bluuebunny/crossref_metadata_2025_split huggingface dataset:
Insufficiency of 40S ribosomal proteins, RPS26 and RPS25, negatively affects biosynthesis of polyglycine-containing proteins in fragile-X associated conditions
Authors: Tutak Katarzyna , Broniarek Izabela , Zielezinski Andrzej , Niewiadomska Daria , Skrzypczak Tomasz , Baud Anna , Sobczak Krzysztof 
Published: March 2024

AbstractExpansion of CGG repeats (CGGexp) in the 5’ untranslated region (5’UTR) of theFMR1gene underlies the fragile X premutation-associated conditions including tremor/ataxia syndrome, a late-onset neurodegenerative disease and fragile X-associated primary ovarian insufficiency. One common pathomechanism of these conditions is the repeat-associated non-AUG-initiated (RAN) translation of CGG repeats of mutantFMR1mRNA, resulting in production of FMRpolyG, a toxic protein containing long polyglycine tract. To identify novel modifiers of RAN translation we used an RNA-tagging system and mass spectrometry-based screening. It revealed proteins enriched on CGGexp-containingFMR1RNA in cellulo, including a ribosomal protein RPS26, a component of the 40S subunit. We demonstrated that depletion of RPS26 and its chaperone TSR2, modulates FMRpolyG production and its toxicity. We also found that the RPS26 insufficiency impacted translation of limited number of proteins, and 5’UTRs of mRNAs encoding these proteins were short and guanosine and cytosine-rich. Moreover, the silencing of another component of the 40S subunit, the ribosomal protein RPS25, also induced repression of FMRpolyG biosynthesis. Results of this study suggest that the two 40S ribosomal proteins and chaperone TSR2 play an important role in noncanonical CGGexp-related RAN translation.
Proliferative cell targeting and epithelial cell turnover fuels hepatitis E virus replication in human intestinal enteroids
Authors: Santos-Ferreira Nanci , Zhang Xin , Corneillie Laura , Van Dycke Jana , Chiu Winston , Montpellier Claire , Neyts Johan , Cocquerel Laurence , Kaptein Suzanne J. F. , Rocha-Pereira Joana 
Published: March 2024

AbstractHepatitis E virus (HEV) is a leading pathogen causing acute viral hepatitis globally. While HEV is primarily spread fecal-orally, the role of the gut in HEV pathogenesis remains largely unexplored, including how HEV disseminates from gut to liver, and whether the gut is an HEV reservoir. We here aimed to illuminate HEV biology in the gut using human intestinal enteroids (HIEs).Three strategies were explored to establish an HEV-HIE model: three-dimensional (3D) HIEs, two-dimensional (2D) HIEs in transwell, and HEV RNA-electroporated HIEs. HEV particles produced by electroporated HIEs were characterized by western blot and gradient centrifugation. The intestinal tropism of HEV was investigated through confocal fluorescent microscopy and gene expression analysis.HEV infection in 3D-HIEs and 2D-HIEs showed limited replication, whereas HIEs electroporation led to a sustained increase in the release of non-enveloped infectious virions. These virions could re-infect new 3D-HIEs, yielding a ∼2 log10increase in HEV RNA. In electroporated HIEs, high expression of the infectious ORF2 capsid form was observed in the supernatant. Importantly, 70% of all HEV-infected cells were identified as proliferative cells. ORF2 staining was also observed in absorptive enterocytes, goblet, and enteroendocrine cells.Overall, we established a robust HEV-HIE model that yields high titers of infectious non-enveloped virions. Proliferative cells and the fast intestinal epithelial cell turnover are important features that facilitate efficient HEV replication, and likely also its dissemination. This study suggests that the gut is an HEV reservoir, capable of producing some of the non-enveloped HEV shed in the feces.SignificanceHepatitis E virus (HEV) causes ∼20 million enterically transmitted cases of viral hepatitis globally. To better understand the largely unexplored role of the gut during an HEV infection, we used human intestinal enteroids (HIEs). We show that HEV-electroporated HIEs produce highly infectious non-envelopedHEV particles, resulting in a more efficient re-infection compared with traditional gold-standard methods. Importantly, we detected HEV in multiple human intestinal epithelial cell types within the human intestinal epithelium, with the majority of proliferative cells infected. This model offers valuable insights into the intestinal tropism of HEV and can serve to further investigate chronic HEV infection in the gut. Moreover, this underscores the importance of using physiologically relevant models to unravel details of HEV infection in the gut.
Astrocytic metabolic control of orexinergic activity in the lateral hypothalamus regulates sleep and wake architecture
Authors: Braga Alice , Chiacchiaretta Martina , Pellerin Luc , Kong Dong , Haydon Philip G. 
Published: March 2024

