bluuebunny/crossref_metadata_2025_split huggingface dataset:
A 4-base pair core-enclosing helix in telomerase RNA is essential and binds to the TERT catalytic protein subunit
Authors: Mefford Melissa A. , Hass Evan P. , Zappulla David C. 
Published: January 2020

ABSTRACTThe telomerase RNP counters the chromosome end-replication problem, completing genome replication to prevent cellular senescence in yeast, humans, and most other eukaryotes. The telomerase RNP core enzyme is composed of a dedicated RNA subunit and a reverse transcriptase (TERT). Although the majority of the 1157-ntSaccharomyces cerevisiaetelomerase RNA, TLC1, is rapidly evolving, the central catalytic core is largely conserved, containing the template, template-boundary helix, pseudoknot, and core-enclosing helix (CEH). Here, we show that 4-base pairs of core-enclosing helix is required for telomerase to be activein vitroand to maintain yeast telomeresin vivo, whereas ΔCEH, 1-bp, and 2-bp alleles do not support telomerase function. Using the CRISPR/dCas9-based “CARRY two-hybrid” assay to assess binding of our CEH mutant RNAs to TERT, we find that the 4-bp CEH RNA binds to TERT, but the shorter-CEH constructs do not, consistent with the telomerase activity andin vivocomplementation results. Thus, the CEH is essential in yeast telomerase RNA because it is needed to bind TERT to form the core RNP enzyme. Although the 8 nucleotides that form this 4-bp stem at the base of the CEH are nearly invariant amongSaccharomycesspecies, our results with sequence-randomized and truncated-CEH helices strongly suggest that this binding interaction with TERT is dictated more by secondary than primary structure. In summary, we have mapped an essential binding site in telomerase RNA for TERT that is crucial to form the catalytic core of this biomedically important RNP enzyme.
Expansion of Functional Regulatory T Cells Using Soluble RAGE Prevents Type 1 Diabetes
Authors: Leung Sherman S. , Borg Danielle J. , McCarthy Domenica A. , Boursalian Tamar E. , Cracraft Justen , Zhuang Aowen , Fotheringham Amelia K. , Flemming Nicole , Watkins Thomas , Miles John J. , Groop Per-Henrik , Scheijen Jean L. , Schalkwijk Casper G. , Steptoe Raymond J. , Radford Kristen J. , Knip Mikael , Forbes Josephine M. 
Published: January 2020

AbstractType 1 diabetes (T1D) is an autoimmune disease with no cure. Therapeutic translation has been hampered by preclinical reproducibility. Here, short-term administration of an antagonist to the receptor for advanced glycation end products (sRAGE) protected against murine diabetes at two independent centers. Treatment with sRAGE increased regulatory T cells (Tregs) within islets, pancreatic lymph nodes and spleen, increasing islet insulin expression and function. Diabetes protection was abrogated by Tregdepletion and shown to be dependent on antagonizing RAGE using knockout mice. Human Tregstreated with a RAGE ligand downregulated genes for suppression, migration and Treghomeostasis (FOXP3, IL7R, TIGIT, JAK1, STAT3, STAT5b, CCR4). Loss of suppressive function was reversed by sRAGE, where Tregsincreased proliferation and suppressed conventional T cell division, confirming that sRAGE expands functional human Tregs. These results highlight sRAGE as an attractive treatment to prevent diabetes, showing efficacy at multiple research centers and in human T cells.
The rifampicin-inactivating mono-ADP-ribosyl transferase ofMycobacterium smegmatissignificantly influences reactive oxygen species levels in the actively growing cells
Authors: Swaminath Sharmada , Pradhan Atul , Nair Rashmi Ravindran , Ajitkumar Parthasarathi 
Published: January 2020

