Biology subjects

Green, E.

Publications and source records attributed to Green, E..

18 recordsLinked to original sources

bioRxiv2026

Spatial atlas highlights contribution of C. difficile in early-stage colorectal cancer

Background and AimsIntratumoral heterogeneity with respect to both host and microbial components has emerged as an important contributor to colorectal cancer (CRC) biology. Although the overall taxonomy and abundance of the CRC tumor microbiome have become well characterized, much less is known about bacterial niches within the tumor microenvironment (TME). Escherichia coli and Fusobacterium nucleatum are highly prevalent and abundant species linked to CRC. Less abundant organisms, like Clostridioides difficile and Enterocloster aldenensis, are emerging as potentially important CRC-associated bacteria. Additionally, many studies are confounded by preoperative oral antibiotics or the inclusion of late-stage cancers, both of which may alter the gut microbiome. To better understand the spatial relationships between bacteria and CRC, we generated a molecular atlas based on surgically resected tissue specimens collected from a unique cohort of early-stage CRC patients in whom oral antibiotics were not administered preoperatively. MethodsFrom 20 CRC specimens, we performed matched histopathological analysis, fluorescence in situ hybridization (FISH), whole exome sequencing (WES), 16S rRNA amplicon bacterial DNA sequencing, codetection by indexing (CODEX) multiplex immunofluorescence, and spatial transcriptomics of 756 regions among the specimens. We used 16S rRNA amplicon sequencing data to design and experimentally validate custom bacterial probes that were applied to the spatial transcriptomics. Human colonic organoids were used to validate the relationship between Clostridioides difficile toxin B (TcdB) and an E-twenty-six family transcription factor, ELF3. ResultsWe identified eight intratumoral bacterial niches consisting of a variety of bacterial species, and each niche was associated with unique tumor features. We further defined tumor gene expression patterns correlating with individual bacterial species and show that C. difficile has a disproportionate impact on the tumor transcriptome given its relatively low abundance. Specifically, TcdB induces nuclear localization of ELF3, increased cytosolic {beta}-catenin protein, and upregulated WNT signaling. ConclusionOur study integrates multi-omics to identify bacterial species with biological and spatial relevance in CRC regardless of abundance. These findings will enable further studies to define diagnostic and therapeutic targets for bacteria-associated CRC.

Cancer Biology↗

bioRxiv2026

G-VEP: GPU-Accelerated Variant Effect Prediction for Clinical Whole-Genome Sequencing Analysis

Whole-genome sequencing (WGS) has transformed clinical diagnostics, yet variant annotation remains a computational bottleneck. The Variant Effect Predictor (VEP) integrates pathogenicity predictors and population databases essential for ACMG/AMP variant classification, but these annotation plugins are fundamentally I/O-bound, consuming over 70% of total pipeline runtime. Here, we present G-VEP, a GPU-accelerated annotation framework built on a custom CUDA kernel that replaces sequential per-variant database lookups with massively parallel binary search across precomputed indices. By executing annotation lookups for all input variants simultaneously, G-VEP reduces plugin runtime from 72 minutes to 4 minutes (17-fold acceleration) and total annotation runtime from 100 minutes to 33 minutes (3-fold acceleration), while maintaining complete concordance with standard VEP output. Benchmarking across 75 clinical WGS samples demonstrated consistent performance, with no annotation discrepancies; validation on samples containing known pathogenic variants confirmed the preservation of all clinically significant findings. The 8.8 GB index footprint fits within consumer-grade 16 GB GPUs. G-VEP addresses an unmet need in clinical WGS analysis, while GPU suites such as NVIDIA Parabricks accelerate alignment and variant calling, they do not provide the Ensembl VEP plugin ecosystem used in clinical interpretation. G-VEP removes this final bottleneck and enables accelerated WGS interpretation. G-VEP is freely available through a web-based user interface with REST API documentation at https://www.phenomeportal.org/gvep, and source code for local installation and deployment at https://github.com/Phenome-Longevity/G-VEP.

bioinformatics↗

bioRxiv2025

Archetype analysis of lung adenocarcinoma premalignancy links heterogeneity in premalignant lesions to diverging features of invasive disease

