bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prenatal exposure to valproic acid reduces synaptic δ-catenin levels and disrupts ultrasonic vocalization in neonates
Authors: Roh Seung Hyun , Mendez-Vazquez Hadassah , Sathler Matheus F. , Doolittle Michael J. , Zaytseva Anastasiya , Brown Hannah , Sainsbury Morgan , Kim Seonil 
Published: December 2023

AbstractValproic acid (VPA) is an effective and commonly prescribed drug for epilepsy and bipolar disorder. However, children born from mothers treated with VPA during pregnancy exhibit an increased incidence of autism spectrum disorder (ASD). Although VPA may impair brain development at the cellular level, the mechanism of VPA-induced ASD has not been completely addressed. A previous study has found that VPA treatment strongly reduces δ-catenin mRNA levels in cultured human neurons. δ-catenin is important for the control of glutamatergic synapses and is strongly associated with ASD. VPA inhibits dendritic morphogenesis in developing neurons, an effect that is also found in neurons lacking δ-catenin expression. We thus hypothesize that prenatal exposure to VPA significantly reduces δ-catenin levels in the brain, which impairs glutamatergic synapses to cause ASD. Here, we found that prenatal exposure to VPA markedly reduced δ-catenin levels in the brain of mouse pups. VPA treatment also impaired dendritic branching in developing mouse cortical neurons, which was reversed by elevating δ-catenin expression. Prenatal VPA exposure significantly reduced synaptic AMPA receptor levels and postsynaptic density 95 (PSD95) in the brain of mouse pups, indicating dysfunctions in glutamatergic synaptic transmission. VPA exposure also significantly altered ultrasonic vocalization (USV) in newly born pups when they were isolated from their nest. Moreover, VPA-exposed pups show impaired hypothalamic response to isolation, which is required to produce animals’ USVs following isolation from the nest. Therefore, these results suggest that VPA-induced ASD pathology can be mediated by the loss of δ-catenin functions.HighlightsPrenatal exposure of valproic acid (VPA) in mice significantly reduces synaptic δ-catenin protein and AMPA receptor levels in the pups’ brains.VPA treatment significantly impairs dendritic branching in cultured cortical neurons, which is reversed by increased δ-catenin expression.VPA exposed pups exhibit impaired communication such as ultrasonic vocalization.Neuronal activation linked to ultrasonic vocalization is absent in VPA-exposed pups.The loss of δ-catenin functions underlies VPA-induced autism spectrum disorder (ASD) in early childhood.
AVENGERS: Analysis of Variant Effects using Next Generation sequencing to EnhanceBRCA2Stratification
Authors: Sahu Sounak , Galloux Melissa , Southon Eileen , Caylor Dylan , Sullivan Teresa , Arnaudi Matteo , Geh Josephine , Chari Raj , Papaleo Elena , Sharan Shyam K. 
Published: December 2023

AbstractAccurate interpretation of genetic variation is a critical step towards realizing the potential of precision medicine. Sequencing-based genetic tests have uncovered a vast array ofBRCA2sequence variants. Due to limited clinical, familial and/or epidemiological data, thousands of variants are considered to be variants of uncertain significance (VUS). To determine the functional impact of VUSs, here we develop AVENGERS: Analysis of Variant Effects using NGs to Enhance BRCA2 Stratification, utilizing CRISPR-Cas9-based saturation genome editing (SGE) in a humanized-mouse embryonic stem cell line. We have categorized nearly all possible missense single nucleotide variants (SNVs) encompassing the C-terminal DNA binding domain ofBRCA2.We have generated the function scores for 6270 SNVs, covering 95.5% of possible SNVs in exons 15-26 spanning residues 2479-3216, including 1069 unique missense VUS, with 81% functional and 14% found to be nonfunctional. Our classification aligns strongly with pathogenicity data from ClinVar, orthogonal functional assays and computational meta predictors. Our statistical classifier exhibits 92.2% sensitivity and 96% specificity in distinguishing clinically benign and pathogenic variants recorded in ClinVar. Furthermore, we offer proactive evidence for 617 SNVs being non-functional and 3396 SNVs being functional demonstrated by impact on cell growth and response to DNA damaging drugs like cisplatin and olaparib. This classification serves as a valuable resource for interpreting unidentified variants in the population and for physicians and genetic counselors assessingBRCA2VUSs in patients.
A molecular toolkit for heterologous protein secretion across Bacteroides species
Authors: Yeh Yu-Hsuan , Kelly Vince W. , Pour Rahman Rahman , Sirk Shannon J. 
Published: December 2023

