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Describes procedures for extracting nematodes from soil and culturing them for study and experimentation. (PB)

Children with vesicoureteral reflux (VUR) are at an increased risk of recurrent urinary tract infections (UTIs) and renal scarring. Gut microbiota are associated with disease phenotypes, but there has been no study that associates urinary microbiota (uMB) and metabolic profiles with VUR pathology. To identify dominant uMB genera and metabolites associated with UTIs in VUR, urine samples collected under sterile conditions underwent 16S ribosomal RNA sequencing (n = 49) and metabolomic analysis by mass spectrometry (n = 96). Alterations in uMB and metabolomic profiles in VUR patients suggest remodeling of urinary bacterial communities after UTIs: Dorea- and Escherichia-dominant uMB profiles were more frequently identified in participants with VUR. Prevotella- and Lactobacillus-dominant uMB profiles were more prevalent in controls (p 0.001). Microbial composition varied based on recurrent febrile UTI status (p = 0.001). A total of 243 urinary metabolites involved in energy, amino acid, nucleotide, and lipid metabolism were altered in VUR patients with UTIs (p 0.05). Importantly, VUR specimens revealed changes in the bacteria-associated metabolic pathways such as glutamate degradation, methyl-citrate cycle, and bile acid metabolism. PATIENT SUMMARY: Differences in urinary commensal bacteria and metabolites exist between children with and without vesicoureteral reflux (VUR). These changes may be utilized to identify patients at risk of VUR-associated kidney damage.

Chronic kidney disease involves disturbances in iron metabolism including anemia caused by insufficient erythropoietin (EPO) production. However, underlying mechanisms responsible for the dysregulation of cellular iron metabolism are incompletely defined. Using the unilateral ureteral obstruction (UUO) model in Irp1(+/+) and Irp1(−/−) mice, we asked if iron regulatory proteins (IRPs), the central regulators of cellular iron metabolism and suppressors of EPO production, contribute to the etiology of anemia in kidney failure. We identified a significant reduction in IRP protein level and RNA binding activity that associates with a loss of the iron uptake protein transferrin receptor 1 (TfR1), increased expression of the iron storage protein subunits H- and L-ferritin, and a low but overall variable level of stainable iron in the obstructed kidney. This reduction in IRP RNA binding activity and ferritin RNA levels suggests the concomitant rise in ferritin expression and iron content in kidney failure is IRP dependent. In contrast, the reduction in the Epo mRNA level in the obstructed kidney was not rescued by genetic ablation of IRP1, suggesting disruption of normal hypoxia-inducible factor (HIF)-2α regulation. Furthermore, reduced expression of some HIF-α target genes in UUO occurred in the face of increased expression of HIF-α proteins and prolyl hydroxylases 2 and 1, the latter of which is not known to be HIF-α mediated. Our results suggest that the IRP system drives changes in cellular iron metabolism that are associated with kidney failure in UUO but that the impact of IRPs on EPO production is overridden by disrupted hypoxia signaling. NEW & NOTEWORTHY This study demonstrates that iron metabolism and hypoxia signaling are dysregulated in unilateral obstructive nephropathy. Expression of iron regulatory proteins (IRPs), central regulators of cellular iron metabolism, and the iron uptake (transferrin receptor 1) and storage (ferritins) proteins they target is strongly altered. This suggests a role of IRPs in previously observed changes in iron metabolism in progressive renal disease. Hypoxia signaling is disrupted and appeared to dominate the action of IRP1 in controlling erythropoietin expression.

The pathogenesis of Ebola virus disease (EVD) is still incomplete, although the non-human primate model has been studied for more than 4 decades. To further investigate EVD pathogenesis, a natural history study has been conducted using 27 Chinese-origin rhesus macaques. Of them, 24 macaques were exposed intramuscularly to Kikwit Ebola virus (EBOV) and euthanized at pre-determined timepoints or when end stage clinical disease criteria were met, while 3 other sham-exposed macaques were euthanized at the study day 0. This study demonstrates for the first time that Ebola virus causes uterine cervicitis, vaginitis, posthitis, and medullary adrenalitis. Not only is Ebola virus detected in the interstitial stromal cells of the genital tract, but it is also present in the epididymal and seminal vesicular tubular epithelial cells, ectocervical and vaginal squamous epithelial cells, and seminal fluid. Furthermore, as early as day 3 after exposure, EBOV replicative intermediate RNA was detected in Kupffer cells and hepatocytes. These findings in the nonhuman model provide additional insight into potential sexual transmission, possible disruption of sympathetic hormone production, and early virus replication sites in human EVD patients.

Long-chain alkenones (LCAs) produced by phylogenetically classified Groups 1 and 3 Isochrysidales are increasingly used for paleotemperature and/or paleosalinity reconstructions in oligohaline lakes and marine environments. However, there are considerable difficulties in the paleoenvironmental interpretation of LCAs from Group 2 Isochrysidales thriving in saline lakes. The biggest challenge lies in our poor understanding of the complexity and ecological niches of individual Group 2 subclades in saline lakes. Here, we perform comprehensive analysis of haptophyte-specific 18S rRNA sequences and distributions of LCAs, and long-chain alkenoates (LCEs) in surface sediments and suspended particulate matter (SPM) from 37 saline lakes in northern China. These lakes span a large salinity gradient from 0.5 to 308‰. Combined with published genomic data of Group 2 Isochrysidales, our phylogenetic analysis reveals three Group 2 subclades occupying distinct ecological niches: one ice-related bloomer Group 2i and two warm-season bloomers Groups 2w1 and 2w2. Group 2i, the earliest seasonal bloomer, frequently co-occurs with Group 2w1 in sediments from saline lakes with relatively low to intermediate salinity waters, whereas Group 2w2 blooms in hypersaline waters. Based on existing data, C39:4 methyl alkenone is a chemotaxonomic biomarker for Group 2i. %C37:4 (relative abundance of C37:4 to the total C37 alkenones) values in the three Group subclades follow the order: Group 2i > Group 2w2 > Group 2w1. The %C37:4 in sediment cores of saline lakes does not directly record past salinity changes, but instead reflects variable contributions in production by these three subclades. This could indirectly and partially reflect overall salinity changes in some lakes dominated by Groups 2i and 2w1, but can be more complicated in lakes dominated by other assemblages. For our sites, we also demonstrate that direct use of C37 alkenone unsaturation indices ( U 37 K , U 37 K ' , and U 37 K ' ' ) for paleotemperature reconstructions in saline lakes is generally not feasible, except for cases where alkenone-producing Isochrysidales are dominated by one single species/subclade and seasonal production effects can be circumvented. We propose two possible alternative proxies for paleotemperature reconstructions in saline lakes: 1) unsaturation ratios of C41 and C42 alkenones, as these compounds are predominantly produced by a limited number of Group 2 species, such as Isochrysis nuda ( Liao et al., 2020 ); 2) C38Et/C36OEt ratio (ratio of C38 ethyl alkenones and C36 ethyl alkenoates), which appears to have similar temperature sensitivity for Groups 2w1 and 2w2, in lakes with no Group 2i inputs. Our study provides new insights into the phylogenetic classifications of Group 2 Isochrysidales and their ecological/environmental niches, which are fundamental for more quantitative and rigorous applications of LCAs and LCEs in saline lakes as paleosalinity and paleotemperature proxies.

