and 3.5% ethanol exposure of A. flavus colonies not only significantly down-regulated the great majority of the aflatoxin cluster genes but also up-regulated important elements of the oxidative stress defense system including the Cat, Cat1, Cat2, CatA, and sod1 genes as well as the oxidative stress response regulator genes ap-1 and msnA . These transcriptional changes coincided with a nearly complete inhibition of aflatoxin B1 production .The remarkable complexity of aflatoxin biosynthesis has been revealed in aflatoxigenic fungi and both biotic and abiotic factors contribute to the fine-tuning of mycotoxin production in these microorganisms . Wang et al. reported on the pivotal role the velvet complex  of Aspergillus ochraceus plays in the maintenance of vegetative growth, asexual sporulation, virulence (on pears), and ochratoxin A production of the fungus and, therefore, elements of the velvet complex seem to be attractive targets for future ochratoxin A control technologies. It is worth noting that LaeA extensively regulates the secondary metabolism of A. ochraceus . Another study by Barda et al. shed light on the remarkable importance of the pH-responsive transcription factor AcPacC on the regulation of fungal growth at neutral/alkaline pH, asexual sporulation, spore germination, gluconic and citric acid productions, ochratoxin A production  and virulence of Aspergillus carbonarius. Importantly, glucose oxidase encoded by Acgox was demonstrated to be a virulence factor of A. carbonarius and its production was also under a strict AcPacC control .The ceaseless demand for the development of novel mycotoxin control technologies also requires further research to be performed on the regulation of mycotoxin biosynthetic gene clusters . In addition to controlling pre-harvest and post-harvest growths of the Aspergilli, the direct inhibitory effects of plant bioactive compounds on aflatoxin biosynthetic pathway seem to be exploitable as well . Various plant extracts containing low molecular weight active ingredients and enzymes have also been shown to be effective in aflatoxin degradation . Cinnamaldehyde, a widely used α,ß-unsaturated aldehyde food additive, appears to be especially promising in control of aflatoxigenic A. flavus e.g., on corn . Considering the antifungal mechanism of action of this compound, cinnamaldehyde triggers a series of apoptotic events in A. flavus, including elevated intracellular Ca2+ and reactive oxygen species levels, various mitochondrial dysfunctions, metacaspase activation, phosphatidylserine externalization, DNA damage and nuclear fragmentation as well as up-regulation of apoptosis-related genes . A methanolic pomegranate peel extract (PPE) acted synergistically with the azole fungicide prochloraz (PRZ) in controlling the growths of the mycotoxigenic fungi A. flavus and Fusarium proliferatum . PPE+PRZ combined treatments delayed conidial germination and hyphal elongation in both fungi, and the combined application of sub-inhibitory doses of PPE and PRZ blocked aflatoxin B1 production by A. flavus . Such combined antifungal treatments may help us to decrease the applied doses of potentially harmful synthetic fungicides like the azole drugs in the agriculture .Bioactive metabolites of plants like phenolic compounds, terpenes and nitrogen-containing compounds may also possess antifungal activities Haidukowski et al.). The characterization of the bio-sorbent properties of the mycelium of the king oyster mushroom (Pleurotus eriyngii) as for Aflatoxin B1 binding capability and the effects of physical and chemical conditions on the binding efficiency is strategic for a sustainable mycotoxin risk minimization in animal health .Adsorbent materials mixed with the feed may protect animals by binding efficiently mycotoxins including aflatoxins. The adsorbents reduce the bioavailability of mycotoxins in the gastro-intestinal tract and thus the diffusion into the bloodstream and transport to the target organs. The use of microbial biomasses as adsorbent seems very promising since less expensive though effective, and environmentally friendly materials (All authors served as co-editors to the Research Topic and also contributed to, critically read and discussed and approved this Editorial.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The Wamide neuropeptide superfamily is of interest due to its distinctive functions in regulating life cycle transitions, metamorphic hormone signaling, and several aspects of digestive system function, from gut muscle contraction to satiety and fat storage. Due to variation among researchers in naming conventions, a global view of Wamide signaling in animals in terms of conservation or diversification of function is currently lacking. Here, I summarize the phylogenetic distribution of Wamide neuropeptides based on current data and describe recent findings in the areas of Wamide receptors and biological functions. Common trends that emerge across Cnidarians and protostomes are the presence of multiple Wamide receptors within a single organism, and the fact that Wamide signaling likely functions across an extensive variety of biological systems, including visual, circadian, and reproductive systems. Important areas of focus for future research are the further identification of Wamide-receptor pairs, confirmation of the phylogenetic distribution of Wamides through largescale sequencing and mass spectrometry, and assignment of different functions to specific subsets of Wamide-expressing neurons. More extensive study of Wamide signaling throughout larval development in a greater number of phyla is also important in order to understand the role of Wamides in hormonal regulation. Defining the evolution and function of neuropeptide signaling in animal nervous systems will benefit from an increased understanding of Wamide function and signaling mechanisms in a wider variety of organisms, beyond the traditional