The development of cognitive knowledge, motor skills, and artistic sense in order to restore lost tooth structure is fundamental for dental professionals. The course of dental anatomy is taught in the initial years of dental school, and is a component of the basic core sciences program in the faculties of dentistry. The learning objectives of the dental anatomy course include identifying anatomical and morphological characteristics of human primary and permanent teeth; identifying and reproducing tooth surface details in order to recognize and diagnose anatomical changes; and developing student’s psychomotor skills for restoring teeth with proper form and function. The majority of dental schools rely on traditional methods to teach dental anatomy, using lectures to convey the theoretical component; whereas the practical component uses two-dimensional drawing of teeth, identification of anatomical features in samples of preserved teeth, and carving of teeth. The aim of the present literature review is to summarize different educational strategies proposed or implemented to challenge the traditional approaches of teaching dental anatomy, specifically the flipped classroom educational model. The goal is to promote this approach as a promising strategy to teaching dental anatomy, in order to foster active learning, critical thinking, and engagement among dental students. Knowledge of tooth morphology and dental anatomy and function is imperative for the practice of modern dentistry. Dental professionals must develop cognitive knowledge, motor skills , and artistic sense, in order to restore the lost tooth structure with different materials ,2. TheseDental anatomy is one of the first courses taught in dental school, and is a component of the basic core sciences program in the faculties of dentistry around the world. The learning objectives of the dental anatomy course include identifying anatomical and morphological characteristics of human primary and permanent teeth; identifying and reproducing tooth surface details in order to recognize and diagnose anatomical changes ; and devAlthough these traditional techniques have been used to teach dental anatomy for decades in dental schools in the U.S. and many countries around the world, several concerns have been raised about their efficacy. One of the main limitations of the dental anatomy course is that it is usually isolated from other courses related to patient care, and present poor emphasis to clinical relevance . LikewisThe aim of the present literature review is to summarize different educational strategies proposed or implemented to challenge the traditional approaches of teaching dental anatomy, specifically the flipped classroom educational model. The goal is to promote this approach as a promising strategy to teaching dental anatomy, in order to foster active learning, critical thinking, and engagement among dental students.In recent years, dental schools around the world have proposed innovative educational strategies and alternatives to the current curricula, challenging the traditional teaching of dental anatomy. Kieser et al. incorporated spontaneous storytelling with problem-based learning, in their dental anatomy course in the University of Otago, New Zealand, in order to stimulate deep learning in a dental anatomy class . The useNew technologies have been designed and implemented to modify traditional aspects of dental anatomy teaching. These technologies include the use of interactive software, three-dimensional (3D) images, and digital atlases to study detailed tooth morphology, virtual patient simulations, and electronic flashcards ; likewisTeaching with digital tools has shown promising results by stimulating interactivity and independence in the learning experience ,17,18. TA flipped classroom  is an instructional strategy and a type of blended learning methodology that reverses the traditional learning environment, combining online learning, and the face-to-face classroom. This methodology leverages computer technology to reverse the traditional learning environment, delivering instructional contents outside the classroom, allowing students to acquire basic knowledge and concepts through the use of audiovisual tools  hosted online ,22. The The flipped classroom has been introduced successfully in the past two decades in different fields of dental education in order to promote critical thinking, problem-solving, and self-directed assessment among dental students . FaraoneThe flipped classroom approach combined with online learning tools have been successfully used to increase student engagement and support student learning in the dental anatomy program ,35,36,37The implementation of a flipped dental anatomy classroom was proposed in order to enhance student’s cognitive and psychomotor skills in human tooth morphology. The dental anatomy course might be offered in the same traditional time  in the first year of dental school. The traditional lecture approach will be substituted by a flipped classroom, incorporating didactic activities and a laboratory component .Students will be required to study the theoretical and foundational knowledge before each class. Content will be delivered using an on-line blackboard; different technological tools might be used, including but not limited to syllabus, course manual (developed by the instructor), instructional videos, short PowerPoint slide presentations (15–30 min voiceover slides with annotations), clinical cases, access to 3D software, dental atlases, and others. A low stakes quiz (5–10 multiple choice questions) addressing the first two levels of Blooms taxonomy, will be completed on-line before the class encounter, in order to ensure that students had reviewed the online material . The insStudent learning/performance will be assessed during each class session. The evaluation will include instructor and peer assessments. Instructor assessment will evaluate students’ attendance to class sessions, active participation in the small-groups discussion, completion of all the laboratory work, and a performance examination using standardized assessment tools. Likewise, after each class, the students will be asked to complete a peer assessment (only for the peers in their group) in order to provide feedback and promote a peer-to-peer learning environment and accountability for preparing prior to classes ,36.Flipping the dental anatomy classroom promises to revolutionize the teaching of tooth morphology in dental school, by articulating cognitive and psychomotor learning, and facilitating the transition to patient care in a clinical scenario ,37. The Although the flipped classroom offers many advantages, several challenges can be expected by both, students and instructors. One of the main challenges is the heavy reliance on student self-motivation and self-discipline to prepare before each face-to-face session. A strategy to minimize this limitation is incorporating a low stakes quiz prior the class to guarantee students compliance. Likewise, concerns have been raised regarding the increase in workload and preparation time for the student. Therefore, careful consideration of student’s workloads needs to be undertaken, balancing the pre-class material, and establishing clear learning goals and desired results ,39,40,41The flipped classroom method has been incorporated successfully in medical and dental education. This model can enhance teaching and maximize the use of time and resources. Flipping the dental anatomy classroom is a promising method that allows students to take control of the development of their cognitive and psychomotor skills; whereas, the instructor facilitates and supports the student learning process. Further long-term studies exploring this educational model in dentistry and dental anatomy, including its implementation, students’ engagement and assessment, and strategies to overcome unresolved limitations and barriers are needed.


Cortical processing is dynamically modulated by different neuromodulators. Neuromodulation of the cerebral cortex is crucial for maintaining cognitive brain functions such as perception, attention and learning. However, we do not fully understand how neuromodulatory projections are organized in the cerebral cortex to exert various functions. The basal forebrain (BF) cholinergic projection and the locus coeruleus (LC) noradrenergic projection are well-known neuromodulatory projections to the cortex. Decades of studies have identified anatomical and physiological characteristics of these circuits. While both cholinergic and noradrenergic neurons widely project to the cortex, they exhibit different levels of selectivity. Here, we summarize their anatomical and physiological features, highlighting selectivity and specificity of these circuits to different cortical regions. We discuss the importance of selective modulation by comparing their functions in the cortex. We highlight key features in the input-output circuits and target selectivity of these neuromodulatory projections and their roles in controlling four major brain functions: attention, reinforcement, learning and memory, sleep and wakefulness. The cerebral cortex is divided into distinct areas that compute specific sensory, motor, or other cognitive information. As the cortex develops into a wide and thick structure, each sub-region of the cortex can work as a module. Depending on the task demand, an animal needs to devote a particular cortical region to process specific information. Neuromodulatory inputs to the cortex are known to play important roles in guiding the transition of cortical processing  and the midbrain cholinergic nuclei. The BF cholinergic neurons send projections to the entire cerebral cortex in both primates and rodents  concentration in the sensory cortex of the anesthetized rat increases when the animal receives sensory stimuli whereas ACh concentration in the medial prefrontal cortex (mPFC) does not show any changes  restricts the diffusion of ACh by enzymatic hydrolysis after the release  are ionotropic receptors that can generate fast excitatory postsynaptic potentials. In the macaque primary visual cortex (V1), nAChRs are found in thalamic axons of excitatory neurons in layer 4c as well as in inhibitory interneurons  regulates a number of brain functions, such as sleep/wakefulness and attention, and it has the potential of modulating wide brain regions including the hippocampus, the amygdala, the thalamus, and the cerebral cortex  of rats induces impairment of the working memory even though the rats detected the visual signal correctly (Chudasama et al., Sara and colleagues have shown the role of noradrenergic modulation of the cortex during learning and memory Sara, . They foAlthough the cholinergic neurons show selective innervation to the cortex, it has also been known that the cholinergic neurons can be involved in modulation of the global brain states during sleep. The BF cholinergic neurons are highly active during wakefulness and paradoxical sleep but show low activity during slow-wave sleep (Lee et al., The LC noradrenergic system is also known to be involved in controlling sleep (Aston-Jones and Bloom, The BF cholinergic and LC noradrenergic systems share common features: broad cortical innervations and regulation of cognitive functions such as arousal, attention, learning, and sleep. However, they clearly show distinct anatomical and physiological characteristics. First, the BF is constructed with multiple sub-nuclei, which project to distinct regions in the brain Figure . The LC,H-JR designed the figures and wrote the manuscript. J-HK wrote the manuscript. S-HL conceived the contents and wrote the manuscript. All authors discussed the contents and revised 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.