PREMISE Reconstructing plant canopy light environment and architecture from the fossil record includes proxies derived from cell wall undulation, cell size, and carbon isotopes. All approaches assume that plant taxa will respond predictably to changes in light environments. However, most species-level studies looking at cell wall undulation only consider "sun" or "shade" leaves, therefore we lack a fully quantitative taxon-specific method. METHODS We quantify the response of cell wall undulation, cell size, and carbon isotopes of Platanus occidentalis using two different experimental setups: (1) two growth chambers at low and high light and (2) a series of outdoor growth experiments using green and black shade cloth at different densities. We then developed and applied a proxy for daily light integral (DLI) to fossil Platanites leaves from two early Paleocene floras from the San Juan Basin in New Mexico. RESULTS All traits responded to light environment. Cell wall undulation was the most useful trait for reconstructing DLI in the geological record. Median reconstructed DLI from early Paleocene leaves was ~44 mol m-2 d-1, with values ranging from 28 - 54 mol m-2 d-1. CONCLUSIONS Cell wall undulation of P. occidentalis is a robust, quantifiable measurement of light environment that can be used to reconstruct paleo-light environment from fossil leaves. The distribution of high DLI values from fossil leaves may provide information on canopy architecture; indicating that either (1) most of the canopy mass is within the upper portion of the crown or (2) leaves exposed to more sunlight are preferentially preserved. This article is protected by copyright. All rights reserved.

Strongyloidiasis represents a major medical and veterinary helminthic disease. Human infection is caused by three species; Strongyloides stercoralis, Strongyloides fuelleborni fuelleborni and Strongyloides fuelleborni kellyi, with S. stercoralis accounting for the majority of cases. Strongyloides f. fuelleborni likely represents a zoonosis acquired from non-human primates (NHPs), while no animal reservoir for S. f. kellyi infection has been found. Whether S. stercoralis represents a zoonosis acquired from dogs and cats remains unanswered. Over the past two decades various tools have been applied to genotype Strongyloides spp. The most commonly sequenced markers have been the hyper-variable regions I and IV of the 18S rRNA gene and selected portions of the cytochrome c oxidase subunit I gene. These markers have been sequenced and compared in Strongyloides from multiple hosts and geographical regions. More recently, a machine learning algorithm multi-locus sequence typing approach has been applied using these markers, while others have applied whole genome sequencing. Genotyping of Strongyloides from dogs, cats, NHPs and humans has identified that S. stercoralis likely originated in dogs and adapted to human hosts. It has also demonstrated that S. stercoralis is distinct from S. f. fuelleborni and S. f. kellyi. Two distinct genetic clades of S. stercoralis exist, one restricted to dogs and another infecting humans, NHPs, dogs and cats. Genotyping of S. f. fuelleborni has identified two separate clades, one associated with African isolates and another Indochinese peninsular clade. This review summarises the history and development of genotyping tools for Strongyloides spp. It describes the findings of major studies to date in the context of the epidemiology and evolutionary biology of these helminths, with a specific focus on human-infecting species.

Small molecules derived from symbiotic microbiota critically contribute to intestinal immune maturation and regulation1. However, little is known about the molecular mechanisms that control immune development in the host–microbiota environment. Here, using a targeted lipidomic analysis and synthetic approach, we carried out a multifaceted investigation of immunomodulatory α-galactosylceramides from the human symbiont Bacteroides fragilis (BfaGCs). The characteristic terminal branching of BfaGCs is the result of incorporation of branched-chain amino acids taken up in the host gut by B. fragilis. A B. fragilis knockout strain that cannot metabolize branched-chain amino acids showed reduced branching in BfaGCs, and mice monocolonized with this mutant strain had impaired colonic natural killer T (NKT) cell regulation, implying structure-specific immunomodulatory activity. The sphinganine chain branching of BfaGCs is a critical determinant of NKT cell activation, which induces specific immunomodulatory gene expression signatures and effector functions. Co-crystal structure and affinity analyses of CD1d–BfaGC–NKT cell receptor complexes confirmed the interaction of BfaGCs as CD1d-restricted ligands. We present a structural and molecular-level paradigm of immunomodulatory control by interactions of endobiotic metabolites with diet, microbiota and the immune system. The symbiotic gut bacterium Bacteroides fragilis produces unique α-galactosylceramides from host dietary branched-chain amino acids, which are presented as CD1d ligands and immunomodulate natural killer T cells.

Strongyloidiasis remains endemic throughout the Island of New Guinea. While many infections are caused by Strongyloides stercoralis, a second human-infecting Strongyloides species, Strongyloides fuelleborni kellyi, is also present. S. f. kellyi infections are most common in infants and young children, and those with high-intensity infections might develop a potentially fatal protein-losing enteropathy, swollen belly syndrome. Surprisingly little work has been performed on S. f. kellyi. Unlike S. stercoralis, S. f. kellyi is passed in faeces as eggs rather than rhabditiform larvae. There is no specific diagnostic test. This review summarises what is currently known about the biology, epidemiology, and clinical impact of S. f. kellyi infections. Features that might be used to differentiate S. f. kellyi from hookworm and S. stercoralis are also discussed. S. f. kellyi remains a neglected, ignored, and unexplored human helminth infection, worthy of further research.

As the outermost layer of plants, the epidermis serves as a critical interface between plants and the environment. During leaf development, the differentiation of specialized epidermal cell types, including stomatal guard cells, pavement cells, and trichomes, occurs simultaneously, each providing unique and pivotal functions for plant growth and survival. Decades of molecular-genetic and physiological studies have unraveled key players and hormone signaling specifying epidermal differentiation. However, most studies focus on only one cell type at a time, and how these distinct cell types coordinate as a unit is far from well-comprehended. Here we provide a review on the current knowledge of regulatory mechanisms underpinning the fate specification, differentiation, morphogenesis, and positioning of these specialized cell types. Emphasis is given to their shared developmental origins, fate flexibility, as well as cell cycle and hormonal controls. Furthermore, we discuss computational modeling approaches to integrate how mechanical properties of individual epidermal cell types and entire tissue/organ properties mutually influence each other. We hope to illuminate the underlying mechanisms coordinating the cell differentiation that ultimately generate a functional leaf epidermis.