ABSTRACTLung adenocarcinoma (LUAD), the most common form of lung cancer, is a heterogeneous disease with highly variable histopathologic features and clinical outcomes. LUAD premalignant lesions (PMLs) are localized proliferative lesions of atypical pneumocytes that proliferate along the alveolar walls. In this study, we characterized the molecular heterogeneity across PMLs, regardless of histologic designation, and identified four unique groups or "archetypes" of PMLs using bulk RNA and exome sequencing data from laser captured microdissected PMLs, normal, and tumor tissue from LUAD resection cases. The archetypes were defined based on expression of recurrent gene co-expression modules discovered using the LUAD PMLs we profiled and three publicly available datasets. One PML archetype, termed "proliferation", was associated with increased expression of genes involved in cell proliferation, a pro-tumor immune environment, and enrichment for EGFR driver mutations. Another archetype, termed "normal-like", was associated with similar features to normal tissue, an anti-tumor immune response, and lacked enrichment for driver mutations compared to other archetypes. We projected the PML archetypes into independent gene expression datasets profiling LUAD and found that tumors closest to the proliferation archetype were enriched for multiple features of aggressive LUAD, including high tumor grade and lymph node invasion, and had significantly lower disease-free survival. Proliferation archetype PMLs may represent a subset of lesions with more aggressive features that could improve patient stratification and suggest novel targets for lung cancer interception.

cancer biology↗

bioRxiv2025

Homozygous kinase-dead Csf1r mutation in outbred mice reveals essential and redundant functions of tissue resident macrophages

The proliferation, differentiation and survival of cells of the macrophage lineage depends on signals from the macrophage colony-stimulating factor receptor (CSF1R). On a C57BL/6J background homozygous kinase-dead Csf1r mutation (Csf1rE631K/E631K - E631Km/m) causes perinatal lethality. Here we demonstrate that E631Km/m mice on a mixed genetic background (C57 x BALB/c F2) are osteopetrotic and growth retarded but viable as adults with no other gross developmental deficits. They lack osteoclasts, microglia and most peripheral tissue resident macrophages and exhibit perturbed hematopoiesis. Although CD169+ tissue resident macrophages in bone marrow are considered an essential component of the hematopoietic niche, CD169 is undetectable in E631Km/m marrow and F4/80+ macrophages are depleted. These changes are associated with expansion of mature and immature granulocytes and reduced B cells, whereas monocytes and stem and progenitor populations are unaffected as a proportion of total cells. Erythropoiesis in bone marrow is maintained in E631Km/m mice, associated with a residual population of CSF1R-independent CD169-ve/F4/80+ macrophages. Nevertheless, splenic extramedullary hematopoiesis in E631Km/m mice indicates a degree of bone marrow insufficiency. Red pulp macrophages are retained but CD169+ marginal metallophil macrophages are absent and CD209b+ (SIGNR1) macrophages are present but disorganized. Circulating white blood cell count is unchanged in E631Km/m mice, but the proportion of neutrophils is greatly increased whilst B cells and monocytes are reduced. This novel model reveals the essential roles of CSF1R-dependent macrophages in hematopoiesis and demonstrates that many developmental and homeostatic functions attributed to murine resident tissue macrophages are redundant and/or specific to inbred mouse strains.

immunology↗

bioRxiv2025

Seasonal assembly of the phyllosphere fungal microbiome of a perennial grass is robust to nutrient addition

The leaf microbiome plays an important role in plant health and defense. Despite its importance, how the assembly of the leaf microbial community is modified by environmental conditions such as nutrient availability remains relatively uninvestigated. Soil nutrient availability may shift the outcome of microbial interactions within a host individual or influence the pool of microbes across the plant community. We hypothesized that leaf microbial diversity would increase across the season as leaves collect additional taxa, and that this seasonal assembly would be sensitive to nutrient addition. To assess this, we tracked the assembly of the fungal phyllosphere microbiome of the grass tall fescue (Lolium arundinaceum) in old-field vegetation over the growing season and experimentally tested whether the seasonality of the microbiome was modified by experimental addition of soil nutrients. Fungal diversity (Shannon diversity index, richness, and evenness) increased early in the season, with most metrics saturating before the end of the season. Community composition as measured by Bray-Curtis dissimilarity also shifted over the early and mid-growing season. Phylogeny-based machine-learning identified fungal lineages that were abundant in different seasons, linking seasonal community shifts to their evolutionary context. Nutrient addition was less important than time of season, but still significantly altered community composition and interacted with time to influence richness, with lowest richness in the low nutrient addition plots early in the season. The clear seasonality of the microbiome provides support for a dynamic phyllosphere microbiome, suggesting further studies manipulating fungal recruitment over the season. Furthermore, it highlights the robustness of seasonal assembly to variation in nutrient availability.