model systems. Neuropeptides are short peptidergic molecules released by animal neurons that act as modulators or hormones to regulate biological processes. These signaling molecules are notable for being present in the nervous system of early metazoans, and for their important functions in regulating animal behavior and physiology. Historically, neuropeptides have been named according to their function, or where function is unknown, according to repetitive conserved sequence motifs found in the precursor peptide. These naming strategies have the unfortunate consequence of often obscuring neuropeptide relationships across species or phyla. The Wamide neuropeptide superfamily is a striking example of this. Wamides are repetitive proneuropeptides that contain multiple cleavage sites flanking short, amidated active peptides with a conserved C-terminal tryptophan (W). This neuropeptide superfamily is of ancient origin, already present in the last common ancestor of cnidarians and protostomes . DependiAchatina fulica , due to their inhibition of ecdysone synthesis in this species (The first known Wamide discovered was locust myoinhibitory peptide (LOM-MIP). LOM-MIP was identified as a suppressor of visceral muscle contraction in hindgut and oviduct in 1991 . Subsequa fulica  and the ntissima . Initial species , althoug species . The fir species .Lottia gigantea reinforced the link between molluscan WWamides and insect ASTB  and L-Phe (F)  , 26. The-Phe (F) , 28. A pTrichoplax adhaerens, an RWamide precursor was found through bioinformatic prediction . This was initially called the sex peptide receptor (SPR) after its first characterized ligand, the  peptide . Whereas mollusc . Followi mollusc , 35, cau mollusc . The con mollusc .Orthologs of the MIP receptor have been biochemically characterized in insects [fruitfly, mosquito , tick 3, kissingPlatynereis dumerilii  responsible for different aspects of Wamide function. It is therefore important for future studies aiming to uncover mechanisms of Wamide signaling to develop methods for manipulating specific subsets of Wamide-expressing neurons. One approach to this is through the development of libraries of reporter constructs driven by different promoters with a range of cell-specificities. Further understanding of Wamide signaling can also be achieved by analysis of genes co-expressed in Wamide- and Wamide receptor-expressing cells and more widespread analyses of Wamide receptor expression. These analyses can indicate mechanisms of signaling, such as autoregulation in cells expressing both Wamide and receptor, or association with specific neurotransmitters, such as GABA or acetylcholine. They can also be used to generate maps of potential signaling cascades, as in insect ecdysis (The most detailed studies of Wamide signaling to date have been carried out in model organisms  ecdysis , through ecdysis , which sAnother aspect of Wamide signaling of importance for future studies is the identification of signaling cascades activated following Wamide release and receptor binding, in terms of which class of G protein is recruited, if indeed the Wamide signal is GPCR-mediated, and which downstream signaling pathways are activated or repressed. Again, single cell RNA-Seq analyses or precise spatial expression mapping can be used to identify which elements of the signaling pathway are present in the target cells of Wamide signaling. With both distinctive and conserved functions, the Wamide superfamily is an excellent model for studying the evolution of neuropeptide signaling and patterns of peptide-receptor coevolution in animal nervous systems.EW conceived the study and wrote the paper.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Morphological diversity and species richness provide insights into biodiversity and have been studied extensively in recent years. Most researchers have found a positive correlation between these factors in many groups at the local community scale; however, this documented relationship has not always been consistent because of diverse niches and the status of an organism in a given ecosystem. Here we propose a new paradigm for the analysis of higher taxa biodiversity based on a cosmopolitan dataset to further investigate this contradiction. The morphological diversity of 1106 buprestid species from around the world was quantified based on the contours of the pronotum and elytron in dorsal view using a geometric morphometric approach. We found a positive correlation between morphological diversity and genus richness while no significance was found in the species-level test. Furthermore, the correlation between morphological diversity and genus richness is higher than it is in the species-level test. Our results demonstrate the superiority of higher taxa in biodiversity, and that the geometric morphometric approach could quite accurately reveal diversity patterns of the family Buprestidae. These conclusions complement the crucial aspect in several disciplines, including biodiversity, phylogeny and evolutionary strategy.A positive correlation between the species richness and morphological diversity of some organisms has been found in almost all studies at the local community scale. However, this documented relationship has not always been consistent because of diverse niches and the status of an organism in an ecosystem. Global taxon sampling, new morphological approaches, and consideration of more taxonomic categories other than species level are possible methods to further investigate this contradiction. In this study, we proposed a new paradigm for higher taxa biodiversity analysis based on a cosmopolitan dataset. A total of 1106 species from around the world representing all subfamilies and 33% genera of Buprestidae (jewel beetles) were selected to test the correlation between morphological diversity (MD) and genus/species richness (GR/SR) among subfamilies. The MD was