bluuebunny/crossref_metadata_2025_split huggingface dataset:
A broad-specificity O-glycoprotease that enables improved analysis of glycoproteins and glycopeptides containing intact complex O-glycans
Authors: Vainauskas Saulius , Guntz Hélène , McLeod Elizabeth , McClung Colleen , Ruse Cristian , Shi Xiaofeng , Taron Christopher H. 
Published: September 2021

AbstractAnalysis of mucin type O-glycans linked to serine/threonine of glycoproteins is technically challenging, in part, due to a lack of effective enzymatic tools that enable their analysis. Recently, several O-glycan-specific endoproteases that can cleave the protein adjacent to the appended glycan have been described. Despite significant progress in understanding the biochemistry of these enzymes, known O-glycoproteases have specificity constrains, such as inefficient cleavage of glycoproteins bearing sialylated O-glycans, high selectivity for certain type of glycoproteins or protein sequence bias, that limit their analytical application. In this study, we examined the capabilities of an immunomodulating metalloprotease (IMPa) from Pseudomonas aeruginosa. The peptide substrate sequence selectivity and its impact on IMPa activity was interrogated using an array of synthetic peptides and their glycoforms. We show that IMPa has no specific P1 residue preference and can tolerate most amino acids at the P1 position, except aspartic acid. The enzyme does not cleave between two adjacent O-glycosites, indicating that O-glycosylated serine/threonine is not allowed at position P1. Glycopeptides with as few as two amino acids on either side of an O-glycosite were specifically cleaved by IMPa. Finally, IMPa efficiently cleaved peptides and proteins carrying sialylated and asialylated O-glycans of varying complexity. We present the use of IMPa in a one-step O-glycoproteomics workflow for glycoprofiling of individual purified glycoproteins granulocyte colony-stimulating factor (G-CSF) and receptor-type tyrosine-protein phosphatase C (CD45) without the need for glycopeptide enrichment. In these examples, IMPa enabled identification of O-glycosites and the range of complex O-glycan structures at each site.
Trans-cellular tunnels induced by the fungal pathogen Candida albicans facilitate invasion through successive epithelial cells without host damage
Authors: Lachat Joy , Pascault Alice , Thibaut Delphine , Le Borgne Rémi , Verbavatz Jean-Marc , Weiner Allon 
Published: September 2021

SummaryThe opportunistic fungal pathogen Candida albicans is normally commensal, residing in the mucosa of most healthy individuals. In susceptible hosts, its filamentous hyphal form can invade epithelial layers leading to superficial or severe systemic infection. Invasion is mainly intracellular, though it causes no apparent damage to host cells. We investigated the invasive lifestyle of C. albicans in vitro using live-cell imaging and the damage-sensitive reporter galectin-3. Quantitative single cell analysis showed that invasion can result in host membrane breaching at different stages of invasion and cell death, or in traversal of host cells without membrane breaching. Membrane labelling and three-dimensional “volume” electron microscopy revealed that hyphae can traverse several host cells within trans-cellular tunnels that are progressively remodelled and may undergo ‘inflations’ linked to host glycogen stores, possibly during nutrient uptake. Thus, C. albicans invade epithelial tissues by either inflicting or avoiding host damage, the latter facilitated by trans-cellular tunnelling.
Long-term persistence of viral RNA and inflammation in the CNS of macaques exposed to aerosolized Venezuelan equine encephalitis virus
Authors: Ma Henry , Albe Joseph R. , Gilliland Theron , McMillen Cynthia M. , Gardner Christina L. , Boyles Devin A. , Cottle Emily L. , Dunn Matthew D. , Lundy Jeneveve D. , Salama Noah , O’Malley Katherine J. , Pandrea Ivona , Teichert Tobias , Barrick Stacey , Klimstra William B. , Hartman Amy L. , Reed Douglas S. 
Published: September 2021

