bluuebunny/crossref_metadata_2025_split huggingface dataset:
Suramin action in African trypanosomes involves a RuvB-like DNA helicase
Authors: Albisetti Anna , Hälg Silvan , Zoltner Martin , Mäser Pascal , Wiedemar Natalie 
Published: October 2022

AbstractSuramin is one of the oldest drugs in use today. It is still the treatment of choice for the hemolymphatic stage of African sleeping sickness caused byTrypanosoma brucei rhodesienseand it is also used for surra in camels, caused byTrypanosoma evansi. Yet despite one hundred years of use, suramin’s mode of action is not fully understood. Suramin is a polypharmacologic molecule that inhibits diverse proteins. Here we demonstrate that a DNA helicase of the pontin/ruvB-like 1 family, termedT. bruceiRuvBL1, is involved in suramin resistance in African trypanosomes. Bloodstream-formT. b. rhodesienseunder long-term selection for suramin resistance acquired a homozygous point mutation, isoleucin-312 to valine, close to the ATP binding site ofT. bruceiRuvBL1. The introduction of this missense mutation, by reverse genetics, into drug-sensitive trypanosomes significantly decreased their sensitivity to suramin. Intriguingly, the corresponding residue ofT. evansiRuvBL1 was found mutated in a suramin-resistant field isolate, in that case to a leucin. RuvBL1 (Tb927.4.1270) is predicted to build a heterohexameric complex with RuvBL2 (Tb927.4.2000). RNAi-mediated silencing of gene expression of eitherT. bruceiRuvBL1 or RuvBL2 caused cell death within 72 h. At 36 h after induction of RNAi, bloodstream-form trypanosomes exhibited a cytokinesis defect resulting in the accumulation of cells with two nuclei and two or more kinetoplasts. Taken together, these data indicate that RuvBL1 DNA helicase is among the primary targets of suramin in African trypanosomes.Abstract ImportanceAfrican trypanosomes cause sleeping sickness in humans, nagana in cattle, and surra in camels – lethal diseases for which there is no vaccine and only few drugs. One of the drugs is suramin, developed by Bayer in 1916. Yet despite 100 years of use, suramin’s mode of action is not fully understood at the molecular level. Here we show that a DNA helicase is involved:Trypanosoma brucei rhodesiense(causative agent of sleeping sickness) selected for suramin resistance acquired a point mutation in the DNA helicase RuvBL1 that, when introduced to wild-type trypanosomes, reduced their sensitivity to suramin. Intriguingly, the same site in RuvBL1 was mutated also in a suramin-resistant field isolate ofT. evansi(causative agent of surra). We further demonstrate that RuvBL1 is essential for proper cell division ofT. brucei. Thus we conclude that inhibition of RuvBL1 contributes to the trypanocidal action of suramin.
Purine and carbohydrate availability driveEnterococcus faecalisfitness during wound infection
Authors: Tan Casandra Ai Zhu , Chong Kelvin Kian Long , Yeong Daryl Yu Xuan , Ng Celine Hui Min , Ismail Muhammad Hafiz , Tay Vanessa Shi Yun , Ali Yusuf , Kline Kimberly A. 
Published: October 2022