SummaryBacteroides species are abundant and prevalent stably colonizing members of the human gut microbiota, making them a promising chassis for developing long-term interventions for chronic diseases. Engineering these bacteria as on-site production and delivery vehicles for biologic drugs or diagnostics, however, requires efficient heterologous protein secretion tools, which are currently lacking. To address this limitation, we systematically investigated methods to enable heterologous protein secretion in Bacteroides using both endogenous and exogenous secretion systems. Here, we report a collection of secretion carriers that can export functional proteins across multiple Bacteroides species at high titers. To understand the mechanistic drivers of Bacteroides secretion, we characterized signal peptide sequence features as well as post-secretion extracellular fate and cargo size limit of protein cargo. To increase titers and enable flexible control of protein secretion, we developed a strong, self-contained, inducible expression circuit. Finally, we validated the functionality of our secretion carriers in vivo in a mouse model. This toolkit should enable expanded development of long-term living therapeutic interventions for chronic gastrointestinal disease.
Genetic landscape of anin vivoprotein interactome
Authors: Besse Savandara , Sakaguchi Tatsuya , Gauthier Louis , Sahaf Zahra , Péloquin Olivier , Gonzalez Lidice , Castellanos-Girouard Xavier , Koçatug Nazli , Matta Chloé , Hussin Julie G. , Michnick Stephen W. , Serohijos Adrian W.R. 
Published: December 2023

SUMMARYProtein-protein interaction (PPI) networks accurately map environmental perturbations to their molecular consequences in cells, but effects of genome-wide genetic variation on PPIs remain unknown. We hypothesized that PPI networks integrate genetic and environmental effects, potentially defining biochemical mechanisms underlying complex polygenic traits. Here, we measured 61 PPIs in inbred strains ofSaccharomyces cerevisiaewith ∼12,000 single-nucleotide polymorphisms (SNPs) across the genome. Unlike mRNA expression and protein abundance that are primarily affected by SNPs local (in “cis”) to a gene, PPIs are predominantly affected by SNPs far (in “trans”) to the genomic loci of the interacting proteins. However, consistent with the PPI network’s small-world characteristic, these transacting SNPs are in neighboring genes in the network. We likewise discovered SNPs in non-coding RNAs and post-transcriptional regulators (3’ UTRs) with, counterintuitively, larger PPI-modulating effects than SNPs within protein-coding regions. Finally, we inferred known and novel mechanisms of action for yeast and human drugs.HIGHLIGHTSProtein-interaction quantitative trait locus (“piQTL”) mapping reveals sensitivity ofin vivoPPIs to polymorphisms across the yeast genomeTrans-piQTLs significantly outnumber and are stronger than cis-piQTLsSNPs in non-coding RNAs and 3’ UTRs have comparable effects to PPI as SNPs in coding regionspiQTL mapping reveals known and novel mechanism of yeast and human drugs
A high-throughput screening approach to discover potential colorectal cancer chemotherapeutics: Repurposing drugs to disrupt 14-3-3 protein-BAD interactions
Authors: He Siyi , Silva Luis Delgadillo , Rutter Guy A. , Lim Gareth E. 
Published: December 2023

AbstractInducing apoptosis in different types of cancer cells is an effective therapeutic strategy. However, the success of existing chemotherapeutics can be compromised by tumor cell resistance and systemic off-target effects. Therefore, the discovery of pro-apoptotic compounds with minimal systemic side-effects is crucial. 14-3-3 proteins are molecular scaffolds that serve as important regulators of cell survival. Our previous study demonstrated that 14-3-3ζ can sequester BAD, a pro-apoptotic member of the BCL-2 protein family, in the cytoplasm and prevent its translocation to mitochondria to inhibit the induction of apoptosis. Despite being a critical mechanism of cell survival, it is unclear whether disrupting 14-3-3 protein:BAD interactions could be harnessed as a chemotherapeutic approach. Herein, we established a BRET-based high-throughput drug screening approach (Z’-score= 0.52) capable of identifying molecules that can disrupt 14-3-3ζBAD interactions. An FDA-approved drug library containing 1971 compounds was used for screening, and the capacity of identified hits to induce cell death was examined in NIH3T3-fibroblasts and colorectal cancer cell lines, HT-29 and Caco-2. Ourin vitroresults suggest that terfenadine, penfluridol, and lomitapide could be potentially repurposed for treating colorectal cancer. Moreover, our screening method demonstrates the feasibility of identifying pro-apoptotic agents that can be applied towards conditions where aberrant cell growth or function are key determinants of disease pathogenesis.
Ecological niche contributes to the persistence of the Western x Glaucous-winged Gull hybrid zone
Authors: Geng Xuewen , Summers Jeremy , Chen Nancy 
Published: December 2023

