Architecture and regulation of a GDNF-GFRα1 synaptic adhesion assembly.

Glial-cell line derived neurotrophic factor (GDNF) bound to its co-receptor GFRα1 stimulates the RET receptor tyrosine kinase, promoting neuronal survival and neuroprotection. The GDNF-GFRα1 complex also supports synaptic cell adhesion independently of RET. Here, we describe the structure of a decameric GDNF-GFRα1 assembly determined by crystallography and electron microscopy, revealing two GFRα1 pentamers bridged by five GDNF dimers. We reconsitituted the assembly between adhering liposomes and used cryo-electron tomography to visualize how the complex fulfils its membrane adhesion function. The GFRα1:GFRα1 pentameric interface was further validated both in vitro by native PAGE and in cellulo by cell-clustering and dendritic spine assays. Finally, we provide biochemical and cell-based evidence that RET and heparan sulfate cooperate to prevent assembly of the adhesion complex by competing for the adhesion interface. Our results provide a mechanistic framework to understand GDNF-driven cell adhesion, its relationship to trophic signalling, and the central role played by GFRα1.

BSHI/BTS guidance on crossmatching before deceased donor kidney transplantation.

All UK H&I laboratories and transplant units operate under a single national kidney offering policy, but there have been variations in approach regarding when to undertake the pre-transplant crossmatch test. In order to minimize cold ischaemia times for deceased donor kidney transplantation we sought to find ways to be able to report a crossmatch result as early as possible in the donation process. A panel of experts in transplant surgery, nephrology, specialist nursing in organ donation and H&I (all relevant UK laboratories represented) assessed evidence and opinion concerning five factors that relate to the effectiveness of the crossmatch process, as follows: when the result should be ready for reporting; what level of donor HLA typing is needed; crossmatch sample type and availability; fairness and equity; risks and patient safety. Guidelines aimed at improving practice based on these issues are presented, and we expect that following these will allow H&I laboratories to contribute to reducing CIT in deceased donor kidney transplantation.

C-Reactive Protein and Covid-19 Severity: A Systematic Review.

BACKGROUND: COVID-19 clinical course has been quite unpredictable and efforts have been made to identify reliable markers that will help in early disease progression, prognosis and severity detection. Objective: This study thus aimed to provide evidence that will guide clinical management by reviewing studies that assessed CRP concentration and COVID-19 severity/outcome. METHODS: Three electronic databases, PubMed/Medline, Google Scholar, and JSTOR were searched to identify studies available online as at 1st September 2020 which assessed COVID-19 clinical outcome and CRP concentration. The search strategy involved words combination like "C-reactive protein" OR "inflammatory markers" OR "acute phase reactants" and "coronavirus 2019" OR ''COVID-19" OR "2019-nCoV" OR "SARS-CoV-2". RESULTS: Sixty-one articles were systematically reviewed out of 812 studies identified after duplicates were removed. The 61 studies comprised 13,891 COVID-19 patients made of 7,840 (56.4%) males and 6,051 (43.6%) females. All the papers revised were observational studies except one case-control and they cut across fifteen countries. The result of the review demonstrated that the severe cases had higher levels of C - reactive protein when compared to the mild cases in all the studies (100%). The increase in C-reactive protein was statistically significant in 78.7% of the cases. CONCLUSION: High levels of CRP are associated with COVID-19 severity. Highlights: Severe cases of COVID-19 is characterized with higher CRP levels. COVID-19 cases should be screened regularly for CRP to monitor severity.