ABSTRACTEnterococcus faecalisis commonly isolated from a variety of wound types. Despite its prevalence, the pathogenic mechanisms ofE. faecalisduring wound infection are poorly understood. Using a mouse wound infection model, we performedin vivo E. faecalistransposon sequencing and RNA sequencing to identify fitness determinants that are crucial for replication and persistence ofE. faecalisduring wound infection. We found thatE. faecalispurine biosynthesis genes are important for bacterial replication during the early stages of wound infection, a time when purine metabolites are rapidly consumed byE. faecaliswithin wounds. We also found that theE. faecalisMptABCD phosphotransferase system (PTS), involved in the import of galactose and mannose, is crucial forE. faecalispersistence within wounds of both healthy and diabetic mice, especially when carbohydrate availability changes throughout the course of infection. Duringin vitrogrowth with mannose as the sole carbohydrate source, shikimate and purine biosynthesis genes were downregulated in the OG1RF ΔmptDmutant compared to the isogenic wild-type strain, suggesting a link between mannose transport, shikimate, and purine biosynthesis. Together, our results suggest that dynamic and temporal microenvironment changes at the wound site affects pathogenic requirements and mechanisms ofE. faecalisand raise the possibility of lowering exogenous purine availability and/or targeting galactose/mannose PTS to control wound infections.IMPORTANCEAlthoughE. faecalisis a common wound pathogen, its pathogenic mechanisms during wound infection are unexplored. Here, combining a mouse wound infection model within vivotransposon and RNA sequencing approaches, we identified theE. faecalispurine biosynthetic pathway and galactose/mannose MptABCD phosphotransferase system as essential forE. faecalisacute replication and persistence during wound infection, respectively. The essentiality of purine biosynthesis and the MptABCD PTS is driven by the rapid consumption of purine metabolites byE. faecalisduring acute replication and changing carbohydrate availability during the course of wound infection. Overall, our findings reveal the importance of the wound microenvironment inE. faecaliswound pathogenesis and how these metabolic pathways can be targeted to better control wound infections.
Interim Analysis of a Phase I Randomized Clinical Trial on the Safety and Immunogenicity of the mRNA-1283 SARS-CoV-2 Vaccine in Adults
Authors: Yassini Patrick , Hutchens Mark , Paila Yamuna D. , Schoch Lorraine , Aunins Anne , Siangphoe Uma , Paris Robert 
Published: October 2022

ABSTRACTBackgroundThis interim analysis of an ongoing phase I randomized clinical trial evaluated the safety, reactogenicity, and immunogenicity of mRNA-1283, a next-generation SARS-CoV-2 messenger RNA (mRNA)-based vaccine encoding 2 segments of the spike protein (ie, receptor binding and N-terminal domains).MethodsHealthy aged adults 18-55 years (n = 104) were randomized (1:1:1:1:1) to receive 2 doses of mRNA-1283 (10, 30, or 100 μg) or mRNA-1273 (100 μg) administered 28 days apart, or a single dose of mRNA-1283 (100 μg). Safety was assessed and immunogenicity was measured by serum neutralizing antibody (nAb) or binding antibody (bAb) responses.ResultsAt the interim analysis, no safety concerns were identified and no serious adverse events, adverse events of special interest, or deaths were reported. Solicited systemic adverse reactions were more frequent with higher dose levels of mRNA-1283 than with mRNA-1273. At day 57, all dose levels of the 2-dose mRNA-1283 regimen (including the lowest dose level [10 μg]) induced robust nAb and bAb responses that were comparable to those of mRNA-1273 (100 μg).ConclusionsmRNA-1283 was generally safe in adults, with all dose levels of the 2-dose regimen (10, 30, and 100 μg) eliciting similar immunogenicity as the 2-dose mRNA-1273 regimen (100 μg).Clinical Trials RegistrationClinicaltrials.gov,NCT04813796
The epigenetic modifier DOT1L regulates gene regulatory networks necessary for cardiac patterning and cardiomyocyte cell cycle withdrawal
Authors: Cattaneo Paola , Hayes Michael G. B. , Baumgarten Nina , Hecker Dennis , Peruzzo Sofia , Kunderfranco Paolo , Larcher Veronica , Zhang Lunfeng , Contu Riccardo , Fonseca Gregory , Spinozzi Simone , Chen Ju , Condorelli Gianluigi , Schulz Marcel H , Heinz Sven , Guimarães-Camboa Nuno , Evans Sylvia M. 
Published: October 2022

ABSTRACTMechanisms by which specific histone modifications regulate distinct gene regulatory networks remain little understood. We investigated how H3K79me2, a modification catalyzed by DOT1L and previously considered a general transcriptional activation mark, regulates gene expression in mammalian cardiogenesis. Early embryonic cardiomyocyte ablation ofDot1lrevealed that H3K79me2 does not act as a general transcriptional activator, but rather regulates highly specific gene regulatory networks at two critical cardiogenic junctures: left ventricle patterning and postnatal cardiomyocyte cell cycle withdrawal. Mechanistic analyses revealed that H3K79me2 in two distinct domains, gene bodies and regulatory elements, synergized to promote expression of genes activated by DOT1L. Surprisingly, these analyses also revealed that H3K79me2 in specific regulatory elements contributed to silencing genes usually not expressed in cardiomyocytes. As DOT1L mutants had increased numbers of postnatal mononuclear cardiomyocytes and prolonged cardiomyocyte cell cycle activity, controlled inhibition of DOT1L might be a strategy to promote cardiac regeneration post-injury.
Identification of RNA virus-derived RdRp sequences in publicly available transcriptomic datasets
Authors: Olendraite Ingrida , Brown Katherine , Firth Andrew E. 
Published: October 2022

