bluuebunny/crossref_metadata_2025 huggingface dataset:
Recommended primary outcomes for clinical trials evaluating hemostatic blood products and agents in patients with bleeding: Proceedings of a National Heart Lung and Blood Institute and US Department of Defense Consensus Conference
Authors: Spinella Philip C. , El Kassar Nahed , Cap Andrew P. , Kindzelski Andrei L. , Almond Christopher S. , Barkun Alan , Gernsheimer Terry B. , Goldstein Joshua N. , Holcomb John B. , Iorio Alfonso , Jensen Dennis M. , Key Nigel S. , Levy Jerrold H. , Mayer Stephan A. , Moore Ernest E. , Stanworth Simon J. , Lewis Roger J. , Steiner Marie E. ,   for the Hemostasis Trials Outcomes Working Group
Published: May 2021

High-quality evidence guiding optimal transfusion and other supportive therapies to reduce bleeding is needed to improve outcomes for patients with either severe bleeding or hemostatic disorders that are associated with poor outcomes. Alongside challenges in performing high-quality clinical trials in patient populations who are at risk of bleeding or who are actively bleeding, the interpretation of research evaluating hemostatic agents has been limited by inconsistency in the choice of primary trial outcomes. This lack of standardization of primary endpoints or outcomes decreases the ability of clinicians to assess the validity of endpoints and compare research results across studies, impairs meta-analytic efforts, and, ultimately, delays the translation of research results into clinical practice. To address this challenge, an international panel of experts was convened by the National Heart Lung and Blood Institute and the US Department of Defense on September 23 and 24, 2019, to develop expert opinion, consensus-based recommendations for primary clinical trial outcomes for pivotal trials in pediatric and adult patients with six categories in various clinical settings. This publication documents the conference proceedings from the workshop funded by the National Heart Lung and Blood Institute and the US Department of Defense that consolidated expert opinion regarding clinically meaningful outcomes across a wide range of disciplines to provide guidance for outcomes of future trials of hemostatic products and agents for patients with active bleeding.
Multi-level multi-state modelling applied to hospital admission in mexican patients with COVID-19
Authors: Martinez Juan Pablo Diaz , Becerril Karen Janik Orozco , Herrada Marco Antonio Gallegos , Garcia Mayra Alejandra Gutierrez , Espin-Garcia Osvaldo , Garcia Ruth Fuentes
Published: May 2021

Since the beginning of the SARS-CoV 2 pandemic, healthcare authorities have made clear that it is crucial to track and identify COVID-19 symptoms and seek medical attention in the presence of the first warning signs, as immediate medical attention can improve the patient’s prognosis. Therefore the present work aims to analyze the risks associated with the time between the patient’s first symptoms and hospitalization followed by death. A cross-sectional study was performed among Mexican population diagnosed with COVID-19 and hospitalized from March to January 2021. Four different Bayesian models were developed to asses the risk associated with different patient trajectories: symptoms-hospitalization and hospitalization-death. Comorbidities that could worsen the patient outcome were included as linear predictions; these analyses were further broken down to the different states of the Mexican Republic and the healthcare providers within. Model III was chosen as the best performance through a validation of leaving one out (LOO). Increased risk for hospitalization was observed at the global population level for chronic renal disease, whereas for death such was the case for COPD and the interaction of diabetes:hypertension:obesity. Our results show that there are differences in mortality between the states without accounting for institution and it is related to the prompt time of death or viceversa. Regarding the 6 healthcare providers included in the analysis differences were also found. While state-managed hospitals and private sector showed lower risks, in contrast the IMSS seems to be the one with the highest risk. The proposed modelling can be helpful to improve healthcare assistance at a regional level, additionally it could inform statistical parameter inference in epidemiological models.
Early transfusion of convalescent plasma improves the clinical outcome in severe SARS-CoV2 infection
Authors: Fodor Eszter , Müller Veronika , Iványi Zsolt , Berki Tímea , Olga Kuten Pella , Ambrus Mira , Sárkány Ágnes , Skázel Árpád , Madár Ágnes , Kardos Dorottya , Kemenesi Gábor , Földes Fanni , Nagy Sándor , Matusovits Andrea , János Nacsa , Tordai Attila , Jakab Ferenc , Lacza Zsombor
Published: May 2021

Plasma harvested from convalescent COVID-19 patients (CCP) has been applied as first-line therapy in the early phase of the SARS-CoV2 pandemic through clinical studies using various protocols. We present data from a cohort of 267 hospitalized, severe COVID-19 patients who received CCP. No transfusion-related complications were reported, indicating the overall safety of CCP therapy. Patients who eventually died from COVID-19 received CCP significantly later (3.95 versus 5.22 days after hospital admission) and had higher interleukin 6 (IL-6) levels (28.9 pg/ml versus 102.5 pg/ml) than those who survived. In addition, CCP-transfusion caused a significant reduction in the overall inflammatory status of the patients regardless of the severity of disease or outcome, as evidenced by decreasing C-reactive protein, IL6 and ferritin levels. We conclude that, CCP-transfusion is a safe and effective supplementary treatment modality for hospitalized COVID-19 patients characterized by better expected outcome if applied as early as possible. We also observed that, IL-6 may be a suitable laboratory parameter for patient selection and monitoring of CCP therapy effectiveness.
On the inference of positive and negative species associations and their relation to abundance
Authors: Rominger Andrew J.
Published: May 2021