Atherosclerosis (AS) is a disorder of large and medium-sized arteries; it consists in the formation of lipid-rich plaques in the intima and inner media, whose pathophysiology is mostly driven by inflammation. Currently available interventions and therapies for treating atherosclerosis are not always completely effective; side effects associated with treatments, mainly caused by immunodepression for anti-inflammatory molecules, limit the systemic administration of these and other drugs. Given the high degree of freedom in the design of nanoconstructs, in the last decades researchers have put high effort in the development of nanoparticles (NPs) formulations specifically designed for either drug delivery, visualization of atherosclerotic plaques, or possibly the combination of both these and other functionalities. Here we will present the state of the art of these subjects, the knowledge of which is necessary to rationally address the use of NPs for prevention, diagnosis, and/or treatment of AS. We will analyse the work that has been done on: (a) understanding the role of the immune system and inflammation in cardiovascular diseases, (b) the pathological and biochemical principles in atherosclerotic plaque formation, (c) the latest advances in the use of NPs for the recognition and treatment of cardiovascular diseases, (d) the cellular and animal models useful to study the interactions of NPs with the immune system cells. Atherosclerotic disease, or simply atherosclerosis (AS), is initially characterized by the formation of fatty streaks, with the accumulation of lipids [primarily cholesterol, but also triglycerides , especially circulating monocytes (MCs) that subsequently differentiate into macrophages (MΦs) and then into foam cells (FCs), going along with plaque formation and evolution  or alternatively activated M2 (anti-inflammatory), evidences have brought to the definitions of additional subtypes  and even “sub-subtypes” ; there could actually be a continuum of specializations, and MΦs could even convert into each other , especially if agglomerated within oxidized low density lipoprotein (oxLDL). Upon this process, efferocytosis efficiency of cells is reduced, they transform into foam cells, and are easily subjected to imbalances in cholesterol influx/efflux process and hypoxia. The final outcome is most often apoptosis or necrosis, which resolve in local accumulation of the lipid content, the major constituent of the inner (possibly necrotic) core of the atherosclerotic plaque  is characteristic of the vulnerable plaque. The presence of activated macrophages leads to the secretion of matrix-components degrading enzymes , inhibits the production of collagen by the SMCs, and finally contributes to vascular wall reshaping. These mechanisms are useful in early lesions, supporting resolution of disrupted endothelial layers and avoiding accumulation of toxic species, as well as for the reduction of atherosclerotic plaques; but, at the same time, they induce fragility in late plaques, with the possible final consequence of plaque rupture  particularly affect production and transport of cholesterol (or other lipids) to the arterial walls  trial  or Gadolinium (Gd), and DiI fluorescence markers to image atherosclerotic plaques in ApoE−/− mice. Alternatively, it has been proposed to target collagen IV, which is present on the vascular basement and exposed when vascular permeability increases , changing their biological identity. PC formation (or “opsonisation”) is often the first step toward the sequestration of NPs by the RES. Various approaches have been considered to avoid such phenomenon. Recent strategies are based on controlling nanoconstruct stiffness, since it was found that deformable particles are less subject to uptake by macrophages in RES in off-target tissues  or exogenous .Especially under this view, an interesting development can arise from a synergy between nanotechnology and personalized medicine  and infused with angiotensin II, promoting inflammation driven by monocytes/macrophages. 2 days post injection the ratio between peripheral pro- and non-inflammatory monocytes subsets decreased substantially, and tissue macrophage polarity was regulated toward the non-inflammatory phenotype M2, with consequent suppression of EMMPRIN/MMP pathway and reduction of plaque destabilization risk.A different, interesting strategy consists on preventively acting on the selective recruitment of monocytes from the precursors of pro-inflammatory macrophages M1 , a key player in recruiting inflamed monocytes to atherosclerotic plaques; they observed reduced PET signals from As reviewed above, fundamental and pre-clinical researches on AI-drugs-loaded NPs against AS are beginning to bloom, and there are also clinical trials for the (separated) use of NPs and AI drugs for regression or consolidation of AS plaques. More fundamental and translational research on the mechanisms underlying their action could inspire and motivate future more efficient clinical trials using AI-drug loaded NPs against AS.Important future steps for the development of new NP-based theranostics strategies can arise from the study of interaction mechanisms between NPs and immune cells, with particular regard to what fate NPs and their components will face upon site deposition and cellular uptake. Investigations on these directions will remarkably improve the development of NPs rational designs aimed at specific accumulation and cargo smart activation.Indeed, where, how and in which proportion NPs are internalized in the various subtypes of ICs is still not clear, also due to the different internalization routes observed even in close phenotypes tools needs a highly interdisciplinary environment, involving e.g. cardiologists, biologists, physicists, chemists, engineers; professionals able to talk with each other, and fostering the formation of multi-faceted scientists.SL conceived the idea. SL and AC wrote the manuscript, with major contributions from RP, RC, and GV, starting from texts written in collaboration with LM, ME, and PD. All authors discussed, edited, and contributed to 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.


Colorectal cancer is a major health burden and despite the recent advances in healthcare and screening programs, a great percentage of patients already present metastases once their disease is found. In those cases, liver surgery has an essential role, but even with neoadjuvant chemotherapy there is a high rate of intrahepatic recurrence. New prognostic factors are needed in order to decide the best surgical approach considering the biological behavior of the tumors in order to tailor the used therapies, moving towards individualized medicine/treatment. However, the majority of the factors described in literature are expensive, time consuming, and difficult to apply on a daily basis. Histological growth patterns have emerged over the past few years as a reproducible characteristic, an easy to apply one, and with very low costs since it only needs the standard Haematoxylin and Eosin stained slides of observation. In this article, we provide a review of the histological growth patterns of liver metastases and their prognostic significance, biological meaning, and therapeutic importance. The liver is a common site for metastatic dissemination and in some regions of the globe, namely, Europe, secondary liver tumors are far more common than primary  ones. ReIn the past few years major advances regarding treatment strategies of colorectal cancer liver metastases (CRCLM), such as more effective chemotherapy regimens, portal vein embolization and staged hepatectomies , and one-stage ultrasound-guided parenchymal preserving resections, all have contributed to extend the limits of oncological resectability , 4. In sRecently, histological growth patterns (GP) have been identified as a practical and reproducible factor of prognosis, easily assessed by Haematoxylin and Eosin (H&E) stained slides by an experienced pathologist. They are defined as expansive (when tumor growth compresses the hepatocytes), desmoplastic (with presence of fibrous tissue in the periphery of the tumor), and replacement  Figures  and 2. OThe correct analysis and consequent accurate classification of the CRCLM growth pattern implies a detailed gross examination with adequate sampling with at least one sample per tumor centimeter, similar to the sampling used for tumor regression grade assessment, for all the lesions , 13; in Other studies have attempted to identify the characteristics that allow for a better or worse prognosis for overall survival such as tumor thickness at the tumor-normal interface , 15 and The reason for this behavior has not been fully understood yet. However, it can represent the complexity of tumor/host interactions, with the pushing pattern described by some authors as more angiogenic  and the The thick band of stroma present in the desmoplastic growth pattern, enriched with collagen, may act as an obstacle to tumor expansion, representing an improved host response with dense lymphocytic infiltration, increase in collagen type IV and integrin blockade, reducing the infiltration of nontumoral parenchyma, therefore demonstrating a more favorable characteristic , 20.The pushing pattern displays biologic properties with increased levels of endothelial cell proliferation fraction , 21 and The information gained by studying the molecular mechanisms underlying the complex tumor/host interactions associated with the distinct GPs may aid in the selection of new therapeutic targets. Growth receptor blockade (with anti-EGFR antibodies) and antiangiogenic agents (with anti-VEGF antibodies) have already demonstrated excellent results in the treatment of advanced metastatic disease.Several studies have addressed the relation of GPs and angiogenesis capacity, and the recent discovery mechanism of vessel cooption vascularization of tumors has explained a possible resistance to antiangiogenic agents. This led to the suggestion of a combined therapeutic as possible approach and linked this biological behavior to a specific GP .Other mechanisms of tumoral survival, such as evading the host immune response, epithelial-mesenchymal transition, or hypoxia-resisting factors, may serve as targets for molecular therapies in the near future, namely, immunotherapy , 28 and By reflecting the tumor-host interaction, the GP of liver metastases can influence overall and disease-free survival. Although GP could influence patient management, being a histopathological characteristic, the GP can only be known after surgical resection. However, imaging techniques could potentially detect different types of growth patterns before surgery.Due to the simplicity of this biological characteristic, other studies have assessed this biological behavior in liver metastases of nonintestinal carcinomas, such as breast cancer  and uveaThe different prognoses of the colorectal cancer liver metastases (CRCLM) may represent a new prognostic characteristic that may be related to the primary tumor properties and may be predicted by preoperative imaging, allowing individualized patient care.The radiological response pattern to chemotherapy, particularly with antiangiogenic drugs, has been previously reported as having implications in the prognosis. In a cohort of 209 patients with CRCLM undergoing hepatectomy after neoadjuvant chemotherapy, the presence of a sharply defined border on preoperative computed tomography (CT) was associated with improved overall and disease-free survival . HoweverMore accurate in tissue analysis than CT, Magnetic Resonance imaging (MRI) could provide important answers in this regard, especially with the use of hepatospecific contrast agents. In fact, the relative tumor enhancement by gadoxetic acid (a contrast agent used in hepatobiliary