AbstractRNA viruses are abundant, highly diverse, and infect all or most eukaryotic organisms. However, only a tiny fraction of the number and diversity of RNA virus species have been catalogued. To cost effectively expand the diversity of known RNA virus sequences we mined publicly available transcriptomic datasets. We developed 77 family-level Hidden Markov Model profiles for the viral RNA dependent RNA polymerase – the only universal “hall-mark” gene of RNA viruses. By using these to search the NCBI Transcriptome Shotgun Assembly database, we identified 5,867 contigs encoding RNA virus RdRps or fragments thereof and analysed their diversity, taxonomic classification, phylogeny and host associations. Our study expands the known diversity of RNA viruses, and the 77 curated RdRp pHMMs provide a useful resource for the virus discovery community.
Prevention of thrombocytopenia and thrombosis in heparin-induced thrombocytopenia (HIT) using deglycosylated KKO: A novel therapeutic?
Authors: Sarkar Amrita , Khandelwal Sanjay , Kim Hyunjun , Gruel Yves , Rollin Jerome , Wool Geoffrey D. , Arepally Gowthami M. , Cines Douglas B. , Rauova Lubica , Poncz Mortimer 
Published: October 2022

AbstractHeparin-induced thrombocytopenia (HIT) is characterized by mild thrombocytopenia associated with a highly prothrombotic state due to the development of pathogenic antibodies that recognize human (h) platelet factor 4 (PF4) complexed with various polyanions. While non-heparin anticoagulants and intravenous immunoglobulin (IVIG) are the mainstay of care, bleeding may develop, and risk of new thromboembolic events remain. We had described a mouse IgGκ2b antibody KKO that mimics the sentinel features of pathogenic HIT antibodies, including binding to the same neoepitope on hPF4:polyanion complexes. KKO, like HIT IgGs, activates platelets through FcγRIIA and induces complement activation. We now asked whether Fc-modified KKO can be used as a novel therapeutic to prevent or treat HIT. Using the endoglycosidase EndoS, we created deglycosylated KKO (DGKKO). DGKKO bound to PF4-polyanion complexes, and blocked FcγRIIA-dependent activation of PF4 treated platelets by KKO, 5B9 (another HIT-like monoclonal antibody), and isolated IgGs from HIT patients. DGKKO also decreased complement activation and deposition of C3c on platelets. Injection of DGKKO into “HIT mice” lacking mouse PF4, but transgenic for hPF4 and FcγRIIA, prevented and reversed thrombocytopenia when injected before or after KKO, 5B9 or HIT IgG, respectively, in a microfluidic system. DGKKO reversed antibody-induced thrombus growth in HIT mice. In contrast, DGKKO was ineffective in preventing thrombosis by IgG from a patient with the HIT-related disorder, vaccine-induced immune thrombotic thrombocytopenia. Thus, DGKKO may represent a new class of therapeutics for targeted treatment of patients with HIT.Key PointsDeglycosylated (DG) KKO can reverse thrombocytopenia in a HIT murine model.DGKKO can prevent/reverse thrombosisin vitroand in a HIT murine model.
Centriolar subdistal appendages promote double strand break repair through homologous recombination
Authors: Rodríguez-Real Guillermo , Domínguez-Calvo Andrés , Prados-Carvajal Rosario , Bayona-Feliú Aleix , Gomes-Pereira Sónia , Balestra Fernando R. , Huertas Pablo 
Published: October 2022