ecology↗

bioRxiv2025

Differential Glutamatergic Inputs to Semilunar Granule Cells and Granule Cells Underscore Dentate Gyrus Projection Neuron Diversity

Semilunar Granule Cells (SGCs) are sparse dentate gyrus projection neurons whose role in the dentate circuit, including pathway specific inputs, remains unknown. We report that SGCs receive more frequent spontaneous excitatory synaptic inputs than granule cells (GCs). Dual GC-SGC recordings identified that SGCs receive stronger medial entorhinal cortex and associational synaptic drive but lack short-term facilitation of lateral entorhinal cortex inputs observed in GCs. SGCs dendritic spine density in proximal and middle dendrites was greater than in GCs. However, the strength of commissural inputs and dendritic input integration, examined in passive morphometric simulations, were not different between cell types. Activity dependent labeling identified an overrepresentation of SGCs among neuronal ensembles in both mice trained in a spatial memory task and task naive controls. The divergence of modality specific inputs to SGCs and GCs can enable parallel processing of information streams and expand the computational capacity of the dentate gyrus.

neuroscience↗

bioRxiv2025

Epithelial miR-149-5p up-regulation is associated with immune evasion in progressive bronchial premalignant lesions

The molecular drivers bronchial premalignant lesion progression to invasive lung squamous cell carcinoma are not well defined. Prior work profiling longitudinally collected bronchial premalignant lesion biopsies by RNA sequencing defined a proliferative subtype, enriched with bronchial dysplasia. We found that a gene co-expression module associated with interferon gamma signaling and antigen processing/presentation was down-regulated in progressive/persistent versus regressive lesions within the proliferative subtype, suggesting a functional impact of these genes on immune evasion. RNA from these same premalignant lesions was profiled by microRNA (miRNA) sequencing and a miRNA-gene network analysis identified hsa-miR-149-5p as a potential regulator of this antigen presentation gene co-expression module associated with lesion progression. hsa-miR-149-5p was found to be predominantly expressed in the epithelium and up-regulated in progressive/persistent versus regressive proliferative lesions while targets of this miRNA, the transcriptional coactivator of MHC-I gene expression, NLRC5, and the genes it regulates were down-regulated. MicroRNA in situ hybridization of hsa-miR-149-5p in tissue from adjacent fixed biopsies showed that hsa-miR-149-5p was increased in areas of bronchial dysplasia in progressive/persistent versus regressive lesions. Imaging mass cytometry showed that NLRC5 protein expression was decreased in progressive/persistent versus regressive lesions within areas of hyperplasia, metaplasia, and dysplasia. Additionally, basal cells with high versus low levels of NLRC5 were found to be in close spatial proximity to CD8 T cells, suggesting that these cells exhibit increased functional MHC-I gene expression in lesions with low hsa-miR-149-5p expression. Collectively, our data suggests a functional role for hsa-miR-149-5p in bronchial premalignant lesions and may serve as a therapeutic target for PML immunomodulation. STATEMENT OF SIGNIFICANCEIntegrative analysis across bronchial premalignant lesions has identified and localized a potential regulator of immune evasion in progressive/persistent lesions that could be a novel therapeutic target.

