bluuebunny/crossref_metadata_2025_split huggingface dataset:
DNA looping mediates cooperative transcription activation
Authors: Han Shu-Jing , Jiang Yong-Liang , You Lin-Lin , Shen Li-Qiang , Yang Feng , Cui Ning , Kong Wen-Wen , Sun Hui , Zhou Ke , Meng Hui-Chao , Chen Zhi-Peng , Chen Yuxing , Zhang Yu , Zhou Cong-Zhao 
Published: October 2022

AbstractTranscription factors respond to multi-level stimuli and co-occupy promoter regions of target genes to activate RNA polymerase (RNAP) in a cooperative manner. To decipher the molecular mechanism, here we report two cryo-electron microscopy structures ofAnabaenatranscription activation complexes (TACs): NtcA-TAC composed of RNAP holoenzyme, promoter and a global activator NtcA, and NtcA-NtcB-TAC comprising an extra context-specific regulator NtcB. Structural analysis showed that NtcA binding makes the promoter DNA bend by ∼50º, which facilitates RNAP to contact NtcB at the distal upstream NtcB box. The sequential binding of NtcA and NtcB induces looping back of promoter DNA towards RNAP, enabling the assembly of a fully activated TAC bound with two activators. Together with biochemical assays, we propose a ‘DNA looping’ mechanism of cooperative transcription activation in bacteria.
Differential innate immune response of endometrial cells to porcine reproductive and respiratory syndrome virus type 1 versus type 2
Authors: Lothong Muttarin , Rukarcheep Dran , Wattanaphansak Suphot , Thammacharoen Sumpun , Deachapunya Chatsri , Poonyachoti Sutthasinee 
Published: October 2022

AbstractModification of cellular and immunological events due to porcine reproductive and respiratory syndrome virus (PRRSV) infection is associated with pathogenesis in lungs. PRRSV also causes female reproductive dysfunction and persistent infection which can spread to fetus, stillbirth, and offspring. In this study, alterations in cellular and innate immune responses to PRRSV type 1 or type 2 infection, including expression of PRRSV mediators, mRNA expression of toll-like receptor (TLRs) and cytokine, and cytokine secretion, were examined in primary porcine glandular endometrial cells (PGE). Cell infectivity as observed by cytopathic effect (CPE), PRRSV nucleocapsid proteins, and viral nucleic acids was early detected at two days post-infection (2 dpi) and persisted to 6 dpi. Higher percentage of CPE and PRRSV positive cells were detected in type 2 infection. PRRSV mediator proteins, CD151, CD163, sialoadhesin (Sn), integrin and vimentin, were upregulated following type 1 and type 2 infection.CD151,CD163andSnwere upregulated by type 2. Both PRRSV types upregulatedTLR1andTLR6.Only type 2 infection upregulatedTLR3,but downregulatedTLR4andTLR8. By contrast, both types upregulated TLR4 and downregulated TLR6 protein expression.Interleukin(IL)-1β,IL-6andtumor necrotic factor(TNF)-αwere upregulated by type 2, butIL-8was upregulated by type 1. Both PRRSV type 1 and 2 stimulated IL-6 but suppressed TNF-α secretion. In addition, IL-1β secretion was suppressed by type 2. These findings reveal one of the important mechanisms underlying the strategy of PRRSV on innate immune evasion in endometrium which is associated with the viral persistence.Author SummaryWidely prevalence of porcine reproductive and respiratory syndrome virus (PRRSV) remains the leading cause of huge economic losses to the global swine industry. Due to an infection of macrophages, PRRSV can persist in animals for extended periods of time associated with long-lasting reproductive disorders. Modification of cellular and immunological responses to PRRSV infection which may be related with the pathogenesis of reproductive disorders remains unclear. Herein, direct PRRSV infection of primary porcine glandular endometrial epithelial cell culture (PGE) demonstrated that PRRSV type 1 and 2 upregulated the protein expression of PRRSV mediators correlated with cell persistence of PRRSV. However, TLR and cytokines gene expression, and cytokine secretion were differentially modulated in response to PRRSV type 1 vs. type 2. Our study provides new insights into the cellular mechanism associating with PRRSV persistence in the endometrial cells and the underlying interaction of virus with the host.
Simultaneous widefield voltage and interferometric dye-free optical mapping quantifies electromechanical waves in human iPSC-cardiomyocytes
Authors: Liu Wei , Han Julie Leann , Tomek Jakub , Bub Gil , Entcheva Emilia 
Published: October 2022