The prevalence of rare species in ecosystems begs the question of how they persist. In a recent paper, Calatayuda et al. (CEA) provided a new hypothesis that rare species, in contrast to common species, share unique microhabitats and/or preferentially engage in mutualistic interactions. CEA support this hypotheses by reconstructing association networks from spatially replicated abundance data finding that rare species are over-representing in positive association networks while common species are over-representing in negative association networks. However, the use of abundance and co-occurrence data to infer true species associations is difficult and often inaccurate. Here, I show that the finding of rare species being more represented in positive association networks can be explained by statistical artifacts in the inference of species associations from abundance data. I caution against the inference of ecological association networks from abundance data alone.
Neural coding of visual objects rapidly reconfigures to reflect sub-trial shifts in attentional focus
Authors: Barnes Lydia , Goddard Erin , Woolgar Alexandra
Published: May 2021

Every day, we respond to the dynamic world around us by flexibly choosing actions to meet our goals. This constant problem solving, in familiar settings and in novel tasks, is a defining feature of human behaviour. Flexible neural populations are thought to support this process by adapting to prioritise task-relevant information, driving coding in specialised brain regions toward stimuli and actions that are important for our goal. Accordingly, human fMRI shows that activity patterns in frontoparietal cortex contain more information about visual features when they are task-relevant. However, if this preferential coding drives momentary focus, for example to solve each part of a task, it must reconfigure more quickly than we can observe with fMRI. Here we used MVPA with MEG to test for rapid reconfiguration of stimulus information when a new feature becomes relevant within a trial. Participants saw two displays on each trial. They attended to the shape of a first target then the colour of a second, or vice versa, and reported the attended features at a choice display. We found evidence of preferential coding for the relevant features in both trial phases, even as participants shifted attention mid-trial, commensurate with fast sub-trial reconfiguration. However, we only found this pattern of results when the task was difficult, and the stimulus displays contained multiple objects, and not in a simpler task with the same structure. The data suggest that adaptive coding in humans can operate on a fast, sub-trial timescale, suitable for supporting periods of momentary focus when complex tasks are broken down into simpler ones, but may not always do so.
LMO2 is critical for early metastatic events in breast cancer
Authors: Sikandar Shaheen , Antony Jane , Gulati Gunsagar S. , Kuo Angera H. , Dang Ho William Hai , Das Soumyashree , Steen Chloé B. , Pereira Thiago Almeida , Qian Dalong , Beachy Philip A. , Dirbas Fredrick , Red-Horse Kristy , Rabbitts Terence H. , Thiery Jean Paul , Newman Aaron M. , Clarke Michael F.
Published: May 2021

Metastasis is responsible for the majority of breast cancer-related deaths, however identifying the cellular determinants of metastasis has remained challenging. Here, we identified a minority population of immatureTHY1+/VEGFA+tumor epithelial cells in human breast tumor biopsies that display angiogenic features and are marked by the expression of the oncogene,LMO2. Higher abundance ofLMO2+basal cells correlated with tumor endothelial content and predicted poor distant recurrence-free survival in patients. UsingMMTV-PyMT/Lmo2CreERT2mice, we demonstrated thatLmo2lineage- traced cells have a higher propensity to metastasize. LMO2 knockdown in human breast tumors reduced lung metastasis by impairing intravasation, leading to a reduced frequency of circulating tumor cells. Mechanistically, we find that LMO2 binds to STAT3 and is required for STAT3 activation by TNFα and IL6. Collectively, our study identifies a population of metastasis-initiating cells with angiogenic features and establishes the LMO2-STAT3 signaling axis as a therapeutic target in breast cancer metastasis.

One sentence summary

LMO2 modulates STAT3 signaling in breast cancer metastasis.
Mechanosensitive Osteogenesis on Native Cellulose Scaffolds for Bone Tissue Engineering
Authors: Latour Maxime Leblanc , Pelling Andrew E.
Published: May 2021

In recent years, plant-derived cellulosic biomaterials have become a popular way to create scaffolds for a variety of tissue engineering applications. Moreover, such scaffolds possess similar physical properties (porosity, stiffness) that resemble bone tissues and have been explored as potential biomaterials for tissue engineering applications. Here, plant-derived cellulose scaffolds were seeded with MC3T3-E1 pre-osteoblast cells. Moreover, to assess the potential of these biomaterials, we also applied cyclic hydrostatic pressure (HP) to the cells and scaffolds over time to mimic a bone-like environment more closely. After one week of proliferation, cell-seeded scaffolds were exposed to HP up to 270 KPa at a frequency of 1Hz, once per day, for up to two weeks. Scaffolds were incubated in osteogenic inducing media (OM) or regular culture media (CM). The effect of cyclic HP combined with OM on cell-seeded scaffolds resulted in an increase of differentiated cells. This corresponded to an upregulation of alkaline phosphatase activity and scaffold mineralization. Importantly, the results reveal that well known mechanosensitive pathways cells which regulate osteogenesis appear to remain functional even on novel plant-derived cellulosic biomaterials.
EVL and MIM/MTSS1 regulate actin cytoskeletal remodeling to promote dendritic filopodia in developing neurons
Authors: Parker SS , Ly KT , Grant AD , Wang A , Parker JD , Roman MR , Padi M , Wolgemuth CW , Langlais PR , Mouneimne G
Published: May 2021