imaging) can accurately predict the response to chemotherapy in treatment-naïve patients with CRCLM . MoreoveOne may raise the question about patients with several metastases; in our study , the majAdvances in imaging could detect distinct GPs, thus impacting patient management. This could lead to a tailoring of the therapy, both in the choice and duration of chemotherapy and in the use of resection and other locoregional techniques.In fact, there are two distinct currents of thought in the scientific literature regarding the surgical management of CRCLM : on the Moreover, knowledge of the GP could also influence the choice and duration of chemotherapy regimens. Although beneficial, preoperative chemotherapy in CRCLM can cause hepatotoxicity and increase postoperative morbidity , 40–42 AHistological GPs are indeed a powerful tool, but in the past, they were described using several designations, thus raising barriers for worldwide harmonisation . The recConcerning the surgical approach, literature is not unanimous regarding the perfect approach to synchronous CRCLM: some advocate the colorectal surgery first , others The possibility of predicting the GP of CRCLM in preoperatory evaluation may allow for an individualized treatment algorithm for each patient. Particular imaging features of the metastatic disease could also expand this information. In this way, neoadjuvant chemotherapy, known to be more effective on the secondary tumor than on the primary  one, couWe hope that further investigation into GPs can help clinicians to choose therapies in a multimodal perspective, not only in cases of CRCLM but also in other indications, such as gastric or breast cancer liver metastases. The consistent report of the CRCLM GP in pathology reports according to the correct consensus  should b


The diagnosis of paucibacillary (PB) leprosy cases remains a challenge because of the absence of a confirmatory laboratory method. While quantitative polymerase chain reaction (qPCR) has been shown to provide reliable sensitivity and specificity in PB diagnoses, a thorough investigation of its efficacy in clinical practice has not yet been published. The present study evaluated patients with suspected leprosy skin lesions by using qPCR to identify PB individuals in the Leprosy Outpatient clinic at the Oswaldo Cruz Foundation in Rio de Janeiro, Brazil.Mycobacterium leprae qPCR-targeting 16S gene) results were analyzed, and a decision regarding multi-drug therapy was made. A year later, the patients were re-examined, and the consensus diagnosis was established.One hundred seventy-two suspected PB cases were included in the study. The patients were evaluated by a dermatologist at three different times. The clinical dermato-neurological examination and collected samples were performed on the first visit. On the second visit, the results of the histopathological analysis and PCR assay (DNA-based In 58% (100/172) of cases, a conclusive diagnosis via histopathological analysis was not possible; however, 30% (30/100) of these cases had a positive PCR. One hundred ten patients (110/172) attended the third visit. The analysis showed that while the sensitivity of the histopathological test was very low (35%), a qPCR alone was more effective for identifying leprosy, with 57% sensitivity.The use of qPCR in suspected PB cases with an inconclusive histology improved the sensitivity of leprosy diagnoses. Mycobacterium leprae DNA direct detection in clinical samples has been developed to support diagnosis. This study used qPCR to differentially diagnose suspected cases of paucibacillary leprosy and to follow them after treatment to confirm the consensus diagnosis and therapeutic approaches. It was concluded that qPCR and histopathological analysis increase clinical diagnosis accuracy, and qPCR has higher sensitivity. The routine use of qPCR laboratory tests could therefore improve the precision of early diagnosis for the achievement of leprosy control.Leprosy is a major public health problem in several low- to middle-developed countries where a consistent number of new cases are diagnosed every year. Irrespective of the use of efficient treatment, diagnosis is late because of the absence of a gold standard method. Thus, the multi-drug therapy (MDT) is unable to control of disease transmission even though early diagnosis, adequate MDT treatment of new cases, and contact surveillance have been considered the pillars of leprosy endemic control. Quantitative polymerase chain reaction (qPCR)-based  Prerequisites for leprosy control, early detection, and adequate treatment are crucial for reducing the risk of disability and interrupting transmission, thereby decreasing the burden of the disease. Leprosy diagnoses are based on clinical manifestations, the presence of acid-fast bacilli (AFB) determined through microscopy in slit skin smears, and histopathological analyses of skin biopsy samples. Nevertheless, diagnosis is challenging, particularly in the early stages and in a paucibacillary (PB) form characterized by the absence of bacilli in slit skin smears. In addition, skin macules or plaques, frequently found in PB leprosy, represent the clinical manifestations of many other dermatoses, such as pityriasis alba, rosacea granulomatosa, granuloma anulare, cutaneous tuberculosis, and sarcoidosis.Histological diagnosis of leprosy is indicated by two cardinal features: neural aggression and bacillar identification . In PB, Indeed, no clinical or histopathology tests are capable of either confirming or rejecting all suspected PB cases. In the last two decades, studies have demonstrated the potential of polymerase chain reaction (PCR) technology for the rapid detection and identification of leprosy in clinical specimens –8. More In the absence of a gold standard, previous studies have used clinical diagnoses or histopathologic analyses as reference standards for evaluating PCR sensitivity and specificity. However, the inherent subjectivity of these methods underscores the need for more reliable criteria for defining reference standards.Since 2012, qPCR has been used as a support in the clinical and histopathological leprosy diagnostic procedures at our clinic. The present study aimed to provide more precise sensitivity and specificity for qPCR in PB leprosy. Suspected PB cases were followed for a year, and consensus diagnoses were established after observation and evaluation of the presence or absence of skin lesions one year later. The consensus diagnosis became the reference standard for analyzing the role of qPCR in PB leprosy diagnoses in clinical practice.This prospective observational study, nested in a clinic cohort, was performed at the Leprosy Outpatient Clinic of the Oswaldo Cruz Foundation (Fiocruz), a national reference center for leprosy in Rio de Janeiro, RJ, Brazil. All individuals referred to the clinic for diagnosis, from January 2012 through December 2016, presenting with suspicious PB leprosy skin lesions were included.Excluded were all cases exhibiting a positive bacteriological index (BI) on a slit skin smear, visualized bacilli in the first histopathological analysis, and suspicion of relapse. Also excluded were those cases suspected of having the pure neural form without skin lesions.Each patient had three evaluations. The first included a clinical evaluation and sample collection (skin biopsy for histology and qPCR). In the second evaluation one month later, the histopathological and PCR results were analyzed, and a decision regarding MDT was made. At the third visit one year later, the consensus diagnosis was established . For eacMycobacterium (M.) leprae DNA detection.All patient clinical-epidemiological data, including age, gender, contact with leprosy patients, and lesion duration, were collected and recorded on the standard outpatient form. The symptoms and the clinical description of the skin lesions, i.e., number, color, size, location, and type (macula or plaque), including lesion sensitivity, were recorded after the dermatological and physiotherapeutic examinations. Slit skin smears were taken from four sites  for bacteriological diagnosis. One skin biopsy was performed (punch 6 mm) from suspected lesions for a histopathological analysis and a PCR assay. These skin samples were divided into two parts. One was stored in formalin for histological study, and the other, in 70% ethanol for One skin biopsy sample was prepared for routine histopathological examination, hematoxilin-eosin, and Wade staining. All the cases were analyzed by an experienced dermatopathologist and were classified as either PB leprosy, other dermatoses, or nonspecific histological features (NSHFs). The cases were categorized as PB leprosy in the presence of an inflammatory lymphohistiocytic or granulomatous infiltrate in the nervous fillets and/or the erector muscle of the hair . If the M. leprae-specific target through the use of the TaqMan amplification assay. All the reactions were performed in triplicate through the same real-time PCR system (Applied Biosystems StepOne). The results were obtained in accordance with the first fluorescent signal detection, cycle threshold (Ct) [For DNA extraction, the DNeasy blood and tissue kit  was used in accordance with the specifications. The qPCR technique is standardized to amplify the 16S rRNA old (Ct) . As recoold (Ct) ,15,16.At the second visit, the dermatologist used the Ministry of Health clinical criteria for leprosy diagnosis  to categoPatients with skin lesion remission on the third visit post-MDT; andPatients who on the third visit had skin lesions without MDT and those whose new biopsy results showed the histopathological characteristics of leprosy or a positive PCR.Patients with skin lesions on the third visit despite receiving MDT;Patients with lesion remission on the third visit after specific treatment for other dermatoses; andPatients with lesion remission on the third visit without MDT or any other medication.All of the patients were invited for a third clinical examination during which the consensus diagnosis was established. The presence or absence of skin lesion and the results of the PCR and histological analysis performed during the first visit were considered. If the lesions remained unchanged, a new biopsy was performed for diagnostic clarification through PCR and histopathological analysis. The cases were classified as either leprosy or non-leprosy (consensus diagnosis) according to the following criteria:After the third visit, the consensus diagnosis (leprosy or non-leprosy) was established as the reference standard.The frequencies for the clinical-epidemiological characteristics were ascertained through the use of proportions for the categorical variables and centrality and dispersion measures for the continuous variables. The proportions of the histological diagnostic  and the PCR results (positive or negative) were calculated and compared by use of the chi-square test (significance level of 5%). The frequency distribution of the cases on the basis of the related qPCR, histopathological tests, and consensus diagnoses was described through a flow chart. The sensitivity and specificity of the histology, the qPCR, and the combination of these two diagnostic methods were also calculated. The confidence intervals were calculated at a level of 95%.This study was approved by the Ethics in Research Committee of the Oswaldo Cruz Foundation CAAE: 38053314.2.0000.5248, number: 976.330–10/03/2015. Written informed consent was received from all the participants and the parents or guardians of the children under 18 years old included in the study.As is shown in The results of the histopathological and PCR tests in the skin samples indicated that it was impossible to reach a leprosy diagnosis via histology in 58% (100/172) of all cases. In addition, among those with NSHFS, 30% had a positive PCR .The