cancer biology↗

bioRxiv2024

Most Beefalo cattle have no detectable bison genetic ancestry

Hybridization is common among lineages in the genus Bos, often mediated through human management for the selection of adaptive or desirable traits. A recent example is the American Beefalo cattle breed, which was developed in the 1970s and defined as a hybrid between American bison (Bison bison) and cattle (Bos taurus). The American Beefalo Association (ABA) typically require [3/8] bison ancestry to qualify as Beefalo. Here, we sought to characterize admixed ancestry among Beefalo as a component of a larger project to understand the role of hybridization in shaping present-day diversity in bison and cattle. We generated genomic data from 50 historical and present-day Beefalo and bison hybrids, including several important founding animals, as well as from 10 bison originating from commercial herds that represent potential sources of bison ancestry in Beefalo. We found that most Beefalo did not contain detectable bison ancestry. No individual Beefalo within our data set satisfies the ancestry requirements specified by the ABA, although several Beefalo had smaller proportions of bison ancestry (2-18%). Some beefalo had detectable zebu cattle ancestry (2-38%), suggesting that hybridization of taurine and zebu cattle may contribute to morphological similarity between some Beefalo and bison. Overall, ancestry profiles of Beefalo and bison hybrid genomes are consistent with repeated backcrossing to either parental species rather than the breeding between hybrids themselves, implying significant barriers to gene flow between bison and cattle. Our results call into question the [3/8] bison ancestry targeted by the breed association and demonstrate the value of genomic information in examining claims of interspecies gene flow among Bos species.

evolutionary biology↗

bioRxiv2024

An off-lattice discrete model to characterise filamentous yeast colony morphology

We combine an off-lattice agent-based mathematical model and experimentation to explore filamentous growth of a yeast colony. Under environmental stress, Saccharomyces cerevisiae yeast cells can transition from a bipolar (sated) to unipolar (pseudohyphal) budding mechanism, where cells elongate and bud end-to-end. This budding asymmetry yields spatially non-uniform growth, where filaments extend away from the colony centre, foraging for food. We use approximate Bayesian computation to quantify how individual cell budding mechanisms give rise to spatial patterns observed in experiments. We apply this method of parameter inference to experimental images of colonies of two strains of S. cerevisiae, in low and high nutrient environments. The initial nutrient concentration and a forking mechanism for pseudohyphal cell proliferation are the key features driving colony morphology. Simulations run with the most likely inferred parameters produce colony morphologies that closely resemble experimental results.

cell biology↗

bioRxiv2024

Direct interrogation of context-dependent GPCR activity with a universal biosensor platform

G protein-coupled receptors (GPCRs) are the largest family of druggable proteins in the human genome, but progress in understanding and targeting them is hindered by the lack of tools to reliably measure their nuanced behavior in physiologically-relevant contexts. Here, we developed a collection of compact ONE vector G-protein Optical (ONE-GO) biosensor constructs as a scalable platform that can be conveniently deployed to measure G-protein activation by virtually any GPCR with high fidelity even when expressed endogenously in primary cells. By characterizing dozens of GPCRs across many cell types like primary cardiovascular cells or neurons, we revealed new insights into the molecular basis for G-protein coupling selectivity of GPCRs, pharmacogenomic profiles of anti-psychotics on naturally-occurring GPCR variants, and G-protein subtype signaling bias by endogenous GPCRs depending on cell type or upon inducing disease-like states. In summary, this open-source platform makes the direct interrogation of context-dependent GPCR activity broadly accessible.

biochemistry↗

bioRxiv2023

Overexpression of Alpha-1 Antitrypsin Increases the Proliferation of Mesenchymal Stem Cells by Upregulation of Cyclin D1 and is Independent of the Wnt Signaling Pathway

Alaph-1 antitrypsin overexpressing mesenchymal stromal/stem cells (AAT-MSCs) showed improved innate properties with a faster proliferation rate when studied for their protective effects in mouse models of diseases. Here, we investigated the potential mechanism(s) by which AAT gene insertion increases MSC proliferation. Human bone marrow-derived primary or immortalized MSCs (iMSCs) or AAT-MSCs (iAAT-MSCs) were used in the study. Cell proliferation was measured by cell counting and cell cycle analysis. Possible pathways involved in the pro-proliferation effect of AAT were investigated by measuring mRNA and protein expression of key cell cycle genes. Interval cell counting showed increased proliferation in AAT-MSCs or iAAT-MSCs compared to their corresponding MSC controls. Cell cycle analysis revealed more cells progressing into the S and G2/M phases in iAAT-MSCs, with a notable increase in the cell cycle protein, Cyclin D1. Moreover, treatment with Cyclin D1 inhibitors showed that the increase in proliferation is due to Cyclin D1 and that the AAT protein is upstream and a positive regulator of Cyclin D1. Furthermore, AATs effect on Cyclin D1 is independent of the Wnt signaling pathway as there were no differences in the expression of regulatory proteins, including GSK3{beta} and {beta}-Catenin in iMSC and iAAT-MSCs. In summary, our results indicate that AAT gene insertion in an immortalized MSC cell line increases cell proliferation and growth by increasing Cyclin D1 expression and consequently causing cells to progress through the cell cycle at a significantly faster rate.