ABSTRACTNeuronal activity undergoes significant changes during vigilance states, accompanied by an accommodation of energy demands. While the astrocyte-neuron lactate shuttle has shown that lactate is the primary energy substrate for sustaining neuronal activity in multiple brain regions, its role in regulating sleep/wake architecture is not fully understood. We manipulated the cell-specific expression of monocarboxylate transporters (MCTs), the major lactate transporters, to examine the involvement of astrocytic lactate supply in maintaining consolidated wakefulness. Our results demonstrate that reduced expression of MCT4 in astrocytes disrupts lactate supply to orexin neurons in the lateral hypothalamus (LH), impairing wakefulness stability. We also show that MCT2-mediated lactate uptake is necessary for maintaining tonic firing of orexinergic neurons and stabilizing wakefulness. Our findings provide bothin vivoandin vitroevidence supporting the critical role of astrocyte-to-orexinergic neuron lactate shuttle in regulating proper sleep/wake stability— a crucial step for maintaining physiological functions and overall well-being.
Characterization of West Nile virus Koutango lineage from Phlebotomine Sandflies in Kenya 2021
Authors: Thiiru Jane Wambui , Langat Solomon , Mulwa Francis , Cinkovich Stephanie , Koka Hellen , Yalwala Santos , Khamadi Samoel , Onguso Justus , Odemba Nicholas , Ngere Francis , Johnson Jaree , Egbo Timothy , Garges Eric , Ojwang Elly , Eyase Fredrick 
Published: March 2024

AbstractThe West Nile virus (WNV), primarily transmitted by mosquitoes, is one of the most widespread flaviviruses globally, with past outbreaks occurring in the USA and Europe. Recent studies in parts of Africa, including Kenya, have identified the West Nile virus Koutango lineage (WN-KOUTV) among phlebotomine sandfly populations, however, our understanding of this virus remains limited. Hence, this study aimed to characterize WN-KOUTV from phlebotomine sandflies. Sandflies were sampled between 12-16thMarch 2021 from six villages in Baringo South, Kenya, using CDC light traps. Female sandflies were taxonomically identified and pooled based on genus. Virus isolation was performed in Vero cells. Viral genome was determined using next-generation sequencing. Phylogenetic and molecular clock analyses were done to decipher the virus’s evolutionary relationships. Comparative analyses of amino acid sequences were performed to determine variations. Protein modeling in Pymol was conducted to elucidate variations in key protein regions. Evolutionary pressure analysis investigated the selection pressures on the virus.In vitroexperiments were done to investigate the virus growth kinetics in mammalian (Vero-E6) and mosquito (C636) cells. We report the isolation of WN-KOUTV from Salabani Baringo South, Kenya. The isolated WN-KOUTV clustered with previously identified WN-KOUTV strains. Comparative analysis revealed unique amino acid at NS5 653. Diversifying pressure was acting NS3 267 of the WN-KOUTV lineage. WN-KOUTV replicates efficiently in Vero-E6 and C636 cells comparable to West Nile virus Lineage 1a, isolated from mosquitoes. The isolation of WN-KOUTV in sandflies points to them as potential vectors, however, vector competence studies would confirm this. The efficient replication in mammalian and mosquito cell lines elucidated its adaptability to host and vector. We speculate the close genetic relationship of WN-KOUTV strains is enabled by the bird migratory route between East and West Africa. If proven, this may point to a potential future pandemic pathway for this virus.
Zika Virus Infection Knowledge and Communication Preferences Among Women of Reproductive Age in Central Brooklyn, New York: A Thematic Analysis
Authors: Dowling Russell , Kolokotronis Sergios-Orestis , Thompson Azure B. 
Published: March 2024