AbstractHybrid zones occur in nature when populations with limited reproductive barriers overlap in space. Many hybrid zones persist over time, and different models have been proposed to explain how selection can maintain hybrid zone stability. More empirical studies are needed to elucidate the role of ecological adaptation in maintaining stable hybrid zones. Here, we investigated the role of exogenous factors in maintaining a hybrid zone between western gulls (Larus occidentalis) and glaucous-winged gulls (L. glaucescens). We used ecological niche models (ENMs) and niche similarity tests to quantify and examine the ecological niches of western gulls, glaucous-winged gulls, and their hybrids. We found evidence of niche divergence between all three groups. Our results best support the bounded superiority model, providing further evidence that exogenous selection favoring hybrids may be an important factor in maintaining this stable hybrid zone.
Metabolic and genomic characterization of two novel butyrate producing Lachnospiraceae isolated from swine feces
Authors: Watkins Hannah R. , Trachsel Julian M. , Bearson Shawn M. D. , Loving Crystal L. , Anderson Christopher L. 
Published: December 2023

AbstractCommensal bacteria from the swine gut microbiome that can be isolated have numerous potential applications in the animal production industry, including mitigation of disease, improving performance, and promoting colonization resistance to human foodborne pathogens. Butyrate-producing bacteria are targets for next-generation probiotics and microbiome-engineering strategies because butyrate is a metabolite of central importance in large intestinal homeostasis and may augment colonization resistance to enteric pathogens. However, relatively few butyrate-producers from swine have been cultured and extensively characterized. Here, we describe the substrate utilization, metabolic profiles, and genomic features of two novel species that produce high concentration of butyratein vitro,Roseburia sp.831b andPetralouisia sp.499, isolated from swine feces. The complete genomes illustrated versatility in carbon metabolism and unique carbohydrate-active enzymes not observed in other species ofRoseburiaandPetralouisiathat encode a combination of glycosidic hydrolases and carbohydrate-binding modules involved in starch and pectin utilization.Roseburia sp.831b fermented a broader range and more complex mono- and polysaccharides thanPetralouisia sp.499. Fecal and cecal metagenomes from eight-week-old pigs challenged with Salmonella revealed thatRoseburia sp.831b increased to detectable abundances in the swine hindgut in most animals at ∼63-70 days of age. Additionally, the abundance ofRoseburia sp.831b in fecal metagenomes correlated with fecal butyrate concentrations in the pigs fed a diet supplemented with a prebiotic resistant potato starch. Together, these findings highlight the probiotic potential and ecological niche in the swine gastrointestinal tract for two novel butyrate-producers.ImportanceAntibiotics have been important for swine production and management of enteric pathogens; however, the Veterinary Feed Directive limits the use of medically important in-feed antibiotics for production purposes. As a result, there is a need for alternatives to antibiotics. Butyrate-producing bacteria can improve colonization resistance to human pathogens within the swine gastrointestinal tract by reinforcing the intestinal barrier, increasing mucus production, and reducing local oxygen and pH levels. Here, we demonstrate the versatile substrate utilization and metabolic potential of two novel species isolated from swine that produce high butyrate concentrationsin vitro. These findings will help develop strategies that increase the abundance of these species and other butyrate producers in the swine gut. Further, isolating and characterizing swine butyrate producers is necessary for controlled studies that provide a mechanistic understanding of how this functional group of bacteria promotes swine gut health and colonization resistance to bacteria of public health concern.
50-color phenotyping of the human immune system with in-depth assessment of T cells and dendritic cells
Authors: Konecny Andrew J. , Mage Peter , Tyznik Aaron J. , Prlic Martin , Mair Florian 
Published: December 2023