cell biology↗

bioRxiv2022

Structural Basis of Prostaglandin Efflux by MRP4

MRP4 is unique among the C family of ATP-binding cassette transporters for its role in translocating prostanoids, an important group of signaling molecules derived from unsaturated fatty acids. Using a reconstituted system, we report that a pair of prostaglandins (PGs) and the sulfonated-sterol DHEA-S preferentially enhance the ATPase activity of MRP4 over other previously proposed physiological substrates such as cyclic nucleotides or leukotrienes. We determined the cryo-EM structures of nanodisc embedded bovine MRP4 in (i) a nucleotide- and substrate-free state, (ii) in complex with PGE1, (iii) PGE2, and (iv) DHEA-S, and (v) a catalytically dead mutant E1202Q bound to ATP-Mg2+. The substrate-bound structures suggest unique features of the MRP4 binding site that distinguish its specificity for prostanoids from that of the related leukotriene transporter MRP1. The ATP-bound structure is in an outward-occluded conformation, revealing a novel state in the proposed alternate-access mechanism of MRP transport. Our study provides insights into the endogenous function of this versatile efflux transporter.

biochemistry↗

bioRxiv2022

Widespread, depth-dependent microstructural damage in the cortex of children with drug resistant focal epilepsy: A quantitative T1 and T2 mapping study

ObjectiveTissue abnormalities in focal epilepsy may extend beyond the presumed focus. The underlying pathophysiology of these broader changes is unclear, and it is not known whether they result from ongoing disease processes, treatment-related side-effects, or whether they emerge earlier. Few studies have focused on the period of onset for most focal epilepsies, childhood. Fewer still have utilised quantitative MRI, which may provide a more sensitive and interpretable measure of tissue microstructural change. Here, we aimed to determine common spatial modes of changes in cortical architecture in children with heterogeneous drug-resistant focal epilepsy and, secondarily, whether changes were related to disease severity. MethodsTo assess cortical microstructure, quantitative T1 and T2 relaxometry (qT1 and qT2) was measured in 43 children with drug-resistant focal epilepsy [age-range=4-18 years] and 46 typically-developing children [age-range=2-18 years]. We assessed depth-dependent qT1 and qT2 values across the neocortex, as well as their gradient of change across cortical depths. We also determined whether global changes seen in group analyses were driven by focal pathologies in individual patients. Finally, as a proof-of-concept, we trained a classifier using qT1 and qT2 gradient maps from patients with radiologically-defined abnormalities (MRI-positive) and healthy controls, and tested if this could classify patients without reported radiological abnormalities (MRI-negative). ResultsWe uncovered depth-dependent qT1 and qT2 increases in widespread cortical areas in patients, likely representing microstructural alterations in myelin or gliosis. Changes did not correlate with disease severity measures, suggesting they may represent antecedent neurobiological alterations. Using a classifier trained with MRI-positive patients and controls, sensitivity was 62% at 100% specificity on held-out MRI-negative patients. SignificanceThese findings suggest the presence of a potential imaging endophenotype of focal epilepsy, detectable irrespective of radiologically-identified abnormalities, and potentially evident pre-symptomatically. Key PointsO_LIWe assessed cortical microstructure in children with focal epilepsy C_LIO_LIQuantitative T1 and T2 relaxometry (qT1 and qT2) was measured in the neocortex C_LIO_LIPatients showed extensive qT1/qT2 increases and intracortical organization changes C_LIO_LIAlterations may appear during cerebral development, prior to disease onset C_LI