AbstractThe 2016 outbreak of Zika virus (ZIKV) infected millions and resulted in thousands of infants born with malformations. Though the clusters of severe birth defects resulting from this outbreak have subsided, ZIKV continues to be a concern throughout much of Latin America and the Caribbean. Travel and sexual intercourse remain the dominant transmission risk factors for women of reproductive age and their partners. This is particularly true for communities in Brooklyn, New York, that comprise large immigrant and foreign-born populations. Practitioners of public health understand little about how women at risk for ZIKV are most likely to receive information about the virus or who they trust most to provide that information. In the context of five focus group discussions, this study explored the knowledge and communication preferences of 20 women of reproductive age in Central Brooklyn. Results derived from a thematic analysis suggest that while most women are familiar with mosquitos as ZIKV vectors, knowledge of sexual transmission is considerably lower. Many respondents believe that only women who are pregnant or trying to become pregnant are at risk, and public health agencies, such as the U.S. Centers for Disease Control and Prevention, remain the most trusted sources of information. These findings can support more effective communication about the risks of ZIKV infection and other vector-borne diseases to women in New York City and similar urban communities.
Adolescent Dating and HIV Perceptions: A Phenomenological Study in N’Djamena, Chad
Authors: Bedingar Esias , Bedingar Ngarossorang , Seli Djimet , Sudfeld Christopher 
Published: March 2024

AbstractIntroductionThe study focuses on understanding the complex interplay between dating behaviors and HIV perceptions among adolescents and young people (ages 15-24 years) in Chad. As adolescents and young people are disproportionately affected by the HIV epidemic, this research aims to uncover how cultural and social influences shape young people’s attitudes towards dating and HIV prevention. The significance of this study lies in its potential to inform targeted, culturally sensitive HIV prevention strategies for the youth in Chad.MethodsA qualitative phenomenological approach was adopted, using 4 focus group discussions (n=12 each) with high school students in N’Djamena, Chad. Participants were divided into beneficiary and non-beneficiary groups based on their exposure to a peer education program about HIV. Data analysis was conducted using ATLAS.ti software and the descriptive Colaizzi method, ensuring a comprehensive understanding of participants’ perspectives.ResultsFindings revealed significant gender differences in dating motivations and partner selection criteria. While males primarily viewed dating as a means to fulfill sexual desires, females often sought emotional connection and companionship. There was a general lack of awareness about PrEP, with more emphasis on condom use for pregnancy prevention rather than HIV or STI protection. Gender roles played a substantial part in decisions regarding safe sex practices, with a notable discrepancy in shared versus individual responsibility. Additionally, risk behaviors like alcohol use and multiple partnerships were identified as prevalent among youth.ConclusionsThe study highlights the need for gender-sensitive educational interventions focusing on the realities of adolescent sexual behavior in Chad. It underscores the disparity in HIV knowledge and prevention awareness among adolescents, pointing out the absence of awareness about PrEP. Future research should emphasize developing HIV prevention strategies that resonate with the unique cultural and social dynamics of Chadian youth, considering their specific dating behaviors and perceptions towards HIV.
Endothelial calcium firing mediates extravasation of metastatic tumor cells
Authors: Peralta Marina , Dupas Amandine , Larnicol Annabel , Lefebvre Olivier , Goswami Ruchi , Stemmelen Tristan , Molitor Anne , Carapito Raphael , Girardo Salvatore , Osmani Naël , Goetz Jacky G. 
Published: March 2024

ABSTRACTMetastatic dissemination is driven by genetical, biochemical and biophysical cues that favor the distant colonization of organs and the formation of life-threatening secondary tumors. We have previously demonstrated that endothelial cells (ECs) actively remodel during extravasation by enwrapping arrested tumor cells (TCs) and extrude them from the vascular lumen while maintaining perfusion. In this work, we dissect the cellular and molecular mechanisms driving endothelial remodeling. Using high-resolution intravital imaging in zebrafish embryos, we demonstrate that the actomyosin network of ECs controls tissue remodeling and subsequent TC extravasation. Furthermore, we uncovered that this cytoskeletal remodeling is driven by altered endothelial- calcium (Ca2+) signaling caused by arrested TCs. Accordingly, we demonstrated that inhibition of voltage-dependent calcium L-type channels impairs extravasation. Lastly, we identified P2X4, TRP, and Piezo1 mechano-gated Ca2+channels as key mediators of the process. These results further highlight the central role of endothelial remodeling during extravasation of TCs and open avenues for successful therapeutic targeting.Graphical Abstract
An evolutionary cell biology perspective into the diverging mechanisms of clathrin-mediated endocytosis in dikarya fungi
Authors: Picco Andrea , Toret Christopher P. , Rivier-Cordey Anne-Sophie , Kaksonen Marko 
Published: March 2024

