bluuebunny/crossref_metadata_2025_split huggingface dataset:
The RgaS-RgaR two-component system promotesClostridioides difficilesporulation through a small RNA and the Agr1 system
Authors: Edwards Adrianne N. , McBride Shonna M. 
Published: June 2023

ABSTRACTThe ability to form a dormant spore is essential for the survival of the anaerobic, gastrointestinal pathogenClostridioides difficileoutside of the mammalian gastrointestinal tract. The initiation of sporulation is governed by the master regulator of sporulation, Spo0A, which is activated by phosphorylation. Multiple sporulation factors control Spo0A phosphorylation; however, this regulatory pathway is not well defined inC. difficile. We discovered that RgaS and RgaR, a conserved orphan histidine kinase and orphan response regulator, function together as a cognate two-component regulatory system to directly activate transcription of several genes. One of these targets,agrB1D1, encodes gene products that synthesize and export a small quorum- sensing peptide, AgrD1, which positively influences expression of early sporulation genes. Another target, a small regulatory RNA now known as SrsR, impacts later stages of sporulation through an unknown regulatory mechanism(s). Unlike Agr systems in many organisms, AgrD1 does not activate the RgaS-RgaR two-component system, and thus, is not responsible for autoregulating its own production. Altogether, we demonstrate thatC. difficileutilizes a conserved two-component system that is uncoupled from quorum-sensing to promote sporulation through two distinct regulatory pathways.AUTHOR SUMMARYThe formation of an inactive spore by the anaerobic gastrointestinal pathogen,Clostridioides difficile, is required for its survival outside of the mammalian host. The sporulation process is induced by the regulator, Spo0A; yet, how Spo0A is activated inC. difficileremains unknown. To address this question, we investigated potential activators of Spo0A. Here, we demonstrate that the sensor RgaS activates sporulation, but not by direct activation of Spo0A. Instead, RgaS activates the response regulator, RgaR, which in turn activates transcription of several genes. We found two direct RgaS- RgaR targets independently promote sporulation:agrB1D1, encoding a quorum-sensing peptide, AgrD1, andsrsR, encoding a small regulatory RNA. Unlike most other characterized Agr systems, the AgrD1 peptide does not affect RgaS-RgaR activity, indicating that AgrD1 does not activate its own production through RgaS-RgaR. Altogether, the RgaS-RgaR regulon functions at multiple points within the sporulation pathway to tightly controlC. difficilespore formation.
Association of Domains of Sedentary Behavior with Cardiovascular Disease Events in The Multi-Ethnic Study of Atherosclerosis
Authors: Mayo Nicole L. , Lopez Daniel A. , Block Robert C. , Shi Hangchuan , Bertoni Alain G. , Diaz Keith M. , Ding Jingzhong , Post Wendy S. , Liu Yongmei , Li Dongmei 
Published: June 2023

Structured AbstractBackgroundSedentary behavior is associated with an increased risk for adverse health outcomes, including cardiovascular disease (CVD), independent of physical activity status. Little is known about this relationship in an ethnically diverse population. The objective of our study is to assess the effects of leisure time and occupational sedentary behavior on multiple cardiovascular outcomes in a multi-ethnic cohort.MethodsThe Multi-Ethnic Study of Atherosclerosis (MESA) includes 2619 Caucasian, 1495 Hispanic, 1891 Black, and 804 Chinese-American adults aged 45-84 years and free of clinical CVD at enrollment, Sedentary behavior was self-reported at baseline. Participants were followed for an average of 13.6 years, and 14 types of cardiovascular outcomes were ascertained. Hazards of each cardiovascular outcome were modeled with adjustment for potential confounders, including physical activity.ResultsEvery one hour per day increase in leisure time sedentary behavior predicts a 6% increase in the adjusted hazards for CVD death (P< 0.05). Every one hour increase in occupational sedentary time predicts a 21% and 20% decrease in the hazard for PVD and other revascularization, respectively (P< 0.05).ConclusionsLeisure time sedentary behavior was associated with increased hazards for CVD death, but occupational sedentary time appears to be protective of peripheral vascular disease and other revascularization.Condensed AbstractSedentary behavior has been consistently associated with an increased risk for adverse health outcomes, including cardiovascular disease (CVD), independent of physical activity status. The Multi-Ethnic Study of Atherosclerosis (MESA) consists of a racially and ethnically diverse cohort of adults age 45-84, free from CVD at baseline. Greater levels of leisure time sedentary behavior predicted increased hazards for PVD and CVD death after an average follow up of 13.6 years whereas occupational sedentary behaviors predicted reduced PVD. These results underscore the importance of reducing time spent sitting in addition to advocating for meeting physical activity targets across ethnicities.
LPCAT2 Regulates CD14 Expression During Macrophage Inflammatory Response to E. coli O111:B4
Authors: Poloamina Victory Ibigo , Abate Wondwossen , Fejer Gyorgy , Jackson Simon K 
Published: June 2023