Contexte: L'évolution clinique du COVID-19 a été assez imprévisible et des efforts ont été faits pour identifier des marqueurs fiables qui aideront à la progression précoce de la maladie, au pronostic et à la détection de la gravité. Objectif : Cette étude visait donc à fournir des preuves qui guideront la gestion clinique en passant en revue les études qui ont évalué la concentration de CRP et la gravité/l'issue du COVID-19. Méthodes: Trois bases de données électroniques, PubMed/Medline, Google Scholar et JSTOR, ont été consultées pour identifier les études disponibles en ligne au 1er septembre 2020 qui évaluaient le résultat clinique du COVID-19 et la concentration de CRP. La stratégie de recherche comportait des combinaisons de mots comme "protéine Créactive" OU "marqueurs inflammatoires" OU "réactifs de phase aiguë" et "coronavirus 2019" OU "COVID-19" OU "2019-nCoV" OU "SARS-CoV-2". Résultats: Soixante et un articles ont été systématiquement examinés sur les 812 études identifiées après suppression des doublons. Les 61 études comprenaient 13 891 patients atteints de COVID-19, dont 7 840 (56,4 %) hommes et 6 051 (43,6 %) femmes. Tous les articles examinés étaient des études d'observation, à l'exception d'un cas-témoin, et ils couvraient quinze pays. Le résultat de l'examen a démontré que les cas graves avaient des niveaux plus élevés de protéine C-réactive par rapport aux cas légers dans toutes les études (100%). L'augmentation de la protéine C-réactive était statistiquement significative dans 78,7% des cas. Conclusion: Des niveaux élevés de CRP sont associés à la sévérité du COVID-19. Mots clés: Protéine C-réactive, COVID-19, SRAS-COV-2 et Coronavirus. Points forts: Les cas graves de COVID-19 sont caractérisés par des niveaux de CRP plus élevés. · Les cas de COVID-19 doivent faire l'objet d'un dépistage régulier de la CRP afin de surveiller la gravité de la maladie.

Three-dimensional visualization as a tool for interpreting locomotion strategies in ophiuroids from the Devonian Hunsrück Slate.

Living brittle stars (Echinodermata: Ophiuroidea) employ a very different locomotion strategy to that of any other metazoan: five or more arms coordinate powerful strides for rapid movement across the ocean floor. This mode of locomotion is reliant on the unique morphology and arrangement of multifaceted skeletal elements and associated muscles and other soft tissues. The skeleton of many Palaeozoic ophiuroids differs markedly from that in living forms, making it difficult to infer their mode of locomotion and, therefore, to resolve the evolutionary history of locomotion in the group. Here, we present three-dimensional digital renderings of specimens of six ophiuroid taxa from the Lower Devonian Hunsrück Slate: four displaying the arm structure typical of Palaeozoic taxa (Encrinaster roemeri, Euzonosoma tischbeinianum, Loriolaster mirabilis, Cheiropteraster giganteus) and two (Furcaster palaeozoicus, Ophiurina lymani) with morphologies more similar to those in living forms. The use of three-dimensional digital visualization allows the structure of the arms of specimens of these taxa to be visualized in situ in the round, to our knowledge for the first time. The lack of joint interfaces necessary for musculoskeletally-driven locomotion supports the interpretation that taxa with offset ambulacrals would not be able to conduct this form of locomotion, and probably used podial walking. This approach promises new insights into the phylogeny, functional morphology and ecological role of Palaeozoic brittle stars.

Effects of Digit Orientation on Gecko Adhesive Force Capacity: Synthetic and Behavioral Studies.

In this study we developed an analytical relationship between adhesive digit orientation and adhesive force capacity to describe the tendencies of climbing organisms that use adhesion for climbing to align their toes in the direction of loading, maximizing adhesive force capacity. We fabricated a multi-component adhesive device with multiple contact surfaces, or digits, to act as a model system mimicking the angular motion of a foot and found the synthetic experiments agree with the developed analytical relationship. In turn, we find that observations of gekkonid lizards climbing on vertical substrates correlate well with our analytical relationship; a reduction in toe spacing is seen on the forelimbs when the animals are facing up. Interestingly, the toes on the hindlimbs tend to have an increase in spacing, possibly a mechanism for stabilization rather than load-bearing.

A Field Guide to Finding Fossils on Mars.

