bluuebunny/crossref_metadata_2025_split huggingface dataset:
Crystal structure and catalytic mechanism of PL35 family glycosaminoglycan lyases with an ultrabroad substrate spectrum
Authors: Wei Lin , Cao Hai-Yan , Zou Ruyi , Du Min , Zhang Qingdong , Lu Danrong , Xu Xiangyu , Xu Yingying , Wang Wenshuang , Chen Xiu-Lan , Zhang Yu-Zhong , Li Fuchuan 
Published: September 2024

AbstractRecently, a new class of glycosaminoglycan (GAG) lyases (GAGases) belonging to PL35 family has been discovered with an ultrabroad substrate spectrum that can degrade three types of uronic acid-containing GAGs (hyaluronic acid, chondroitin sulfate and heparan sulfate) or even alginate. In this study, the structures of GAGase II fromSpirosoma fluvialeand GAGase VII fromBacteroides intestinalisDSM 17393 were determined at 1.9 and 2.4 Å resolution, respectively, and their catalytic mechanism was investigated by the site-directed mutant of their crucial residues and molecular docking assay. Structural analysis showed that GAGase II and GAGase VII consist of an N-terminal (α/α)6toroid multidomain and a C-terminal two-layered β-sheet domain with Mn2+. Notably, although GAGases share similar folds and catalytic mechanisms with some GAG lyases and alginate lyases, they exhibit higher structural similarity with alginate lyases than GAG lyases, which may present a crucial structural evidence for the speculation that GAG lyases with (α/α)ntoroid and antiparallel β-sheet structures arrived by a divergent evolution from alginate lyases with the same folds. Overall, this study not only solved the structure of PL35 GAG lyases for the first time and investigated their catalytic mechanism, especially the reason why GAGase III can additionally degrade alginate, but also provided a key clue in the divergent evolution of GAG lyases that originated from alginate lyases.
RNase III inSalmonellaEnteritidis enhances bacterial virulence by reducing host immune responses
Authors: Chan Bill Kwan-wai , Li Yingxue , Ni Hongyuhang , Chan Edward Wai-chi , Deng Xin , Huang Linfeng , Chen Sheng 
Published: September 2024

ABSTRACTSalmonellais an important foodborne pathogen which comprises strains that exhibit varied virulence phenotypes and the capability of causing invasive human infection. In this study, the gene expression profile of foodborne and clinicalSalmonellastrains that exhibit high- and low-level virulence was investigated, with results showing that the expression level of a number of genes, including therncgene which encodes the RNase III ribonuclease, were exceptionally high in the high virulence strains. Investigation of the role ofrncin mediating expression of virulence phenotypes inSalmonellashowed that the product of this gene could enhance expression of the superoxide dismutase SodA, which is an essential determinant of survival fitness ofSalmonellaunder the oxidative stress elicited by the host immunity. On the other hand, we also discovered that the double-stranded RNA (dsRNA) released fromSalmonellacould trigger immune response of the host, and that the high-level expression of therncgene enabledSalmonellato evade the host immunity by reducing the amount of dsRNA accumulated in the bacterial cell. These findings provide insightful understanding of the regulation ofSalmonellavirulence and facilitate development of novel antimicrobial treatments through suppression of virulence expression and survival fitness of this important pathogen.
Developmental changes in diffusion markers of neurite vary across the hippocampus and covary with the cellular composition of hippocampal tissue
Authors: Kember J. , Gracia-Tabuenca Z. , Patel R. , Chakravarty M. , Chai X.J. 
Published: September 2024