AbstractLPCAT2 is a lipid-modifying enzyme that co-localises in lipid rafts with TLR4 and regulates macrophage inflammatory response; however, its effect on TLR4 co-receptor–CD14 is unknown. RAW264.7 cells, a common murine macrophage experimental model, were used to study the effect of LPCAT2 on CD14 expression. The expression of LPCAT2 in RAW264.7 cells was silenced using RNA interference and treated with 100ng/ml of various lipopolysaccharide chemotypes. We found that CD14 expression induced by smooth lipopolysaccharide was significantly decreased (p < 0.05) in RAW264.7 macrophages with LPCAT2 silenced. This study suggests that LPCAT2 regulates CD14 gene and protein expression. This implies that LPCAT2 can regulate CD14-dependent cellular activities.
Decline of DNA damage response along with myogenic differentiation
Authors: Sutcu Haser Hasan , Rassinoux Phoebe , Donnio Lise-Marie , Neuillet Damien , Vianna François , Gabillot Olivier , Mari Pierre-Olivier , Baldeyron Céline , Giglia-Mari Giuseppina 
Published: June 2023

ABSTRACTDNA integrity is incessantly confronted to endogenous and exogenous agents inducing DNA lesions, which are harmful for cellular homeostasis. Luckily all organisms are equipped with a network of DNA damage response (DDR) mechanisms that will repair DNA lesions and restore the proper cellular activities. Despite DNA repair mechanisms have been revealed in vitro and in replicating cells, still little is known on how DNA lesions are repaired and consequently how cellular homeostasis is maintained in post-mitotic cells. Muscle fibers are highly specialised post-mitotic cells organized in syncytia and, they are vulnerable to age-related degeneration and atrophy following radiotherapy treatment. We have here studied in detail the DNA repair capacity of muscle fibers nuclei and compared it with the one measured in proliferative myoblasts. We focused on the DNA repair mechanisms that correct ionizing radiation (IR)-induced lesions, namely the base excision repair (BER), the non-homologous end joining (NHEJ) and the homologous recombination (HR). We found that in the most differentiated myogenic cells, myotubes, all of these DNA repair mechanisms present weakened kinetics of recruitment of DNA repair proteins to IR-damaged DNA. For BER and HR, this decline can be linked to reduced steady state levels of key proteins involved in these processes, probably since nuclei within muscle fibers no longer replicate their DNA.SIGNIFICANCEAlthough skeletal muscle tissue has been considered as resistant to injury caused by ionizing radiations (IR), in the long term, muscle surrounding irradiated regions of the body following radiotherapy has physiological consequences such as atrophy and muscle wasting. To prevent and/or counteract the harmful effects of IR in muscle fibers, it is essential, to evaluate the DNA repair capacity of post-mitotic myonuclei. By using fluorescently tagged DNA repair proteins expressed in myoblasts and myotubes, we were able to assess the kinetics of the DNA damage response in these cells and determine that DNA damage response is weakened during the process of myofibrillogenesis.
Dual-specific autophosphorylation of kinase IKK2 enables phosphorylation of substrate IκBα through a phosphoenzyme intermediate
Authors: Borar Prateeka , Biswas Tapan , Chaudhuri Ankur , Pallavi Rao T , Raychaudhuri Swasti , Huxford Tom , Chakrabarti Saikat , Ghosh Gourisankar , Polley Smarajit 
Published: June 2023