Dendritic spines are the postsynaptic compartment of a functional neuronal synapse, and are critical for synaptic connectivity and plasticity. The developmental precursor to dendritic spines, dendritic filopodia, are highly motile protrusions that facilitate synapse formation by sampling the environment for suitable axon partners during development and learning. Despite the significance of the actin cytoskeleton in driving these protrusions, the actin remodeling factors involved in this process are not fully characterized. In this work, we identify a critical function for the Ena/VASP protein EVL in the regulation of dendritic filopodia. Amongst the Ena/VASP proteins, EVL is uniquely required for the characteristic morphology and dynamics of dendritic filopodia. Using a combination of genetic and optogenetic manipulations, we demonstrate that EVL promotes protrusive motility through membrane-direct actin polymerization at dendritic filopodia tips. EVL forms a complex at nascent protrusions and dendritic filopodia tips with MIM/MTSS1, an I-BAR protein recently discovered to be important for initiation of dendritic filopodia. We propose a model in which EVL cooperates with MIM to elongate and coalesce branched actin filaments, establishing the dynamic lamellipodia-like architecture of dendritic filopodia in developing neurons.
The parasitic nematode Strongyloides ratti exists as populations of long-lived asexual lineages
Authors: Cole Rebecca , Holroyd Nancy , Tracey Alan , Berriman Matt , Viney Mark
Published: May 2021

Nematodes are important parasites of people and animals and in natural ecosystems they are a major ecological force. Strongyloides ratti is a common parasitic nematode of wild rats and we have investigated its population genetics using single worm, whole genome sequencing. We find that S. ratti populations in the UK consist of mixtures of asexual lineages that are widely dispersed across a host population. These parasite lineages are likely very old and may have originated in Asia from where rats originated. Genes that underly the parasitic phase of the parasite’s life cycle are hyperdiverse, compared with the rest of the genome, and this may allow the parasites to maximise their fitness in a diverse host population. These patterns of parasitic nematode population genetics have not been found before and may also apply to Strongyloides spp. that infect people, which will affect how we should approach their control.
Sir3 Heterochromatin Protein Promotes NHEJ by Direct Inhibition of Sae2
Authors: Bordelet Hélène , Costa Rafaël , Brocas Clémentine , Dépagne Jordane , Veaute Xavier , Busso Didier , Batté Amandine , Guérois Raphaël , Marcand Stéphane , Dubrana Karine
Published: May 2021

Heterochromatin is a conserved feature of eukaryotic chromosomes, with central roles in gene expression regulation and maintenance of genome stability. How heterochromatin proteins regulate DNA repair remains poorly described. In Saccharomyces cerevisiae, the Silent Information Regulator (SIR) complex assembles heterochromatin-like chromatin at subtelomeres. SIR-mediated repressive chromatin limits double strand break (DSB) resection protecting damaged chromosome ends during HR. As resection initiation marks the cross-road between repair by non-homologous end joining (NHEJ) or HR, we asked whether SIR- mediated heterochromatin regulates NHEJ. We show that SIRs promote NHEJ through two pathways, one depending on repressive chromatin assembly, and the other relying on Sir3 in a manner that is independent of its heterochromatin-promoting function. Sir3 physically interacts with Sae2 and this interaction impairs Sae2-dependent MRX functions. As a consequence, Sir3 limits Mre11-mediated resection, delays MRX removal from DSB ends and promotes NHEJ.
TnCentral: A Prokaryotic Transposable Element Database and Web Portal for Transposon Analysis
Authors: Ross Karen , Varani Alessandro M. , Snesrud Erik , Huang Hongzhan , Alvarenga Danillo Oliveira , Zhang Jian , Wu Cathy , McGann Patrick , Chandler Mick
Published: May 2021

We describe here the structure and organization of TnCentral (https://tncentral.proteininformationresource.org/). a web resource for prokaryotic transposable elements (TE). TnCentral currently contains ~400 carefully annotated TE, including transposons from the Tn3, Tn7, Tn402 and Tn554 families, compound transposons, integrons and associated insertion sequences (IS). These TE carry passenger genes, including genes conferring resistance to over 25 classes of antibiotics and nine types of heavy metal as well as genes responsible for pathogenesis in plants, toxin/antitoxin gene pairs, transcription factors and genes involved in metabolism. Each TE has its own entry page providing details about its transposition genes, passenger genes, and other sequence features required for transposition as well as a graphical map of all features. TnCentral content can be browsed and queried through text and sequence-based searches with a graphic output. We describe three use cases, which illustrate how the search interface, results tables, and entry pages can be used to explore and compare TEs.

TnCentral also includes downloadable software to facilitate user-driven identification, with manual annotation, of certain types of TE in genomic sequences. Through the TnCentral homepage, users can also access TnPedia which provides comprehensive reviews of the major TE families including an extensive general section, and specialised sections with descriptions of insertion sequence and transposon families. TnCentral and TnPedia are intuitive resources that can be used by clinicians and scientists to assess TE diversity in clinical, veterinary and environmental samples.