AbstractThe hippocampus is a critical brain structure supporting memory encoding and retrieval, yet the development of its microstructure in humans remains unknown. Understanding this development may provide insight into the mechanisms underlying memory and their disruption in disease. To address this, we non-invasively estimated the density and branching complexity of neurite (dendrites, axons, glial processes) using diffusion-weighted MRI in 364 participants aged 8–21. With development, we observed large increases in neurite density and branching complexity that persisted until approximately 15 years of age before stabilizing at adult-like values. Increases in neurite density were relatively homogenous across hippocampal axes, whereas increases in branching complexity were heterogeneous; increasing primarily in CA1, SRLM, subiculum, and anterior hippocampus. To assess whether this development may be attributable to specific cell-types, we tested for spatial overlap between age-related change in neurite and the cell-type composition of hippocampal tissue via cross-reference with an out-of-sample gene-expression atlas. We found age-related changes in neurite density spatially overlapped with a granule cell component; whereas age-related changes in neurite branching complexity overlapped with a pyramidal neuron component. These results provide the first glimpse at the nonlinear maturation of hippocampal microstructure and the cell-type composition of hippocampal tissue underlying these changes.
Dynamics in thePhytophthora capsicieffector AVR3a11 confirm the core WY domain fold
Authors: Tolchard James , Chambers Vicki S. , Boutemy Laurence S. , Banfield Mark J. , Blumenschein Tharin M. A. 
Published: September 2024

AbstractOomycete pathogens cause large economic losses in agriculture through diseases such as late blight (Phytophthora infestans), and stem and root rot of soybean (Phytophthora sojae). The effector protein AVR3a, fromP. infestans, and its homologue AVR3a11 fromP. capsici, are examples of host-translocated effectors that interact with plant proteins to evade defence mechanisms and enable infection. Both proteins belong to the family of RXLR effectors and contain an N-terminal secretion signal, an RXLR motif for translocation into the host cell, and a C-terminal effector domain. Within this family, a large number of proteins have been predicted to contain one or more WY domains as their effector domain, and this domain is proposed to encompass a conserved minimal core fold containing three helices, further stabilised by additional helices or dimerization. In AVR3a11, a helical N-terminal extension to the core fold forms a four-helix bundle, as determined by X-ray crystallography. For a complete picture of the dynamics of AVR3a11, we have determined the solution structure of AVR3a11, and studied its dynamics in the fast timescale (ns-ps, from NMR relaxation parameters) and in the slow timescale (seconds to minutes, from hydrogen/deuterium exchange experiments). Hydrogen/deuterium exchange showed that the N-terminal helix is less stable than the other three helices, confirming the core fold originally proposed. Relaxation measurements confirm that AVR3a11 undergoes extensive conformational exchange, despite the uniform presence of fast motions in the spectral density function throughout most of its sequence. As functional residues are located in the more mobile regions, this flexibility in the slow/intermediate timescale may be functionally important.Lay SummaryThe effector protein AVR3a11, from the plant pathogenPhytophthora capsici, shows conformational flexibility in solution, particularly in the N-terminal helix, and in an intermediate timescale (ms-s). This confirms the core fold originally proposed and suggests that flexibility may be functionally important.
Copper resistance inLegionella pneumophila: role of genetic factors and host cells
Authors: Cameron Gillian , Faucher Sebastien P. 
Published: September 2024

AbstractCopper is frequently found in drinking water due to its presence in the natural environment and the widespread usage of copper pipes. This toxic metal has a well-known antimicrobial activity, an activity harnessed in copper-silver ionization (CSI) to eliminate the opportunistic pathogenLegionella pneumophilafrom engineered water systems. Despite utilizing the antimicrobial properties of copper inLegionellacontrol, little is known about how copper containing environments affectL. pneumophilapopulations. The goal of this study is to understand howL. pneumophilaresponds to copper within a hot water distribution system (HWDS) environment. To answer this question, different sequence types and regulatory mutants were exposed to copper to compare their survival.L. pneumophilaisolates of 4 sequence types from 3 different HWDSs exhibited a wide diversity of phenotypes after copper stress. The ΔletAand ΔletSmutants were sensitive to copper, indicating that the LetAS two component system is important for copper resistance. Additionaly, transmissive phase cultures were more resistant to copper than replicative phase cultures. Therefore, the regulation of entry into transmissive phase by the LetAS system is essential forL. pneumophila’s ability to survive copper stress. In a water system,L. pneumophilareplicates within eukaryotic hosts. When cocultured with the host ciliateTetrahymena pyriformis,L. pneumophilawas more resistant to copper than when the bacteria were in a monoculture. No difference inL. pneumophilareplication inside of hosts in cocultures with or without copper was observed. This result confirms that the presence of host cells protectsL. pneumophilafrom copper stress. Therefore, presence of host cells in water system may limit the efficacy of copper-based control strategies.
Sweet science: Exploring the impact of fructose and glucose on brown adipocyte differentiation using optical diffraction tomography
Authors: Anantha Pooja , Wu Xiangdong , Elsaid Salaheldeen , Raj Piyush , Barman Ishan , Tee Sui Seng 
Published: September 2024