AbstractRapid and high-fidelity phosphorylation of two serines (S32 and S36) of IκBα by a prototype Ser/Thr kinase IKK2 is critical for fruitful canonical NF-κB activation. Here, we report that IKK2 is a dual specificity Ser/Thr kinase that autophosphorylates itself at tyrosine residues in addition to its activation loop serines. Mutation of one such tyrosine, Y169, located in proximity to the active site, to phenylalanine, renders IKK2 inactive for phosphorylation of S32 of IκBα. Surprisingly, auto-phosphorylated IKK2 relayed phosphate group(s) to IκBα without ATP when ADP is present. We also observed that mutation of K44, an ATP-binding lysine conserved in all protein kinases, to methionine renders IKK2 inactive towards specific phosphorylation of S32 or S36 of IκBα, but not non-specific substrates. These observations highlight an unusual evolution of IKK2, in which autophosphorylation of tyrosine(s) in the activation loop and the invariant ATP-binding K44 residue define its signal-responsive substrate specificity ensuring the fidelity of NF-κB activation.
Acyl-CoA thioesterase-2 facilitates β-oxidation in glycolytic skeletal muscle in a lipid supply dependent manner
Authors: Bekeova Carmen , Han Ji In , Xu Heli , Kerr Evan , Blackburne Brittney , Lynch Shannon C. , Mesaros Clementina , Murgia Marta , Vadigepalli Rajanikanth , Beld Joris , Leonardi Roberta , Snyder Nathaniel W. , Seifert Erin L. 
Published: June 2023

ABSTRACTAcyl-Coenzyme A (acyl-CoA) thioesters are compartmentalized intermediates that participate in in multiple metabolic reactions within the mitochondrial matrix. The limited availability of free CoA (CoASH) in the matrix raises the question of how the local acyl-CoA concentration is regulated to prevent trapping of CoASH from overload of any specific substrate. Acyl-CoA thioesterase-2 (ACOT2) hydrolyzes long-chain acyl-CoAs to their constituent fatty acids and CoASH, and is the only mitochondrial matrix ACOT refractory to inhibition by CoASH. Thus, we reasoned that ACOT2 may constitutively regulate matrix acyl-CoA levels.Acot2deletion in murine skeletal muscle (SM) resulted in acyl-CoA build-up when lipid supply and energy demands were modest. When energy demand and pyruvate availability were elevated, lack of ACOT2 activity promoted glucose oxidation. This preference for glucose over fatty acid oxidation was recapitulated in C2C12 myotubes with acute depletion ofAcot2, and overt inhibition of β-oxidation was demonstrated in isolated mitochondria fromAcot2-depleted glycolytic SM. In mice fed a high fat diet, ACOT2 enabled the accretion of acyl-CoAs and ceramide derivatives in glycolytic SM, and this was associated with worse glucose homeostasis compared to when ACOT2 was absent. These observations suggest that ACOT2 supports CoASH availability to facilitate β-oxidation in glycolytic SM when lipid supply is modest. However, when lipid supply is high, ACOT2 enables acyl-CoA and lipid accumulation, CoASH sequestration, and poor glucose homeostasis. Thus, ACOT2 regulates matrix acyl-CoA concentration in glycolytic muscle, and its impact depends on lipid supply.
Establishment of a novel non-integrated human pluripotent stem cell-based gastruloid model
Authors: Yuan Gege , Wang Jiachen , Liu Zhaode , Chen Mengqi , Zhu Pinmou , Zhang Hao , Hu Zhibin , Cui Yiqiang , Yuan Yan , Sha Jiahao 
Published: June 2023

AbstractEmbryo loss and pregnancy disorders are prevalent worldwide, with both conditions critically associated with dysfunctioning gastrulation processes. Gastrulation and post-gastrulation organogenesis are crucial stages of embryonic development that establish the blueprint for body part formation. These processes involve the sequential generation of three germ layer cells and primordial germ cells, as well as the assembly of the precursor tissues for body parts. However, due to ethical limitations associated with studying human embryogenesis, a more detailed understanding of gastrulation and post-gastrulation organogenesis remains elusive. To ensure that the knowledge obtained from gastruloids is biologically meaningful and clinically relevant, it is critical to create high-fidelity human embryo models that closely mimic embryogenesisin vivo. Here, we developed a two-stage derivation gastruloidsin vitrobased on human pluripotent stem cells. Morphological tracking mimicks the developmental processes of models from Carnegie Stage 4 (CS4) to early CS7. Our gastruloids exhibit key structures characteristic of human embryos, including amniotic cavity, embryonic disc, primitive streak, primary yolk sac, secondary yolk sac, and blood islets. Comparison of our cell lineage development maps showed that gastruloids closely resembled human natural CS7 gastrula. Our gastruloids exhibited transcriptional characteristics that mimicked the molecular pathways observed in natural embryos development. Importantly, we found that in our model, extraembryonic mesoderm originates from the yolk sac and primordial germ cells originate from the posterior epiblast of the embryonic disc. Moreover, we found that thalidomide affects the differentiation of three germ layer cells, resulting in the arrest of human gastruloid development. In conclusion, by establishing a human gastruloid, we were able to gain valuable insights into the mechanisms responsible for human gastrulation and shed light on the causes of early embryo loss and pregnancy disorders.
Synergistic effects of PARP inhibitors by Schlafen 11 and BRCA2-deficiency through accumulation of single-strand DNA gaps behind a fork
Authors: Onji Hiroshi , Tate Sota , Sakaue Tomohisa , Onishi Nobuyuki , Matsumoto Takashi , Sugiyama Takashi , Higashiyama Shigeki , Murai Junko 
Published: June 2023