Importance

The ability of bacteria to undergo rapid evolution and adapt to changing environmental circumstances drives the public health crisis of multiple antibiotic resistance as well as outbreaks of disease in economically important agricultural crops and animal husbandry. Prokaryotic transposable elements (TE) play a critical role in this. Many carry “passenger genes” (not required for the transposition process) conferring resistance to antibiotics or heavy metals or causing disease in plants and animals. Passenger genes are spread by the normal TE transposition activities, by insertion into plasmids which then spread via conjugation within and across bacterial populations. Thus, an understanding of TE composition and transposition mechanisms is key to developing strategies to combat bacterial pathogenesis. Toward this end, we have developed TnCentral, a bioinformatics resource dedicated to describing and exploring the structural and functional features of prokaryotic TE and whose use is intuitive and accessible to users with or without bioinformatics expertise.
Neuron-radial glial cell communication via BMP/Id1 signaling maintains the regenerative capacity of the adult zebrafish telencephalon
Authors: Zhang Gaoqun , Lübke Luisa , Chen Fushun , Beil Tanja , Takamiya Masanari , Diotel Nicolas , Strähle Uwe , Rastegar Sepand
Published: May 2021

The mechanisms of the management of the neural stem cell (NSC) pool underlying the regenerative capacity of the adult zebrafish brain are not understood. We show that Bone Morphogenetic Proteins (BMPs) which are exclusively expressed by neurons in the adult telencephalon and the helix-loop-helix (HLH) transcription co-regulator, Inhibitor of differentiation 1 (Id1), control quiescence of NSCs. Upon injury, lack ofid1function leads to an initial over-proliferation and subsequent loss of NSCs and the regenerative capacity. BMP/Id1 signaling up-regulates the transcription factorher4.1which is also a target of Notch signaling mediating short-range control of NSC quiescence. Hence, the two signaling systems converge onto Her4.1. Our data show that neurons feedback on NSC proliferation. BMP1/Id1 signaling appears as the predominant safeguard of the NSC pool under regenerative conditions while Notch signaling is sufficient to maintain NSCs under homeostatic baseline neurogenesis in the uninjured animal.
Genome-wide functional analysis reveals key roles for kinesins in the mammalian and mosquito stages of the malaria parasite life cycle
Authors: Zeeshan Mohammad , Rashpa Ravish , Ferguson David J , Abel Steven , Chahine Zeinab , Brady Declan , Vaughan Sue , Moores Carolyn A. , Le Roch Karine G. , Brochet Mathieu , Holder Anthony A. , Tewari Rita
Published: May 2021

Kinesins are microtubule-based motors important in cell division, motility, polarity, and intracellular transport in many eukaryotes. However, they are poorly studied in the divergent eukaryotic pathogens-Plasmodium spp.,the causative agents of malaria, which manifest atypical aspects of cell division and plasticity of morphology throughout the lifecycle in both mammalian and mosquito hosts. Here we describe a genome-wide screen ofPlasmodiumkinesins, revealing diverse subcellular locations and functions in spindle assembly, axoneme formation and cell morphology. Surprisingly, only kinesin-13 is essential for growth in the mammalian host while the other eight kinesins are required during the proliferative and invasive stages of parasite transmission through the mosquito vector. In-depth analyses of kinesin-13 and kinesin-20 revealed functions in microtubule dynamics during apical cell polarity formation, spindle assembly, and axoneme biogenesis. These findings help us to understand the importance of microtubule motors and may be exploited to discover new therapeutic interventions against malaria.
Plasmodium falciparumand helminth coinfections increase IgE and parasite-specific IgG responses
Authors: Santano Rebeca , Rubio Rocío , Grau-Pujol Berta , Escola Valdemiro , Muchisse Osvaldo , Cuamba Inocência , Vidal Marta , Cisteró Pau , Ruiz-Olalla Gemma , Aguilar Ruth , Demontis Maria , Jamine Jose Carlos , Cossa Anélsio , Sacoor Charfudin , Cano Jorge , Izquierdo Luis , Chitnis Chetan E , Coppel Ross L , Chauhan Virander , Cavanagh David , Dutta Sheetij , Angov Evelina , Gaur Deepak , van Lieshout Lisette , Zhan Bin , Muñoz José , Moncunill Gemma , Dobaño Carlota
Published: May 2021

Coinfection withPlasmodium falciparumand helminths may impact the immune response to these parasites since they induce different immune profiles. We studied the effects of coinfections on the antibody profile in a cohort of 715 Mozambican children and adults using the Luminex technology with a panel of 16 antigens fromP. falciparumand 11 antigens from helminths (Ascaris lumbricoides, hookworm,Trichuris trichiura,Strongyloides stercoralisandSchistosomaspp.) and measured antigen-specific IgG and total IgE responses. We compared the antibody profile between groups defined byP. falciparumand helminth previous exposure (based on serology) and/or current infection (determined by microscopy and/or qPCR). In multivariable regression models adjusted by demographic, socioeconomic, water and sanitation variables, individuals exposed/infected withP. falciparumand helminths had significantly higher total IgE and antigen-specific IgG levels, magnitude (sum of all levels) and breadth of response to both types of parasites compared to individuals exposed/infected with only one type of parasite (p≤ 0.05). There was a positive association between exposure/infection toP. falciparumand exposure/infection to helminths or the number of helminth species, andvice versa(p≤ 0.001). In addition, children coexposed/coinfected tended (p= 0.062) to have higherP. falciparumparasitemia than those single exposed/infected. Our results suggest that an increase in the antibody responses in coexposed/coinfected individuals may reflect higher exposure and be due to a more permissive immune environment to infection in the host.