Background Telomere maintenance is increasingly recognized as being fundamental to glioma oncogenesis with longer leukocyte telomere length (LTL) reported to increase risk of glioma. To gain further insight into the relationship between telomere genetics and risk of glioma, we conducted several complementary analyses, using genome-wide association studies data on LTL (78 592 individuals) and glioma (12 488 cases and 18 169 controls). Methods We performed both classical and summary Mendelian randomization (SMR), coupled with heterogeneity in dependent instruments tests, at genome-wide significant LTL loci to examine if an association was mediated by the same causal variant in glioma. To prioritize genes underscoring glioma-LTL associations, we analyzed gene expression and DNA methylation data. Results Genetically increased LTL was significantly associated with increased glioma risk, random-effects inverse variance weighted ORs per 1 SD unit increase in the putative risk factor (odds ratio [OR]SD) 4.79 (95% confidence interval: 2.11-10.85; P = 1.76 × 10−4). SMR confirmed the previously reported LTL associations at 3q26.2 (TERC; PSMR = 1.33 × 10−5), 5p15.33 (TERT; PSMR = 9.80 × 10−27), 10q24.33 (STN1 alias OBFC1; PSMR = 4.31 × 10−5), and 20q13.3 (STMN3/RTEL1; PSMR = 2.47 × 10−4) glioma risk loci. Our analysis implicates variation at 1q42.12 (PSMR = 1.55 × 10−2), 6p21.3 (PSMR = 9.76 × 10−3), 6p22.2 (PSMR = 5.45 × 10−3), 7q31.33 (PSMR = 6.52 × 10−3), and 11q22.3 (PSMR = 8.89 × 10−4) as risk factors for glioma risk. While complicated by patterns of linkage disequilibrium, genetic variation involving PARP1, PRRC2A, CARMIL1, POT1, and ATM-NPAT1 was implicated in the etiology of glioma. Conclusions These observations extend the role of telomere-related genes in the development of glioma.

Enhancing host defences through induced resistance, disease tolerance, and/or escape, in combination with current disease management regimes may be a valuable strategy to reduce pesticide use. Since both ‘on-farm’ seed priming (OSP) and chitosan priming (CHP) have been reported to confer varying levels of host defence, this study sought to investigate their potential to deliver disease control as a strategy for sustainable management of foliar pathogens in winter barley. Field experiments were conducted to determine the effects of OSP and CHP at two different field sites using three different cultivars under fungicide/non-fungicide regimes. Overall, no evidence was found to suggest that CHP or OSP can induce effective resistance in temperate field conditions. However, these field trials enabled the identification of candidate traits to deliver disease tolerance (and escape) for the primary and secondary spread of powdery mildew, i.e., large canopies and rapid stem elongation respectively. Thus, these seed treatments may deliver disease tolerance and escape traits, but these benefits are dependent upon successful establishment and vigour first. The integration of seed treatments into sustainable crop protection may be better undertaken with spring crops or in semi-arid agriculture where the added vigour at emergence can help compensate for negative environmental interactions.

Age-related cardiac amyloidosis results from deposits of wild-type tranthyretin amyloid (ATTRwt) in cardiac tissue. ATTR may play a role in carpal tunnel syndrome (CTS) and in spinal stenosis (SS), indicating or presaging systemic amyloidosis. We investigated consecutive patients undergoing surgery for SS for ATTR deposition in the resected ligamentum flavum (LF) and concomitant risk of cardiac amyloidosis. Each surgical specimen (LF) was stained with Congo red, and if positive, the amyloid deposits were typed by mass spectrometry. Patients with positive specimens underwent standard of care evaluation with fat pad aspirates, serum and urine protein electrophoresis with immunofixation, free light-chain assay

Background: Spinocerebellar ataxias (SCA) are a group of neurodegenerative disorders characterized by neuronal degeneration leading to loss of motor coordination. A number of different mutations gives rise to different types of SCA with characteristic ages of onset, symptomatology, and rates of progression. SCA type 34 (SCA34) is an age-related cerebellar neurodegenerative disorder caused by mutations in the fatty acid elongase-4 (ELOVL4). The ELOVL4 is an essential enzyme that mediates biosynthesis of Very Long Chain Saturated and Polyunsaturated Fatty Acids (VLC-SFA and VLC-PUFA, resp., ≥28 carbons) that are critical for the normal function of brain, skin, retina, Meibomian glands, and testes in which ELOVL4 is expressed. Global deletion or homozygous expression of truncated mutant ELOVL4 that lack VLC-SFA and VLC-PUFA biosynthesis cause severe skin disorders, seizures and neonatal mortality in rodents and humans. Methods: To understand role of ELOVL4 and its products in neuronal function and to evaluate the consequences of ELOVL4 mutations in pathogenesis of age-related SCA34, we generated a rat model of SCA34 by knock-in of the SCA34-causing 736T>G (p.W246G) ELOVL4 mutation. We performed biochemical, neuroanatomical and behavioral analyses by rotorod to measure motor function. We used electrophysiological recordings from cerebellar slices to determine the impact of the W246G mutation on neuronal function. Results were analyzed using GraphPad Prism Statistical software. Results: Heterozygous and homozygous rats carrying the W246G mutation developed impaired motor deficits by two months of age. To understand the mechanism of these motor deficits, we performed electrophysiological studies using cerebellar slices from rats homozygous for W246G mutant ELOVL4 and found marked reduction of long-term potentiation at parallel fiber synapses and long-term depression at climbing fiber synapses onto Purkinje cells. Neuroanatomical analysis of the cerebellum up 6 months of age showed normal cytoarchitectural organization despite the early-onset motor deficits and defects in synaptic plasticity. Conclusions: Our results point to ELOVL4 and its products being essential for motor function and cerebellar synaptic plasticity. The results further suggest that in SCA34 patients, ataxia arises from primary impairment of synaptic plasticity and cerebellar network desynchronization that precedes cerebellar degeneration and loss of motor coordination with aging.

While most testicular germ cell tumours (TGCTs) exhibit exquisite sensitivity to platinum chemotherapy, ~10% are platinum resistant. To gain insight into the underlying mechanisms, we undertake whole exome sequencing and copy number analysis in 40 tumours from 26 cases with platinum-resistant TGCT, and combine this with published genomic data on an additional 624 TGCTs. We integrate analyses for driver mutations, mutational burden, global, arm-level and focal copy number (CN) events, and SNV and CN signatures. Albeit preliminary and observational in nature, these analyses provide support for a possible mechanistic link between early driver mutations in RAS and KIT and the widespread copy number events by which TGCT is characterised., Most testicular germ-cell tumours are exquisitely sensitive to platinum-based chemotherapies, but little is known about why 10% are resistant. Here, the authors explore the potential underlying mechanisms by probing the genomic landscape of platinum-resistant disease.