frequency distribution for the qPCR results, histopathology diagnosis, and consensus diagnosis is presented for the 110 cases that made the third visit (110/172). The consensus diagnosis of almost half of this group (54/110) was leprosy .Only one of the five cases with a false positive PCR  was 22% higher than histopathology sensitivity (35%) .The results of this study made clear that for some patients whose clinical and histological changes were insufficient for reaching a diagnosis, qPCR aided in establishing leprosy. In the cases in which the PB leprosy was suspected, the simultaneous use of qPCR and histology increased the sensitivity of a PB diagnosis without a loss of specificity.A majority of the cases evaluated in this study had one skin lesion, and most of these were the macula type. Moreover, most patients reported having noticed their lesions one year or less before their initial visit. Nayar et al. affirmed the near impossibility of visualizing inflammation in the nervous fillets by histopathology during the first year of the disease . The resM. leprae detection by qPCR in patients with a negative bacillary load and NSHFs would be of great value for limiting subjectivity in the diagnosis of PB leprosy.Studies have found qPCR sensitivity ranges of 34%–80% in PB patients –11. HoweM. leprae [One of the five apparently false positive PCR cases had tuberculoid granulomas. Because the histopathological diagnosis was granulomatous rosacea, the patient was treated with doxycycline at a general dermatological clinic. Surprisingly, the rosacea plaque cleared only after seven months of daily use of the antibiotic. Doxycycline and minocycline are antibiotics in the tetracycline group with the same mechanism of action. For decades, minocycline, which has a strong bactericidal action against . leprae , was use. leprae –21. A poThree explanations are possible for the four positive PCR cases in which the lesions cleared without MDT or any other medication. It should be noted that studies using qPCR-targeting 16S and other genes have described a 100% specificity for PB leprosy ,10. In aLeprosy is a slow-progressing disease for which a gold standard for diagnosis does not exist. Therefore, defining the consensus diagnosis after observing the progression of the cases, as was done in the present study, was an additional step forward in providing higher precision for the PCR sensitivity and specificity measures. The sample size could be considered a limitation even though it appears that no other study has yet analyzed more than 100 PB patients for molecular diagnosis .The present study clearly demonstrated that even for experienced dermatologists and pathologists, the difficulties encountered in interpreting the clinical and histopathological differential diagnosis of PB cases may be responsible for many delayed diagnoses. Because of the integration of leprosy control actions into the delivery of general health services in Brazil and other endemic countries, patients will continue to escape detection because few clinicians have the skills, experience, or expertise to diagnose the disease, especially in the early stages .In addition, studies have highlighted that any declaration of the elimination of leprosy as a public health problem results in the neglect of ongoing disease control strategies, such as household surveillance, community awareness campaigns, and training for health professionals. Consequently, the lower the incidence, the less prepared will be clinicians and pathologists to identify leprosy in their clinical practice ,26. BrazM. leprae DNA detection; however, reference centers are still necessary.In the last decade, real-time PCR has proved to be particularly useful in three areas: the identification of the drug-resistant bacilli used for treating leprosy , differeIn conclusion, the use of qPCR in suspected PB cases with a non-conclusive histology improved the sensitivity of leprosy diagnoses. Because early diagnosis is crucial to avoiding transmission and incapacitating neural lesions and achieving the goal of a leprosy-free world, the need for tools for the detection and confirmation of the disease among suspected individuals wherever they may live is indeed urgent.S1 Checklist(DOC)Click here for additional data file.


With the purpose of estimating the source contributions of nonmethane hydrocarbons (NMHC) to the atmosphere at three different sites in the Mexico City Metropolitan Area, 92 ambient air samples were measured from February 23 to March 22 of 1997. Light- and heavy-duty vehicular profiles were determined to differentiate the NMHC contribution of diesel and gasoline to the atmosphere. Food cooking source profiles were also determined for chemical mass balance receptor model application. Initial source contribution estimates were carried out to determine the adequate combination of source profiles and fitting species. Ambient samples of NMHC were apportioned to motor vehicle exhaust, gasoline vapor, handling and distribution of liquefied petroleum gas (LP gas), asphalt operations, painting operations, landfills, and food cooking. Both gasoline and diesel motor vehicle exhaust were the major NMHC contributors for all sites and times, with a percentage of up to 75%. The average motor vehicle exhaust contributions increased during the day. In contrast, LP gas contribution was higher during the morning than in the afternoon. Apportionment for the most abundant individual NMHC showed that the vehicular source is the major contributor to acetylene, ethylene, pentanes, n-hexane, toluene, and xylenes, while handling and distribution of LP gas was the major source contributor to propane and butanes. Comparison between CMB estimates of NMHC and the emission inventory showed a good agreement for vehicles, handling and distribution of LP gas, and painting operations; nevertheless, emissions from diesel exhaust and asphalt operations showed differences, and the results suggest that these emissions could be underestimated.


This ancient and highly conserved enzymatic complex from the family of heme-copper oxidase possesses two copper sites: CuA and CuB. Biosynthesis of the oxidase is a complex, stepwise process that requires a high number of assembly factors. In this review, we summarize the state-of-the-art in the assembly of COX, with special emphasis in the assembly of copper sites. Assembly of the CuA site is better understood, being at the same time highly variable among organisms. We also discuss the current challenges that prevent the full comprehension of the mechanisms of assembly and the pending issues in the field.Copper is an essential cofactor for aerobic respiration, since it is required as a redox cofactor in Cytochrome  First-row transition metal ions are essential for the viability of all living organisms. Organisms from the three domains of life have adopted the utilization of different metal ions as cofactors for challenging enzymatic processes. Taking advantage of their particular chemical properties, transition metal ions are key players in essential metabolic processes like condensation and hydrolysis pathways and (particularly) in redox events based on the different oxidation states displayed by them .These redox properties make them essential but also toxic when their levels are not adequately regulated ,3. For eIn order to maintain and regulate the homeostasis of transition metal ions, organisms have developed scavenging, storage and metal efflux strategies that are regulated depending on the environmental metal availability ,9,10. BaEukaryotes have evolved more sophisticated regulatory pathways. From a cellular point of view, metal homeostasis depends on mechanisms similar to those present in bacteria. Metal uptake, storage and efflux are tightly regulated ,18. CellAdditionally, superior eukaryotes ensure the systemic metal homeostasis by several regulation mechanisms. First, the incorporation of dietary iron and copper has to be regulated in order to limit the uptake of these metals to trace amounts and avoid toxicity. Second, the metal ions are transported through the blood system by specific metal carriers to ensure the delivery of metals to the cells. And last, the metal ion incorporation is regulated by the expression of carrier interacting proteins in the surface of the cells, to mediate metal ion delivery to the required cells .+1/Cu+2. This one-electron reaction is used in nature in different catalytic processes that span a wide range of redox potentials (200 to 1000 mV) based on the influence of the metal ligands and the protein environment that modulate the electronic structure of the copper site and, consequently, its reactivity features. For example, in the oxygen cycle, copper is involved both in the reduction of oxygen (aerobic respiration), but also in the formation of molecular oxygen in the photosynthetic pathway ,73Strept-hairpin  73,74]..StreptomS. lividans Sco is devoid of thiol-reductase activity, but is able to act as the copper chaperone for the CuA site, being critical for copper capture from the extracellular medium and subsequent transfer to Cox II. However, at the moment, it is not clear yet whether Sco simply sequesters the copper ions from the medium or ECuC is a copper donor for Sco, acting upstream in the mechanism of copper acquisition. S. lividans Sco is critical for morphological development and copper homeostasis, suggesting that this protein also plays regulatory roles . T. TA asseA assembly in two important model organisms from this group: Paracoccus denitrificans and Bradyrhizobium japonicum.Among gram-negative microorganisms, alpha-proteobacteria are particularly relevant for the study of Cox assembly factors based on their evolutionary relationship with eukaryotic mitochondria and provide a simple working model. Next, we will review the state-of-the-art and the perspectives regarding CuParacoccus denitrificans expresses two Sco orthologs (ScoA and ScoB), and two periplasmic copper chaperones related to pCuAC (pCu1 and pCu2) [P. denitrificans COX activity under normal copper levels. However, when bacteria are grown on low-copper media, ScoB becomes critical for CuA assembly. This suggests that ScoB is the copper chaperone for CuA site  . DeletioCuA site e. Despitnditions ,79. FurtBradyrhizobium japonicum is more complex. This gram-negative bacteria encodes two periplasmic pCuAC-like proteins, pCuA and pCuC, located in a copper-inducible operon that also contains a membrane copper receptor and a cytoplasmic transporter [B. japonicum expresses a membrane-anchored thioredoxin-like protein, TlpA, and a Sco ortholog, ScoI. TlpA was originally annotated as a thiol-reductase since its discovery [A site in B. japonicum remains to be elucidated, the obvious candidates being ScoI and the two periplasmic chaperones pCuA and pCuC  that lack these structural features and still can complement the lack of other Sco proteins is very surprising. This highlights that these proteins are also performing a different role, apart from the typical thioredoxin/metallochaperone activities. Further research is required to assess which is this function.The Sco family includes the most important players in the assembly of the CuOne of the most significant challenges is to identify the biochemistry of each protein separately, and to describe how they cooperate to fulfill their function. In this review, we have discussed the high diversity and complexity of the mechanisms of assembly of COX copper sites, even when the COX structural subunits are highly conserved. This diversity encompasses changes in the required assembly factors upon exogenous conditions, or even in different tissues of the same organism. This picture discloses how cells have evolved to provide alternative routes to guarantee the assembly of essential copper sites under different  conditions.