AbstractThe centrosome is a cytoplasmic organelle with roles in microtubule organization which has also been proposed to act as a hub for cellular signaling. Some centrosomal components are required for full activation of the DNA Damage Response. However, if the centrosome regulates specific DNA repair pathways is not known. Here, we show that centrosomes presence is required to fully activate recombination, specifically to completely license its initial step, the so-called DNA end resection. Furthermore, we identify a centriolar structure, the subdistal appendages, and a specific factor, CEP170, as the critical centrosomal component involved in the regulation of recombination and resection, albeit it does not control end-joining repair. Cells lacking centrosomes or depleted for CEP170 are, consequently, hyper-sensitive to DNA damaging agents. Moreover, low levels of CEP170 in multiple cancer types correlate with an increase of the mutation burden associated with specific mutational signatures and a better prognosis, suggesting that changes in CEP170 can act as a mutation driver but also could be targeted to improve current oncological treatments.
Unravelling the regulation pathway of photosynthetic AB-GAPDH
Authors: Marotta Roberto , Del Giudice Alessandra , Gurrieri Libero , Fanti Silvia , Swuec Paolo , Galantini Luciano , Falini Giuseppe , Trost Paolo , Fermani Simona , Sparla Francesca 
Published: October 2022

Oxygenic phototrophs perform carbon fixation through the Calvin–Benson cycle. Different mechanisms adjust the cycle and the light-harvesting reactions to rapid environmental changes. Photosynthetic glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a key enzyme in the cycle. In land plants, different photosynthetic GAPDHs exist: the most abundant isoform is formed by A2B2 heterotetramers and the least abundant by A4 homotetramers. Regardless of the subunit composition, GAPDH is the major consumer of photosynthetic NADPH and its activity is strictly regulated. While A4-GAPDH is regulated by CP12, AB-GAPDH is autonomously regulated through the C-terminal extension (CTE) of its B subunits. Reversible inhibition of AB-GAPDH occurs via the oxidation of a cysteine pair located in the CTE and the substitution of NADP(H) with NAD(H) in the cofactor-binding site. These combined conditions lead to a change in the oligomerization state and enzyme inhibition. SEC–SAXS and single-particle cryo-EM analysis were applied to reveal the structural basis of this regulatory mechanism. Both approaches revealed that spinach (A2B2)
n
-GAPDH oligomers with n = 1, 2, 4 and 5 co-exist in a dynamic system. B subunits mediate the contacts between adjacent tetramers in A4B4 and A8B8 oligomers. The CTE of each B subunit penetrates into the active site of a B subunit of the adjacent tetramer, which in turn moves its CTE in the opposite direction, effectively preventing the binding of the substrate 1,3-bisphosphoglycerate in the B subunits. The whole mechanism is made possible, and eventually controlled, by pyridine nucleotides. In fact, NAD(H), by removing NADP(H) from A subunits, allows the entrance of the CTE into the active site of the B subunit, hence stabilizing inhibited oligomers.
The #MeToo legacy and “the Collective Us”: conceptualising accountability for sexual misconduct at work
Authors: Goncharenko Galina 
Published: October 2022

PurposeThis study aims to analyse how the collective processing of the #MeToo legacy in the form of community discourses and activism conceptualises organisational accountability for sexual misconduct at work and enhances the development of new accountability instruments.Design/methodology/approachThe study draws on social movement theory and the intellectual problematics of accountability, together with the empirical insights from two research engagement projects established and facilitated by the author.FindingsThe study reveals multiple dimensions of how post-#MeToo community activism impacted the conceptualisation of organisational accountability for sexual misconduct at work. The movement enhanced discourses prompting a new societal sense of accountability for sexual wrongdoings. This in turn facilitated public demands for accountability that pressured organisations to respond. The accountability crisis created an opportunity for community activists to influence understanding of organisational accountability for sexual misconduct at work and to propose new accountability instruments advancing harassment reporting technology, as well as an enhancing the behavioural consciousness and self-assessment of individuals.Originality/valueThe study addresses a topic of social importance in analysing how community activism arising from a social movement has transformed accountability demands and thus both advanced the conceptualisation of organisational accountability for sexual misconduct at work and established socially desirable practices for it. The study contributes to theory by revealing the emancipatory potential of community activism to influence organisational accountability practices and to propose new instruments at a moment of organisational hesitation and crisis of accountability.
When work and life boundaries are blurred: the effect of after-hours work communication through communication technology on employee outcomes
Authors: Kim Katie Haejung , Chon Myoung-Gi 
Published: October 2022