The Martian surface is cold, dry, exposed to biologically harmful radiation and apparently barren today. Nevertheless, there is clear geological evidence for warmer, wetter intervals in the past that could have supported life at or near the surface. This evidence has motivated National Aeronautics and Space Administration and European Space Agency to prioritize the search for any remains or traces of organisms from early Mars in forthcoming missions. Informed by (1) stratigraphic, mineralogical and geochemical data collected by previous and current missions, (2) Earth's fossil record, and (3) experimental studies of organic decay and preservation, we here consider whether, how, and where fossils and isotopic biosignatures could have been preserved in the depositional environments and mineralizing media thought to have been present in habitable settings on early Mars. We conclude that Noachian-Hesperian Fe-bearing clay-rich fluvio-lacustrine siliciclastic deposits, especially where enriched in silica, currently represent the most promising and best understood astropaleontological targets. Siliceous sinters would also be an excellent target, but their presence on Mars awaits confirmation. More work is needed to improve our understanding of fossil preservation in the context of other environments specific to Mars, particularly within evaporative salts and pore/fracture-filling subsurface minerals.

The effect of source of supplemental dietary calcium and magnesium in the peripartum period, and level of dietary magnesium postpartum, on mineral status, performance, and energy metabolites in multiparous Holstein cows.

The objective of this study was to determine the effects of feeding different supplemental sources of Ca and Mg in the peripartum period, and different dietary levels of Mg postpartum, on plasma mineral status, performance, and aspects of energy metabolism in transition dairy cows. Multiparous Holstein cows (n = 41) were used in a completely randomized design with a 2 × 2 factorial arrangement of treatments starting at 28 d before expected parturition. Main effects were source assignments (CS = common sources of supplemental Ca and Mg, or MA = a blend of common and commercial mineral sources with supplemental minerals primarily from a commercial Ca-Mg dolomite source; MIN-AD, Papillon Agricultural Company Inc., Easton, MD) beginning at 21 d before due date; cows were further randomized within source treatments to 1 of 2 levels of Mg supplementation (LM = formulated postpartum diet Mg at 0.30% of dry matter (DM), or HM = formulated postpartum diet Mg at 0.45% of DM) beginning within 1 d after parturition. Final treatment groups included the following: common source, low Mg (CS-LM, n = 11); common source, high Mg (CS-HM, n = 11); MIN-AD, low Mg (MA-LM, n = 10); and MIN-AD, high Mg (MA-HM, n = 9). Treatment diets were fed and data collected through 42 d in milk. Postpartum plasma Mg concentrations tended to be higher for cows fed HM and cows fed CS, but no effects were observed on peripartum plasma Ca concentrations. Peripartum plasma P concentrations were higher for cows fed MA. Dry matter intake (DMI) in the prepartum period was higher for cows fed MA (CS = 15.9 vs. MA = 16.8 kg/d) and postpartum DMI was higher in some groups depending on week. Plasma nonesterified fatty acid concentrations were lower for cows fed MA during both the prepartum and postpartum periods. A source by level interaction was observed for postpartum plasma ß-hydroxybutyrate (BHB) concentrations such that cows fed CS-LM had numerically higher BHB and cows fed MA-LM had numerically lower BHB (geometric means; CS-LM = 7.9, CS-HM = 6.9, MA-LM = 6.3, and MA-HM = 7.3 mg/dL) than cows fed the other 2 treatments. Higher milk fat yield, milk fat content, and fat- and energy-corrected yield during wk 1 for cows fed MA resulted in source by week interactions for these outcomes. This study demonstrated that varying supplemental Ca and Mg sources and feeding rates had minimal effect on plasma Ca status despite differences in plasma Mg and P concentrations. Effects on DMI and plasma energy metabolites suggest an opportunity for strategic use of mineral sources in the transition period to promote metabolic health.

Transitional B lymphocytes are associated with protection from kidney allograft rejection: a prospective study.