quantified by the contours of the pronotum and elytron in dorsal view based on a geometric morphometric approach. The positive correlation between MD and GR was found in all test combinations, but was irrelevant in the species-level test. Interestingly, the correlation between MD and GR was higher than MD and SR in both pronotum and elytron measurements. Additionally, the MD of the pronotum is obviously higher than the MD of the elytron. Our results demonstrate that the geometric morphometric approach could quite accurately reveal diversity patterns of the family Buprestidae. Future studies on different groups, using more characters, more analyses and detailed biological interpretations, are required to fully understand the relationship between MD and SR. Biodiversity plays a host of important roles in biosphere operations, which is usually quantified via proxy indices , and can be used to reveal changes in ecosystems. The use of different biodiversity indices highlights the disparate nature of biodiversity research ,2. AmongThe beetle family Buprestidae (jewel beetles) was selected as a test group because it is one of the most successful beetle clades, with more than 14,000 described species, and has a fairly complex life history, thus improving their adaptability to the environment ,26. OwinThe morphological diversity was quantified by the shape in dorsal view of the selected species, represented by the outline of the pronotum and elytra, based on a geometric morphometric approach. Different from the previous studies which focused on the mandibles, antenna, genitalia, hind wing, etc., that strongly reflect the selective pressure of the environment  ,32,33,34The aim of this study was to explore whether the correlation between morphological diversity and genus/species richness in higher taxa was consistent with species-level studies of local communities. Test indices were explored based on a large dataset covering 1106 species of Buprestidae, including all 6 extant subfamilies and 173 genera, representing 8% of all known species. The aim of this paper was not only to reveal the expression of biodiversity according to all the subfamilies of Buprestidae around the world as a whole, but also to pave a new way for research on other groups and the relationship between MD and SR.This study analyzed 1106 species (1215 specimens) from all six extant subfamilies   coveringThe genus/species richness value was obtained by counting the number of genera and species in different test subfamilies . The objThe morphological diversity was quantified based on the shape of the pronotum and elytra, which represent two major parts of the body structure in dorsal view and have typically been used as an index for body size, and both of which contain evolutionary information . The heaThe studied images were collected from the collections of the IZAS , NMPC , MNHN , NHML , USNM , and sources in the literature ,48,49,50Two curves were extracted from the left contours of the pronotum and elytron to represent their external forms . Curve OThe first three principal components (PCs) accounted for 88.109% of observed shape variation for the pronotum and 91.348% of observed shape variation of the elytra . Along tAlong the positive direction of its first PC axis, the half contour of the test elytron became more prominent, the posterior margin protruded, and the anterior margin stretched horizontally. The shrinking smaller scutellum causes the anterior margin to become longer while the base of the lateral margin becomes protruding outward originally and the rear part becomes contracted inward. Along the positive direction of the second elytron PC axis, the overall morphology of the elytron became narrow and long. This pattern was contrary to the previous one: the posterior border of the elytron expanded laterally, while the anterior angle broadened outward. The expansion of the scutellum edge led to a more inward trend of the anterior margin of the elytron. Along the positive direction of the third elytron PC axis, the smaller scutellum flattened the front margin and, at the same time, the posterior border of the elytron contracted inward, leading the overall morphology of the elytron to become an inverted triangle.The Agrilinae exhibited both the highest pronotum shape diversity (0.0187) and elytron shape diversity (0.0062), with the highest genus richness in our study . SimilarIn addition, morphological diversity was also found to vary between the pronotum and elytron of the same taxon. The changing trends of characters and species richness values were not always highly associated: elytron diversity of the Julodinae was relatively high compared with Galbellinae despite showing the lowest test elytron diversity.p < 0.05 in pronotum-genus test and p is close to 0.05 in elytron-genus test, respectively), while in the species-level test of the pronotum and the elytron, the correlation test result was non-significant (p > 0.05). Secondly, the correlation between morphological diversity and species richness in the pronotum test was found to be weaker than it in the elytron test. The coefficient value was 0.714 in pronotum (genus-level) test and became 0.771 in elytron (genus-level) test. It was 0.371 in the pronotum (species-level) test and 0.543 in the elytron (species-level) test. Thirdly, the degree of correlation between parameters was found to be superior in the higher taxonomic category . In addition, four best-fit lines  used through ordinary least squares (OLS) regression analysis were processed to macroscopically visualize the correlation coefficient situation based on the regular aggregation degree of the test groups  [Macrozamia communis and the larvae feed within the frond-stalks of cycads [This study has demonstrated the positive correlation between MD and GR, based on the global sampling of the beetle family Buprestidae, for the first time. Our results confirmed the previous