PurposeAs new work environments have been created and accelerated due to technological advances, after-hours work communication has become one of the prominent features of the technology-enabled work environment. Grounded in the conservation of resources theory, this study aims to uncover the effects of after-hours work communication through communication technologies on employees' burnout and extra-role behaviors.Design/methodology/approachThe study conducted an online survey of 315 US full-time employees from a wide range of industrial sectors.FindingsThe findings of the study show that after-hours work communication through communication technologies increases employees' negative extra-role behaviors, specifically counterproductive work behavior and negative word-of-mouth. Furthermore, emotional exhaustion partially mediated the relationship between after-hours work communication and negative extra-role behaviors.Practical implicationsThe study provides practical implications for an organization's effective optimization of work-related communication to overcome adverse consequences for employees and organizations. Corporate leaders and communication professionals are encouraged to establish clear guidelines related to the use of communication technologies after hours and exercise leadership practices to prevent after-hours communication through communication technologies from contributing to employees' burnout.Originality/valueThe findings enhance the understanding of the role of after-hours work communication through communication technologies in the workplace by revealing how the effects of after-hours work communication enabled by communication technologies on employees' psychological well-being spills over to organizational effectiveness.
The SECURE STAIRS framework: preliminary evaluation of formulation developments in the Children and Young People’s Secure Estate
Authors: McKeown Annette , Martin Aisling , Farooq Romana , Wilson Amy , Addy Chelsea , Kennedy Patrick J. 
Published: October 2022

Purpose
The purpose of this paper is to evaluate pre- and post-formulation findings with multi-disciplinary staff within two secure children’s homes (SCHs) in the North East of England.

Design/methodology/approach
Multi-disciplinary staff teams involved in formulation across two SCHs were administered pre- and post-formulation questionnaires. The pre- and post-formulation questionnaires focused on five domains: knowledge; confidence; motivation; understanding; and satisfaction with treatment plan.

Findings
A total of 338 pre- and post-formulation questionnaires were administered across 43 formulation meetings. The highest proportion of formulation attendees were: residential staff (44%); mental health staff (17%); case managers (12%); and education staff (9%). Paired samples t-tests showed significant post-formulation improvements across all domains including: knowledge [t(337) = 22.65, p < 0.001]; confidence [t(337) = 15.12, p < 0.001]; motivation [t(337) = 8.27, p < 0.001]; understanding [t(337) = 19.13, p < 0.001]; and satisfaction [t(337) = 18.81, p < 0.001].

Research limitations/implications
The SECURE STAIRS framework has supported formulation developments across the Children and Young People’s Secure Estate. Preliminary findings within two SCHs suggest multi-disciplinary staff teams find psychologically informed formulation beneficial. Future directions are considered including future evaluation of young person involvement in formulation meetings.

Originality/value
There is a notable lack of existing research within the child and young people secure estate evaluating the impact of SECURE STAIRS trauma-informed care developments including the impact of team formulation. This paper adds to the evidence base.
Impact of internal corporate social responsibility: a parallel mediation analysis
Authors: Ranjan Sushant , Dash Sanket 
Published: October 2022

PurposeWorkplace deviant behaviors (WDBs) have a significant negative impact on firms. Present study explores the role of employees’ perception of firms’ internal corporate social responsibility (internal CSR) in reducing their intention to engage in WDB. Social exchange theory (SET) and job demand-resource (JD-R) model form the conceptual underpinning of the study.Design/methodology/approachHypotheses were developed based on a comprehensive literature review and tested on employees working in various public and private sector organizations in India. AMOS and SPSS PROCESS macro were used to test the conceptual model.FindingsEmployees’ perception of firms’ internal CSR reduced their intention to engage in WDB. Occupational strain was confirmed as a mediator in the above mentioned relationship. Further, the study also establishes internal CSR as an antecedent to increased perceptions of procedural justice.Practical implicationsManagers may leverage internal CSR communication as a tool to minimize WDB at the workplace. Moreover, it may also be used to reduce occupational strain and strengthen the perceptions of fairness among employees.Originality/valueVery limited research is available on internal CSR and WDB. Through this study authors contribute to the nascent literature by affirming the negative relationship between internal CSR and WDB using the SET and JD-R model.