Recent cross-sectional studies suggest an important role for transitional B lymphocytes (CD19 + CD24hiCD38hi) in promoting transplant tolerance, and protecting from late antibody-mediated rejection (ABMR). However, prospective studies are lacking. This study enrolled 73 de novo transplant recipients, and collected serial clinical, immunological and biochemical information over 48 ± 6 months. Cell phenotyping was conducted immediately prior to transplantation, and then on five occasions during the first year posttransplantation. When modeled as a time-dependent covariate, transitional B cell frequencies (but not total B cells or "regulatory" T cells) were associated with protection from acute rejection (any Banff grade; HR: 0.60; 95% CI: 0.37-0.95; p = 0.03). No association between transitional B cell proportions and either de novo donor-specific or nondonor-specific antibody (dnDSA; dnNDSA) formation was evident, although preserved transitional B cell proportions were associated with reduced rejection rates in those patients developing dnDSA. Three episodes of ABMR occurred, all in the context of nonadherence, and all associated with in vitro anti-HLA T cell responses in an ELISPOT assay (p = 0.008 versus antibody-positive patients not experiencing ABMR). This prospective study supports the potential relevance of transitional ("regulatory") B cells as a biomarker and therapeutic intervention in transplantation, and highlights relationships between humoral immunity, cellular immunity and nonadherence.

Meeting report: 3rd international transplant conference: how much risk can you take?

The 3rd International Transplant Conference took place on 31st October and 1st November 2014 at the University of Warwick, Coventry, UK. Key focal points of the meeting were the exploration of the molecular basis of antibody-antigen interactions and their relation to clinical practice and to share experiences and knowledge regarding strategies to transplant the 'high-risk' patient. In addition, lively debate sessions were hosted where controversial clinical and immunological themes were discussed by leading experts in the field.

Fighting rabies in Eastern Europe, the Middle East and Central Asia--experts call for a regional initiative for rabies elimination.

MEEREB is an informal network of rabies experts from the Middle East, Eastern Europe and Central Asia, seeking to eliminate rabies from the region. They met for the second time to review the current rabies situation, both globally and in their respective countries, highlighting current rabies control problems and potential solutions. Success stories in Latin America, in Western Europe, in some Asian countries, as well as in Croatia and Serbia prove that elimination of human rabies is achievable in the MEEREB region. It requires political willingness and cooperation of all stakeholders, including Ministries of Health and of Agriculture; adequate management of animal bites through post-exposure prophylaxis; pre-exposure prophylaxis for populations at high risk of rabies exposure, animal vaccination and humane control of stray dog populations. MEEREB members called for a regional initiative for rabies elimination in Eastern Europe and the Middle East. They are confident that the elimination of human rabies of canine origin can be achieved in the region through adopting a One Health approach, and that campaigns for rabies elimination will have significant benefit for public health, including strengthening the structure for control of other zoonoses.

Influence of preformed donor-specific antibodies and C4d on early liver allograft function.

INTRODUCTION. The impact of preformed donor-specific antibodies (DSA) is incompletely understood in liver transplantation. The incidence and impact of preformed DSA on early post liver transplant were assessed and these were correlated with compliment fragment C4d on allograft biopsy. METHODS. Pretransplant serum from 41 consecutive liver transplant recipients (brain dead donors; DBD = 27 and cardiac death donors; DCD = 14) were tested for class-specific anti-human leukocyte antigen (HLA) and compared against donor HLA types. Liver biopsies were taken during cold storage (t-1) and post-reperfusion (t0) stained with C4d and graded for preservation-reperfusion injury (PRI). RESULTS. Of the 41 recipients, 8 (20%) had anti-HLA class I/II antibodies pretransplant, 3 (7%) were confirmed preformed DSA; classes I and II (n=1) and class I only (n=2). No biopsies showed definite evidence of antibody-mediated rejection. Graft biopsies in overall showed only mild PRI with ischemic hepatocyte C4d pattern similar in both positive and negative DSA patients. One DSA-positive (33%) compared with four DSA-negative patients (10%) had significant early graft dysfunction; severe PRI causing graft loss from primary nonfunction was seen only in DSA-negative group. Allograft biopsy of preformed DSA-positive patient demonstrated only minimal PRI; however, no identifiable cause could be attributed to graft dysfunction other than preformed DSA. CONCLUSION. Preformed DSA are present in 5-10% liver transplant recipients. There is no association between anti-HLA DSA and PRI and C4d, but preformed DSA may cause early morbidity. Larger studies on the impact of DSA with optimization of C4d techniques are required.