conclusion that the morphological diversity was strongly related to genus category . Based oelidini)  whereby f cycads . The funAgrilus with more than 3000 species [Aphanisticus, Sambus, Habroloma and Trachys. Many groups are only distributed in a single region. This may indicate that most genera of this subfamily have evolved relatively special morphologies to adapt to different plant communities in different biogeographic regions, occupying different niches. At the same time, the larvae of this subfamily exhibit three major feeding habits: xylophagous type, stem-miners and leaf-miners [Julodis algirica, Julodis angolensis, Neojulodis bequaerti). The wide distribution and different geographical environments have presumably led to the complex morphological structure of this group, which is reflected in the morphological diversity test of the pronotum. Galbellinae has only one genus with 84 species from around the world, which is mainly distributed in the Afrotropical region, resulting in the low morphological diversity of the elytron. The larvae of Buprestinae bore into weak wood or dead plant parts, and the adults feed mainly on flowers or leaves. Members of this subfamily are also distributed in the major seven geographic regions, but, with the exception of a few taxa , most genera are limited to one geographic region [Acmaeoderella and Acmaeodera, whilst others feed on leaves. Furthermore, significant morphological changes of body shape were also found in some taxa (such as Paratrachys) [Different morphologies are associated with diverse functional aspects of niches ,64 and b species , which if-miners ,65,67. Tf-miners ,69. Specf-miners . Moreovec region ,65, whictrachys) ,28,49. IBy using geometric morphometrics, we have here found that a large amount of deformation occurs on specific parts of the pronotum and elytra. In particular, the posterior angle of the pronotum and the scutellum edge attached to the anterior ledge of elytron, which showed the relevance of the morphological function of these above-mentioned traits in morph-space. Excluding the influence of size and orientation factors through Procrustes fit, we found that the test traits cover a large number of variable factors in morphology whilst the morphological diversity obtained based on these deformation factors was also closely related to genus richness. Therefore, these morphological measurements of shape have the potential to become an important part of morphological diversity indices. Nevertheless, for most statistical morphometric measurement-design strategies, the acquisition of two-dimensional linear distances from extremal points of specimens is still the main method ,71. BiosIt is true that linear distances play a certain role in reflecting the growth rate and evolution of organisms, whilst it is scalar magnitudes that can be used to represent size, but contain no shape information whatsoever . In addiSome scholars believe that higher taxonomic levels are questionable and contain certain subjective elements, and the inconsistent level boundaries make the taxa defined in this framework lack actual evolutionary significance ,80. HoweIn this study, we report a positive correlation between MD and GR based on similar sampling proportions in based on test jewel beetles. However, our results also highlighted the unconformity between species richness and the change trend of morphological diversity in certain groups , which suggests that the inconsistent correlation between MD and GR/SR could be caused by the influence of other categories, such as tribe rank.Due to the variable ecological niches influences by the latitudinal and elevational gradients throughout world ecosystems, the species richness of organisms decreases from the equator to the north and south poles ,91. At tIt is widely accepted that biodiversity research based on large-scale patterns will be influenced by environmental factors  ,100,101;In this study, we showed that there was a positive correlation between morphological diversity and genus richness by regarding the Buprestidae (jewel beetles) as a whole test group. We also found that the geometric morphological method could extract the deformations in the morph-space to improve the morphological diversity data, rather than just relying on the traditional morphometric measurements of features. Furthermore, we found that the pronotum morphological diversity was higher than it was in the elytron and correlations between the genus richness and morphological diversity were stronger than the species level tests.


The retina transmits light signals to the brain via a complex structure composed of photoreceptor cells, neurons including ganglion cells, glial cells such as astrocytes and Mueller cells, as well as retinal blood vessels that feed the retina. The retina performs such high-level physiological function and maintains homeostasis effectively through interactions among the cells that form the neurovascular units (NVUs). Furthermore, as a component of the blood‒retinal barrier (BRB), the vascular structure of the retina is functionally based on the NVUs, in which the nervous system and the vascular tissues collaborate in a mutually supportive relationship. Retinal neurons such as ganglion cells and amacrine cells are traditionally considered to be involved only in visual function, but multiple functionality of neurons attracted attention lately, and retinal neurons play an important role in the formation and function of retinal blood vessels. In other words, damage to neurons indirectly affects retinal blood vessels. Diabetic macular edema is the leading cause of vision loss in diabetic retinopathy, and this type of edema results in neurological and vascular disorders. In this article, the regulatory mechanism of retinal capillaries in diabetic macular edema is reviewed from the viewpoint of NVU. Diabetic macular edema is the leading cause of visual impairment in