ABSTRACTClathrin-mediated endocytosis is an ancient eukaryotic trafficking pathway, which transports plasma membrane and associated cargo into the cell and is involved in numerous cell- and tissue-level processes. Cargo selection and clathrin-coated vesicle formation is mediated by over 60 proteins that assemble in a regular and sequential manner at the plasma membrane. Decades of endocytosis studies have followed the tenet that uncovering the conserved core molecular mechanisms is sufficient to understand a cellular process. However, this approach also revealed a number of cell type or species-related variations that challenge a universal conserved, core mechanism. In this paper, we refocus on the endocytic diversity to understand how evolution shapes endocytic mechanisms. We define a comparative evolutionary cell biology approach that uses dikarya fungi as a model clade and live-cell fluorescence microscopy to study endocytosis dynamics in three species:Saccharomyces cerevisiae,Schizosaccharomyces pombeandUstilago maydis. Our results quantitatively define several phenotypic differences between the species. We uncover several differences that impact the endocytic early phase, the protein assembly order, actin regulation, membrane invagination and scission. These findings demonstrate a mosaic evolution of endocytic traits, suggest ancestral states and direction of changes. We also investigate the phenotypic plasticity and robustness against environmental conditions. Lastly, we demonstrate that relatively minor evolutionary changes can majorly impact endocytic phenotypes. These studies force an appreciation of endocytic variation as not auxiliary, but vital to mechanistic understanding of this conserved cellular pathway.
Plasma Proteomics of Genetic Brain Arteriosclerosis and Dementia Syndrome Identifies Signatures of Fibrosis, Angiogenesis, and Metabolic Alterations
Authors: Keller Jonah N. , Radabaugh Hannah , Karvelas Nikolaos , Fitzsimons Stephen , Treiman Scott , Palafox Maria F. , McDonnell Lisa , Quiroz Yakeel T. , Lopera Francisco J. , Banerjee Debarag , Wang Michael M. , Arboleda-Velasquez Joseph F. , Meschia James F. , Ferguson Adam R. , Elahi Fanny M. 
Published: March 2024

AbstractCerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common monogenic form of vascular cognitive impairment and dementia. A genetic arteriolosclerotic disease, the molecular mechanisms driving vascular brain degeneration and decline remain unclear. With the goal of driving discovery of disease-relevant biological perturbations in CADASIL, we used machine learning approaches to extract proteomic disease signatures from large-scale proteomics generated from plasma collected from three distinct cohorts in US and Colombia: CADASIL-Early (N= 53), CADASIL-Late (N= 45), and CADASIL-Colombia (N= 71). We extracted molecular signatures with high predictive value for early and late-stage CADASIL and performed robust cross- and external-validation. We examined the biological and clinical relevance of our findings through pathway enrichment analysis and testing of associations with clinical outcomes. Our study represents a model for unbiased discovery of molecular signatures and disease biomarkers, combining non-invasive plasma proteomics with clinical data. We report on novel disease-associated molecular signatures for CADASIL, derived from the accessible plasma proteome, with relevance to vascular cognitive impairment and dementia.
Exhaustion profile on classical monocytes after LPS stimulation in patients with Crohn’s disease
Authors: Oliveira Lucas Pires Garcia , Xavier Rafaela Gomes , Nora Claudia Concer Viero , Mangueira Cristóvão Luis Pitangueira , Rosseto Eliane Aparecida , Aloia Thiago , Gil Jaime Zaladek , Neto Arceu Scanavini , Lopes Filipa Blasco Tavares Pereira , Carvalho Karina Inacio 
Published: March 2024

AbstractCrohn’s disease is an inflammatory bowel disease that induces diarrhea, abdominal pain, weight loss, and even susceptibility to developing tumors. The immune system is pivotal in the gastrointestinal tract, promoting tolerance against commensal antigens and food. However, Crohn’s disease manifests by a breakdown in the mechanism of immune tolerance and the consequent development of exacerbated chronic inflammatory responses. The involvement of the immune system is pivotal in Crohn’s disease, with a wide range of immune cells being altered, which may include monocytes. Taking the lack of knowledge regarding monocytes in Crohn’s disease, we ought to elucidate the cytokine production and activation profile of monocyte subsets in the pathophysiology. We used multiparametric flow cytometry, quantified gene expression using qPCR, and made a correlation matrix regarding flow cytometry data and qPCR using a bioinformatic approach to examine monocyte status. The Corhn’s patients show a decrease in all subsets of monocytes.In contrast, classical monocytes show an exhaustion profile with increased expression of CD38 and decreased production of IL-1β after LPS stimulation in the patients’ group. These results indicate that monocyte subsets are differentially involved in the pathophysiology. These findings may suggest that monocytes favor disease chronicity and lack immune response resolution.
Proteome dynamics reveal Leiomodin 1 as a key regulator of myogenic differentiation
Authors: Späth Ellen , Schüler Svenja C. , Heinze Ivonne , Dau Therese , Minetti Alberto , Hofmann Maleen , Maltzahn Julia von , Ori Alessandro 
Published: March 2024