neuroscience↗

bioRxiv2022

Automated assembly of high-quality diploid human reference genomes

The current human reference genome, GRCh38, represents over 20 years of effort to generate a high-quality assembly, which has greatly benefited society1, 2. However, it still has many gaps and errors, and does not represent a biological human genome since it is a blend of multiple individuals3, 4. Recently, a high-quality telomere-to-telomere reference genome, CHM13, was generated with the latest long-read technologies, but it was derived from a hydatidiform mole cell line with a duplicate genome, and is thus nearly homozygous5. To address these limitations, the Human Pangenome Reference Consortium (HPRC) recently formed with the goal of creating a collection of high-quality, cost-effective, diploid genome assemblies for a pangenome reference that represents human genetic diversity6. Here, in our first scientific report, we determined which combination of current genome sequencing and automated assembly approaches yields the most complete, accurate, and cost-effective diploid genome assemblies with minimal manual curation. Approaches that used highly accurate long reads and parent-child data to sort haplotypes during assembly outperformed those that did not. Developing a combination of all the top performing methods, we generated our first high- quality diploid reference assembly, containing only [~]4 gaps (range 0-12) per chromosome, most within + 1% of CHM13s length. Nearly 1/4th of protein coding genes have synonymous amino acid changes between haplotypes, and centromeric regions showed the highest density of variation. Our findings serve as a foundation for assembling near-complete diploid human genomes at the scale required for constructing a human pangenome reference that captures all genetic variation from single nucleotides to large structural rearrangements.

genomics↗

bioRxiv2019

Prevalence and distribution of antimicrobial resistance determinants of Escherichia coli isolates obtained from meat in South Africa

ObjectiveThis study aimed to characterise antibiotics resistance of Escherichia coli isolates from the formal meat sector (FMS) and informal meat sectors (INMS).\n\nMethodA total of 162 and 102 E. coli isolates from the FMS, and INMS respectively were isolated by standard culture-based, and biochemical reactions. The isolates were further confirmed by polymerase chain reaction (PCR). The disc diffusion method was used to screen for antimicrobial susceptibility against 19 different antibiotics. The presence of class 1-2 integrons in each E. coli isolates was assessed using [Formula] and [Formula] regions specific primers.\n\nResultAmong the 19 antimicrobials, resistance to tetracyclines, aminoglycosides, cephalosporins, and nitrofurans were found to be more frequent than carbapenems and phenicol with a noticeable increase in the number of multi-drug resistance ranging from three to ten antimicrobials. A total of 20 resistance determinants were assessed with their prevalence and distributions obtained as follows for FMS and INMS respectively; [aminoglycosides: aadA (40.6%; 31.9%), aacC2 (21.4%; 31%), aphA1 (20.8%; 15.1%), aphA2 (37.7%; 18.9%) and strA (6.5%; 9.4%)], [{beta}-lactams: ampC (20%; 45%), blaTEM, (4.4%; 13.3), and blaZ (8.9%; 2.2%)], [Chloramphenicol: catI (1.7%; 1.7%), and cmIA1 (1.7%; 1.7%)] and [tetracyclines: tetA (7.7%; 15.4%), tetB (11.5%; 24%), and tetM, (1.9%; 8.7%)], and [sulfonamides: sul1 (22.2%; 26.7%), sul2 (17.8%; 6.7%)].\n\nConclusionMultiple antibiotic resistance (MAR) indexes ranged from 0.2 - 0.5. The results reveal a high prevalence of multidrug-resistant E. coli isolates and resistance determinants suggesting that consumers and handlers of such meat are at risk of contracting antibiotic resistant E. coli-related foodborne disease.

microbiology↗

bioRxiv2018

Epidemiological impact of hepatitis B vaccination in Monastir Tunisia (2000-17)