ABSTRACTThe thermogenic capacity of brown adipose tissue (BAT) has garnered much attention for its potential to regulate systemic energy balance. BAT depot size and function need to be tightly to prevent loss of metabolic homeostasis due to energy dissipation via non-shivering thermogenesis. While adipocyte-intrinsic mechanisms controlling thermogenesis are critical, an increasing appreciation for the role of the BAT microenvironment is emerging. For example, changes in circulating hexoses due to dietary intake have shown to impact BAT function. Here, we show that murine BAT preadipocytes metabolism is impacted when fructose is used as the sole carbon source. Similarly differentiation medium containing only fructose yield mature adipocytes with fewer lipid droplets, with a concomitant decrease in adipogenic genes. These deficiencies are also observed in human BAT preadipocytes, where cutting-edge optical imaging modalities show a decrease in total cell mass and lipid mass in fructose-only medium. Taken together, the metabolic microenvironment significantly impacts BAT growth and function, with implications for the role of diets potentially mitigating the efficacy of BAT-targeted therapies.
Characterization and bioinformatic filtering of ambient gRNAs in single-cell CRISPR screens using CLEANSER
Authors: Liu Siyan , Hamilton Marisa C. , Cowart Thomas , Barrera Alejandro , Bounds Lexi R. , Nelson Alexander C. , Doty Richard W. , Allen Andrew S. , Crawford Gregory E. , Majoros William H. , Gersbach Charles A. 
Published: September 2024

AbstractRecent technological developments in single-cell RNA-seq CRISPR screens enable high-throughput investigation of the genome. Through transduction of a gRNA library to a cell population followed by transcriptomic profiling by scRNA-seq, it is possible to characterize the effects of thousands of genomic perturbations on global gene expression. A major source of noise in scRNA-seq CRISPR screens are ambient gRNAs, which are contaminating gRNAs that likely originate from other cells. If not properly filtered, ambient gRNAs can result in an excess of false positive gRNA assignments. Here, we utilize CRISPR barnyard assays to characterize ambient gRNA noise in single-cell CRISPR screens. We use these datasets to develop and train CLEANSER, a mixture model that identifies and filters ambient gRNA noise. This model takes advantage of the bimodal distribution between native and ambient gRNAs and includes both gRNA and cell-specific normalization parameters, correcting for confounding technical factors that affect individual gRNAs and cells. The output of CLEANSER is the probability that a gRNA-cell assignment is in the native distribution over the ambient distribution. We find that ambient gRNA filtering methods impact differential gene expression analysis outcomes and that CLEANSER outperforms alternate approaches by increasing gRNA-cell assignment accuracy.Graphical Abstract
Extracellular vesicle bioactivity and potential clinical utility is determined by mesenchymal stromal cell clonal subtype
Authors: Ioannou Savvas , Kay Alasdair G. , Stone Andrew P. , Rand Emma , Elberfeld Samuel , Bolton William , Larson Tony , Crossland Rachel E. , Kehoe Oksana , Mentlak David A. , Wang Xiao-Nong , MacDonald Chris , Genever Paul G. 
Published: September 2024