Contrast opacification of the vascular system has become the mainstay of radiological examinations. This brief illustrated and chronological historical overview commences with the vascular opacification on a corpse via direct puncture of the aorta at both the thoracic and the abdominal levels and ends with the analogous exams on computed tomography (CT) and magnetic resonance imaging (MRI) via peripheral venous puncture. Victory has many fathers, but defeat is always an orphan.—John F Kennedy To know where you go to, it helps to know where you come from. Contrast opacification of the vascular system has become the mainstay of radiological examinations. This brief illustrated and chronological historical overview commences with the vascular opacification on a corpse via direct puncture of the aorta at both the thoracic and the abdominal levels and ends with the analogous exams on computed tomography (CT) and magnetic resonance imaging (MRI) via peripheral venous puncture. The major improvements applied to both the equipment and the techniques over more than a century are highlighted, with a special emphasis for the people who have pioneered and modified these approaches. While therapeutic and diagnostic approaches are increasingly distinctive radiological domains, both highly rely on a precise anatomical assessment of macroscopic blood vessels and the microvasculature. The road from the simple radiographer to the actual interventional practitioner has been long, with intricate paths that mainly include the contrast media, the vascular access, and the image capture.In January 1896, Eduard Haschek and Otto Theodor Lindenthal of Vienna opacified—by means of a mixture of chalk, cinnabar (mercury sulfide), and Vaseline—the vessels of an amputated hand  and Jacques Forestier (Aix-en-Provence) injected 5 cc of Lipiodol, an oil-based contrast agent, into the femoral vein to visualize the cardio-vascular system of a living dog. They observed that the contrast agent was scattered by the right ventricle and formed multiple pulmonary emboli that disappeared in 10–12 minutes ..4].The need for contrast agents suitable to more recent imaging techniques such as ultrasound (US) and magnetic resonance imaging (MRI) has led to the development of microbubbles and gadolinium/manganese-based contrast agents, respectively. The bio-distribution of MRI contrast agents parallels those of X-ray contrast agents, as they diffuse to the interstitial space from the vascular space and vice versa, whereas US microbubbles are limited to the vascular space. Current research efforts are directed towards increasing these contrast agents’ specificity for diseases.In 1929, Werner Forssmann, working at the Surgery Hospital of Berlin, performed the first cardiac catheterization on himself Figure  5]. He . He 5]. In 1931, Egas Moniz, Lopo de Carvalho, and Almeida Lima, from Lisbon, published an article titled “Angiopneumography”, a term which they applied to techniques of opacification of the pulmonary vessels by the means of sodium iodine .In 1933, Peter Rousthöi, at the University of Stockholm, established and published a new technique of angiocardiography via retrograde catheterization Figure  of the aThat same year, Henri Reboul  and Maurice Racine  studied, through trans-ventricular puncture Figure  of an anIn 1936, direct puncture of the thoracic aorta was first performed on humans by Innocenzo Nuvoli , who succeeded in demonstrating aneurysms, tortuosity, and other abnormalities. He utilized a bone marrow needle to perforate the skin, the subcutaneous tissue, and the sternum in the midline at the level of the second intercostal space [In 1938, George Porter Robb, of the College of Medicine University of New York, and Israël Steinberg, of Harvard University, described a new method of angiocardiography. They used ether and cyanide circulation time as a guide Figure  for timiA decade later, in 1948, Stig Radner  published a new technique  set out to develop a simpler and more reliable catheterization method Figure  that cou18Lastly, the important role played by the industry designing the catheters that later allowed for the development of interventional procedures such as embolization and drug infusion should be examined .The advent of radio-cinema is an important milestone in vascular imaging. A gastro-intestinal radiologist from Brussels, Dr. Van de Maele, developed an apparatus between 1935 and 1938 that allowed 16 radiographic images per second Figure . This fo21In 1949, in California, Merell Sisson  proposed a simple and manual system of a radiological cassettes changer for arteriography Figure  that wouIn Sweden, angiography was emphasised from the very beginning. In 1949, Ake Gidlund of the St Eriks Hospital (Stockholm) developed an apparatus Figure  that allIn 1957, Alan Thal and colleagues described a manually compressed injector Figure  with a sIn 1960, Kurt Amplatz  described a cardiovascular injector powered by carbon dioxide cartridges Figure , such asFor decades, only a catheter-based angiography was able to display accurate anatomic detail of the blood vessels for both diagnosis and therapy. Today, computerized tomography (CT), magnetic resonance imaging (MRI), and ultrasound (US) often serve as definitive angiographic studies, replacing invasive catheterization. In most cases, using these techniques only requires a peripheral venipuncture. With these techniques, interest has also shifted from macroscopic angiography to microvasculature, with the aim to precisely describe the distribution of the contrast agent at a microscopic level. This enables, for example, the evaluation of tumour microvasculature at different locations and in response to specific therapeutic agents to refine the treatment and the prognosis of patients with cancer, as these responses are far more sensitive than the change in size.Recent improvements with all these techniques have gone towards better tissue characterization and faster scanning and processing times. For example, during the past decade, CT has seen the development of several technologies, such as the slip-ring gantry, the multi-row detector arrays, high-load X-ray tubes, and faster computers. Altogether, these progresses enabled dual-energy scanning, which allows extracting and reconstructing for the spectra related to each element of the periodic table. Applying this to iodine realistically paves the way not only for the easiest non-invasive angiographies, but also advanced applications such as perfusion. While we are about to reach this level of sophistication and comfort for the patient and the staff, it has to be stressed that our ancestors were pioneers who deserve our full consideration and respect.


The Topic focused on recent research conducted in the field of metabolism and growth, and aimed to address key questions about the interplay between nutritional, environmental, or other external factors  and the endocrine system, as well as the modulation of signals involved in the control of feed intake, regulating these processes.Metabolism and growth are under the control of the endocrine system that, working in cooperation with the nervous system, regulates these functions. The objective of this Research Topic was to provide a multidisciplinary approach of cutting-edge research on metabolism and growth in different experimental models, including farmed species. The Research Topic contains 12 contributions, comprising 7 original research articles, 3 reviews, and 2 minireviews. These works include a wide range of cellular and Pérez-Sánchez et al. reviewed the evolution of the GH, prolactin, and somatolactin family of peptides and their sub-functionalization in marine fish species, and revisited the direct and indirect effects of GH and IGF-I on growth and development. In addition to growth, the GH/IGF-I system controls other physiological functions including metabolism and mineral balance, and, to properly grow and meet metabolic demands, hormones produced from the brain and peripheral tissues contribute regulating feeding  of different genetic background, demonstrating that initial body reserves affect whether or not GH resistance is acquired under catabolic physiological conditions. In another study in pregnant women treated with Lycium barbarum L. polysaccharides (LBLP), a relation between insulin resistance and secretion, lipid profiles, and miR-33 levels was reported, showing the beneficial effects of LBLP improving the symptoms of gestational diabetes mellitus . Moreover, the close relationship between growth and reproduction has been discussed in a manuscript focusing on the low levels of leptin associated with the overexpression of GH in transgenic common carp (Cyprinus carpio) and the resulting delay in puberty onset .From fish to mammals, the growth hormone (GH)/insulin-like growth factor I (IGF-I) axis is the major endocrine system stimulating growth , 2, indi feeding , 4. The Bertucci et al.). Secondly, the recent knowledge achieved concerning the interaction of stress with the different neurosensing mechanisms and peripheral endocrine signals such as leptin that regulate food intake in fish have been revised . Then, considering the effects of specific nutrients on metabolism, the physiological and epigenetic changes induced by methionine restriction have been reviewed encompassing both, mammalian and fish models . Next, specifically in gilthead seabream (Sparus aurata), the effects of dietary supplementation with the aminoacid derivative creatine on muscle growth and quality, have been explored . Increasing dietary creatine enlarged dorsal cross-sectional muscle area and modulated the expression of genes related to muscle growth  and texture , although further studies are required to better clarify these effects in fish.Among the external signals modulating growth and metabolism, diet availability and composition are the most notable. In line with this, one review within the Topic has explored the nutritional factors that through influencing the endocrine system control feeding and growth in fish, describing the nutrients as well as the brain and peripheral hormones that regulate food intake , that a wide thermal range is associated with significant increases in gene transcription of circadian-clock related genes and monoamines hormone levels, which decrease aggressive behavior, and positively influence stress, welfare, and growth, whereas a restricted thermal range showed the opposite effects. Moreover, since climate change is a major challenge that humanity is facing nowadays, a study investigated in gilthead sea bream, the effects of three rearing temperatures  on growth and lipid metabolism using in vivo and in vitro approaches . Increasing temperatures caused unfavorable musculoskeletal growth conditions due to reduced expression of GH/IGF-I system members and specific MRFs genes. Moreover, to cope with the increased energy needs, lipid metabolism was induced in the muscle although not efficiently.Considering environmental signals, two articles within the Topic have explored the effects of temperature on metabolism and growth in different fish species. Maradonna and Carnevali). Moreover, the effects of several EDCs individually and in mixture were tested on male fathead minnows  through analysis of liver transcriptomes by expression microarrays and subsequent quantitative PCR, identifying distinct modes of action for the different chemicals as well as pinpointing a number of new biomarkers that can potentially be used for environmental screening .Besides abiotic factors, environmental contaminants are known to cause adverse health effects, impairing not only reproduction but also metabolism and development in wildlife and humans, since there is evidence that endocrine disruptive chemicals (EDCs) can interact with a variety of hormones and/or hormone receptors, exerting actions as agonists or antagonists , 6. In tTaken together, these studies have demonstrated the effects of dietary and environmental factors on metabolic processes, and the complex interactions of all these factors on the control of the endocrine system regulating growth and metabolism mainly in model organisms and farmed species.OC and EC drafted and critically reviewed the article.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.