GRAPHICAL ABSTRACT
Population genomics of wall lizards reflects the dynamic history of the Mediterranean Basin
Authors: Yang Weizhao , Feiner Nathalie , Salvi Daniele , Laakkonen Hanna , Jablonski Daniel , Pinho Catarina , Carretero Miguel A. , Sacchi Roberto , Zuffi Marco A. L. , Scali Stefano , Plavos Konstantinos , Pafilis Panayiotis , Poulakakis Nikos , Lymberakis Petros , Jandzik David , Schulte Ulrich , Aubret Fabien , Badiane Arnaud , i de Lanuza Guillem Perez , Abalos Javier , While Geoffrey M. , Uller Tobias
Published: May 2021

The Mediterranean Basin has experienced extensive change in geology and climate over the past six million years. Yet, the relative importance of key geological events for the distribution and genetic structure of the Mediterranean fauna remains poorly understood. Here, we use population genomic and phylogenomic analyses to establish the evolutionary history and genetic structure of common wall lizards (Podarcis muralis). This species is particularly informative because, in contrast to other Mediterranean lizards, it is widespread across the Iberian, Italian, and Balkan peninsulas, and in extra-Mediterranean regions. We found strong support for six major lineages within P. muralis, which were largely discordant with the phylogenetic relationship of mitochondrial DNA. The most recent common ancestor of extant P. muralis was likely distributed in the Italian Peninsula, and experienced an “Out-of-Italy” expansion following the Messinian salinity crisis (~5 Mya), resulting in the differentiation into the extant lineages on the Iberian, Italian and Balkan peninsulas. Introgression analysis revealed that both inter- and intraspecific gene flow have been pervasive throughout the evolutionary history of P. muralis. For example, the Southern Italy lineage has a hybrid origin, formed through admixture between the Central Italy lineage and an ancient lineage that was the sister to all other P. muralis. More recent genetic differentiation is associated with the onset of the Quaternary glaciations, which influenced population dynamics and genetic diversity of contemporary lineages. These results demonstrate the pervasive role of Mediterranean geology and climate for the evolutionary history and population genetic structure of extant species.
Dissociating the impact of movement time and energy costs on decision-making and action initiation in humans
Authors: Lunazzi Clara Saleri , Reynaud Amélie J. , Thura David
Published: May 2021

Recent theories and data suggest that adapted behavior involves economic computations during which multiple trade-offs between reward value, accuracy requirement, energy expenditure and elapsing time are solved so as to obtain rewards as soon as possible while spending the least possible amount of energy. However, the relative impact of movement energy and duration costs on perceptual decision-making and movement initiation is poorly understood. Here, we tested 31 healthy subjects on a perceptual decision-making task in which they executed reaching movements to report probabilistic choices. In three distinct blocks of trials, the reaching time and energy costs were independently varied while decision difficulty was maintained similar at the block level. Participants also performed a fully instructed delayed-reaching (DR) task in each motor condition. Results in that DR task show that time-consuming movements extended reaction times (RTs) in most subjects, whereas energy-consuming movements led to mixed effects on RTs. In the choice task, about half of the subjects decreased their decision durations (DDs) in the time consuming condition, while the impact of energy costs on DDs were again mixed across subjects. Decision accuracy was overall similar across motor conditions. These results indicate that movement duration and, to a lesser extent, energy expenditure, idiosyncratically affect perceptual decision-making and action initiation. We propose that subjects who shortened their decisions in the time consuming condition of the choice task did so to limit a drop of their rate of reward.
Purification of immunostimulatory glucose homopolymer from broccoli (Brassica oleracea var. italica)
Authors: Miyashita Atsushi , Kataoka Keiko , Tsuchida Toshio , Ogasawara Akihiko Ano , Hamamoto Hiroshi , Takahashi Megumu , Sekimizu Kazuhisa
Published: May 2021

We prepared broccoli (Brassica oleracea var. italica) neutral polysaccharides (flow-through fractions of anion exchange column chromatography from hot water extracts) from different broccoli cultivars and compared their immunostimulatory effects in the silkworm muscle contraction assay. The cultivars showed a wide range of activity, with the cultivar ‘Winter dome’ showing the highest specific activity (more than 100 times higher than curdlan). Furthermore, the active substance was purified by gel filtration column chromatography. The active substance showed heterogeneous molecular weights of more than 270 kDa. Sugar composition analysis of the purified fraction revealed that more than 95% of its sugar component was glucose, suggesting that the immunostimulatory neutral polysaccharide from broccoli cultivar ‘Winter dome’ was a homopolymer of glucose. The purified fraction also induced TNFα production in cultured mouse macrophage cells. These results suggest that the glucose homopolymer in broccoli has an immunostimulatory effect on both arthropod and mammalian immune system.
Control of spinal motor neuron terminal differentiation through sustained Hoxc8 gene activity
Authors: Catela Catarina , Weng Yifei , Wen Kailong , Feng Weidong , Kratsios Paschalis
Published: May 2021