AbstractWe report the development of an optimized 50-color spectral flow cytometry panel designed for the in-depth analysis of the immune system in human blood and tissues, with the goal of maximizing the amount of information that can be collected using currently available flow cytometry platforms. We established and tested this panel using peripheral blood mononuclear cells (PBMCs), but included CD45 to enable its use for the analysis of human tissue samples. The panel contains lineage markers for all major immune cell subsets, and an extensive set of phenotyping markers focused on the activation and differentiation status of the T cell and dendritic cell (DC) compartment.We outline the biological insight that can be gained from the simultaneous measurement of such a large number of proteins and propose that this approach provides a unique opportunity for the comprehensive exploration of the immune status in tissue biopsies and other human samples with a limited number of cells. Of note, we tested the panel to be compatible with cell sorting for further downstream applications. Furthermore, to facilitate the wide-spread implementation of such a panel across different cohorts and samples, we established a trimmed-down 45-color version which can be used with different spectral cytometry platforms.Finally, to generate this panel, we utilized not only existing panel design guidelines, but also developed new metrics to systematically identify the optimal combination of 50 fluorochromes and evaluate fluorochrome-specific resolution in the context of a 50-color unmixing matrix.
BMI and BPH correlate with urinary microbiome diversity and lower urinary tract symptoms in men
Authors: Bowie Kate R. , Garzotto Mark , Orwoll Eric , Karstens Lisa 
Published: December 2023

AbstractSeveral studies have identified bacteria and other microbes in the bladder and lower urinary tract in the absence of infection. In women, the urinary microbiome has been associated with lower urinary tract symptoms (LUTS), however, similar studies have not been undertaken in large cohorts of men. Here we examine the urinary microbiome and its association with LUTS in a subset of 500 men aged 65 to 90 years from the Osteoporotic Fractures in Men (MrOS) study. We identified significant associations between benign prostatic hyperplasia (BPH), age, and body mass index (BMI) with several diversity metrics. Our analysis revealed complex relationships between BMI, BPH, LUTS, and alpha diversity which give insight into the intricate dynamics of the urinary microbiome. By beginning to uncover the interrelationships of BPH, BMI, LUTS, and the urinary microbiome, these results can inform future study design to better understand the heterogeneity of the male urinary microbiome.
SpatialPPI: three-dimensional space protein-protein interaction prediction with AlphaFold Multimer
Authors: Hu Wenxing , Ohue Masahito 
Published: December 2023

AbstractThe rapid advancement of protein sequencing technology has resulted in a gap between proteins with identified sequences and those with mapped structures. Although sequence-based predictions offer insights, they can be incomplete due to the absence of structural details. On the other hand, structure-based methods face challenges with newly sequenced proteins. AlphaFold emerges as a potential solution, especially in predicting protein-protein interactions. This research delves into using deep neural networks, specifically analyzing protein complex structures as predicted by AlphaFold Multimer. By transforming atomic coordinates and utilizing sophisticated image processing techniques, the study extracts vital 3D structural details from protein complexes. Recognizing the significance of evaluating residue distances in protein interactions, the study leverages image recognition approaches, notably integrating DenseNet and ResNet within 3D convolutional networks for protein 3D structure analysis. When benchmarked against leading protein-protein interaction prediction methods like SpeedPPI, D-script, DeepTrio, and PEPPI, our proposed method, named SpatialPPI, exhibits notable efficacy, emphasizing the promising role of 3D spatial processing in advancing the realm of structural biology.Abstract Figure
Potassium dependent structural changes in the selectivity filter of HERG potassium channels
Authors: Lau Carus H.Y. , Flood Emelie , Hunter Mark J. , Williams-Noonan Billy J , Corbett Karen M. , Ng Chai-Ann , Bouwer James C. , Stewart Alastair G. , Perozo Eduardo , Allen Toby W. , Vandenberg Jamie I. 
Published: December 2023