Kinetoplastida trypanosomatidae microorganisms are protozoan parasites exhibiting a developmental stage in the gut of insect vectors and tissues of vertebrate hosts. During the vertebrate infective stages, these parasites alter the differential expression of virulence genes, modifying their biological and antigenic properties in order to subvert the host protective immune responses and establish a persistent infection. One of the hallmarks of kinetoplastid parasites is their evasion mechanisms from host immunity, leading to disease chronification. The diseases caused by kinetoplastid parasites are neglected by the global expenditures in research and development, affecting millions of individuals in the low and middle-income countries located mainly in the tropical and subtropical regions. However, investments made by public and private initiatives have over the past decade leveraged important lines of intervention that if well-integrated to health care programs will likely accelerate disease control initiatives. This review summarizes recent advances in public health care principles, including new drug discoveries and their rational use with chemotherapeutic vaccines, and the implementation of control efforts to spatially mapping the kinetoplastid infections through monitoring of infected individuals in epidemic areas. These approaches should bring us the means to track genetic variation of parasites and drug resistance, integrating this knowledge into effective stewardship programs to prevent vector-borne kinetoplastid infections in areas at risk of disease spreading. Protozoan infections are one of the most devastating causes of human death worldwide. These infections are caused by protozoan parasites, microorganisms originally classified in the Kingdom Protozoa, which comprises a diverse group of unicellular eukaryotes . AlthougTrypanosoma brucei [human African trypanosomiasis (HAT) or sleeping sickness], Trypanosoma cruzi (Chagas disease), and Leishmania spp. (leishmaniasis)  , 4, whicaniasis)  5)..TrypanosAlthough the advances in the development of drug therapies and vector control agents against kinetoplastid diseases , new strIn spite of the low priority of the investments received, studies in the past decades have pointed out the paths for developing effective actions to control human kinetoplastid protozoan infections through a better understanding of the pathogen–vertebrate host interactions during their life cycles, disease pathogenesis in the hosts, and methods that allow the diagnosis, even in the acute phase of the infection , 4. ThisTreatment approaches based on drug therapies in kinetoplastid infections are scarce and have been considered to be highly toxic . HoweverT. cruzi parasitism in infected patients. Furthermore, its use in combination with benzanidazole is more effective as compared to monotheraphy-based protocols  is covered by a dense coat of glycoconjugates that play important roles in the evasion of host immune responses  and TSA-1 (Trypomastigote surface antigen-1) by a global consortium (In the case of Chagas disease, initial studies have pointed out potential antigens in experimental mice models capable of inducing host protective immune responses. They were also effective in reducing cardiac parasite loads and disease pathology, increasing host survival indices. Those studies have proposed the use of Tc24 (calcium binding protein associated with flagellar pocket of nsortium –42. ThesTo eradicate kinetoplastid infections in humans, we must gain a better understanding of variants of pathogens and their vectors, the transmission models among their hosts, and efficacious preventive vaccine approaches, to understand and design strategies to intervene in the real dynamics of disease spreading in epidemic areas . To achiThe GIS technology is able to compile multi-analysis related to public health data collected from endemic urban and wild areas, including analyses of disease dispersal, genetic variation of parasites, vector and host habitats, and their relationship with microclimates in affected areas. These metadata analyses can be correlated with geographic positioning system to establish interventions of potential infections, thus allowing for a rational use of medications to prevent the emergence of drug resistance in the context of disease surveillance , 45. NowLeishmania spp. together with surveillance has been demonstrated to be of great value in predicting areas of risk. Epidemiological analysis of the transmission cycles in Europe, and in recent kala-azar outbreaks in Nepal, Kenya, and Brazil, have helped to precisely define that the priority of intervention is in the vector control with the use of insecticides, in order to block the transmission cycle of disease in potential risk areas, identified to be priority to reduce the spread of infection (In kinetoplastid infections, spatial clustering using GIS technology when applied with accurate diagnostic test, analysis of the phylogenetic distribution of nfection –49.Neglected tropical diseases caused by kinetoplastid protozoan parasites are considered endemic in lower-middle-income economies, although in many countries they are in process of epidemiological containment and elimination. However, the eradication of these diseases is a complex issue that must involve health management policies, coordinated among the affected countries, such as the development of public policies to combat parasitic diseases and improvement of social conditions. This will only be possible through the cooperation of local governments in a participatory search for bilateral relations between public and private interests capable of fostering translational studies that guarantee the access of the population to appropriate treatments. The development of accurate diagnosis capable of identifying these diseases in the acute phase of infection, as well as the identification of parasite polymorphisms and variants of drug resistance, together with application of health care models based on GIS technologies will enable both preventive and therapeutic actions based on new generations of drugs and vaccines currently developed for these neglected diseases.AM conceived the manuscript. AAF and AM wrote the manuscript. AAF, KG-P, MPN, KZ, LF, LP, DON, LC, and AM participated in the preparation 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.


Previous studies have reported the use of complementary therapies to reduce the risk of gout attacks. In this study, we assessed the effectiveness of cherries in reducing uric acid levels associated with gout.  We searched for relevant studies on PubMed, Embase, and the Cochrane Library without restrictions on language from inception until August 15, 2019. The risk of bias was evaluated using the PRISMA statement and checklist, and the methodological quality was assessed using the Cochrane Collaboration tool.  The six studies included in this systematic review reported decreases in the incidence and severity of gout following the ingestion of cherries. Gout patients regularly ingesting cherry extract/juice reported fewer gout flare ups than those patients who did not supplement their diets with cherry products. Overall, we observed a positive correlation between the consumption of tart cherry juice and a decrease in serum uric acid concentration.  Current evidence supports an association between cherry intake and a reduced risk of gout attacks. Note however that we were unable to conduct effective meta-analysis due to a lack of relevant studies and a high degree of variation in the methodologies and metrics used in previous studies. Further comprehensive trials or long-term follow-up studies will be required to evaluate the efficacy of cherry intake in treating patients with gout or hyperuricemia. Gout refers to the crystallization of uric acid in or around the joints. It is a prevalent consequence of inflammatory arthritis , commonlCherry is a nutritious fruit containing a variety of chemical compounds, which have proven beneficial to patients with gout, insomnia, and sore muscles. It has also proven beneficial to patients with chronic diseases, such as cardiovascular disease, diabetes, and cancer . From thThere is evidence indicating that the consumption of cherry can reduce uric acid concentrations; however, more substantial and integrative results are required. Our objective in the current systematic review was to assess the effectiveness of cherry products in reducing uric acid levels and mediating the incidence of gout attack.We searched through PubMed, Embase, and the Cochrane Library for relevant studies (without language limitations) from inception until August 15, 2019. These databases cover most of the research articles pertaining to this topic. Eligible studies were identified by scanning electronic databases using various combinations of Medical Subject Headings (MeSH) and non-MeSH terms.The search process was extended by (i) perusing the reference section of all relevant studies and (ii) manually searching through the abstracts of key journals and papers published at major annual meetings. The search terms included the following: (gout OR hyperuricemia) AND (cherry or cherry juice or cherry extract) AND (efficacy or effectiveness). We also checked the reference list of screening studies to identify other similar studies. The search strategy excluding manual search methods is shown in A data extraction form was used to obtain the following data from the included studies: first author, publication year, country, database used, study duration, study design, study subjects, mean age of study subjects, assigned groups, and outcomes. An assessment of methodological quality was performed independently by the authors Pei-En Chen and Tao-Hsin Tung. The quality assessment included the following items: allocation generation and concealment, blinding, follow-up duration, loss follow-up (%), and data-analysis method (intention-to-treat or per protocol). Discrepancies were resolved through discussion and consensus. Two authors also independently reviewed the titles and abstracts after all of the references from relevant studies had been imported to EndNote. After a thorough appraisal of selected publications, we indexed the full text and subsequently assessed the risk of bias using the Cochrane Handbook for Systematic Reviews of Interventions . The hanμmol/L), the number of gout flare incidents, the risk of gout attacks (OR), serum urate levels, and serum uric acid concentrations.The outcomes of the selected studies were qualitatively assessed, with a focus on plasma urate level  in affected joints [β secretion [The ingestion of cherry has proven effective in lowering urate levels, and previous studies have attributed the suppression of gout-related inflammation to the anti-inflammatory properties of cherry . Key enzd joints . A failud joints . These fd joints  and Coliecretion . Note thβ and TNF-α, which are used as biomarkers in monitoring acute gout flare ups [The strength of the study resides in the quality of the review. We separately evaluated the studies using assessment tools, and it appears that we covered most of the articles dealing with the relationship between cherry consumption and gout. The studies included in our review demonstrate the influence of time in the relationship between gout attacks or plasma uric acid levels and cherry intake. Numerous studies have investigated the relationship between cherry and gout or uric acid; however, the focus has been on the anti-inflammatory effects of IL-1 lare ups , 10, 19.Several limitations need to be addressed. First, the small number of included studies calls into question the reliability of the findings and the strength of the conclusions. Second, it was difficult to conduct effective meta-analysis due to considerable heterogeneity in the design of the studies, the outcome variables, and the biomarkers used to assess the risk of bias . Third, Current evidence supports an association between cherry intake and a reduced risk of gout attacks. Note however that we were unable to conduct effective meta-analysis due to a lack of relevant studies and a high degree of variation in the methodologies and metrics used in previous studies. Furthermore, there are at present an insufficient number of studies presenting the quantitative result required to conduct effective meta-analysis. We suggest that researchers conduct randomized control trials to further assess this correlation. We also recommend research on a wider range of populations  to provide more comprehensive and generalizable findings.