ABSTRACTVenezuelan equine encephalitis virus (VEEV) is a positively-stranded RNA arbovirus of the genus Alphavirus that causes encephalitis in humans. Cynomolgus macaques are a relevant model of the human disease caused by VEEV and are useful in exploring pathogenic mechanisms and the host response to VEEV infection. Macaques were exposed to small-particle aerosols containing virus derived from an infectious clone of VEEV strain INH-9813, a subtype IC strain isolated from a human infection. VEEV-exposed macaques developed a biphasic fever after infection similar to that seen in humans. Maximum temperature deviation correlated with the inhaled dose, but fever duration did not. Neurological signs, suggestive of virus penetration into the CNS, were predominantly seen in the second febrile period. Electroencephalography data indicated a statistically significant decrease in all power bands and circadian index during the second febrile period that returned to normal after fever resolved. Intracranial pressure increased late in the second febrile period. On day 6 post-infection macaques had high levels of MCP-1 and IP-10 chemokines in the CNS, as well as a marked increase of T lymphocytes and activated microglia. More than four weeks after infection, viral genomic RNA was found in the brain, cerebrospinal fluid and cervical lymph nodes. Pro-inflammatory cytokines & chemokines, infiltrating leukocytes and pathological changes were seen in the CNS tissues of macaques euthanized at these times. These data are consistent with persistence of virus replication and/or genomic RNA and potentially, inflammatory sequelae in the central nervous system after resolution of acute VEEV disease.
Identification of a stereotypic molecular arrangement of endogenous glycine receptors at spinal cord synapses
Authors: Maynard Stephanie A , Rostaing Philippe , Schaefer Natascha , Gemin Olivier , Candat Adrien , Dumoulin Andréa , Villmann Carmen , Triller Antoine , Specht Christian G 
Published: September 2021

AbstractPrecise quantitative information about the molecular architecture of synapses is essential to understanding the functional specificity and downstream signaling processes at specific populations of synapses. Glycine receptors (GlyRs) are the primary fast inhibitory neurotransmitter receptors in the spinal cord and brainstem. These inhibitory glycinergic networks crucially regulate motor and sensory processes. Thus far the nanoscale organization of GlyRs underlying the different network specificities has not been defined. Here, we have quantitatively characterized the molecular arrangement and ultra-structure of glycinergic synapses in spinal cord tissue using quantitative super-resolution correlative light and electron microscopy (SR-CLEM). We show that endogenous GlyRs exhibit equal receptor-scaffold occupancy and constant packing densities of about 2000 GlyRs μm-2at synapses across the spinal cord and throughout adulthood, even though ventral horn synapses have twice the total copy numbers, larger postsynaptic domains and more convoluted morphologies than dorsal horn synapses. We demonstrate that this stereotypic molecular arrangement is maintained at glycinergic synapses in theoscillatormouse model of the neuromotor disease hyperekplexia despite a decrease in synapse size, indicating that the molecular organization of GlyRs is preserved in this hypomorph. We thus conclude that the morphology and size of inhibitory postsynaptic specializations rather than differences in GlyR packing determine the postsynaptic strength of glycinergic neurotransmission in motor and sensory spinal cord networks.
Global population genomics of two subspecies ofCryptosporidium hominisduring 500 years of evolution
Authors: Tichkule Swapnil , Cacciò Simone M. , Robinson Guy , Chalmers Rachel M. , Mueller Ivo , Emery-Corbin Samantha J. , Eibach Daniel , Tyler Kevin M. , van Oosterhout Cock , Jex Aaron R. 
Published: September 2021

AbstractCryptosporidiosis is a major global health problem and a primary cause of diarrhoea, particularly in young children in low- and middle-income countries (LMICs). The zoonoticCryptosporidium parvumand anthroponoticC. hominiscause most human infections. Here, we present a comprehensive whole-genome study ofC. hominis, comprising 114 isolates from 16 countries within five continents. We detect two lineages with distinct biology and demography, which diverged circa 500 years ago. We consider these lineages two subspecies and propose the namesC. hominis hominisandC. hominis aquapotentis(gp60subtype IbA10G2). In our study,C. h. hominisis almost exclusively represented by isolates from LMICs in Africa and Asia and appears to have undergone recent population contraction. In contrast,C. h. aquapotentiswas found in high-income countries, mainly in Europe, North America and Oceania, and appears to be expanding. Notably,C. h. aquapotentisis associated with high rates of direct human-to-human transmission, which may explain its success in countries with well-developed environmental sanitation infrastructure. Intriguingly, we detected genomic regions of introgression following secondary contact between the subspecies. This resulted in high diversity and divergence in genomic islands of putative virulence genes (GIPVs), includingmuc5(CHUDEA2_430) and a hypothetical protein (CHUDEA6_5270). This diversity is maintained by balancing selection, suggesting a coevolutionary arms race with the host. Lastly, we find that recent gene flow fromC. h. aquapotentistoC. h. hominis, likely associated with increased human migration, may be driving evolution of more virulentC. hominisvariants.
Simultaneous Triple-Parametric Optical Mapping of Transmembrane Potential, Intracellular Calcium and NADH for Cardiac Physiology Assessment
Authors: George Sharon A , Lin Zexu , Efimov Igor R 
Published: September 2021