The avocado (Persea americana Mill.) is highly valued worldwide for its desirable nutritional properties and broader applications in the oil and pharmaceutical industry. Despite northwestern Mexico is not considered as a region of the avocado origin, it possesses a high morphological diversity of avocado fruits and trees. In an effort to provide more information about the diversity present in this region and at the same time, to support a more efficient production of lowland avocado in a tropical semi desert climate, this study characterized the genetic diversity of avocado trees growing at northwestern Mexico. Genetic diversity was estimated by using eight microsatellite loci. 45 seed-derived trees, with contrasting fruit morphology were identified. Results showed a high level of genetic diversity with 11.5 alleles per locus, a polymorphic index content of 0.75, and observed and expected heterozygosity values of 0.58 and 0.79, respectively, these values were similar to those observed in germplasm banks and genotypes from centers of origin. Clustering analysis assigned local genotypes to six clusters but did not provide a clear separation among avocado races, suggesting that local genotypes are a result of racial hybridization. Interestingly, avocados from the Mexican race and the commercial variety Hass clustered into two different groups. Despite not being considered a center of origin, results confirm an extensive diversity in northwestern Mexico, encouraging further exploration and preservation of genotypes with desirable traits to future breeding programs for the selection of local genotypes adapted to a lowland tropical climate.

RNA exosomopathies, a growing family of diseases, are linked to missense mutations in genes encoding structural subunits of the evolutionarily conserved, 10-subunit exoribonuclease complex, the RNA exosome. This complex consists of a three-subunit cap, a six-subunit, barrel-shaped core, and a catalytic base subunit. While a number of mutations in RNA exosome genes cause pontocerebellar hypoplasia, mutations in the cap subunit gene EXOSC2 cause an apparently distinct clinical presentation that has been defined as a novel syndrome SHRF (short stature, hearing loss, retinitis pigmentosa, and distinctive facies). We generated the first in vivo model of the SHRF pathogenic amino acid substitutions using budding yeast by modeling pathogenic EXOSC2 missense mutations (p.Gly30Val and p.Gly198Asp) in the orthologous S. cerevisiae gene RRP4. The resulting rrp4 mutant cells show defects in cell growth and RNA exosome function. Consistent with altered RNA exosome function, we detect significant transcriptomic changes in both coding and noncoding RNAs in rrp4-G226D cells that model EXOSC2 p.Gly198Asp, suggesting defects in nuclear surveillance. Biochemical and genetic analyses suggest that the Rrp4 G226D variant subunit shows impaired interactions with key RNA exosome cofactors that modulate the function of the complex. These results provide the first in vivo evidence that pathogenic missense mutations present in EXOSC2 impair the function of the RNA exosome. This study also sets the stage to compare exosomopathy models to understand how defects in RNA exosome function underlie distinct pathologies.

Purpose The endoplasmic reticulum membrane complex (EMC) is a highly conserved, multifunctional 10-protein complex related to membrane protein biology. In seven families, we identified 13 individuals with highly overlapping phenotypes who harbor a single identical homozygous frameshift variant in EMC10. Methods Using exome, genome, and Sanger sequencing, a recurrent frameshift EMC10 variant was identified in affected individuals in an international cohort of consanguineous families. Multiple families were independently identified and connected via Matchmaker Exchange and internal databases. We assessed the effect of the frameshift variant on EMC10 RNA and protein expression and evaluated EMC10 expression in normal human brain tissue using immunohistochemistry. Results A homozygous variant EMC10 c.287delG (Refseq NM_206538.3, p.Gly96Alafs*9) segregated with affected individuals in each family, who exhibited a phenotypic spectrum of intellectual disability (ID) and global developmental delay (GDD), variable seizures and variable dysmorphic features (elongated face, curly hair, cubitus valgus, and arachnodactyly). The variant arose on two founder haplotypes and results in significantly reduced EMC10 RNA expression and an unstable truncated EMC10 protein. Conclusion We propose that a homozygous loss-of-function variant in EMC10 causes a novel syndromic neurodevelopmental phenotype. Remarkably, the recurrent variant is likely the result of a hypermutable site and arose on distinct founder haplotypes.

Spinal muscular atrophy is a progressive, recessively inherited monogenic neurologic disease, the genetic root cause of which is the absence of a functional survival motor neuron 1 gene. Onasemnogene abeparvovec (formerly AVXS-101) is an adeno-associated virus serotype 9 vector-based gene therapy that delivers a fully functional copy of the human survival motor neuron gene. We report anti–adeno-associated virus serotype 9 antibody titers for patients with spinal muscular atrophy when they were screened for eligibility in the onasemnogene abeparvovec clinical trials (intravenous and intrathecal administration) and managed access programs (intravenous). Through December 31, 2019, 196 patients and 155 biologic mothers were screened for anti–adeno-associated virus serotype 9 binding antibodies with an enzyme-linked immunosorbent assay. Of these, 15 patients (7.7%) and 23 biologic mothers (14.8%) had titers >1:50 on their initial screening tests. Eleven patients (5.6%) had elevated titers on their final screening tests. The low percentage of patients with exclusionary antibody titers indicates that most infants with spinal muscular atrophy type 1 should be able to receive onasemnogene abeparvovec. Retesting may identify patients whose antibody titers later decrease to below the threshold for treatment, and retesting should be considered for patients with anti–adeno-associated virus serotype 9 antibody titers >1:50., Graphical abstract, Day and colleagues report that 5.6% of patients screened across the onasemnogene abeparvovec clinical development program and US MAPs were excluded because of elevated anti-adeno-associated virus serotype 9 binding antibody titers, indicating that most patients with spinal muscular atrophy should be able to receive adeno-associated virus serotype 9-based therapy.

Patients with end stage kidney disease receiving in-center hemodialysis (ICHD) have had high rates of SARS-CoV-2 infection. Following infection, patients receiving ICHD frequently develop circulating antibodies to SARS-CoV-2, even with asymptomatic infection. Here, we investigated the durability and functionality of the immune responses to SARS-CoV-2 infection in patients receiving ICHD. Three hundred and fifty-six such patients were longitudinally screened for SARS-CoV-2 antibodies and underwent routine PCR-testing for symptomatic and asymptomatic infection. Patients were regularly screened for nucleocapsid protein (anti-NP) and receptor binding domain (anti-RBD) antibodies, and those who became seronegative at six months were screened for SARS-CoV-2 specific T-cell responses. One hundred and twenty-nine (36.2%) patients had detectable antibody to anti-NP at time zero, of whom 127 also had detectable anti-RBD. Significantly, at six months, 71/111 (64.0%) and 99/116 (85.3%) remained anti-NP and anti-RBD seropositive, respectively. For patients who retained antibody, both anti-NP and anti-RBD levels were reduced significantly after six months. Eleven patients who were anti-NP seropositive at time zero, had no detectable antibody at six months; of whom eight were found to have SARS-CoV-2 antigen specific T cell responses. Independent of antibody status at six months, patients with baseline positive SARS-CoV-2 serology were significantly less likely to have PCR confirmed infection over the following six months. Thus, patients receiving ICHD mount durable immune responses six months post SARS-CoV-2 infection, with fewer than 3% of patients showing no evidence of humoral or cellular immunity., Graphical abstract