Spinal motor neurons (MNs) constitute cellular substrates for several movement disorders. Although their early development has received much attention, how spinal MNs become and remain terminally differentiated is poorly understood. Here, we determined the transcriptome of mouse brachial MNs at embryonic and postnatal stages. We found that genes encoding homeodomain (HOX, LIM) transcription factors (TFs), previously implicated in early MN development, continue to be expressed postnatally, suggesting later functions. To test this, we inactivated Hoxc8 at successive stages of MN development. We found that Hoxc8 is not only required to establish but also maintain expression of several MN terminal differentiation markers. Furthermore, we uncovered novel TFs with continuous MN expression, a Hoxc8 dependency for maintained expression of Iroquois (Irx) homeodomain TFs, and a new role for Irx2 in MN development. Our findings dovetail recent observations in C. elegans MNs, pointing toward an evolutionarily conserved role for Hox in neuronal terminal differentiation.
Variable Induction of Pro-inflammatory Cytokines by Commercial SARS CoV-2 Spike Protein Reagents: Potential Impacts of LPS on In Vitro Modeling and Pathogenic Mechanisms In Vivo
Authors: Ouyang Weiming , Xie Tao , Fang Hui , Gao Chunling , Stantchev Tzanko , Clouse Kathleen A. , Yuan Kun , Ju Tongzhong , Frucht David M.
Published: May 2021

Proinflammatory cytokine production following infection with severe acute respiratory syndrome coronavirus 2 (SARS CoV-2) is associated with poor clinical outcomes. Like SARS CoV-1, SARS CoV-2 enters host cells via its spike protein, which attaches to angiotensin-converting enzyme 2 (ACE2). As SARS CoV-1 spike protein is reported to induce cytokine production, we hypothesized that this pathway could be a shared mechanism underlying pathogenic immune responses. We herein compared the capabilities of Middle East Respiratory Syndrome (MERS), SARS CoV-1 and SARS CoV-2 spike proteins to induce cytokine expression in human peripheral blood mononuclear cells (PBMC). We observed that only specific commercial lots of SARS CoV-2 induce cytokine production. Surprisingly, recombinant SARS CoV-2 spike proteins from different vendors and batches exhibited different patterns of cytokine induction, and these activities were not inhibited by blockade of spike protein-ACE2 binding using either soluble ACE2 or neutralizing anti-S1 antibody. Moreover, commercial spike protein reagents contained varying levels of endotoxin, which correlated directly with their abilities to induce cytokine production. The lipopolysaccharide (LPS) inhibitor, polymyxin B, blocked this cytokine induction activity. In addition, SARS CoV-2 spike protein avidly bound soluble LPS in vitro, rendering it a cytokine inducer. These results not only suggest caution in monitoring the purity of SARS CoV-2 spike protein reagents, but they indicate the possibility that interactions of SARS CoV-2 spike protein with LPS from commensal bacteria in virally infected mucosal tissues could promote pathogenic inflammatory cytokine production.
Extracellular electron transfer increases fermentation in lactic acid bacteria via a hybrid metabolism
Authors: Tejedor-Sanz Sara , Stevens Eric T. , Finnegan Peter , Nelson James , Knoessen Andre , Light Samuel H. , Ajo-Franklin Caroline M. , Marco Maria L.
Published: May 2021

Energy conservation in microorganisms is classically categorized into respiration and fermentation, however recent work shows some species can use mixed or alternative bioenergetic strategies. We explored the utility of a flavin-based extracellular electron transport (FLEET) system for energy conservation within diverse lactic acid bacteria (LAB), microorganisms that mainly rely on fermentative metabolism and are important in food fermentations. The LAB Lactiplantibacillus plantarum uses extracellular electron transfer to increase its NAD+/NADH ratio, generate more ATP through substrate-level phosphorylation and accumulate biomass more rapidly. This novel, hybrid metabolism was dependent on a type-II NADH dehydrogenase (Ndh2) and conditionally required a flavin-binding extracellular lipoprotein (PplA) in the FLEET system to confer increased fermentation yield, metabolic flux, and environmental acidification in both laboratory media and food fermentation. The discovery of a single pathway that blends features of fermentation and respiration expands our knowledge of energy conservation metabolism and provides immediate biotechnology applications.
Evidence of neutralizing antibodies against SARS-CoV-2 in domestic cats living with owners with a history of COVID-19 in Lima – Peru
Authors: Jara Luis M. , Ferradas Cusi , Schiaffino Francesca , Sánchez-Carrión Camila , Martinez Ana , Ulloa Alexandra , Isasi-Rivas Gisela , Montalván Angela , Sarmiento Luis Guevara , Fernández Manolo , Zimic Mirko
Published: May 2021

SARS-CoV-2 can infect a variety of wild and domestic animals worldwide. Of these, domestic cats are highly susceptible species and potential viral reservoirs. As such, it is important to investigate disease exposure in areas with active community transmission and high disease prevalence. In this report we demonstrate the presence of serum neutralizing antibodies against the receptor binding-domain (RBD) of the SARS-CoV-2 in cats whose owners had been infected with SARS-CoV-2 in Lima, Peru, using a commercial competitive ELISA SARS-CoV-2 Surrogate Virus Neutralization Test. Out of 41 samples, 17.1% (7/41) and 31.7% (13/41) were positive, using the cut-off inhibition value of 30% and 20%, respectively. Not all cats living in a single house had detectable neutralizing antibodies showing that heterogenous exposure and immune among cohabiting animals. This is the first report of SARS-COV-2 exposure of domestic cats in Lima, Peru. Further studies are required to ascertain the prevalence of SARS-COV-2 exposure among domestic cats of Lima, Peru.
Phage-Derived Depolymerase as an Antibiotic Adjuvant Against Multidrug-Resistant Acinetobacter Baumannii
Authors: Chen Xi , Liu Miao , Zhang Pengfei , Xu Miao , Yuan Weihao , Bian Liming , Liu Yannan , Xia Jiang , Leung Sharon S.Y.
Published: May 2021