AbstractInvestigation of the complex relationships and dependencies of multiple cellular processes that govern cardiac physiology and pathophysiology requires simultaneous dynamic assessment of multiple parameters. In this study, we introduce triple-parametric optical mapping to simultaneously image metabolism, electrical excitation, and calcium signaling from the same field of view and demonstrate its application in the field of drug testing and cardiovascular research. We applied this metabolism-excitation-contraction coupling (MECC) methodology to test the effects of blebbistatin, 4-aminopyridine and verapamil on cardiac physiology. While blebbistatin and 4-aminopyridine alter multiple aspects of cardiac function suggesting off-target effects, the effects of verapamil were on-target and it altered only one of ten tested parameters. Triple-parametric optical mapping was also applied during ischemia and reperfusion; and we identified that metabolic changes precede the effects of ischemia on cardiac electrophysiology.
Targeting the Cx26/NANOG/Focal Adhesion Kinase Complex via Cell-Penetrating Peptides in Triple-Negative Breast Cancers
Authors: Mulkearns-Hubert Erin E. , Rhoades Emily Esakov , Ben-Salem Salma , Bharti Rashmi , Hajdari Nicole , Johnson Sarah , Myers Alex , Smith Iris Nira , Bandyopadhyay Smarajit , Eng Charis , Downs-Kelly Erinn , Lathia Justin D. , Reizes Ofer 
Published: September 2021

AbstractTriple-negative breast cancer (TNBC) represents the most lethal and treatment-resistant breast cancer subtype with limited treatment options. We previously identified a protein complex unique to TNBC cancer stem cells composed of the gap junction protein connexin 26 (Cx26), the pluripotency transcription factor NANOG, and focal adhesion kinase (FAK). We sought to determine whether a peptide mimetic of Cx26 designed to target the complex attenuated tumor growth in pre-clinical models. Histological assessment was employed to verify expression of complex members. We designed peptides based on Cx26 juxtamembrane domains and performed binding experiments with NANOG and FAK using surface plasmon resonance. Peptides with high affinity were engineered with a cell-penetrating sequence and assessed in functional assays including cell proliferation, self-renewal, and in vivo tumor growth, and downstream signaling changes were measured. Binding studies revealed that the Cx26 C-terminal tail and intracellular loop bound to NANOG and FAK with submicromolar-to-micromolar affinity and that a 5-amino acid sequence in the C-terminal tail of Cx26 (RYCSG) was sufficient for binding. The Cx26 C-terminal tail was tagged with an antennapedia cell-penetrating peptide sequence and intracellular localization was confirmed. The cell-penetrating Cx26 peptide (aCx26-pep) disrupted self-renewal as assessed by tumorsphere formation assay while reducing nuclear FAK and NANOG and inhibiting NANOG target gene expression in TNBC cells but not luminal mammary epithelial cells. In vivo, aCx26-pep reduced tumor growth and proliferation and induced cell death. We provide proof-of-concept that a Cx26 peptide-based strategy inhibits growth and alters NANOG activity in TNBC.
Computational drug repurposing against SARS-CoV-2 reveals plasma membrane cholesterol depletion as key factor of antiviral drug activity
Authors: Barsi Szilvia , Papp Henrietta , Urbelz Alberto Valdeolivas , Tóth Dániel J. , Kuczmog Anett , Madai Mónika , Hunyady László , Várnai Péter , Saez-Rodriguez Julio , Jakab Ferenc , Szalai Bence 
Published: September 2021