ABSTRACTA classic example of antibiotic inactivating function in bacteria is theMycobacterium smegmatis(Msm) encoded rifampicin-inactivating mono-ADP-ribosyl transferase (arr). Since its probable biological role has been proposed to be in DNA damage response, which is inflicted by reactive oxygen species (ROS), in the present study, we examined whetherMsmArr influences ROS levels. For this purpose, the levels of the ROS, hydroxyl radical and superoxide, were determined in the mid-log phase (MLP) cells ofMsm arrknockout (arr-KO) strain, in comparison to those in the equivalently grownMsm arr+wild-type (WT) strain. The MLParr-KO cells generated significantly elevated levels of superoxide and hydroxyl radical, unlike the equivalently grown WT MLP cells. Complementation ofarr-KO witharr, but not with empty vector, restored the ROS levels comparable to those in the WT strain. Elevated ROS levels in thearr-KO strain enabled selection of rifampicin-resistant mutants at 10-7cfu/ml from the rifampicin-unexposed MLP cells ofarr-KO, which is one-log10higher than that for WT cells (10-8). Upon prolonged exposure to rifampicin, the susceptibility, persister formation, generation of elevated levels of hydroxyl radical by the persisters, rifampicin-resister generation frequency of the persisters and regrowth of the rifampicin-resistant mutants from the respective persisters were all comparable between thearr-KO and WT strains. These observations revealed that Arr influences ROS levels in the actively growingM. smegmatiscells but not in the rifampicin-exposed cells. We proposed the probable pathway through which Arr might be influencing ROS levels in the actively growingM. smegmatiscells.IMPORTANCEDiverse genera of bacteria consisting of pathogens, opportunistic pathogens and non-pathogens, possess Arr-type activities that confer equally efficient rifampicin resistance, thereby posing serious health hazard. Acquisition of this function by other bacteria through horizontal gene transfer enhances the hazard posed by the bacteria possessing it.M. smegmatisis an opportunistic human pathogen that causes infections of skin and soft tissues. Moreover,M. smegmatisis a genetically tractable model organism forM. tuberculosiswith the potential to function even as tuberculosis vaccine. In view of these significant aspects of Arr andM. smegmatis, the study to find out the natural physiological role of Arr inM. smegmatis, gains importance for designing strategies to prevent antibiotic inactivation and to target the cellular function to contain the bacterium. Above all, the three-dimensional structure ofM. smegmatisArr reveals significant structural homology with eukaryotic ADP-ribosyltransferases and bacterial toxins, thereby giving the study broad significance.
Visible Light Plasmon Excitation of Silver Nanoparticles Against Antibiotic-ResistantPseudomonas aeruginosa
Authors: da Silva Rafael T. P. , Petri Marcos V. , Valencia Estela Y. , Camargo Pedro H. C. , de Torresi Susana I. C. , Spira Beny 
Published: January 2020

ABSTRACTThe interaction of metallic nanoparticles with light excites a local surface plasmon resonance (LSPR). This phenomenon enables the transfer of hot electrons to substrates that release Reactive Oxygen Species (ROS). In this context, the present study was aimed at enhancing the antibacterial effect of citrate-covered silver nanoparticles (AgNPs), which already possess excellent antimicrobial properties, via LSPR excitation with visible LED againstPseudomonas aeruginosa, one of the most refractory organisms to antibiotic treatment. The Minimum Inhibitory Concentration (MIC) of AgNPs was 10 μg/ml under dark conditions and 5 μg/ml under light conditions. The combination of light and AgNPs led to 100% cell death after 60 minutes. Quantification of ROS via flow cytometry showed that LSPR stimulated AgNPs increased intracellular ROS concentration by 4.8-fold, suggesting that light-exposed AgNPs caused cell death via ROS production. Light exposition caused a small release of silver ions (0.4%) reaching a maximum after 6 hours. This indicates that silver ions play at most a secondary role inP. aeruginosadeath. Overall, the results presented here show that LSPR generation from AgNPs by visible light enhances the antimicrobial activity of silver nanoparticles and can be an alternative for the treatment of topic infections caused by antibiotic-resistant bacteria such asP. aeruginosa.
Mechanisms ofIgf2inhibition in thymic epithelial cells infected by coxsackievirus CV-B4
Authors: Michaux Hélène , Halouani Aymen , Trussart Charlotte , Renard Chantal , Jaïdane Hela , Martens Henri , Hober Didier , Geenen Vincent 
Published: January 2020