The vast majority of plant viruses are transmitted by insect vectors, with many crucial aspects of the transmission process being mediated by key protein-protein interactions. Still, very few vector proteins interacting with viruses have been identified and functionally characterized. Potato leafroll virus (PLRV) is transmitted most efficiently by Myzus persicae, the green peach aphid, in a circulative, non-propagative manner. Using affinity purification coupled to high-resolution mass spectrometry (AP-MS), we identified 11 proteins from M. persicaedisplaying a high probability of interaction with PLRV and an additional 23 vector proteins with medium confidence interaction scores. Three of these aphid proteins were confirmed to directly interact with the structural proteins of PLRV and other luteovirid species via yeast two-hybrid. Immunolocalization of one of these direct PLRV-interacting proteins, an orthologue of the human innate immunity protein complement component 1 Q subcomponent-binding protein (C1QBP), shows that MpC1QBP partially co-localizes with PLRV in cytoplasmic puncta and along the periphery of aphid gut epithelial cells. Artificial diet delivery to aphids of a chemical inhibitor of C1QBP leads to increased PLRV acquisition by aphids and subsequently increased titer in inoculated plants, supporting a role for C1QBP in the acquisition and transmission efficiency of PLRV by M. persicae. This study presents the first use of AP-MS for the in vivo isolation of a functionally relevant insect vector-virus protein complex. MS data are available from ProteomeXchange.org using the project identifier PXD022167.

Composting has been used to dispose of animal mortalities and infected materials, such as manure and feed, during major animal disease outbreaks. In this study, we adapted the plastic-wrapped mortality composting system developed by the Canadian Food Inspection Agency during the 2004 highly pathogenic avian influenza outbreak to compost swine mortalities. The goals of the study were to evaluate the performance of the plastic-wrapped composting system to dispose of swine mortalities and to field test its ability to eliminate the spread of airborne pathogens through the aeration ducts. Two cover materials, ground cornstalks and woodchips, were tested using passively and actively aerated composting sheds. The mortalities were inoculated with Salmonella spp. and vaccine strains of Bovine herpesvirus-1 and Bovine viral diarrhea virus. Air samples collected from the upper aeration duct (air outlet) during the first 10 d of composting were negative for Salmonella and the viruses tested, which indicated that aerosol transmission of the pathogens was limited. The aeration plenum placed under the mortalities helped to keep conditions aerobic, as O2 concentrations of both passively and actively aerated test units were above 11%. Actively aerated cornstalks had the highest degree-hours (1,462 °C h d-1 ), which was followed by passively aerated cornstalks (1,312 °C h d-1 ), actively aerated woodchips (1,303 °C h d-1 ), and passively aerated woodchips (1,062 °C h d-1 ). After a 7-wk composting period, all three pathogens were inactivated based on quantitative polymerase chain reaction test results. The mortalities were not inoculated with the African swine fever virus, but temperature data showed that if they were, the system had the potential to eliminate this virus.

Isolated lateral compartment osteoarthritis of the knee is a rare condition affecting approximately 1% of the population, which is ten times less common than osteoarthritis affecting only the medial compartment. Unicompartmental knee arthroplasty (UKA) has many potential advantages over total knee arthroplasty. The benefits of UKA include a smaller incision, preservation of more native tissue (including cruciate ligaments and bone), decreased blood loss, and better overall proprioception. When UKA was first introduced in the 1970s, the outcomes of medial UKA (MUKA) were poor, but the few cases of lateral UKA (LUKA) showed promise. Since that time, there has been a relative paucity of literature focused specifically on LUKA given it is a rare procedure. Refinements in patient selection criteria, implant design, and surgical technique have been made leading to increased popularity. A review of the recent literature reveals that LUKA is associated with excellent long-term clinical outcomes and implant survivorship when performed in properly selected patients. Implant design options include fixed vs mobile bearing as well as metal backed vs all polyethylene tibial component, with improved outcomes noted with fixed bearing designs. Three reasons cited for revision (i.e., fracture of the femoral component, fracture of the tibial component, and valgus malalignment) had been reported in past literature but not recently. Presently, while rare, the most common cause of failure and need for revision are osteoarthritis progression and aseptic loosening. Despite the need for an occasional revision procedure, the survivorship of LUKA is comparable to MUKA, although it should be noted that outcomes of MUKA have been notably varied. Continued pursuit of improved techniques and implant designs will continue to show LUKA to be an excellent procedure for appropriately indicated patients.

Understanding host use by psyllids (Hemiptera: Psylloidea) benefits from comparative studies of behavior on host and nonhost plant species. While most psyllid species develop on one or a few closely related plant species, some species are generalized enough to develop on species across plant families. We used electropenetography (EPG) technology to compare probing activities of an oligophagous psyllid (Bactericera cockerelli (Šulc)) and a host-specialized psyllid (Bactericera maculipennis) on two species of Solanaceae (potato, Solanum tuberosum L. and matrimony vine, Lycium barbarum L.) and two species of Convolvulaceae (field bindweed, Convolvulus arvensis L. and sweet potato, Ipomoea batatas). Bactericera cockerelli develops on all four species, albeit with longer development times on Convolvulaceae. Bactericera maculipennis develops only on Convolvulaceae. Bactericera cockerelli fed readily from phloem of all four species, but the likelihood of entering phloem and duration of time in phloem was reduced on suboptimal hosts (Convolvulaceae) relative to behavior on Solanaceae. We observed instances of cycling between bouts of phloem salivation and ingestion in assays of optimal (Solanaceae) hosts not observed on Convolvulaceae. The Convolvulaceae-specialized B. maculipennis (Crawford) failed to feed from phloem of nonhosts (Solanaceae). Both psyllid species readily ingested from xylem of all plant species, irrespective of host status. Our finding that phloem feeding by B. maculipennis did not occur on potato has implications for understanding epidemiology of phloem-limited psyllid-vectored plant pathogens. Our results also showed that EPG assays detect subtle variation in probing activities that assist in understanding host use by psyllids.

Purpose: The therapeutic utility of Cannabis in cancer is a topic of intense interest. Dronabinol is synthetic Δ(9)-tetrahydrocannabinol (THC), the primary psychoactive component of Cannabis sativa, and is approved for treating refractory chemotherapy-induced nausea and vomiting. Little is known about dronabinol prescribing in children and young adults, and no published concentration data are available. This study evaluated national level dronabinol use and assessed concentrations of THC and its primary metabolites in patients with cancer

Background The movement ecology of mutton snapper Lutjanus analis is poorly understood despite their ecological and economic importance in the Caribbean. Passive acoustic telemetry was used to determine home ranges of six adult L. analis, including diel patterns, in Brewers Bay, St. Thomas, US Virgin Islands. Understanding long-term space use, including site fidelity and habitat usage, is necessary to implement effective and appropriate management actions for a species with extensive space and resource needs. Results Individual L. analis were tracked over an average period of 316 days (range 125–509 days) and showed high site fidelity to relatively small home ranges (mean ± SD: 0.103 ± 0.028 km2, range 0.019–0.190 km2) and core use areas with low overlap among individuals. Most home ranges had a habitat composition dominated by seagrass and to a lesser degree, coral reef and/or pavement. Nighttime activity spaces were distinct from but contained within daytime areas. Conclusions Mutton snapper showed strong site fidelity to home ranges in Brewers Bay. Two individuals that were absent from the array for more than a few hours were detected at separate arrays at spawning aggregation sites. This study expands upon knowledge of mutton snapper home range characteristics, highlights the importance of maintaining adjacent high-quality habitat types in any spatial management plan, and encourages the adoption of other types of management strategies, particularly for transient-aggregating species.