AbstractComparing SARS-CoV-2 infection-induced gene expression signatures to drug treatment-induced gene expression signatures is a promising bioinformatic tool to repurpose existing drugs against SARS-CoV-2. The general hypothesis of signature based drug repurposing is that drugs with inverse similarity to a disease signature can reverse disease phenotype and thus be effective against it. However, in the case of viral infection diseases, like SARS-CoV-2, infected cells also activate adaptive, antiviral pathways, so that the relationship between effective drug and disease signature can be more ambiguous.To address this question, we analysed gene expression data fromin vitroSARS-CoV-2 infected cell lines, and gene expression signatures of drugs showing anti-SARS-CoV-2 activity. Our extensive functional genomic analysis showed that both infection and treatment within vitroeffective drugs leads to activation of antiviral pathways like NFkB and JAK-STAT. Based on the similarity - and not inverse similarity - between drug and infection-induced gene expression signatures, we were able to predict thein vitroantiviral activity of drugs. We also identified SREBF1/2, key regulators of lipid metabolising enzymes, as the most activated transcription factors by severalin vitroeffective antiviral drugs. Using a fluorescently labeled cholesterol sensor, we showed that these drugs decrease the cholesterol levels of plasma-membrane. Supplementing drug-treated cells with cholesterol reversed thein vitroantiviral effect, suggesting the depleting plasma-membrane cholesterol plays a key role in virus inhibitory mechanism.Our results can help to more effectively repurpose approved drugs against SARS-CoV-2, and also highlights key mechanisms behind their antiviral effect.Abstract Figure
Women and Alcohol Consumption in Fascist Italy
Authors: Ferris Kate 
Published: September 2021

AbstractThis article examines discourses and practices around women's drinking in Fascist Italy. The history of alcohol production and consumption in Italy during the fascist dictatorship has only recently received attention; alcohol's gendered dimensions, especially women's drinking, have been hitherto overlooked. While the production of legislation, rhetoric and propaganda on alcohol consumption was dominated by men, women were identified as key constituents whose alcohol‐related practices could make or break the causes of fascist propagandists, ‘anti‐alcohol’ campaigners and alcohol industry associations. The article explains how Italian women were imagined and addressed by regime propagandists, alcohol industry producers and temperance campaigners as (a) simultaneously the principal victims of and responsibility bearers for male excess alcohol consumption, (b) potential ‘crisis‐women’ whose unpatriotic drinking choices (whether English tea, French champagne or American cocktails) denoted their prioritising of fashion over fascist values and (c) gatekeepers of family alcohol consumer practices and consumers of alcohol in their own right. It then moves to examine sources left by interwar Italian women to explore what, how and when they drank. Ultimately, it argues that despite attempts to construct women's drinking in archly nationalistic terms, the discourses and actual practices of Italian women around alcohol consumption operated within profoundly transnational frames.
Cardiac geometry, function, and remodeling patterns in patients under maintenance hemodialysis and peritoneal dialysis treatment
Authors: Alexandrou Maria‐Eleni , Sarafidis Pantelis , P. Theodorakopoulou Μarieta , Sachpekidis Vasileios , Papadopoulos Christodoulos , Loutradis Charalampos , Kamperidis Vasileios , Boulmpou Aristi , Bakaloudi Dimitra‐Rafailia , Faitatzidou Danai , Pateinakis Panagiotis , Papagianni Aikaterini 
Published: September 2021