AbstractPoly (ADP-ribose) polymerase (PARP) inhibitors (PARPis) induce synthetic lethality in breast cancer gene (BRCA)-deficient tumors. Besides the original model proposed by accumulation of double-strand DNA breaks due to the impaired homologous recombination, accumulation of single-strand DNA (ssDNA) gaps due to impaired BRCA-mediated Okazaki fragment processing has emerged as an alternative mechanism of synthetic lethality. Accordingly, PARPis induce ssDNA gaps behind a replication fork in BRCA-deficient cells. Schlafen 11 (SLFN11), a member of the SLFN family, binds replication protein A (RPA)-coated ssDNA gaps and sensitizes cancer cells to DNA-damaging anticancer agents. These facts motivated us to examine the combinational effects of SLFN11 and BRCA-deficiency on PARPis sensitivity. Here, we show that SLFN11 and BRCA2-deficiency synergistically increased sensitivity to PARPis (talazoparib, niraparib, olaparib, and veliparib) at specific concentrations, where SLFN11 alone showed marginal effects. Using chromatin-bound proteins and alkaline BrdU comet assays in human cancer cells, we revealed that ssDNA gaps induced by PARPis were increased by SLFN11 or BRCA2-deficiency and that the combination of the two had the greatest effect. SLFN11 was recruited to and colocalized with chromatin-bound RPA2 under PARPis. SLFN11 recruited around a fork under DNA damage blocked replication, whereas SLFN11 recruited behind a fork under PARPis did not. Chromatin recruitment of SLFN11 and RPA2 was attenuated by the MRE11 inhibitor mirin. Hence, our studies showed that BRCA2-deficiency increased ssDNA gaps behind a fork under PARPis treatment, where SLFN11 bound and further increased the gaps. Our findings provide a mechanistic understanding of favorable responses to PARPis in SLFN11-proficient and BRCA-deficient tumors.SignificanceThis study reveals how SLFN11 enhances the antitumor effects of PARP inhibitors in BRCA2-deficient cancer cells and highlights the importance of analyzing SLFN11 expression in addition to BRCA analysis in clinical practice.
FISHtoFigure: An easy-to-use tool for rapid, multi-target partitioning and analysis of sub-cellular mRNA transcripts in smFISH data
Authors: Bentley-Abbot Calum , Heslop Rhiannon , Pirillo Chiara , Sinton Matthew C. , Chandrasegaran Praveena , McConnell Gail , Roberts Ed , Hutchinson Edward , MacLeod Annette 
Published: June 2023

AbstractSingle molecule fluorescencein situhybridisation (smFISH) has become a valuable tool to investigate the mRNA expression of single cells. However, it requires a considerable amount of bioinformatic expertise to use currently available open-source analytical software packages to extract and analyse quantitative data about transcript expression. Here, we present FISHtoFigure, a new software tool developed specifically for the analysis of mRNA abundance and co-expression in QuPath-quantified, multi-labelled smFISH data. FISHtoFigure facilitates the automated spatial analysis of transcripts of interest, allowing users to analyse populations of cells positive for specific combinations of mRNA targets without the need for bioinformatics expertise. As a proof of concept and to demonstrate the capabilities of this new research tool, we have validated FISHtoFigure in multiple biological systems. We used FISHtoFigure to identify an upregulation of T-cells in the spleens of mice infected with influenza A virus, before analysing more complex data showing crosstalk between microglia and regulatory B-cells in the brains of mice infected withTrypanosoma brucei brucei. These analyses demonstrate the ease of analysing cell expression profiles using FISHtoFigure and the value of this new tool in the field of smFISH data analysis.
Female smallholder farmers’ preferences for digital and conventional credit attributes: evidence from Madagascar
Authors: Wahbi Annkathrin , Sarfo Yaw , Musshoff Oliver 
Published: June 2023