Background: In 2016, the first global health sector strategy on viral hepatitis was endorsed with the goal of eliminating viral hepatitis as a public health threat by 2030. In Tunisia, effective vaccines for hepatitis B (HBV) have been added to the expanded programme of immunization (EPI) since July 1995 for new borns. We expected to have a decreasing trend in the prevalence rate of reported HBV. Our study aimed to address the epidemiological profile of HBV, to assess trends by age and gender in Monastir governorate over a period of 18 years according to immunization status and to estimate the burden (years lived with disability YLDs) of this pathology. Methods: We performed a descriptive cross sectional study of declared HBV from January 1, 2000 to December 31, 2017 defined as having positive serologic markers for HBs Ag. All declared patients were residents of Monastirs Governorate. EPI included two periods, the first between 1995 and 2006 following a three-dose schedule (3, 4, 9 months). The second PI cohort after 2006 following a three-dose schedule (0, 2, 6 months). Results: During 18 years, 1526 cases of HBV were declared in Monastir with a mean of 85 cases per year. We estimated a mean of 1699 declared cases per year of HBV in Tunisia. CPR was 16.85/100,000 inh being the higher in age group of 20-39 years and in men .ASR was 15.99/100,000 inh, being 35.5 in men and 8.69 in women. During the study period, declared cases among presumed immunized (PI) person against HBV were 32(2.0%). Among PI cases, 29 were from the first period and 3 were from the second. We established a negative trend over 18 years of hepatitis B. The age group of 20 to 39 was the most common with a sharply decline. Presumed not immunized (PNI) HBV cases are decreasing by years with a prediction of 35 cases in 2024. Reported HBV contributed to 1.26 YLDs per 100,000 inh. The highest rate of YLDs occurred at the age 20-39 (2.73 YLDs per 100,000 inh). During 18 years, YLDs were 114.45 in Monastir with a mean of 2293.65 YLDs of HBV in Tunisia. Conclusion, this study showed a law prevalence rate and a decreasing trend of HBV during 18 years showing an efficacy of immunization and confirming that the universal hepatitis B vaccination in Tunisia has resulted in progress towards the prevention and control of hepatitis B infection. These findings should be demonstrated in other Tunisian regions with a standardized serological profile.

immunology↗

bioRxiv2018

The structural basis for activation of voltage sensor domains in an ion channel TPC1

Voltage sensing domains (VSDs) couple changes in transmembrane electrical potential to conformational changes that regulate ion conductance through a central channel. Positively charged amino acids inside each sensor cooperatively respond to changes in voltage. Our previous structure of a TPC1 channel captured the first example of a resting-state VSD in an intact ion channel. To generate an activated state VSD in the same channel we removed the luminal inhibitory Ca2+-binding site (Cai2+), that shifts voltage-dependent opening to more negative voltage and activation at 0 mV. Cryo-EM reveals two coexisting structures of the VSD, an intermediate state 1 that partially closes access to the cytoplasmic side, but remains occluded on the luminal side and an intermediate activated state 2 in which the cytoplasmic solvent access to the gating charges closes, while luminal access partially opens. Activation can be thought of as moving a hydrophobic insulating region of the VSD from the external side, to an alternate grouping on the internal side. This effectively moves the gating charges from the inside potential to that of the outside. Activation also requires binding of Ca2+ to a cytoplasmic site (Caa2+). An X-ray structure with Caa2+ removed and a near-atomic resolution cryo-EM structure with Cai2+ removed define how dramatic conformational changes in the cytoplasmic domains may communicate with the VSD during activation. Together four structures provide a basis for understanding the voltage dependent transition from resting to activated state, the tuning of VSD by thermodynamic stability, and this channels requirement of cytoplasmic Ca2+-ions for activation.

biophysics↗

bioRxiv2017

TOWARDS A GLOBAL SUPPORT OF CORE DATA RESOURCES FOR THE LIFE SCIENCES

On November 18-19, 2016, the Human Frontier Science Program Organization (HFSPO) hosted a meeting of senior managers of key data resources and leaders of several major funding organizations to discuss the challenges associated with sustaining biological and biomedical (i.e., life sciences) data resources and associated infrastructure. A strong consensus emerged from the group that core data resources for the life sciences should be supported through a coordinated international effort(s) that better ensure long-term sustainability and that appropriately align funding with scientific impact. Ideally, funding for such data resources should allow for access at no charge, as is presently the usual (and preferred) mechanism. Below, the rationale for this vision is described, and some important considerations for developing a new international funding model to support core data resources for the life sciences are presented.

scientific communication and education↗