Background Epidemiological studies of adult glioma have identified genetic syndromes and 25 heritable risk loci that modify individual risk for glioma, as well increased risk in association with exposure to ionizing radiation and decreased risk in association with allergies. In this analysis, we assess whether there is a shared genome-wide genetic architecture between glioma and atopic/autoimmune diseases. Methods Using summary statistics from a glioma genome-wide association studies (GWAS) meta-analysis, we identified significant enrichment for risk variants associated with gene expression changes in immune cell populations. We also estimated genetic correlations between glioma and autoimmune, atopic, and hematologic traits using linkage disequilibrium score regression (LDSC), which leverages genome-wide single-nucleotide polymorphism (SNP) associations and patterns of linkage disequilibrium. Results Nominally significant negative correlations were observed for glioblastoma (GB) and primary biliary cirrhosis (rg = −0.26, P = .0228), and for non-GB gliomas and celiac disease (rg = −0.32, P = .0109). Our analyses implicate dendritic cells (GB pHM = 0.0306 and non-GB pHM = 0.0186) in mediating both GB and non-GB genetic predisposition, with GB-specific associations identified in natural killer (NK) cells (pHM = 0.0201) and stem cells (pHM = 0.0265). Conclusions This analysis identifies putative new associations between glioma and autoimmune conditions with genomic architecture that is inversely correlated with that of glioma and that T cells, NK cells, and myeloid cells are involved in mediating glioma predisposition. This provides further evidence that increased activation of the acquired immune system may modify individual susceptibility to glioma.

A low-cost technique named ‘on-farm’ seed priming is increasingly being recognized as an effective approach to maximize crop establishment. It consists of anaerobically soaking seeds in water before sowing resulting in rapid and uniform germination, and enhanced seedling vigour. The extent of these benefits depends on the soaking time. The current determination of optimal soaking time by germination assays and mini-plot trials is resource-intensive, as it is species/genotype-specific. This study aimed to determine the potential of the seed respiration rate (an indicator of metabolic activity) and seed morphological changes during barley priming as predictors of the priming benefits and, thus, facilitate the determination of optimal soaking times. A series of germination tests revealed that the germination rate is mostly attributable to the rapid hydration of embryo tissues, as the highest gains in the germination rate occurred before the resumption of respiration. Germination uniformity, however, was not significantly improved until seeds were primed for at least 8 h, that is, after a first respiration burst was initiated. The maximum seedling vigour was attained when the priming was stopped just before the beginning of the differentiation of embryonic axes (20 h) after which vigour began to decrease (‘over-priming’). The onset of embryonic axis elongation was preceded by a second respiration burst, which can be used as a marker for priming optimization. Thus, monitoring of seed respiration provides a rapid and inexpensive alternative to the current practice. The method could be carried out by agricultural institutions to provide recommended optimal soaking times for the common barley varieties within a specific region.

It is currently unknown how post-COVID-19 syndrome (PCS) may affect those infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This longitudinal study includes healthcare staff who tested positive for SARS-CoV-2 between March and April 2020, with follow-up of their antibody titers and symptoms. More than half (21 of 38) had PCS after 7–8 months. There was no statistically significant difference between initial reverse-transcription polymerase chain reaction titers or serial antibody levels between those who did and those who did not develop PCS. This study highlights the relative commonality of PCS in healthcare workers and this should be considered in vaccination scheduling and workforce planning to allow adequate frontline staffing numbers.

The glycoprotein uromodulin (UMOD) is the most abundant protein in urine, and N-glycans are critical for many biological functions of UMOD. Comprehensive glycan profiling of UMOD provides valuable information to understand the exact mechanisms of glycan-regulated functions. To perform comprehensive glycosylation analysis of UMOD from urine samples with limited volumes, we developed a streamlined workflow that included UMOD isolation from 5 mL of urine from 6 healthy adult donors (3 males and 3 females) and a glycosylation analysis using a highly sensitive and reproducible nanoLC-MS/MS based glycomics approach. In total, 212 N-glycan compositions were identified from the purified UMOD, and 17% were high-mannose glycans, 2% were afucosylated/asialylated, 3% were neutral fucosylated, 28% were sialylated (with no fucose), 46% were fucosylated and sialylated, and 4% were sulfated. We found that isolation of UMOD resulted in a significant decrease in the relative quantity of high-mannose and sulfated glycans with a significant increase of neutral fucosylated glycans in the UMOD-depleted urine relative to the undepleted urine, but depletion had little impact on the sialylated glycans. To our knowledge, this is the first study to perform comprehensive N-glycan profiling of UMOD using nanoLC-MS/MS. This analytical workflow would be very beneficial for studies with limited sample size, such as pediatric studies, and can be applied to larger patient cohorts not only for UMOD interrogation but also for global glycan analysis.

Snow roots are a very special type of roots that counteract geotropism to grow upward into long-lasting snow fields. They develop under snow at near 0oC, a phenomenon that had previously only been reported from plant shoots (Korner et al. 2019). Up to now, this intriguing and spectacular looking plant structure has been discovered and studied in only a single species, i.e. the vernal forb, Corydalis conorhiza Ledeb. (poppy family, Papaveraceae) at an elevation of almost 3 km in the mountains of the northern Caucasus, Russia by Onipchenko et al. (2009a, 2014), who reported results from a snow-field labeling experiment with the natural nitrogen (N) isotope 15 N, accompanied with anatomical and structural root trait measurements.

Interleukin-22 (IL-22) signaling in the intestines is critical for promoting tissue-protective functions. However, since a diverse array of cell types (absorptive and secretory epithelium as well as stem cells) express IL-22Ra1, a receptor for IL-22, it has been difficult to determine what cell type(s) specifically respond to IL-22 to mediate intestinal mucosal host defense. Here, we report that IL-22 signaling in the small intestine is positively correlated with Paneth cell differentiation programs. Our Il22Ra1fl/fl;Lgr5-EGFP-creERT2-specific knockout mice and, independently, our lineage-tracing findings rule out the involvement of Lgr5+ intestinal stem cell (ISC)-dependent IL-22Ra1 signaling in regulating the lineage commitment of epithelial cells, including Paneth cells. Using novel Paneth cell-specific IL-22Ra1 knockout mice (Il22Ra1fl/fl;Defa6-cre), we show that IL-22 signaling in Paneth cells is required for small intestinal host defense. We show that Paneth cell maturation, antimicrobial effector function, expression of specific WNTs, and organoid morphogenesis are dependent on cell-intrinsic IL-22Ra1 signaling. Furthermore, IL-22 signaling in Paneth cells regulates the intestinal commensal bacteria and microbiota-dependent IL-17A immune responses. Finally, we show ISC and, independently, Paneth cell-specific IL-22Ra1 signaling are critical for providing immunity against Salmonella enterica serovar Typhimurium. Collectively, our findings illustrate a previously unknown role of IL-22 in Paneth cell-mediated small intestinal host defense.