ABSTRACTEpidemiological studies have evidenced a link between type 1 diabetes (T1D) and infections by enteroviruses, especially with coxsackievirus B4 (CV-B4). CV-B4 is able to infect human and murine thymic epithelial cells (TECs) and, in a murine TEC line, we have shown that the diabetogenic strain CV-B4 E2 decreases transcription of insulin-like growth factor 2 gene (Igf2), coding for the self-peptide of the insulin family. Here we show that in CV-B4 infection of mice altersIgf2transcripts isoforms in TECs, followed by a decrease of pro-IGF2 precursor in the thymus. CV-B4 infection of a murine TEC line decreasesIgf2P3 promoter activity by targeting the region −68 to −22 upstream of the transcription start site (TSS) whereasIgf2transcripts stability is not affected, pointing towards a regulation ofIgf2transcription. Our data also show that CV-B4 decreases IL-6/STAT3 signalingin vitro. This study provides new knowledge about the regulation of intrathymicIgf2transcription by CV-B4 and reinforces the hypothesis that CV-B4 infection of the thymus could break central self-tolerance of the insulin family by decreasingIgf2transcription and IGF2 presentation in thymus epithelium.IMPORTANCECoxsackievirus B4 represents one of the most important environmental factors associated to type 1 diabetes, autoimmune disease for which no curative treatment exist. The diabetogenic strain Coxsackievirus B4 E2 was previously shown to decreaseIgf2expression, important player for central tolerance towards insulin, in a thymic epithelial cell line. The understanding ofIgf2regulation mechanisms during coxsackievirus B4 infection represents an interest for the understanding of central tolerance development but also forIgf2transcriptional regulation itself, still poorly understood.Here we demonstrate that, some transcripts isoforms ofIgf2are also decreased in thymic epithelial cellsin vivo. Moreover, we show that this decrease is induced by an alteration of specific regions ofIgf2P3 promoter and may be linked by a decrease of STAT3 signaling.In finewe hope that this work could lead to future therapies leading to reprogramming central tolerance towards β cells antigens viaIgf2expression.
NRF1 and NRF3 complementarily maintain a basal proteasome activity in cancer cells through CPEB3-mediated translational repression
Authors: Waku Tsuyoshi , Katayama Hiroyuki , Hiraoka Miyako , Hatanaka Atsushi , Nakamura Nanami , Tanaka Yuya , Tamura Natsuko , Watanabe Akira , Kobayashi Akira 
Published: January 2020

AbstractProteasomes are protease complexes essential for cellular homeostasis, and their activity is crucial for cancer cell growth. However, the mechanism of how proteasome activity is maintained in cancer cells has remained unclear. The CNC family transcription factor NRF1 induces the expression of almost all proteasome-related genes under proteasome inhibition.NRF1and its phylogenetically closest homologNRF3are both highly expressed in several types of cancers, such as colorectal cancer. Herein, we demonstrate that NRF1 and NRF3 complementarily maintain basal proteasome activity in cancer cells. A double knockdown ofNRF1andNRF3impaired the basal proteasome activity in cancer cells and the cancer cell resistance to a proteasome inhibitor anticancer drug bortezomib by significantly reducing basal expression of seven proteasome-related genes, includingPSMB3, PSMB7, PSMC2, PSMD3, PSMG2, PSMG3, andPOMP. Interestingly, the molecular basis behind these cellular consequences was that NRF3 repressed NRF1 translation by the gene induction of translational regulator CPEB3, which binds toNRF1-3′UTR and decreases polysome formation onNRF1mRNA. Consistent results were obtained from clinical analysis, wherein patients with cancer having higherCPEB3/NRF3-expressing tumors exhibit poor prognosis. These results provide the novel regulatory mechanism of basal proteasome activity in cancer cells through an NRF3-CPEB3-NRF1 translational repression axis.
Macromolecular and cytological changes in fission yeast G0 nuclei
Authors: Tan Zhi Yang , Cai Shujun , Paithankar Saayli A. , Nie Xin , Shi Jian , Gan Lu 
Published: January 2020

ABSTRACTWhen starved of nitrogen, fission yeastSchizosaccharomyces pombecells enter a quiescent “G0” state with smaller nuclei and transcriptional repression. The genomics ofS. pombeG0 cells has been well studied, but much of its nuclear cell biology remains unknown. Here we use confocal microscopy, immunoblots, and electron cryotomography to investigate the cytological, biochemical, and ultrastructural differences betweenS. pombeproliferating, G1-arrested, and G0 cell nuclei, with an emphasis on the histone acetylation, RNA polymerase II fates, and macromolecular complex packing. Compared to proliferating cells, G0 cells have lower levels of histone acetylation, nuclear RNA polymerase II, and active transcription. The G0 nucleus has similar macromolecular crowding yet fewer chromatin-associated multi-megadalton globular complexes. Induced histone hyperacetylation in G0 results in cells that have larger nuclei and therefore less compact chromatin. However, these histone-hyperacetylated G0 cells remain transcriptionally repressed with similar nuclear crowding. Canonical nucleosomes – those that resemble the crystal structure – are rare in proliferating, G1-arrested, and G0 cells. Our study therefore shows that extreme changes in nucleus physiology are possible without extreme reorganisation at the macromolecular level.SummaryWe use multiple cell-biological techniques to compare proliferating and quiescent G0 fission yeast cell nucleus transcription, histone acetylation, macromolecular packing.
Scalable generation of mesenchymal stem cells and adipocytes from human pluripotent stem cells
Authors: Karam Manale , Younis Ihab , Abdelalim Essam M. 
Published: January 2020