Due to the merits of Lamb wave to Structural Health Monitoring (SHM) of composite, the Lamb wave-based damage detection and identification technology show a potential solution for the insulation condition evaluation of large generator stator. This was performed in order to overcome the problem that it is difficult to effectively identify the stator insulation damage the using single feature of Lamb wave. In this paper, a damage identification method of stator insulation based on Lamb wave multi-feature fusion is presented. Firstly, the different damage features were extracted from time domain, frequency domain, and fractal dimension of lamb wave signals, respectively. The features of Lamb wave signals were extracted by Hilbert transform (HT), power spectral density (PSD), fast Fourier transform (FFT), and wavelet fractal dimension (WFD). Then, a machine learning method based on support vector machine (SVM) was used to fuse and reconstruct the multi-features of Lamb wave and furtherly identify damage type of stator insulation. Finally, the effect of typical stator insulation damage identification is verified by simulation and experiment. Stator insulation of large generators has a laminated composite structure. In long-term service, stator insulation is exposed to a combination of electrical, thermal, and mechanical stresses. They gradually cause the aging of insulation structure, resulting in internal or surface insulation damages, such as void, delamination, puncture crack, and surface crack, etc. These damages eventually can lead to failure and breakdown of large generator stator insulation ,2,3,4. TImplement of condition monitoring (CM) on stator insulation holds the promise of enhancing the safety and economic operation of a large generator. Structural Health Monitoring (SHM) is a promising method for the non-destructive defect detection of composites . Its purThe type of stator insulation damage can reflect the state of stator insulation aging to a certain extent. So the degree of stator insulation aging can be evaluated by identifying typical stator insulation damage categories. Due to the multi-mode and dispersion of Lamb wave, it is difficult to explain the damage detection signal, which seriously affects the effect of damage identification. In addition, the stator insulation of large motor is a composite material structure, and the Lamb wave propagation process is very complex, the reflection and scattering of Lamb wave make it difficult to extract the damage features, thus causing great obstacles to the identification of damage types. In particular, using a single damage feature for damage identification can also reduce the accuracy of stator insulation damage type identification. Moreover, the traditional damage classification method often depends on manual intervention and experience, and the results of damage identification are quite different. With the rapid development of computer technology, especially machine learning algorithms, intelligent damage identification has become an inevitable trend. At present, among the common machine learning algorithms, clustering algorithm (CL)  classifiThis paper is organized as follows. In Lamb wave is a kind of guided wave propagating in plate structure. When there is damage in stator insulation structure, Lamb wave will reflect and scatter with the damage, which will have different effects on Lamb wave signal. By analyzing Lamb wave signal, the existence and degree of insulation damage can be detected. The principle of damage identification based on Lamb wave is shown in The feature extraction of stator insulation damage is an important part for damage identification. In this paper, the stator insulation damage features are extracted from time domain, frequency domain and fractal dimension respectively. These methods are described as follows.x(t) in time domain, its HT definition is shown by the following [The energy distribution characteristic of Lamb waves signals can be obtained by Hilbert transform ,16. The ollowing :(1)H(t)The envelope of Hilbert transform can represent the energy distribution feature of the signalin time domain, and the damage characteristic of stator insulation can be reflected by extracting the amplitude of the envelope. In order to obtain the envelope of Lamb wave signal in time domain. The analytic signal can be expressed as:By extracting frequency components and amplitude characteristics of lamb wave signal in frequency domain, which can be used for identifying the damage of stator insulation. Based on frequency domain analysis theory, the Fast Fourier transform (FFT) ,19 and pAccording to the symmetry and periodicity of The results of previous research  indicateDue to the fractal dimension can quantitatively characterize the irregularity and self-similarity features of signals , which hD can be obtained as:Firstly, the range of each frequency band is determined by wavelet decomposition, the Lamb wave signal is decomposed by wavelet transform. The frequency range of detail component Based on the extraction of damage characteristics, multi-feature fusion and damage identification of stator insulation can be further carried out.Damage classification method of stator insulation based on single feature can result in the problem of low rate of damage identification. Multi-feature fusion identification is a comprehensive processing of target multi-feature information, which can obtain more accurate and comprehensive estimation and judgment than single feature. In order to identify the type of stator insulation damage effectively, Hilbert transform (HT), power spectral density (PSD) analysis, fast Fourier transform (FFT) and wavelet fractal dimension (WFD) methods were respectively used to extract features of Lamb wave signals received in stator insulation damage detection. In addition, since these damage features are directly related to the geometric dimension of insulation damage, the effect of stator insulation damage identification can be further improved if the geometric dimension of insulation damage and the extracted multi-features are comprehensively considered. So support vector regression (SVR) algorithm is used to predict the damage size based on the extracted damage features in this paper, then the geometric dimension and the extracted features of stator insulation damage are fused to further identify the type of stator insulation damage.The design of classifier is an important part of insulation damage identification. In practical application, it is very difficult to obtain sufficient typical damage samples of stator insulation. Therefore, in the case of small samples, the design of classifier is very critical, which directly determines the effect of stator insulation damage identification. Because SVM has obvious advantages in solving small sample, non-linear and high-dimensional pattern recognition problems  and has n is the number of training samples, d is the dimension of each training sample, and b is the scalar threshold. Lagrange principle is used to solve the optimization problem and the optimal separating hyperplane is obtained as:M is the number of support vectors. The general form of SVM classifier can be expressed as:x can be determined by the positive or negative of classifier The classical support vector machine (SVM) is mainly designed for binary classification problem, and its design principle is as follows. It is assumed that the given training data sample set is  follows :(10)y(x)n is the number of training samples, d is the dimension of each training sample, and N is the category of stator insulation damage. The classification output function of the kth  is used to fuse the damage geometric dimensions and the extracted damage features. Combining the geometric dimension information of stator insulation damage and the extracted damage features information, the SVR training samples are constructed and the SVR regression model is trained. Finally, by fusing the damage features with the regression predicted geometric dimensions, the categories of the stator insulation typical damage testing samples can be identified effectively.In this study, the finite element method (FEM) software ABAQUS is used to establish the stator insulation damage model, and the effectiveness of the stator insulation damage identification method based on Lamb wave multi-feature fusion is verified by simulation analysis data.3, and Poisson’s ratio of 0.2. Moreover, PZT wafers are surface-attached on the insulation of stator bar using an adhesive epoxy to establish the active sensor network. PZT wafers are configured in the middle of widths (x direction) and 150 mm apart in z direction, as illustrated in The three-dimensional FEM model of the stator bar is established with geometry of 60 mm in width (x direction), 30 mm in height (y direction) and 1200 mm in length (z direction), as shown in The typical stator insulation damage of large generator mainly includes four kinds, i.e., surface crack, longitudinal crack, inner delamination, and inner cavity. From the perspective of industrial application, insulation damage may occur at any location of stator bar. In this paper, a PZT sensor network is constructed to realize excitation and reception of Lamb, so as to acquire insulation damage detection signals for stator insulation damage identification. In PZT sensor networks, any PZT can either excite Lamb waves or receive Lamb waves. Therefore, in practical applications, there are multiple paths for Lamb wave excitation and reception . In this way, the possible damage at any location can be detected through sensor networks. In this study, only one example is given to illustrate the relationship between excitation and reception. The working modes of all paths are similar. According to the According to ,9,30, whAs shown in It is seen from To overcome the problem of low identification rate of single feature, the type of stator insulation damage is identified based on multi-feature fusion. According to the four damage features, the insulation damage sizes In order to evaluate the quality of the regression model predicting results, a relative error evaluation method is used to this issue in this paper. The relative error calculation formula is as follows:According to The predicted insulation damage sizes are fused with the damage features to identify the insulation damage. Compared with the identification effect based on damage features of Lamb wave, the results are shown in To verify the damage identification effect of large generator stator insulation based on Lamb wave multi-feature fusion, a damage identification experiment is performed. The experimental system is set up, as shown in In the system, an arbitrary waveform generator (AFG3022B) is used to generate Lamb wave excitation waveforms. A power amplifier (7602M) is used to drive PZT actuator. An oscilloscope (DPO3014) can acquire Lamb wave signals. DELL T7600 workstation is used for signal processing. PZT wafers are attached to the surface of the stator insulation. PZT A9 acts as an actuator to excite the Lamb wave signal, and PZT A1~A8 act as sensors to receive the Lamb wave propagation signals. In the experiment, the arbitrary waveform generator generates a preset excitation signal to PZT A9, and PZT A1~A8 receive the Lamb wave signals which can be collected and saved by oscilloscope for further analysis and processing.The stator bar specimens used in this experiment are taken from a large generator with a rate of 18 kV/300 MW. As shown in Similar to the numerical simulation analysis, the features of received Lamb wave signals are extracted separately, and the type of insulation damage is identified by SVM classifier and ANN classifier. The damage identification results based on single damage feature and multi-feature fusion are shown in From In this paper, a damage identification method of stator insulation based on Lamb wave multi-feature fusion is proposed. From the simulation and experimental results, the following conclusions can be drawn:The damage features of stator insulation extracted by HT, PSD, FFT, and WFD vary with the different size of damage. It indicates that the damage features extracted can effectively characterize the damage features of stator insulation, which lays a foundation for damage identification.According to the stator insulation damage features, the type of stator insulation damage can be identified by SVM classifier. In addition, the classification effect of SVM is better than that of ANN classifier under the same conditions.The accuracy of stator insulation damage identification using Lamb wave multi-feature fusion is higher than that of single feature. Furthermore, the insulation damage identification accuracy can be further improved by predicting the damage geometry size and fusing with Lamb wave features, which can provide effective reference information for stator insulation condition assessment of large generators.