Prevalence of asthma and respiratory symptoms in children in a low socio-economic status area of Nigeria.

SETTING: Warri and environs in the Niger Delta, in the Southern region of Nigeria. OBJECTIVE: To investigate the burden of respiratory illness in children in the Niger Delta. DESIGN: A cross-sectional survey of 1397 schoolchildren aged 7-14 years in areas of low socio-economic status (SES), using written questionnaires in English based on the International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire, with explanation of symptoms. RESULTS: The prevalence of doctor-diagnosed asthma was 0.9% (95%CI 0.4-1.3), and wheeze in the last 12 months was 5.4% (95%CI 4.3-6.6). The prevalence of other respiratory symptoms was high, in particular night cough 23.3% (95%CI 21.1-25.5), rhinitis 19.2% (95%CI 17.1-21.3) and phlegm production 16.6% (95%CI 14.7-18.6). Risk of wheeze in the younger age groups was twice (OR 2.09, 95%CI 1.29-3.39) that of older ages after controlling for sex and geographical area. Risk of asthma in urban children was reduced (OR 0.15, 95%CI 0.05-0.50) compared to rural children after controlling for age and sex. Urban children also had a lower risk of rhinitis, exercise limitations and absenteeism due to respiratory illness than rural children. CONCLUSION: The prevalence of symptoms of allergic disease was lower than in the African centres in the ISAAC surveys conducted in urban centres. Doctor-diagnosed asthma is likely to be a poor indicator of asthma in low SES areas of developing countries due to health care access issues.

Eliminating canine rabies, the principal source of human infection: what will it take?

More than 50,000 people die of rabies each year; most are children in developing countries, and almost all have been bitten by dogs. Eliminating canine rabies throughout the world would save thousands of lives and would reduce the economic impact of the disease by dramatically reducing the requirement for postexposure prophylaxis (PEP). Lengthy experience in the industrialized countries and ongoing programs in Latin America, Africa, and Asia have shown that the elimination of rabies in dogs is an achievable goal. The presence of canine rabies in developing countries is associated with poverty, and most deaths occur in the lowest socioeconomic sectors. To be successful, national rabies control programs should share responsibility with local communities for prevention and control activities and maintaining disease-free status. Legislation should be adapted to local conditions and the realities of dog ownership. While the provision of PEP to all bite victims is affordable in many countries, it is usually beyond the capacity of impoverished nations, which deal with many other health priorities. Ministries of health should provide PEP, either free or with a charge preferably at a subsidized price, replacing the current system in many countries, in which biologics are sold by government-owned and private clinics at a cost beyond the means of bite victims. The public health sector should assume responsibility when animal control strategies are not effectively implemented or when PEP is not administered correctly or is not available. A global strategy is needed to identify gaps in surveillance and diagnosis, improve access to PEP and enhance canine immunization and population management. Such approaches based on a "One Health" model should be coordinated across regions, and should extend control efforts to other dog-related zoonoses. This article introduces a symposium in Antiviral Research on the elimination of canine rabies.

Significant IgG subclass heterogeneity in HLA-specific antibodies: Implications for pathogenicity, prognosis, and the rejection response.