PurposeDigital credit is spreading rapidly across Sub-Saharan Africa and holds potential for financial inclusion and female financial autonomy. Women in developing economies have long been targeted by microfinance institutions due to the women’s reliability and positive spillover effects. Yet, adoption rates for digital financial innovations remain moderate among rural women in Sub-Saharan Africa. The authors explore whether female preferences for digital and conventional credit differ from males.Design/methodology/approachThe authors conduct a Discrete Choice Experiment with 420 smallholder farmers in central Madagascar, one of the region's poorest countries, to assess preferences for selected digital and conventional credit attributes.FindingsResults of the mixed logit model and the comparison of the willingness-to-pay via Poe-test suggest high general demand for both credit forms. The demand of female respondents is higher than that of males, suggesting that they might be underserved. This holds for both credit forms. However, differences in willingness to pay for the credit attributes are mostly not statistically significant, indicating that designing gender-specific services may not be advisable.Originality/valueThis article is believed to be the first to assess and compare gendered willingness to pay for digital and conventional credit. The study’s findings give valuable insights to decision-makers in development politics as well as the fintech industry.
Using accounting theory to develop a theoretical model for credit card rewards programme transactions
Authors: Brink Sophia , Steenkamp Gretha 
Published: June 2023

PurposeAfter the effective date of International Financial Reporting Standard (IFRS) 15, the accounting treatment of credit card rewards programmes (CCRPs) is no longer explicitly prescribed. Uncertainty regarding what constitutes faithful representation, and the inconsistent accounting practices observed, has created a need for guidance on the appropriate accounting treatment of CCRP transactions. Accounting theory has the potential to provide the foundation for this guidance. As a result, the objective of this study was to develop a theoretical model for the accounting treatment of CCRP transactions using accounting theory.Design/methodology/approachThis non-empirical qualitative conceptual study utilised document analysis, focussing specifically on accounting theory, to construct an accounting treatment model.FindingsApplying the relevant accounting theory (International Accounting Standards Board's (IASB's) Conceptual Framework), a theoretical model for the accounting treatment of CCRP transactions was developed, which emphasises the importance of understanding the economic phenomenon (the CCRP transaction) and determining how management views the transaction (in isolation as marketing or as an integral part of the credit card transaction).Originality/valueAddressing the problem of accounting for CCRP transactions with reference to accounting theory (which is the main element of scholarly activity in accounting) distinguishes this study from previous research on the topic. The CCRP accounting treatment theoretical model could assist CCRP management in faithfully accounting for a CCRP transaction and reduce uncertainty and inconsistency in practice. Moreover, this study identified the procedures to be employed when using accounting theory to determine the appropriate accounting treatment of business transactions. These procedures could be employed by accountants when faced with other transactions not covered by specific accounting standards.
Current earnings, persistence and schooling returns
Authors: Andini Corrado , Santos José Eusébio 
Published: June 2023

PurposeThe aim is to study the impact of schooling on between-groups wage inequality beyond the lens of the standard approach in the literature.Design/methodology/approachSimple econometric theory is used to make the main point of the paper. Supporting empirical evidence is also presented.FindingsDisregarding the persistence of current earnings implies a bias in the estimation of the wage return to schooling both at labour-market entry and in the rest of the working life.Research limitations/implicationsThe use of current earnings as a dependent variable in wage-schooling models may be problematic and requires specific handling.Social implicationsThe impact of schooling on the between-groups dimension of wage inequality may be different than previously thought.Originality/valueThe paper is the first to show that, when current earnings are used as a dependent variable, the identification of a wage-schooling model with the standard (time-invariant external instrument-variable) approach may lead to misleading conclusions.
Factors affecting financial decisions of university students: evidence from Pakistan
Authors: Khurshid Muzammil , Zahid R.M. Ammar , Nisa Meher Un 
Published: June 2023

PurposeThis study examined the factors affecting university students' financial decisions in Pakistan.Design/methodology/approachStructural equation models were used to analyze data from 300 university students using a questionnaire. Students' financial decisions were used as the dependent variable, while financial literacy, money ethics, money attitude, time preference, financial experience, and financial specialization agents were the independent variables.FindingsResultantly, power, personal financial literacy, achievement, financial behavior, avoidance, reward for efforts, financial experience, financial attitude, financial socialization agents, and time preference influence the students' financial decisions.Practical implicationsThe findings are useful for financial and educational institutions and policymakers who design academic courses.Originality/valueThis study measured the effects of several critical contextual areas regarding financial literacy and students' decisions in Pakistani universities.