AbstractMesenchymal stromal cells (MSCs) are a promising source of therapeutic extracellular vesicles (EVs), however it is not clear how heterogeneity within a non-clonal MSC population will affect the collective EV pool. Here we used immortalised clonal MSC lines, termed Y201 and Y202, to examine how MSC phenotype influences EV character and function.Although morphologically similar, Y201 EVs were more abundant in EV biomarkers versus Y202 EVs, with an enhanced miRNA and proteomic content, predicted to contribute to an elaborate EV corona particularly abundant in RGD-containing proteins fibronectin and MFG-E8. We demonstrated that Y201 EVs, but not Y202 EVs, significantly increased the proliferation of articular chondrocytes and that the proliferative effect was mediated at least in part via an RGD (integrin)-FAK-ERK1/2 axis. Both Y201 and Y202 EV subsets significantly reduced proliferative index scores of activated T cells. However, only Y201 EVs, not Y202 EVs, suppressed disease activity compared to controls in differentin vivomodels of inflammatory peritonitis and arthritis.EVs released by closely related MSC subtypes within the same heterogeneous population differ significantly in terms of cargo abundance, bioactivity, and pre-clinicalin vivoefficacy. Analysis of defined EV subsets will aid mechanistic understanding and prioritisation for EV therapeutics.
The dark side of work-life policies: the influence of co-workers taking childcare leave on work anxiety
Authors: Osso Katia , Halinski Michael 
Published: September 2024

PurposeWhile work-life policies (WLPs) are tools that employees may draw on to better manage and balance their work and life demands, there is growing evidence that suggests the usage of WLPs may negatively impact other employees. Drawing from the theory of role dynamics and social role theory, we examine the indirect effect of co-workers taking childcare leave (CTCL) on work anxiety via work-role overload, as well as the impact of gender on this indirect relationship.Design/methodology/approachWe used Prolific Academic to recruit 236 employees to participate in a three-wave study. These data were analyzed as a hypothesized structural equation modelling (SEM) using SPSS AMOS.FindingsFindings reveal: (1) CTCL positively relates to work anxiety via work-role overload, and (2) gender moderates this indirect effect such that this positive relationship is stronger for men than women.Practical implicationsWork-life policy makers should take note of the “hidden costs” associated with work-life policy usage on other employees. Managers should work with policy users to mitigate the negative effects of policy usage on others.Originality/valueIn contrast to broader WLP research, which focuses on the benefits of policy usage on the policy user, this research shows the negative implications of work-life policies on others’ work anxiety via work-role overload. In doing so, it becomes the first study to showcase a crossover effect of CTCL on employees’ work anxiety.
The augmenting role of digital banking in reconstructing women's economic empowerment
Authors: Rohatgi Swati , Gera Navneet 
Published: September 2024

PurposeThe purpose of this study is to identify and assess the role of predictors to women’s economic empowerment (WEE). Moreover, the mediating role of digital banking usage (DBU) between financial literacy (FL) and WEE is empirically tested. The study also examines the moderation effect of educational level (EL) and employment sector (ES) on WEE.Design/methodology/approachUsing a mixed-method approach, a comprehensive questionnaire was used to collect data of 482 women working in the formal ESs of Delhi-NCR. Partial least square structural equation modeling using SmartPLS-4 was used to test the explanatory and predictive power of the proposed model. This was followed by semi-structured interviews to collect qualitative data from 14 respondents.FindingsThe results present the following important findings: first, DBU, FL, women’s agency (WA) and workplace human resource policies (HR) significantly impact WEE, whereas government support (GS) and FL significantly impact DBU; second, DBU significantly mediates the relationship between FL and WEE; and third, ES significantly moderates the relationship between DBU and WEE.Practical implicationsThis research also shares significant findings for practitioners and organizations by holistically identifying factors affecting WEE. These findings apply to both the human resource department of the employment sectors and the management of the banking sector.Originality/valueThe present study adds value to the scarce literature on the impact of DBU on WEE and highlights the mediating role of DBU along with the moderation effect of EL and ES. The study model incorporates novel constructs that impact WEE and offers new insights to various stakeholders in enhancing WEE. In addition, qualitative method was used to complement the quantitative findings.
Ethical mindset and the efficacy of tax ethical awareness initiatives
Authors: Danysh Hashemi Shannon , Daneshfar Alireza 
Published: September 2024

Purpose
This study delves into the impact of an ethical mindset on the efficacy of ethical awareness within the tax profession and aims to ascertain whether the presence of an ethical mindset can account for the discrepancies in the literature and enhance the effectiveness of ethical awareness initiatives.