IgG subclasses differ in their ability to fix complement and bind Fc receptors. This study describes a detailed analysis of the distribution of HLA-specific IgG subclasses in order to define how this varies in sensitised waiting-list patients. We found significant variation in the level, presence and combinations of each HLA-specific IgG subclass between and within individuals and this is influenced by the type of sensitising event. Graft failure in particular provokes higher levels of IgG1 (vs transfusion, p=0.071 and pregnancy, p=0.042), IgG2 (vs transfusion, p=0.001 and pregnancy, p=0.016), and IgG4 (vs transfusion, p=0.052). Both graft failure and pregnancy tend to stimulate multiple IgG subclass responses against HLA, whereas transfusion stimulated antibodies are dominated by responses limited to IgG1 (p=0.033) and have a low incidence of IgG4 (p=0.046). In marked contrast, IgG4 characterised nearly all HLA DQ-specific antibodies stimulated by graft rejection (p=0.006). Such widely varying IgG subclass heterogeneity is likely to be due to underlying immunological processes dependent on the route of sensitisation. This diversity, which implies functional variation, may help explain why HLA-specific antibodies are an obstacle to transplantation in some circumstances but not others. The subclass association with rejection has potential as a biomarker for chronic rejection.

Human leukocyte antigen antibody incompatible renal transplantation.

Anti-human leukocyte antigen (HLA) antibodies are recognized as an important problem in organ transplant recipients. This is because antibodies formed against a graft months or years after implantations are the major cause of late allograft failure, and also because protocols allow the transplantation of some grafts across pre-formed HLA antibodies. Advances in our understanding of anti-HLA antibody- mediated rejection (AMR) have occurred because of a better understanding of the histological findings during AMR; more sensitive and specific methods to measure anti-HLA antibodies; and through clinical investigation of patients transplanted across an HLA barrier. Despite advances in therapy and investigation, AMR remains a major problem and treatment protocols often fail to treat it successfully. This review aims to describe the issues in each of these areas and to suggest how clinicians may be able to improve the management of patients with anti-HLA antibodies.

Increased systolic blood pressure in rat offspring following a maternal low-protein diet is normalized by maternal dietary choline supplementation.

An adverse prenatal environment may induce long-term metabolic consequences, in particular hypertension and cardiovascular disease. A maternal low-protein (LP) diet is well known to result in increased blood pressure (BP) in offspring. Choline has been shown to have direct BP-reducing effects in humans and animals. It has been suggested that endogenous choline synthesis via phosphatidylcholine is constrained during maternal LP exposure. The present study investigates the effect of choline supplementation to mothers fed a LP diet during pregnancy on systolic BP (SBP) in offspring as measured by tail-cuff plethysmography. Wistar rats were assigned to one of three diets to be fed ad libitum throughout pregnancy: (1) control diet (CONT, 20% protein); (2) an LP diet (9% protein); and (3) LP supplemented with choline (LP + C). Dams were fed the CONT diet throughout lactation and offspring were fed the CONT diet from weaning for the remainder of the trial. At postnatal day 150, SBP and retroperitoneal fat mass was significantly increased in LP offspring compared with CONT animals and was normalized in LP + C offspring. Effects of LP + C reduction in SBP were similar in both males and females. Plasma choline and phosphatidylcholine concentrations were not different across treatment groups, but maternal choline supplementation resulted in a significant reduction in homocysteine concentrations in LP + C offspring compared with LP and CONT animals. The present trial shows for the first time that maternal supplementation with dietary choline during periods of LP exposure can normalize increased SBP and fat mass observed in offspring in later life.

Diet-induced obesity attenuates fasting-induced hyperphagia.

Obesity impairs arcuate (ARC) neuropeptide Y (NPY)/agouti-releated peptide (AgRP) neuronal function and renders these homeostatic neurones unresponsive to the orexigenic hormone ghrelin. In the present study, we investigated the effect of diet-induced obesity (DIO) on feeding behaviour, ARC neuronal activation and mRNA expression following another orexigenic stimulus, an overnight fast. We show that 9 weeks of high-fat feeding attenuates fasting-induced hyperphagia by suppressing ARC neuronal activation and hypothalamic NPY/AgRP mRNA expression. Thus, the lack of appropriate feeding responses in DIO mice to a fast is caused by failure ARC neurones to recognise and/or respond to orexigenic cues. We propose that fasting-induced hyperphagia is regulated not by homeostatic control of appetite in DIO mice, but rather by changes in the reward circuitry.