AbstractThe fine tuning of biological electrical signaling is mediated by variations in the rates of opening and closing of gates that control ion flux through different ion channels. Human ether-a-go-go related gene (HERG) potassium channels have uniquely rapid inactivation kinetics which are critical to the role they play in regulating cardiac electrical activity. Here, we have exploited the K+sensitivity of HERG inactivation to determine structures of both a conductive and non-conductive selectivity filter structure of HERG. We propose that inactivation is the result of a high propensity for flipping of the selectivity filter valine carbonyl oxygens. Molecular dynamics simulations point to a low energy barrier, and hence rapid kinetics, for flipping of the valine 625 carbonyl oxygens facilitated by a previously unrecognized interaction between S620 and Y616 that stabilizes the transition state between conducting and non-conducting structures. Our model represents a new mechanism by which ion channels fine tune their activity that explains the uniquely rapid inactivation kinetics of HERG.HighlightsStructures of a conductive and non-conductive HERG selectivity filter have been determined.Reduced potassium causes flipping of selectivity filter valine carbonyl oxygens.The sidechain of S620 on the pore helix coordinates distinct sets of interactions between conductive, non-conductive, and transition states.
Acute Neuropixels recordings in the marmoset monkey
Authors: Dotson Nicholas M. , Davis Zachary W. , Jendritza Patrick , Reynolds John H. 
Published: December 2023

ABSTRACTHigh-density linear probes, like Neuropixels, provide an unprecedented opportunity to understand how neural populations within specific laminar compartments contribute to behavior. Marmoset monkeys, unlike macaque monkeys, have a lissencephalic (smooth) cortex that enables recording perpendicular to the cortical surface, thus making them an ideal animal model for studying laminar computations. Here we present a method for acute Neuropixels recordings in the common marmoset (Callithrix jacchus). The approach replaces the native dura with an artificial silicon-based dura that grants visual access to the cortical surface, which is helpful in avoiding blood vessels, ensures perpendicular penetrations, and could be used in conjunction with optical imaging or optogenetic techniques. The chamber housing the artificial dura is simple to maintain with minimal risk of infection and could be combined with semi-chronic microdrives and wireless recording hardware. This technique enables repeated acute penetrations over a period of several months. With occasional removal of tissue growth on the pial surface, recordings can be performed for a year or more. The approach is fully compatible with Neuropixels probes, enabling the recording of hundreds of single neurons distributed throughout the cortical column.
Metacell-based differential expression analysis identifies cell type specific temporal gene response programs in COVID-19 patient PBMCs
Authors: O’Leary Kevin , Zheng Deyou 
Published: December 2023

AbstractBackgroundBy resolving cellular heterogeneity in a biological sample, single cell RNA sequencing (scRNA-seq) can detect gene expression and its dynamics in different cell types. Its application to time-series samples can thus identify temporal genetic programs active in different cell types, for example, immune cells’ responses to viral infection. However, current scRNA-seq analysis need improvement. Two issues are related to data generation. One is that the number of genes detected in each cell is relatively low especially when currently popular dropseq-based technology is used for analyzing thousands of cells or more. The other is the lack of sufficient replicates (often 1-2) due to high cost of library preparation and sequencing. The third issue lies in the data analysis –-usage of individual cells as independent sampling data points leads to inflated statistics.MethodsTo address these issues, we explore a new data analysis framework, specifically whether “metacells” that are carefully constructed to maintain cellular heterogeneity within individual cell types (or clusters) can be used as “replicates” for statistical methods requiring multiple replicates. Toward this, we applied SEACells to a time-series scRNA-seq dataset from peripheral blood mononuclear cells (PBMCs) after SARS-Cov-2 infection to construct metacells, which were then used in maSigPro for quadratic regression to find significantly differentially expressed genes (DEGs) over time, followed by clustering analysis of the expression velocity trends.ResultsWe found that metacells generated using the SEACells algorithm retained greater between-cell variance and produced more biologically meaningful results compared to metacells generated from random cells. Quadratic regression revealed significant DEGs through time that have been previously annotated in the SARS-CoV2 infection response pathway. It also identified significant genes that have not been annotated in this pathway, which were compared to baseline expression and showed unique expression patterns through time.ConclusionsThe results demonstrated that this strategy could overcome the limitation of 1-2 replicates, as it correctly identified the known ISG15 interferon response program in almost all PBMC cell types. Its application further led to the uncovering of additional and more cell type-specific gene expression programs that potentially modulate different levels of host response after infection.