AbstractHuman pluripotent stem cells (hPSCs) can provide unlimited supply for mesenchymal stem cells (MSCs) and adipocytes that can be used for therapeutic applications. Here we developed a simple and highly efficient All-trans-retinoic acid (RA)-based method for generating an off-the-shelf and scalable number of human pluripotent stem cell (hPSC)-derived MSCs with enhanced adipogenic potential. We showed that short exposure of multiple hPSC lines (hESCs/ hiPSCs) to a high RA concentration (10 μM) dramatically enhances embryoid body (EB) formation through regulation of genes activating signaling pathways associated with cell proliferation, survival and adhesion, among others. Disruption of cell adhesion induced the subsequent differentiation of the highly expanded RA-derived EB-forming cells into a pure population of multipotent MSCs (up to 1542-fold increase in comparison to RA-untreated counterparts). Interestingly, the RA-derived MSCs displayed enhanced differentiation potential into adipocytes. Thus, these findings present a novel RA-based approach for providing an unlimited source of MSCs and adipocytes that can be used for regenerative medicine, drug screening, and disease modeling applications.
Evaluating complementary synergies in integrated crop-livestock systems in Ghana
Authors: Asante Bright O. , Villano Renato A. , Battese George E. 
Published: January 2020

PurposeThe purpose of this paper is to investigate the characteristics of crop-livestock diversification among smallholders in the forest-savannah agroecological zone of Ghana.Design/methodology/approachThe study uses a stochastic input distance function to investigate the evidence of economies of diversification and its effect on determining diversification decisions of smallholders in Ghana. Furthermore, this study also explores evidence of scale economies in integrated crop-livestock systems among smallholder farmers.FindingsThe empirical results show that economies of diversification are significant in determining diversification decisions of smallholders. The economies of scale are evident in integrated crop-livestock systems in Ghana, suggesting that opportunities exist to expand crop-livestock outputs without employing additional inputs or improved production technologies. Crop-livestock diversification is a desirable strategy for improving overall farm productivity among smallholders in Ghana. Economies of diversification were found to be significant among the output combinations, maize with other crops and other crops with livestock, in integrated crop-livestock production systems.Research limitations/implicationsThis study is based on a cross-sectional data set. Hence, the findings may be subject to some limitations; however, the authors have sought to minimise such limitations.Practical implicationsThe results imply that crop-livestock diversification is a desirable strategy for improving overall farm productivity. Therefore, the findings are useful for policymakers to design appropriate strategies for enhancing the production of specific output combinations in crop-livestock diversification systems among smallholders in Ghana. Specifically, such policies should promote the production and integration of crops such as legumes with other crops, and livestock with other crops in diversified farming systems to enhance overall farm productivity. This will reduce food insecurity and poverty among rural farm households and the entire rural population.Originality/valueThese results indicate that to improve crop-livestock productivity through diversification, and reduce the effect of climate variability, it is imperative to provide the enabling environment that will facilitate and encourage production of these output combinations among smallholders in Ghana. The empirical results enhance the literature by providing empirical evidence of the complementary synergies and economies of diversification in integrated farming systems and, hence, make recommendations for improving these farming systems.Peer reviewThe peer review history for this paper is available at:https://publons.com/publon/10.1108/IJSE-04-2019-0274.
Social courage fosters both voice and silence in the workplace
Authors: Howard Matt C. , Holmes Philip E. 
Published: January 2020