diabetic retinopathy, and current treatments are not adequately effective. Edema results from diverse causes, and it involves various factors such as physical and mechanical factors represented by the vitreomacular traction syndrome, as well as factors associated with inflammation such as vascular endothelial growth factor (VEGF). The macula and retina possess defense mechanisms against inflammation and immune reactions, and the blood‒retinal barrier (BRB) is a representative mechanism. The BRB is composed of an inner barrier and an outer barrier; they are responsible for selectively and actively transporting two-thirds and one-third, respectively, of substances across retinal capillaries to extracellular tissues. The inner BRB maintains homeostasis by preserving normal functions of the capillaries in the retina ((3). This article reviews the relationship between the BRB and the mechanisms that causes edema in the macula and retina, focusing on the NVUs.In Japan, more than 12,505 people are estimated to become adventitiously blind every year, and diabetic retinopathy, affecting one-third of the patients with diabetes, is the most common causative disease e retina , and its(4).Although the timing of retinal blood vessel formation differs between mice and humans, the formation and distribution of retinal blood vessels are similar. Retinal blood vessels mainly form three layers, the superficial vascular plexus, intermediate vascular plexus, and deep vascular plexus, and these three layers of vascular networks are inter-connected, feeding the intra-retinal tissues. Retinal vascular endothelium is lined by fenestrated and non-fenestrated endothelial cells. Retinal capillaries do not possess adventitia or media but consist of a single layer of vascular endothelial cells. Pericytes are embedded in the perivascular space, and the basement membrane around and between the vascular endothelial cells and pericytes forms the lumen of the blood vessel. The vascular permeability of BRB is regulated by junctions between endothelial cells and binding between endothelial cells and extracellular matrix. For the vascular endothelial cell-cell junction, tight junction is established via tight junction proteins including occludin, VE-cadherin, and claudin present in the intercellular space. In addition, the mechanism ensures that transcellular transport hardly occurs and that the substances inside blood do not pass readily into tissues. As a result, passage of most of the soluble components in blood and immune cells including leukocytes is blocked. In diabetic retinopathy, degeneration and loss of vascular endothelial cells and pericytes, as well as thickening of basement membrane, occur. As a result, the blood vessel wall becomes fragile, and the BRB breaks down, leading to edema. Macular edema is caused by an increase in vascular permeability, due to breakdown of the inner BRB and leakage of intravascular plasma components into the extracellular space. In diabetic macular edema, the inner nuclear layer and outer plexiform layer are more susceptible to edema In the retina, vascular endothelial cells and pericytes of the capillaries, together with neurons, astrocytes, Mueller cells, and glial cells, such as microglia adjoining the capillaries, form a unit, and crosstalk between these cells regulates various functions of the retina. In mouse retina, astrocytes and glial cells such as microglia form the NVUs in the superficial capillary plexus, while neurites of amacrine cells and glial cells such as microglia form the NVUs in the intermediate plexus , and neu(5), although more recent studies revealed that not only glial cells but neurons also play a role. Neurons such as ganglion cells and amacrine cells were originally thought to be involved only in visual function, but they are now known to be important cells constituting the NVUs. Therefore, neuronal damage may indirectly affect retinal blood vessels and glial cells, and researchers speculate that all the cells constituting the NVUs are involved in BRB breakdown in some ways. The capillaries in the intermediate and deep plexuses of the retina are the main capillaries that constitute the inner BRB. In the intermediate plexus of the retina, amacrine cells and microglia are the major cells that form the NVUs both structurally and functionally, as mentioned above (7). In other words, the three components, amacrine cells, microglia and capillaries of the intermediate plexus, form the NVUs as a part of the inner BRB , and abnormal OP has been confirmed also in diabetic model eyes. This OP originated from the amacrine cells present in the inner layers of the retina (9). Findings of various studies suggested that amacrine cells are impaired and capillaries in the inner layers of the retina are also injured in diabetic retinopathy. These findings include thinning of the inner nuclear layer where amacrine cells are present and the inner plexiform layer where neurites of amacrine cells accumulate in diabetic retinopathy (13); reduction of blood flow in the retinal capillaries from before the onset of diabetic retinopathy (14); decreased blood flow in retinal capillaries of the parafoveal region in macular edema (15); and OCT angiography demonstrating impairment of amacrine cells and damage of retinal capillaries in the deep plexus in diabetic retinopathy ((21).Among the above-mentioned eye diseases that cause damage to BRB and NVU, diabetic macular edema will be described as an example. In diabetic retinopathy, oscillatory potential (OP) originating from amacrine cells is impaired from an early stage inopathy  (16),  are primarily damaged by the above-mentioned conditions is possible, and then retinal vascular endothelial cells become secondarily damaged . However(22). Most of the cells that constitute the retina  produce VEGF, and amacrine cells that control the capillaries in the intermediate plexus of retina also produce VEGF (7). While edema is reduced by intravitreal injection of anti-VEGF antibody, edema is not totally resolved in many cases, suggesting the involvement of mechanisms other than VEGF. Indeed, high concentrations of other cytokines including IL-8, MCP-1, IP-10, and Mig were detected in the vitreous humor of diabetic retinopathy (24) and IL-8 and Mig in ischemic retinopathy (25). These humoral factors may induce inflammation, promote chemotaxis and infiltration of inflammatory cells, and disrupt the BRB (26). Activated inflammatory cells express cell adhesion factors and secrete matrix metallopeptidases (MMPs) that possess the function of degrading extracellular matrix proteins, thereby destroying the basement membrane and vascular endothelial cells that constitute the BRB. In particular, MMP-9 is speculated to degrade not only the tight junction proteins occludin and claudins but also type IV collagen that constitutes the basement membrane (27). In addition, apart from MMPs, inflammatory cytokines such as TNF-α, IFN-γ, and IL-1 can also disrupt the BRB (28) When the BRB is disrupted, infiltration of inflammatory cells from the damaged site is facilitated. In addition, microglia adjoining the endothelial cells also respond to external stimuli and promptly elicit inflammatory responses (29). However, in intraocular inflammatory diseases such as uveitis, although macular edema may occur as the main inflammation, not all types of uveoretinitis cause macular edema. The prevalence of macular edema is the highest in ocular sarcoidosis and Behcet disease but is low in Vogt-Koyanagi-Harada disease (data not shown). These findings suggest that macular edema due to BRB breakdown is not simply due to inflammatory cytokines. Anti-VEGF agents show certain efficacy for various types of macular edema, including diabetic macular edema and retinal vein occlusion, indicating that VEGF is involved in the pathology of macular edema. VEGF not only promotes division of vascular endothelial cells but also enhances vascular permeability. The mechanism of action has been reported to be activation of the protein kinase C pathway through phosphorylation of occludin that is localized at the tight junction (30), and that in diabetic macular edema, anti-VEGF agents are less effective in patients with lower density of capillaries in the intermediate or deep plexus of the retina. These phenomena can be explained rationally by adopting the concept of the NVU. Furthermore, VEGF demonstrates a neuroprotective effect. From the viewpoint of neurovascular protection, studies showed that continuous administration of anti-VEGF antibodies for age-related macular degeneration causes atrophy of the retina In this review, we highlighted the current understanding of how the neurons and vascular systems interact to ensure proper diabetic macular edema. With the advent of anti-VEGF agents, a paradigm shift occurred in the treatment of diabetic macular edema and macular edema due to retinal vein occlusion, but the treatment effect for macular edema remains unsatisfactory. One of the reasons is that focus is currently placed on treatments that target the retinal capillaries, despite the fact that the cause is microcirculatory failure due to breakdown of the inner BRB: that is, disruption of NVUs consisting of both the retinal capillaries and the nervous system. In diabetic macular edema, dysfunction of neurons such as amacrine cells is speculated to cause macular edema. This aspect requires further study as a potential target for new drug development, and aiming at restoration of the damaged functional network of glial cells and neurons adjoining retinal blood vessels (neurovascular remodeling) is necessary. For this reason, research and development from a complex perspective are required, and the final endpoint is likely to be a long way away. Since research with an integrated concept of NVU in the retina started not so long ago, many new findings did not pass the speculative stage, and instead, they await further validation. Nevertheless, the notion that macular edema arises from dysfunction of NVUs is a rational hypothesis.An advanced high-throughput approach such as proteomic, transcriptomic, and metabolomic technologies may apply, to extend our understanding of molecular changes in NVU in the retina and to provide potential biomarkers for diabetic macular edema, highlighting the importance of future research in this field of ophthalmology.This article is based on the study, which received the Medical Research Encouragement Prize of The Japan Medical Association in 2019.Yoshihiko Usui received research grants from Bayer Yakuhin, Ltd.This work was supported by Grand-in-Aid for Scientific Research (C) grant numbers 16K11330, 19K09981, and 19K09959 from the Ministry of Education, Culture, Sports, Science and Technology of Japan, Research grants from Public Interest Foundation for the Elderly Eye Diseases Research Foundation.Conceptualization and design of the study was conducted by Y.U. The experiments were performed and analyzed by Y.U. The original draft of the manuscript was prepared by Y.U. Funding acquisition was conducted by Y.U.None