Introduction AKCEA-TTR-LRx is a ligand-conjugated antisense (LICA) drug in development for the treatment of hereditary transthyretin amyloidosis (hATTR), a fatal disease caused by mutations in the transthyretin (TTR) gene. AKCEA-TTR-LRx shares the same nucleotide sequence as inotersen, an antisense medicine approved for use in hATTR polyneuropathy (hATTR-PN). Unlike inotersen, AKCEA-TTR-LRx is conjugated to a triantennary N-acetylgalactosamine moiety that supports receptor-mediated uptake by hepatocytes, the primary source of circulating TTR. This advanced design increases drug potency to allow for lower and less frequent dosing. The NEURO-TTRansform study will investigate whether AKCEA-TTR-LRx is safe and efficacious, with the aim of improving neurologic function and quality of life in hATTR-PN patients. Methods/Design Approximately 140 adults with stage 1 (independent ambulation) or 2 (requires ambulatory support) hATTR-PN are anticipated to enroll in this multicenter, open-label, randomized, phase 3 study. Patients will be assigned 6:1 to AKCEA-TTR-LRx 45 mg subcutaneously every 4 weeks or inotersen 300 mg once weekly until the prespecified week 35 interim efficacy analysis, after which patients receiving inotersen will receive AKCEA-TTR-LRx 45 mg subcutaneously every 4 weeks. All patients will then receive AKCEA-TTR-LRx through the remainder of the study treatment period. The final efficacy analysis at week 66 will compare the AKCEA-TTR-LRx arm with the historical placebo arm from the phase 3 trial of inotersen (NEURO-TTR). The primary outcome measures are between-group differences in the change from baseline in serum TTR, modified Neuropathy Impairment Score + 7, and Norfolk Quality of Life—Diabetic Neuropathy questionnaire. Conclusion NEURO-TTRansform is designed to determine whether targeted delivery of AKCEA-TTR-LRx to hepatocytes with lower and less frequent doses will translate into clinical and quality-of-life benefits for patients with hATTR-PN. Trial Registration The study is registered at ClinicalTrials.gov (NCT04136184) and EudraCT (2019-001698-10). Supplementary Information The online version contains supplementary material available at 10.1007/s40120-021-00235-6., Plain Language Summary Hereditary transthyretin amyloidosis with peripheral neuropathy (hATTR-PN for short) is a rare inherited condition.In hATTR-PN, a protein called transthyretin (TTR for short) builds up and damages nerves throughout the body.This neuropathy causes symptoms such as weakness, loss of sensation, and pain.Currently available medicines can slow disease progression, but researchers are looking for more effective treatments with fewer side effects.AKCEA-TTR-LRx is an investigational treatment for hATTR-PN.AKCEA-TTR-LRx prevents the liver from making TTR, reducing the amount that causes disease progression.It is similar to an existing treatment called inotersen, but designed for better delivery to the liver and is more potent.This article describes the NEURO-TTRansform study that will evaluate how effective AKCEA-TTR-LRx is for treating hATTR-PN.Around 140 adults with hATTR-PN from the USA, Canada, and Europe will be able to take part in this study.The study treatment period will be 85 weeks long. People will receive injections underneath the skin of either:AKCEA-TTR-LRx every 4 weeks, orInotersen once a week for 35 weeks, followed by a switch to AKCEA-TTR-LRx every 4 weeks.People may continue to receive AKCEA-TTR-LRx after the study treatment period ends.In this study, researchers will compare results from people who received AKCEA-TTR-LRx to results from people who received no active ingredients (called placebo) in a similar study (called NEURO-TTR).Researchers will measure the differences in peoples’:Neuropathy symptoms.Quality of life.TTR protein levels in the blood. Supplementary Information The online version contains supplementary material available at 10.1007/s40120-021-00235-6.

Aquaculture policy often promotes production of low-trophic level species for sustainable industry growth. Yet, the application of the trophic level concept to aquaculture is complex, and its value for assessing sustainability is further complicated by continual reformulation of feeds. The majority of fed farmed fish and invertebrate species are produced using human-made compound feeds that can differ markedly from the diet of the same species in the wild and continue to change in composition. Using data on aquaculture feeds, we show that technical advances have substantially decreased the mean effective trophic level of farmed species, such as salmon (mean TL = 3.48 to 2.42) and tilapia (2.32 to 2.06), from 1995 to 2015. As farmed species diverge in effective trophic level from their wild counterparts, they are coalescing at a similar effective trophic level due to standardisation of feeds. This pattern blurs the interpretation of trophic level in aquaculture because it can no longer be viewed as a trait of the farmed species, but rather is a dynamic feature of the production system. Guidance based on wild trophic position or historical resource use is therefore misleading. Effective aquaculture policy needs to avoid overly simplistic sustainability indicators such as trophic level. Instead, employing empirically derived metrics based on the specific farmed properties of species groups, management techniques and advances in feed formulation will be crucial for achieving truly sustainable options for farmed seafood.

Serodiagnosis of SARS-CoV-2 infection is impeded by immunological cross-reactivity among the human coronaviruses (HCoVs): SARS-CoV-2, SARS-CoV-1, MERS-CoV, OC43, 229E, HKU1, and NL63. Here we report the identification of humoral immune responses to SARS-CoV-2 peptides that may enable discrimination between exposure to SARS-CoV-2 and other HCoVs. We used a high-density peptide microarray and plasma samples collected at two time points from 50 subjects with SARS-CoV-2 infection confirmed by qPCR, samples collected in 2004–2005 from 11 subjects with IgG antibodies to SARS-CoV-1, 11 subjects with IgG antibodies to other seasonal human coronaviruses (HCoV), and 10 healthy human subjects. Through statistical modeling with linear regression and multidimensional scaling we identified specific peptides that were reassembled to identify 29 linear SARS-CoV-2 epitopes that were immunoreactive with plasma from individuals who had asymptomatic, mild or severe SARS-CoV-2 infections. Larger studies will be required to determine whether these peptides may be useful in serodiagnostics., Mishra, Huang et al. identify 29 linear SARS-CoV-2 epitopes that are immunoreactive with the plasma from individuals who had asymptomatic, mild, or severe SARS-CoV-2 infections. This study suggests the possibility of using these peptides to discriminate the exposure to SARS-CoV-2 and other human coronaviruses.