Topology, in its many forms, describes relations. It has thus long been a central concept in neuroscience, capturing structural and functional aspects of the organization of the nervous system and their links to cognition. Recent advances in computational topology have extended the breadth and depth of topological descriptions. This Focus Feature offers a unified overview of the emerging field of topological neuroscience and of its applications across the many scales of the nervous system from macro-, over meso-, to microscales. From the early drawings of Ramon y Cajal to today, topological descriptions have played a central role in neuroscience. In recent years, thanks to advancements in both mathematical tools and data availability, the range and diversity of such descriptions are expanding rapidly, spanning theoretical, computational, and experimental approaches to brain connectivity. This Focus Feature on “Topological Neuroscience” aims at presenting the breadth of applicability of topological data analysis (TDA) methods in neuroscience across scales and modalities.Computational topology offers new frameworks for both the analytical description and the understanding of brain function. A common denominator to these new tools is their ability to find meaningful simplifications of high-dimensional data. As such, TDA aims to capture mesoscale patterns of disconnectivity and explicitly encode higher order interactions, that is, interactions between more than two regions or components (Giusti, Ghrist, & Bassett, Recent seminal research has shown the potential and impact of topological approaches. Topological differences have been found at the population and individual levels in functional connectivity (Lee, Chung, Kang, Kim, & Lee, This Focus Feature offers a unified overview of this emerging field of topological neuroscience and of its applications across many scales of the nervous system from macro-, over meso-, to microscales. First, Sizemore, Phillips-Cremins, Ghrist, and Bassett  provide It is now high time to put topological neuroscience center stage and to bring together the growing but often separate communities involved in applied topological analysis. Still, numerous challenges and questions remain before TDA methods become widely accepted and can come to realize their full potential. Notably, more research is needed both in terms of contextualization and functional interpretation of topological features (Lord et al.,


Frontiers in Behavioral Neuroscience, Frontiers in Immunology, and Frontiers in Neurology, we have collected original works and perspectives that provide new insights on the clinical relevance of immune-to-brain and brain-to-immune communications.Experimental and clinical evidence demonstrates an intense crosstalk among the nervous, endocrine and immune systems Dantzer, . The ceninflammation-associated depression  in response to an inflammatory challenge. Although inflammation lowered mood, negative self-schema was not significantly impacted. This indicates that negative self-schemata might not be a mechanism by which inflammation leads to depression. In their review, Lacourt et al. provide new insights on the mechanisms underlying chronic fatigue. They propose that the chronic activation of inflammatory processes at a low-grade state, such as observed in chronic fatigue syndrome , a demyelinating disease affecting optic nerve and spinal cord, exhibited higher plasma levels of pro-inflammatory cytokines and of eosinophil chemoattractants, which was, in turn, associated with the number of relapses. The authors suggest that inflammatory activity could contribute to the pathophysiology of NMOSD by increasing eosinophil chemoattractants, and thus favoring eosinophil infiltration, which is a specific feature of NMOSD , but not in patients with mild cognitive impairment with negative CSF AD's biomarkers and in healthy controls. This indicates a potential role of the adaptive immune system in the pathophysiology of AD.As highlighted by Herz et al. and Tadros et al. reveal the importance of the immune system for the central nervous system during the neonatal period. Based on previous research  neurons is a mechanism by which an immune challenge during the neonatal period leads to heighten inflammatory pain later in life. The authors found that frequency of spontaneous excitatory synaptic currents of SDH neurons was increased in the group of rats having received the neonatal immune challenge compared to saline. This reflects a hyper-excitability of the SDH neuronal network, which could contribute to the altered nociceptive signaling observed after a neonatal immune challenge.The studies from Bajic et al. describe how the gut-brain axis is likely to be involved in the psychological and neurological complications of chemotherapy. Chemotherapy induces mucositis, associated with an inflammatory response, changes in the gut microbiome, increased intestinal permeability, and damages to the nerves of the myenteric plexus. These changes cause a disruption in the gut-brain axis, which is known to alter mood and cognition. Langgartner et al. show that rectal transplantation of feces from non-stressed mice partially reversed the physiological  and behavioral (anxiety) effects of chronic psychosocial stress (induced by housing with a dominant CD-1 mouse). This indicates a potential role of gut microbiota in some of the physiological and behavioral changes induced by chronic psychosocial stress.Gut immunity and the gut-brain axis play an important role in modulating CNS functions  presented a specific inflammatory profile that was not observed in resilient mice. These findings show a strong interaction between stress vulnerability and immunological consequences.Dysregulation of peripheral innate immune responses is observed in stress-associated psychiatric disorders, such as major depression  outcomes. Mindfulness has been shown to improve quality of life, depression and fatigue in MS patients, although the underlying mechanisms remain unknown. The authors provide data supporting immunomodulatory effects of this treatment, and call for the measurement of immunological and neuroimaging biomarkers in studies assessing the effect of mindfulness in MS.Lastly, and animal studies, along with review articles reflecting on the current knowledge, in the quest for a better comprehension of the brain-immune mechanisms and their clinical relevance.The articles combined in this special issue highlight the central role of immune-to-brain and brain-to-immune communications in various clinical pathologies, including, but not limited to, depression, anxiety, fatigue, pain, stress-related disorders, neurological diseases, and inflammatory diseases. This special issue also reveals the importance of clinical JL and MH have written the draft of this editorial. MS and ML have provided critical feedback. All authors approve the final 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.


Managing hyperphosphatemia is one of the most significant challenges in the treatment of individuals with chronic kidney disease (CKD) particularly those on dialysis. Decreased excretion of phosphorus in individuals with CKD results in phosphorus overload, a condition that stimulates the synthesis of fibroblast growth factor 23 (FGF-23) and parathyroid hormone (PTH), two factors involved in the development of chronic kidney disease-mineral and bone disorder (CKD-MBD). CKD-MBD increases the risk for cardiovascular disease, death, and deteriorates quality of life.Organic and inorganic phosphorus can be found in a wide variety of foods. Animal-based foods, such as meat and dairy products, are among the main sources of organic phosphorus in omnivorous diets. The phosphorus present in these foods is more easily digested than the phosphorus from plant-based foods , in which most phosphorus is bound to phytate, a carbohydrate that cannot be digested by the enzymes in the human gastrointestinal tract, thus hampering the absorption of phosphorus.Despite the data presented above, the intake of sources of phosphorus by patients with CKD in Brazil requires further analysis. In this context, the study performed by Nerbass et al.The authors used a food frequency questionnaire designed to assess phosphorus intake and noted that, in general terms, the most frequent reported items were milk and beans. The frequency score for food sources of organic phosphorus was higher than the score for inorganic phosphorus, but only the inorganic phosphorus scores were associated with hyperphosphatemia, an indication of the potential negative effects of phosphorus-containing additives on individuals with CKD. When the two States were compared, Santa Catarina had a higher prevalence of hyperphosphatemia than Tocantins, although the patients underwent the same clinical protocol to manage CKD-MBD. Therefore, although the study did not include a quantitative assessment of phosphorus intake, differences in the food items reported in each State may explain the difference in serum phosphate levels seen in serum phosphate levels. Pork, dairy products, and sausages were more commonly eaten in Santa Catarina, whereas in Tocantins the main food items consumed were foods were beef and beans. The results reported by the authors suggest that the intake of inorganic phosphorus-containing food additives present in sausages and probably in dairy products was higher in Santa Catarina than in Tocantins.The strategy adopted by Nerbass et al. to assess food intake is in line with current research practice current research practice that attempts to comprehend the relationship between diet and clinical outcomes based on food groups and eating habits instead of isolated nutrients. This approach has advantages since it considers synergies between foods, nutrients, and bioactive compounds, in addition to facilitating the communication of dietary advices between healthcare workers and patients. Furthermore, the assessment of isolated nutrients may be more susceptible to measurement errors, due to the limited information on phosphorus contents in food composition tables and processed food labels, which lead to the underestimation of phosphorus intake.The study indicates that nutrition education strategies for the management of to hyperphosphatemia should be based on the particular eating habits of each individual population. Therefore, studies based on qualitative assessment of food intake that support the production of scientific evidence to guide more effective dietary interventions in the management of hyperphosphatemia, are still necessary.