PurposeOne of the strongest and most important outcomes of trait social courage is employee voice, but researchers have only studied this relationship with unidimensional conceptualizations of voice. The purpose of this paper is to apply Van Dyneet al.’s (2003) three-dimensional conceptualization of voice, which also distinguishes three dimensions of silence, to provide a nuanced understanding of the relationship of social courage with voice and silence. The authors also test for the moderating effect of three contextual influences: top management attitudes toward voice and silence, supervisor attitudes toward voice and silence, as well as communication opportunities.Design/methodology/approachThe authors conducted a four-timepoint survey with each measurement occasion separated by one week. A total of 134 participants completed all four timepoints.FindingsThe results support that social courage positively relates to prosocial voice and silence, whereas it negatively relates to defensive voice and silence as well as acquiescent voice and silence. In other words, social courage positively relates to beneficial voice and silence as well as negatively relates to detrimental voice and silence. The results also failed to support any moderating effects, suggesting that the relationships of social courage are very resilient to outside forces.Practical implicationsThese findings both test prior results and discover new relationships of social courage, which can further stress the importance of courage. The authors also draw direct connections between the influence of social courage on the surrounding workplace environment – as well as the influences of the environment on social courage. While the current paper provides insights into social courage, it also directs future researchers toward new insights of their own.Originality/valueCourage is an emergent research topic within organizations. While many authors have assumed that courage is important to work, the current paper is among the few to empirically support this notion.
The Problematic Development of the Stalking Protection Order
Authors: Kelly Rory 
Published: January 2020

AbstractIn 2019, Parliament enacted the Stalking Protection Act. The Act introduces the stalking protection order (SPO); a civil measure the breach of which is an offence. The role of courts in assessing whether similar behaviour orders are penalties has attracted significant scholarly attention. This article examines instead the roles of Government and Parliament in developing the stalking protection order. My central contention is that the Home Office undertook a problematic consultation and the issues to which it gave rise were not addressed in later parliamentary debates. The result was the enactment of a coercive measure of unclear purpose and questionable efficacy. Assessing the roles of the executive and legislature in developing the SPO also allows for fresh insight into wider discussions of behaviour orders. Specifically, I question the language of ‘prevention’ that is ever‐present in such discussions and describe an important development for debates on whether behaviour orders are penalties.
Texture, physicochemical and sensory properties of artisanal Adobera cheese from Los Altos de Jalisco, a genuine Mexican cheese
Authors: Ruvalcaba‐Gómez José M , Ruiz‐Espinosa Héctor , Arteaga‐Garibay Ramón I , Rojas‐López Marlon , Amador‐Espejo Genaro G , Anaya‐Esparza Luis M , Delgado‐Macuil Raúl J 
Published: January 2020

Physicochemical and sensory properties of Adobera, a genuine, understudied Mexican raw‐milk cheese, were explored by analysing commercial samples from different manufacturers and seasons. Composition‐wise, Adobera could be considered a fresh cheese with a high moisture content (42.5%), although its relatively low water activity (0.953) and pH (5.14) and high free amino acid content (0.46 mmol/g, dry basis) could indicate otherwise. Instrumental texture corresponded to that of a semifirm cheese, while its colour was whitish‐to‐ivory. Both texture and composition were significantly affected by sampling season and cheese composition, while some attributes of sensory acceptability significantly varied with brand.
Development of a novel metric of timely care access to primary care services
Authors: Batten Adam J. , Augustine Matthew R. , Nelson Karin M. , Kaboli Peter J. 
Published: January 2020

AbstractObjectiveTo develop a model for identifying clinic performance at fulfilling next‐day and walk‐in requests after adjusting for patient demographics and risk.Data SourceUsing Department of Veterans Affairs (VA) administrative data from 160 VA primary care clinics from 2014 to 2017.Study DesignUsing a retrospective cohort design, we applied Bayesian hierarchical regression models to predict provision of timely care, with clinic‐level random intercept and slope while adjusting for patient demographics and risk status. Timely care was defined as the provision of an appointment within 48 hours of any patient requesting the clinicʼs next available appointment or walking in to receive care.Data Collection/Extraction MethodsWe extracted 1 841 210 timely care requests from 613 263 patients.Principal FindingsAcross 160 primary care clinics, requests for timely care were fulfilled 86 percent of the time (range 83 percent‐88 percent). Our model of timely care fit the data well, with a Bayesian R2 of .8. Over the four years of observation, we identified 25 clinics (16 percent) that were either struggling or excelling at providing timely care.ConclusionStatistical models of timely care allow for identification of clinics in need of improvement after adjusting for patient demographics and risk status. VA primary care clinics fulfilled 86 percent of timely care requests.