AbstractCardiovascular disease is the leading cause of mortality in patients with end‐stage‐kidney disease. Evidence on the possible echocardiographic differences between hemodialysis and peritoneal dialysis (PD) is scarce. This study aimed to evaluate differences in left (LA) and right atrial (RA), left (LV) and right ventricular (RV) geometry, systolic and diastolic function in hemodialysis, and PD patients. Thirty‐eight hemodialysis and 38 PD patients were matched for age, sex, and dialysis vintage. Two‐dimensional and tissue‐Doppler echocardiography, and lung ultrasound were performed during an interdialytic day in hemodialysis and before a programmed follow‐up visit in PD patients. Vena cava diameter (11.09 ± 4.53 vs. 14.91 ± 4.30 mm; p < 0.001) was significantly lower in hemodialysis patients. Indices of LA, RA, LV, and RV dimensions were similar between the two groups. LVMi (116.91 [38.56] vs. 122.83 [52.33] g/m2; p = 0.767) was similar, but relative wall thickness was marginally (0.40 [0.14] vs. 0.45 [0.15] cm; p = 0.055) lower in hemodialysis patients. LV hypertrophy prevalence was similar between groups (73.7% vs. 71.1%; p = 0.798), but hemodialysis patients presented eccentric and PD patients concentric LVH. Regarding ventricular systolic function, stroke volume (p = 0.030) and cardiac output (p = 0.036) were higher in hemodialysis, while RV systolic pressure (RVSP) (20.37 [22.54] vs. 27.68 [14.32] mm Hg; p = 0.009) was higher in PD. No significant differences were evidenced in diastolic function indices and lung water excess between the two groups. A moderate association was noted between ultrasound B‐lines score and LA volume index (r = 0.465, p < 0.001), RVSP (r = 0.431, p < 0.001), and E/e′ ratio (r = 0.304, p = 0.009). Hemodialysis and PD patients present largely similar echocardiographic indices reflecting cardiac geometry, systolic, and diastolic function, but different patterns of abnormal LV remodeling.
The impact of primary human papillomavirus screening on negative loop excision histology following biopsy‐proven high‐grade cervical intra‐epithelial lesions: A review from a large tertiary colposcopy unit
Authors: Xie Huan , Morton Rhett , Balendran Jananie , Philp Shannon , Saidi Samir , Farrell Rhonda , Anderson Lyndal , Pather Selvan 
Published: September 2021

BackgroundThe renewed National Cervical Screening Program incorporating primary human papillomavirus (HPV) screening was implemented in Australia in December 2017. In a previous study conducted in the UK, primary HPV screening was found to be associated with a 25% reduction in the incidence of negative histology following loop electrosurgery excision procedure (LEEP).AimTo examine the change in incidence and associated risk factors for a negative LEEP with introduction of primary HPV screening.Materials and MethodsA retrospective review of the records of all patients undergoing a LEEP excision for biopsy‐proven high‐grade cervical intra‐epithelial lesions between 1 January 2014 and 30 June 2019 in a specialised centre.ResultsThere were 1123 patients who underwent a LEEP included in the analysis. The incidence of a negative LEEP specimen was 7.5% (59/784) and 5.3% (18/339) in the pre‐ and post‐HPV screening cohort. More patients in the post‐HPV screening group had low‐grade cytology on referral (P < 0.001), smaller cervical lesions on colposcopy (P = 0.012) and longer biopsy to treatment interval (P = 0.020). Primary HPV screening was associated with a significant reduction in the incidence of a negative LEEP specimen in a propensity matched cohort (11.2% to 5.1%, P = 0.006) and a 41% (P = 0.045) decreased relative risk of a negative LEEP on multivariate analysis.ConclusionsPrimary HPV screening results in a lower incidence of negative LEEP histology, despite a longer biopsy to treatment wait time and higher proportion of low‐grade cytology at triage.
Better performance of medical students on pharmacotherapy knowledge and skills tests is associated with practising with e‐learning program P‐scribe
Authors: Kalfsvel Laura , Versmissen Jorie , van Doorn Adriaan , van den Broek Walter , van der Kuy Hugo , van Rosse Floor 
Published: September 2021

AimsJunior doctors write most hospital prescriptions, yet are more than twice as likely to make an error in their prescriptions compared to senior doctors. A possibility to enhance pharmacotherapy education is through the use of e‐learning modules. The aim of this study was to determine whether P‐scribe, as the chosen e‐learning resource, helps students in passing their pharmacotherapy assessments.MethodsThis retrospective study was undertaken in the Erasmus Medical Center, the Netherlands. All 270 medical students who started their master's curriculum in the academic session of 2017–2018 were included. Data were analysed to identify the frequency of student's use per e‐learning module, total time students spent on e‐learning modules and timing of the use of e‐learning modules in relation to their assessments. The results of the assessments were analysed to identify possible correlations between the time students spent using P‐scribe, their timing of use and their assessment results.ResultsStudents who passed their knowledge‐based assessment first time had a mean practice time of five more hours than students who did not pass first time (P < .05, 95% CI: 3.4–6.6). These students practised on average six e‐learning modules more (P < .05, 95% CI: 4.1–7.0) than students who failed their first attempt. Students who passed their skill‐based prescription test first time, practised on average five more e‐learning modules (P = .006, 95% CI: 1.4–8.3) than students who failed their first attempt.ConclusionStudents who passed their pharmacotherapy assessments first time spent more time, and practised more frequently, with e‐learning modules.