Wild-type transthyretin (ATTRwt) amyloid deposits have been found in the ligamentum flavum of patients undergoing surgery for spinal stenosis. The relationship between ATTRwt and ligamentum flavum thickness is unclear. We used pre-operative magnetic resonance imaging (MRI) to analyze ligamentum flavum thickness in lumbar spinal stenosis patients with and without ATTRwt amyloid.We retrospectively identified 178 patients who underwent lumbar spine surgery. Ligamentum flavum thickness of 253 specimens was measured on T2-weighted axial MRI. Amyloid presence was confirmed through Congo red staining of specimens, and ATTRwt was confirmed using mass-spectrometry and gene sequencing.Twenty four of the 178 patients (13.5%) were found to have ATTRwt in the ligamentum flavum. Forty ATTRwt specimens and 213 non-ATTRwt specimens were measured. Mean ligamentum flavum thickness was 4.92 (±1.27) mm in the ATTRwt group and 4.00 (±1.21) mm in the non-ATTRwt group (p 0.01). The ligamentum flavum was thickest at L4-L5, with a thickness of 5.15 (±1.27) mm and 4.23 (±1.29) mm in the ATTRwt and non-ATTRwt group, respectively (p = 0.007). There was a significant difference in ligamentum flavum thickness between ATTRwt and non-ATTRwt case for both patients younger than 70 years (p = 0.016) and those older than 70 years (p = 0.004). ATTRwt patients had greater ligamentum flavum thickness by 0.83 mm (95% confidence interval (CI): 0.41-1.25 mm, p 0.001) when controlled for age and lumbar level.Patients with ATTRwt had thicker ligamentum flavum compared to patients without ATTRwt. Further studies are needed to investigate the pathophysiology of ATTRwt in ligamentum flavum thickening.

Mendelian and early-onset severe psychiatric phenotypes often involve genetic variants having a large effect, offering opportunities for genetic discoveries and early therapeutic interventions. Here, the index case is an 18-year-old boy, who at 14 years of age had a decline in cognitive functioning over the course of a year and subsequently presented with catatonia, auditory and visual hallucinations, paranoia, aggression, mood dysregulation, and disorganized thoughts. Exome sequencing revealed a stop-gain mutation in RCL1 (NM_005772.4:c.370 C > T, p.Gln124Ter), encoding an RNA 3′-terminal phosphate cyclase-like protein that is highly conserved across eukaryotic species. Subsequent investigations across two academic medical centers identified eleven additional cases of RCL1 copy number variations (CNVs) with varying neurodevelopmental or psychiatric phenotypes. These findings suggest that dosage variation of RCL1 contributes to a range of neurological and clinical phenotypes.

Horses are used mainly for sports and leisure activities and are predominantly kept in closed individual straw-bedded stalls for a long time during the day, especially in cold weather. Horses are may therefore be exposed to high concentrations of particulate matter and microorganisms from the bedding and fodder material. The dangers posed by exposure the horses to high pollutant concentrations for a long period are respiratory diseases. Thus, the aim of the present research was to detect the potential PM and mold emitting sources in typical horse stables and to identify the concentrations of the particles in the stable air on the basis of quantitative and qualitative studies for the PM and mold concentrations. Long-term measurements of particles and molds were conducted in two horse stables located in Rottgen city, Germany. Collected data from two stables were checked for possible correlations between particulate matter and mold spores. Simulation experiments for the emitted particulates during feeding hay were conducted with a particulate chamber. High diurnal fluctuations in concentrations were observed due to the daily activities in the stables. Fine particles were dominant in the air, especially with a size 0.3–0.4 μm. Mold concentrations in the stables ranged from 3.9 to 5.9 × 105 CFU/m3. The most common mold genera were Eurotium, Wallemia, and Cladosporium. The latter two were found in the straw and hay. A limited prognosis occurred of the mold fungus concentration in horse stables on the basis of airborne particle concentration.

Neutralizing antibody function provides a foundation for the efficacy of vaccines and therapies1–3. Here, using a robust in vitro Ebola virus (EBOV) pseudo-particle infection assay and a well-defined set of solid-phase assays, we describe a wide spectrum of antibody responses in a cohort of healthy survivors of the Sierra Leone EBOV outbreak of 2013–2016. Pseudo-particle virus-neutralizing antibodies correlated with total anti-EBOV reactivity and neutralizing antibodies against live EBOV. Variant EBOV glycoproteins (1995 and 2014 strains) were similarly neutralized. During longitudinal follow-up, antibody responses fluctuated in a ‘decay–stimulation–decay’ pattern that suggests de novo restimulation by EBOV antigens after recovery. A pharmacodynamic model of antibody reactivity identified a decay half-life of 77–100 days and a doubling time of 46–86 days in a high proportion of survivors. The highest antibody reactivity was observed around 200 days after an individual had recovered. The model suggests that EBOV antibody reactivity declines over 0.5–2 years after recovery. In a high proportion of healthy survivors, antibody responses undergo rapid restimulation. Vigilant follow-up of survivors and possible elective de novo antigenic stimulation by vaccine immunization should be considered in order to prevent EBOV viral recrudescence in recovering individuals and thereby to mitigate the potential risk of reseeding an outbreak., In many survivors of Ebola virus infection, antibody responses show long-term patterns of decline followed by restimulation, possibly owing to recrudescence of persisting virus.

Per- and polyfluoroalkyl substances (PFAS) are environmentally persistent, ubiquitous pollutants. It is important to continuously monitor the presence of PFAS contamination, utilizing both legacy and new sentinels. In this study, environmental PFAS levels were evaluated using ticks as a sentinel model due to their world-wide distribution, hematophagous nature, and ease of collection and sampling. Hematophagy in discrete blood meals, from a suite of vertebrates, allows ticks to sample dozens of species of consumers and bioaccumulation across communities. Four different species of ticks, across two states (NY, n = 28 in mid-April of 2020 and FL, n = 32 between 2015 and 2020) with two sampling sites in each state were analyzed for the presence of 53 PFAS. The total PFAS concentration in ticks was the lowest at Newburgh (NY), a site that has been undergoing remediation efforts, while the highest total PFAS concentrations were measured in ticks at the Sweetwater site, a wastewater treatment wetland. Detection of PFAS and the potential for variation between tick species and between locations are necessary to establish the utility of ticks as sentinels, in addition to assessing additional environmental factors, such as other wildlife, water, or soil.

HIV-infected children and adolescents are at increased risk for tuberculosis (TB). Antiretroviral therapy (ART) reduces TB risk in HIV-infected adults, but its effectiveness in HIV-infected children and adolescents is unknown. We analyzed data from 7 integrated pediatric HIV/TB centers in 6 countries in sub-Saharan Africa. We used a Bayesian mixed-effect model to assess association between ART and TB prevalence and used adaptive lasso regression to analyze risk factors for adverse TB outcomes. The study period encompassed 57,525 patient-years and 1,160 TB cases (2,017 cases/100,000 patient-years). Every 10% increase in ART uptake resulted in a 2.33% reduction in TB prevalence. Favorable TB outcomes were associated with increased time in care and early ART initiation, whereas severe immunosuppression was associated with death. These findings support integrated HIV/TB services for HIV-infected children and adults and demonstrate the association of ART uptake with decreased TB incidence in high HIV/TB settings.