Uterine leiomyosarcoma (uLMS) is a rare and aggressive malignancy with poor clinical outcomes. Even when localized, uLMS is associated with high rates of local and distant recurrences that are usually fatal. Common sites of recurrence are lung, liver, pelvic lymph nodes, and vertebral and long bones, though atypical patterns of recurrence have been described. Among them, intracranial recurrence appears as a rare finding, almost exceptional in skull and dura. We describe the case of a solitary skull metastasis from uLMS in a 39-year-old woman, which represents the third reported case of skull recurrence in literature. After multidisciplinary discussion, the patient underwent surgery and received adjuvant radiotherapy. After 4 months, she is currently alive, without evidence of extracranial disease. This case highlights the importance of suspecting and recognizing atypical and extremely rare metastasis to this region. We encourage the need for large case series in order to provide further information about cranial recurrences of uLMS taking into account the paucity of data currently available in literature and the frequently unpredictable behavior of this rare and highly lethal disease. Uterine leiomyosarcoma (uLMS) is a rare malignancy accounting for 1–3% of all uterine cancers and approximately for 65% of all uterine sarcomas (US) . CurrentWe herein report a case of cranial recurrence of uLMS in a young woman, representing to our knowledge the third case of uLMS skull metastasis reported in literature.In January 2018, a 39-year-old woman presented to her gynecologist with menorrhagia and pelvic pain; her past medical history included uterine multinodular fibromatosis. A transvaginal ultrasound showed the presence of an intrauterine mass, for which she underwent a total laparoscopic hysterectomy, bilateral adnexectomy, pelvic and paraaortic lymph node biopsies, and omentectomy. The pathology report described a 7 cm diameter, stage IB uterine mass consistent with grade 3 uLMS. Histology showed smooth muscle actin (SMA) positive and desmin positive spindle cells with marked cytological atypia and 15 mitoses per 10 high power fields (HPF). Tumor necrosis was below 50% and no tumoral invasion was found in the uterine serous and omentum. Postoperatively, the patient received adjuvant chemotherapy with epirubicine plus ifosfamide.Clinical and imaging follow up showed no evidence of recurrence until June 2019, when a total body computed tomography (CT) scan revealed an osteolytic lesion in the right temporal-occipital region, 8 mm in diameter and positive to bone scintigraphy. Physical and neurological examination were normal. The subsequent cranial magnetic resonance imaging (MRI) confirmed the presence of the osteolytic temporal-occipital lesion, with epidural extension and partial compression of the underlying brain parenchyma . MRI shoIn October 2019, she underwent a right craniotomy with removal of the lesion and the involved scalp; epidural extension of the tumor was confirmed. Although surgery resulted in removal of all macroscopic disease, microscopic margins were positive (R1). Histological exam showed the presence of spindle cells with moderate pleomorphism, 15 mitotic figures per 10 HPF , and strAlthough hysterectomy offers the best chance of prolonged survival and it is considered as the standard treatment for uLMS, post-operative recurrence rate ranges from 50 to 70%, with a very poor prognosis . Lungs rTo the best of our knowledge, two cases of skull recurrence of uLMS have been reported in literature to date , 17 Tab. In the Signs and symptoms of skull metastases appears as highly non-specific, depending on tumor extension. This may lead to differential diagnosis, especially after several years of disease-free survival . In thisIf metastatic uLMS is certainly associated with poor prognosis, from a theoretical point of view solitary metastasis may show better clinical outcomes. All the patients reported in literature to date, including our case, showed solitary skull recurrence in the absence of other metastatic foci of uMLS. Given the rarity of metastasis to this region, there is a lack of evidence supporting a specific treatment of intracranial metastases from uLMS, whether metastatic site is brain or skull . ManagemDiagnosis of skull recurrence of uLMS requires a high index of suspicion. Its early recognition can be extremely difficult, especially in asymptomatic, healthy patients in which cranial CT scan is often not performed during post-surgery follow-up. There is lack of evidence for choosing the best therapeutic strategy in these patients and appropriate treatment of intracranial recurrences is yet to be defined.The presented report is the third documented case of skull metastasis from uLMS suggesting the importance of improving awareness of this extremely rare occurrence.Therefore, we encourage the collection of larger series in order to provide further information about cranial recurrences of uLMS, taking into account the paucity of data currently available in literature and the frequently unpredictable behavior of this rare and highly lethal disease.All datasets generated for this study are included in the article/supplementary material.Written informed consent was obtained from the individual(s) for the publication of any potentially identifiable images or data included in this article.AR and MCN have made substantial contributions to conception of the study, interpreted the patient data, and drafted the manuscript. VR performed surgery and has been involved in drafting the manuscript. CG performed the histological and pathologic examination of the surgical tissue and has been involved in drafting figures. AP and PD have been involved in drafting and revising the manuscript. GF, DB, and MD performed radiotherapy and have been involved in drafting and revising the manuscript. JM performed imaging and has been involved in drafting the manuscript. MP has made substantial contributions to conception of the study, interpreted the patient data, and drafted the manuscript. MN was the major contributor in concept and designed and revised the manuscript. All authors read and approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Mainstream next generation electronic devices with miniaturized structures and high levels of performance are needed to meet the characteristic requirements of electronics with flexible and stretchable capabilities. Accordingly, several applied fields of innovative electronic component techniques, such as wearable devices, foldable curtain-like displays, and flexible hybrid electronic (FHE) biosensors, are considered. This study presents a novel inspection system with