Bacteriophage-encoded depolymerases are responsible for degrading capsular polysaccharides (CPS), lipopolysaccharides (LPS) and exopolysachcharides (EPS) of the host bacteria during phage invasion. They have been considered as promising antivirulence agents in controlling bacterial infections, including those caused by drug-resistant bacteria. This feature inspires a hope of utilizing these enzymes to disarm the polysaccharide capsid of the bacterial cells, which then strengthens the action of antibiotics. Here we have identified, cloned, and expressed a depolymerase Dpo71 from a bacteriophage specific for the gram-negative (G-ve) bacterium Acinetobacter baumannii in the heterologous host E. coli. Dpo71 sensitizes the multidrug-resistant (MDR) A. baumannii to the host immune attack, and also acts as an adjuvant to assist or boost the action of antibiotics, for example colistin. Specifically, Dpo71 at 10 µg/ml enables a complete bacterial eradication by human serum at 50% volume ratio. Dpo71 inhibits biofilm formation and disrupts the pre-formed biofilm. Combination of Dpo71 could significantly enhance the antibiofilm activity of colistin, and improve the survival rate of A. baumannii infected Galleria mellonella. Dpo71 retains the strain-specificity of the parent phage from which Dpo71 is derived: the phage-sensitive A. baumannii strains respond to Dpo71 treatment, whereas the phage-insensitive strains do not. This indicates that Dpo71 indeed is responsible for the host specificity of bacteriophages. In summary, our work demonstrates the feasibility of using recombinant depolymerases as an antibiotic adjuvants to supplement the development of new antibacterials and to battle against MDR pathogens.
Genomic signatures of recent adaptation in a wild bumblebee
Authors: Colgan Thomas J. , Arce Andres N. , Gill Richard J. , Rodrigues Ana Ramos , Kanteh Abdoulie , Duncan Elizabeth J. , Li Li , Chittka Lars , Wurm Yannick
Published: May 2021

Behavioral experiments and analyses of observation records have shown that environmental changes threaten insect pollinators, creating risks for agriculture and ecosystem stability. Despite their importance, we know little about how wild insects or other animals can adapt in response to environmental pressures. To understand the genomic bases of adaptation in an ecologically important pollinator, we analyzed genomes ofBombus terrestrisbumblebees collected across Great Britain. We reveal extensive genetic diversity within this population, and strong signatures of recent adaptation throughout the genome. More specifically, we find that selection recently affected key processes underpinning environmental interactions, including neurobiology, wing development, and response to xenobiotics. We also discover unusual features of the genome, including a 53-gene region that lacks genetic diversity in many bee species, and a horizontal gene transfer from aWolbachiabacteria. The genetic diversity and gene flow we observe for this species could support its resilience to ongoing and future challenges. Overall, we provide important insight on the genetic health of an ecologically and economically important pollinator and reveal mechanisms by which it has recently adapted. The approach we used could help to understand how species differ in their adaptive potential, and to develop conservation strategies for those most at risk.
Induction of an immortalized songbird cell line allows for gene characterization and knockout by CRISPR-Cas9
Authors: Biegler Matthew T. , Fedrigo Olivier , Collier Paul , Mountcastle Jacquelyn , Haase Bettina , Tilgner Hagen U. , Jarvis Erich D.
Published: May 2021

The zebra finch is a powerful model for several biological fields, particularly neuroscience and vocal communication. However, this species lacks a robust cell line for molecular biology research and reagent optimization. Here we describe a cell line, CFS414, generated from zebra finch embryonic fibroblasts using the SV40 large and small T antigens. This cell line demonstrates an improvement over previous songbird cell lines through continuous and density-independent growth, allowing for indefinite culture and monoclonal line derivation. Cytogenetic, genomic, and transcriptomic profiling established the provenance of this cell line and identified the expression of genes relevant to ongoing songbird research. Using this cell line, we demonstrated a stress-dependent localization response of the zebra finch song nuclei specialized gene, SAP30L, and disrupted endogenous gene sequences usingS.aureusCas9. The utility of this cell line enhances the molecular potential of the zebra finch and validates cell immortalization strategies in a songbird species.
Targeted chromosomal Escherichia coli:dnaB exterior surface residues regulate DNA helicase behavior to maintain genomic stability and organismal fitness
Authors: Behrmann Megan S. , Perera Himasha M. , Hoang Joy M. , Venkat Trisha A. , Trakselis Michael A.
Published: May 2021