Design/methodology/approach
The research used a tax experiment involving both treatment and control groups. Both groups were presented with a tax-related scenario, with the treatment group subjected to a specific ethical awareness intervention. To gauge the participants’ ethical mindsets, they were divided into strong self-interest and mild self-interest mindset groups based on their Machiavellian scores. The analysis was conducted utilizing ANOVA to scrutinize the results.

Findings
The key findings shed light on the fact that while ethical awareness endeavors can enhance the likelihood of individuals making ethical choices in tax decisions, their effectiveness varies significantly depending on the individual’s ethical mindset. Furthermore, results show that gender affected the relationship between ethical mindset and ethical awareness effectiveness, and males with mild self-interest score reacted more to the ethical awareness intervention. Results support that individuals’ ethical mindset, measured as strong self-interest and mild self-interest, is pivotal in determining the effectiveness of ethical awareness efforts.

Originality/value
This study is unique because it evaluates the effect of ethical mindsets to provide a novel way to improve tax ethical awareness initiatives.
Unlocking the creative potential of health-care employees: a serial mediation model
Authors: Azila-Gbettor Edem M. , Nutsugah Francis Fonyee , Novixoxo Jewel Dela , Glate Stanley Nelvis , Honyenuga Ben Q. 
Published: September 2024

Purpose
This study aims to investigate the mediating roles of servant leadership and employee vitality in the relationship between psychological ownership and employee creativity among healthcare workers in Ghana.

Design/methodology/approach
A sample of 736 public and private healthcare respondents was selected using a convenience sampling technique. Data collected using a self-reported questionnaire was analyzed via partial least square structural equation modeling.

Findings
The findings reveal that psychological ownership directly improves employee creativity, while servant leadership and employee vitality mediate the relationship between psychological ownership and employee creativity separately and complementarily.

Research limitations/implications
The research used self-reported data, increasing the potential for common method variance. However, sufficient care was taken to minimize these limitations.

Practical implications
This research makes valuable contributions to the field of healthcare practice literature. The findings suggest that management of health care entities should focus on creating a workplace culture that cultivates psychological ownership among employees and policies that enhance employee vitality and promote servant behavior to foster employee creativity.

Originality/value
This study represents one of the earliest attempts to examine a theoretical framework that connects servant leadership, employee vitality, employee creativity and psychological ownership within the context of the health service industry.
Exploring the potential impact of big data on the collection of sufficient, appropriate audit evidence: insights from auditors in the UAE
Authors: Huang Yiting , Ndiweni Esinath , Barghathi Yasser 
Published: September 2024

Purpose
This paper aims to understand the impact of big data on the UAE audit profession. Mainly exploring whether the emergence of big data threatens the reliability of audit standards and whether audit standards need to be improved. Also, exploring the impact of big data on the collection of audit evidence.

Design/methodology/approach
Semistructured interviews were used to collect data, mainly targeting the audit-related workers of the Big Four and Non-Big Four audit firms in the UAE. Thematic analysis is adopted to analyze the original data, and the main factors affecting the audit standard and audit evidence collection.

Findings
This study found that the reliability of audit standards and the way audit evidence is collected can be affected by big data. It concludes that audit standards need to be improved and strengthened to include detailed essential elements associated with big data to ensure audit reliability, legitimacy and regularity. The results also identify the impact of big data on audit evidence in terms of adequacy, appropriateness, authenticity, consistency and reliability, as well as the impact on the validity and completeness of evidence collection. The research highlights the importance of big data skills and knowledge education, the contribution and challenges of big data to auditing, and the use of big data in future auditing.

Originality/value
This research provides specific empirical evidence from both Big Four and Non-Big Four audit firms in the UAE, which is lacking in the literature on the use of big data technology by auditors to assist audit works in UAE. It may serve as a reference for other researchers or those interested in relevant research.