multifunctions of stressing tensile and bending mechanical loads to acquire the stretchable and rollable characteristics of soft specimens. The performance of the proposed measurement platform using samples of three different geometric types is evaluated in terms of its stretchability. The results show a remarkable enhancement of mechanical reliability when the sine wave geometric structure is used. A symmetrical sine wave-shaped sample is designed to measure performance under cyclic rolling. The proposed measurement platform of flexible electronics meets the testing requirements of mechanical reliability for the development of future flexible electronic components and FHE products. With the advancement in intelligence and Internet of Things (IoT) applications, including transportation, logistics, medical care, and cloud smart life environments, suitable electronic components must have small dimensions and many features. The structural designs of smart wearable and handheld electronic devices, including active-matrix, organic light-emitting diode (AMOLED) displays ,2,3,4,5 In this research, a measured instrument of flexible electronics was developed. The designed concept is shown in Two major action modes, namely, stretchable and rollable loads, are considered in the proposed measurement platform. As shown in The measured components of apparatus are labeled in As mentioned in the designed probe device, manual adjustment is preferred because different kinds of specimens without standard specifications are most likely to be considered. Consequently, the electrical measurement probe is crucial during dynamic loading. The designs of the dual- and tri-axis probe devices are presented in Another important part of the measurement system is the human–machine interface, because all the initial conditions and testing data of various loading modes need to be separately collected and analyzed. As shown on the left side of TM were regarded as the testing specimens of the experimental setup in TM polyimide (PI) substrate. A small contact angle, i.e., less than 20°, was acquired to enhance the spreading ability when the optimum RF treatment for 180 min was conducted through the examination of various process conditions using Taguchi method. The given strain magnitude during measurements did not exceed the yield point of graphene ink and PI flexible substrate. The interconnect design of FHE is the key to maintaining long-term mechanical reliability. Therefore, two major loading modes, namely, bend and roll, were applied to estimate the loading ability of the measurement platform when different geometrical patterns of FHE interconnects based on the sensing requirements were considered. In this study, three kinds of geometrical shapes, namely, straight line, transition line, and sine wave types, were utilized. The detailed specifications of each interconnect pattern for validating the stretchable function are shown in Several shape designs of layout patterns for graphene ink conductive traces, including straight line, transition line, and sine wave sharp types, were considered in the stretchable and rolling tests. The change trajectory in electrical resistance during the entire testing process was monitored in real time. Accordingly, the graphene-ink interconnects printed on the flexible substrate of ITRI FlexUpThe specimens with a horseshoe line pattern shown in Experimental samples for dynamic stretchable and rolling tests were designed and fabricated in this research to verify the functions of the proposed measurement platform. The analytical results are described in detail in the following sections. The testing results of the specimens with three geometrical designs are shown in The ability to implement a highly rollable feature while maintaining in soft electronics is important. An apparatus to meet the requirements of suitable testing conditions needed to be developed. Accordingly, the proposed measurement platform provides rollable loading function to extract the cycling fatigue lifetime of the specimen in question. In this research, a PI substrate-printed graphene ink interconnect, as depicted in Various styles of FHE architectures and corresponding interconnect designs have successively been developed in response to the widespread utilization of FHEs in IoT. To meet the mechanical reliability requirements of soft electronics, this study proposed a powerful measurement platform integrated with a variable clamping device, a multiaxis probe, and human–computer interaction for movement settings, as well as monitoring software for the measured data. The major functions of the applied mechanical loads, including dynamic stretching and rolling modes, are provided by the proposed apparatus. During the measurement duration prior to the achievement of the loading target, changes in electrical resistance are recorded and collected in real time. Accordingly, the mechanical reliability of the concerned interconnect design may immediately be determined through the assistance of the monitoring software. Several kinds of graphene ink conductive interconnect/PI substrate specimens were considered and fabricated to perform the stretchable and rollable tests to verify the workability and reliability of the proposed measurement platform. The experimental results indicated that the horseshoe line type of interconnect has excellent capabilities in terms of resisting mechanical deformation through its geometrical stress/strain-compliant mechanism. The edges of the graphene ink interconnect after a fatigue rolling loading of more than 1400 cycles became rougher, compared with the sudden break failure of brittle ITO film. Furthermore, multiple freedom designs for soft electronics are crucial, given the increasing usage of FHEs. Consequently, continued refinements, especially for the clamping of torsional mode, will be introduced into the manifold of the proposed measurement platform.