Helicase regulation is vital for replisome progression, where the helicase enzyme functions to unwind duplex DNA and aids in the coordination of replication fork activities. Currently, mechanisms for helicase regulation that involve interactions with both DNA strands through a steric exclusion and wrapping (SEW) model and conformational shifts between dilated and constricted states have been examined in vitro. To better understand the mechanism and cellular impact of helicase regulation, we used CRISPR-Cas9 genome editing to study four previously identified SEW-deficient mutants of the bacterial replicative helicase DnaB. We discovered that these four SEW mutations stabilize constricted states, with more fully constricted mutants having a generally greater impact on genomic stress, suggesting a dynamic model for helicase regulation that involves both excluded strand interactions and conformational states. These dnaB mutations result in increased DNA damage and chromosome complexity, less stable genomes, and ultimately less viable and fit strains. Notably, while two mutations stabilized fully constricted states, they have distinct effects on genomic stability, suggesting a complex relationship between helicase regulation mechanisms and faithful, efficient DNA replication. This work explores the genomic impacts of helicase dysregulation in vivo, supporting a combined dynamic regulatory mechanism involving SEW and conformational changes and relates current mechanistic understanding to functional helicase behavior.
SARS-CoV-2 inactivation by human defensin HNP1 and retrocyclin RC-101
Authors: Kudryashova Elena , Zani Ashley , Vilmen Geraldine , Sharma Amit , Lu Wuyuan , Yount Jacob S. , Kudryashov Dmitri S.
Published: May 2021

Severe acute respiratory syndrome coronavirus (SARS-CoV)-2 is an enveloped virus responsible for the COVID-19 respiratory disease pandemic. While induction of adaptive antiviral immunity via vaccination holds promise for combatting the pandemic, the emergence of new potentially more transmissible and vaccine-resistant variants of SARS-CoV-2 is an ever-present threat. Thus, it remains essential to better understand innate immune mechanisms that are active against the virus. One component of the innate immune system with broad anti-pathogen, including antiviral, activity is a group of cationic immune peptides termed defensins. The defensins’ ability to neutralize enveloped and non-enveloped viruses and to inactivate numerous bacterial toxins correlate with their ability to promote the unfolding of thermodynamically pliable proteins. Accordingly, we found that human neutrophil a-defensin HNP1 and retrocyclin RC-101 destabilize SARS-CoV-2 Spike protein and interfere with Spike-mediated membrane fusion and SARS-CoV-2 infection in cell culture. We show that HNP1 binds to Spike with submicromolar affinity. Although binding of HNP1 to serum albumin is more than 20-fold weaker, serum reduces the anti-SARS-CoV-2 activity of HNP1. At high concentrations of HNP1, its ability to inactivate the virus was preserved even in the presence of serum. These results suggest that specific a- and 8-defensins may be valuable tools in developing SARS-CoV-2 infection prevention strategies.
Actin-related protein 5 functions as a novel modulator of MyoD and MyoG in skeletal muscle and in rhabdomyosarcoma
Authors: Morita Tsuyoshi , Hayashi Ken’ichiro
Published: May 2021

Myogenic regulatory factors (MRFs) are pivotal transcription factors in myogenic differentiation. MyoD commits cells to the skeletal muscle lineage by inducing myogenic genes through recruitment of chromatin remodelers to its target loci. This study showed that Actin-related protein 5 (Arp5) acts as an inhibitory regulator of MyoD and MyoG by binding to their cysteine-rich (CR) region, which overlaps with the region essential for their epigenetic functions. Arp5 expression was faint in skeletal muscle tissues. Excessive Arp5 in mouse hind limbs caused skeletal muscle fiber atrophy. Further, Arp5 overexpression in myoblasts inhibited myotube formation by diminishing myogenic gene expression, whereas Arp5 depletion augmented myogenic gene expression. Arp5 disturbed MyoD-mediated chromatin remodeling through competition with the three-amino-acid-loop-extension-class homeodomain transcription factors the Pbx1–Meis1 heterodimer for binding to the CR region. This antimyogenic function was independent of the INO80 chromatin remodeling complex, although Arp5 is an important component of that. In rhabdomyosarcoma (RMS) cells, Arp5 expression was significantly higher than in normal myoblasts and skeletal muscle tissue, probably contributing to MyoD and MyoG activity dysregulation. Arp5 depletion in RMS partially restored myogenic properties while inhibiting tumorigenic properties. Thus, Arp5 is a novel modulator of MRFs in skeletal muscle differentiation.
Employer brand experience and organizational citizenship behavior: mediating role of employee engagement
Authors: Gupta Sakshi , Bhasin Jaya , Mushtaq Shahid
Published: May 2021

Purpose

The purpose of this paper is to investigate how employer brand experience (EBE) impacts organizational citizenship behavior (OCB). In addition, it aims to identify the mediating role of employee engagement (EE) in relationship between EBE and OCB.

Design/methodology/approach

To test the research hypotheses, a web questionnaire was developed and data were collected from 426 respondents working in the Indian banking sector. Hypotheses were tested using structural equational modeling.

Findings

EBE was positively related to OCB. The predicted mediating role of EE in the relationship between EBE and OCB was also supported.

Research limitations/implications

The study is confined to the banking sector only, which limits the generalization of the findings.

Practical implications

The results imply that firms should leverage on various dimensions of employer brand (EB) i.e. compensation, work–life balance, working environment, training and corporate social responsibility to enhance EE and OCB.

Originality/value

The research is among the very few to confirm the role of EBE vis-à-vis current employees especially in a collectivist society like India. The study also confirmed the mediating role of EE between EBE and OCB which have not been studied previously.