AbstractDuring myogenic differentiation the cellular architecture and proteome of muscle stem cells and myoblasts undergo extensive remodeling. These molecular processes are only partially understood and display alterations in disease conditions as well as during aging resulting in impaired regeneration. Here, we used mass spectrometry to quantify the temporal dynamics of more than 6000 proteins during myogenic differentiation. We identified the actin nucleator leiomodin 1 (LMOD1) among a restricted subset of cytoskeletal proteins increasing in abundance in early phases of myogenic differentiation. We show that LMOD1 is already expressed by muscle stem cellsin vivoand displays increased abundance during skeletal muscle regeneration, especially during early regeneration suggesting that LMOD1 is important for induction of myotube formation. Of note, knockdown of LMOD1 in primary myoblasts and during skeletal muscle regeneration severely affects myogenic differentiation, while overexpression accelerates and improves the formation of myotubes suggesting that LMOD1 is a critical component regulating myogenic differentiation. Mechanistically, we show that LMOD1 physically and functionally interacts with the deacetylase sirtuin1 (SIRT1), a regulator of myogenic differentiation, especially at the onset of myogenic differentiation. We demonstrate that LMOD1 influences SIRT1 localization and the expression of a subset of its target genes. Consistently, depletion or pharmacological inhibition of SIRT1 partially rescues the impairment of myogenic differentiation observed after knockdown of LMOD1. Our work identifies a new regulator of myogenic differentiation that might be targeted to improve muscle regeneration in aging and disease.
Comparison of the tolerability of161Tb- and177Lu-labeled somatostatin analogues in the preclinical setting
Authors: Busslinger Sarah D. , Mapanao Ana Katrina , Kegler Kristel , Bernhardt Peter , Flühmann Fabienne , Fricke Julia , Zeevaart Jan Rijn , Köster Ulli , van der Meulen Nicholas P. , Schibli Roger , Müller Cristina 
Published: March 2024

AbstractPurpose[177Lu]Lu-DOTATATE is an established somatostatin receptor (SSTR) agonist for the treatment of metastasized neuroendocrine neoplasms, while the SSTR antagonist [177Lu]Lu-DOTA-LM3 has only scarcely been employed in clinics. Impressive preclinical data obtained with [161Tb]Tb-DOTA-LM3 in tumor-bearing mice indicated the potential of terbium-161 as an alternative to lutetium-177. The aim of the present study was to compare the tolerability of161Tb- and177Lu-based DOTA-LM3 and DOTATATE in immunocompetent mice.MethodsDosimetry calculations were performed based on biodistribution data of the radiopeptides in immunocompetent mice. Treatment-related effects on blood cell counts were assessed on Days 10, 28 and 56 after application of [161Tb]Tb-DOTA-LM3 or [161Tb]Tb-DOTATATE at 20 MBq per mouse. These radiopeptides were also applied at 100 MBq per mouse and the effects compared to those observed after application of the177Lu-labeled counterparts. Bone marrow smears, blood plasma parameters and organ histology were assessed at the end of the study.ResultsThe absorbed organ dose was commonly higher for the SSTR antagonist than for the SSTR agonist and for terbium-161 over lutetium-177. Application of a therapeutic activity level of 20 MBq [161Tb]Tb-DOTA-LM3 or [161Tb]Tb-DOTATATE was well tolerated without major hematological changes. The injection of 100 MBq of the161Tb- and177Lu-based somatostatin analogues affected the blood cell counts, however. The lymphocytes were 40-50% lower in treated mice compared to the untreated controls on Day 10 irrespective of the radionuclide employed. At the same timepoint, thrombocyte and erythrocyte counts were 30–50% and 6–12% lower, respectively, after administration of the SSTR antagonist (p<0.05) while changes were less pronounced in mice injected with the SSTR agonist. All blood cell counts were in the normal range on Day 56. Histological analyses revealed minimal abnormalities in the kidneys, liver and spleen of treated mice. No correlation was observed between the organ dose and frequency of the occurrence of abnormalities.ConclusionHematologic changes were more pronounced in mice treated with the SSTR antagonist than in those treated with the SSTR agonist. Despite the increased absorbed dose delivered by terbium-161 over lutetium-177, [161Tb]Tb-DOTA-LM3 and [161Tb]Tb-DOTATATE should be safe at activity levels that are recommended for their respective177Lu-based analogues.