ABSTRACTCoupled electro-mechanical waves define heart’s function in health and disease. Genetic abnormalities, drug-triggered or acquired pathologies can disrupt and uncouple these waves with potentially lethal consequences. Optical mapping of electrical waves using fluorescent dyes or genetically-encoded sensors in human induced pluripotent stem cell derived cardiomyocytes (iPSC-CMs) offers mechanistic insights into cardiac conduction abnormalities. Interferometric dye-free/label-free wave mapping (without specific sensors) presents an alternative, likely capturing the mechanical aspects of cardiac conduction. Because of its non-invasive nature and spectral flexibility (not restricted to a specific excitation wavelength), it is an attractive chronic imaging tool in iPSC-CMs, as part of all-optical high-throughput platforms. In this study, we developed simultaneous widefield voltage and interferometric dye-free optical imaging methodology that was used: 1) to validate dye-free optical mapping for quantification of cardiac wave properties in human iPSC-CMs; 2) to demonstrate low-cost optical mapping of electromechanical waves in hiPSC-CMs using recent near-infrared (NIR) voltage sensors and orders of magnitude cheaper miniature CMOS cameras; 3) to uncover previously underexplored frequency- and space-varying parameters of cardiac electromechanical waves in hiPSC-CMs. We find similarity in the frequency-dependent responses of electrical (NIR fluorescence imaged) and mechanical (dye-free imaged) waves, with the latter being more sensitive to faster rates and showing steeper restitution and earlier appearance of wave-front tortuosity. During regular pacing, the dye-free imaged conduction velocity and the electrical wave velocity are correlated; both modalities being sensitive to pharmacological uncoupling and both dependent on gap-junctional protein (connexins) determinants of wave propagation. We uncover strong frequency-dependence of the electromechanical delay (EMD) locally and globally in hiPSC-CMs on a rigid substrate. The presented framework and results offer new means to track the functional responses of hiPSC-CM inexpensively and non-invasively for counteracting heart disease and aiding cardiotoxicity testing and drug development.
A key GPCR phosphorylation motif discovered in arrestin2•CCR5 phosphopeptide complexes
Authors: Isaikina Polina , Petrovic Ivana , Jakob Roman P. , Sarma Parishmita , Ranjan Ashutosh , Baruah Minakshi , Panwalkar Vineet , Maier Timm , Shukla Arun K. , Grzesiek Stephan 
Published: October 2022

SummaryThe two non-visual arrestin isoforms, arrestin2 and arrestin3 recognize and bind hundreds of G protein-coupled receptors (GPCRs) with different phosphorylation patterns leading to distinct functional outcomes. The impact of phosphorylation on arrestin interactions has been well studied only for very few GPCRs. Here we have characterized the interactions between the phosphorylated CC chemokine receptor 5 (CCR5) and arrestin2. We detected several new CCR5 phosphorylation sites, which are necessary for stable complex formation with arrestin2. Crystal structures of arrestin2 in apo form and in complexes with CCR5 C-terminal phosphopeptides together with NMR spectroscopy, biochemical and functional assays revealed three phosphoresidues in a pXpp motif that are essential for the arrestin2 interactions and activation. The same phosphoresidue cluster is present in other receptors, which form stable complexes with arrestin2. We propose that the identified pXpp motif is responsible for robust arrestin2 recruitment in many GPCRs. An analysis of available sequences, structural and functional information on other GPCR•arrestin interactions suggests that a particular arrangement of phosphoresidues within the GPCR intracellular loop 3 and C-terminal tail determines arrestin2 and 3 isoform specificity. Taken together, our findings demonstrate how multi-site phosphorylation controls GPCR•arrestin interactions and provide a framework to probe the intricate details of arrestin activation and signaling.One-sentence summaryA structural and functional analysis of arrestin2 in apo form and complexes with several CCR5 phosphopeptides reveals key phosphorylation sites responsible for stable GPCR•arrestin interactions and their contributions to CCR5-arrestin2 function.
Structure of the Human ATAD2 AAA+ Histone Chaperone Reveals Mechanism of Regulation and Inter-subunit Communication
Authors: Cho Carol , Song Ji-Joon 
Published: October 2022

ABSTRACTATAD2 is a non-canonical ATP-dependent histone chaperone and a major cancer. Despite widespread efforts to design drugs targeting the ATAD2 bromodomain, little is known about the overall structural organization and AAA+ domains of ATAD2. Here, we present the 3.1 Å cryo-EM structure of human ATAD2 in the ATP state, showing a shallow hexameric spiral that binds a peptide substrate at the central pore. The spiral conformation is locked by an N-terminal linker domain (LD) that wedges between the seam subunits, thus limiting ATP-dependent symmetry breaking of the AAA+ ring. In contrast, a structure of the ATAD2-histone H3H4 complex shows the LD undocked from the seam, suggesting that H3H4 binding unlocks the AAA+ spiral by allosterically releasing the LD. These findings, together with the discovery of an inter-subunit signaling mechanism, reveal a unique regulatory mechanism for ATAD2 and lay the foundation for developing new ATAD2 inhibitors.
Perceptions and predictors of COVID-19 vaccine hesitancy among health care providers across five countries in sub-Saharan Africa
Authors: Madzorera Isabel , Abokyi Livesy Naafoe , Apraku Edward , Azemraw Temesgen , Boudo Valentin , James Christabel , Wang Dongqing , Mapendo Frank , Millogo Ourohiré , Assefa Nega , Chukwu Angela , Workneh Firehiwot , Lankoande Bruno , Hemler Elena C. , Ismail Abbas , Abubakari Sulemana , Asante Kwaku Poku , Berhane Yemane , Killewo Japhet , Oduola Ayoade , Sie Ali , Soura Abdramane , Mwanyika-Sando Mary , Vuai Said , Smith Emily , Baernighausen Till , Tajudeen Raji , Fawzi Wafaie W 
Published: October 2022

AbstractThe African continent has some of the world’s lowest COVID-19 vaccination rates. While the limited availability of vaccines is a contributing factor, COVID-19 vaccine hesitancy among health care providers (HCP) is another factor that could adversely affect efforts to control infections on the continent. We sought to understand the extent of COVID-19 vaccine hesitancy among HCP, and its contributing factors in Africa. We evaluated COVID-19 vaccine hesitancy among 1,499 HCP enrolled in a repeated cross-sectional telephone survey in Burkina Faso, Ethiopia, Nigeria, Tanzania and Ghana. We defined COVID-19 vaccine hesitancy among HCP as self-reported responses of definitely not, maybe, unsure, or undecided on whether to get the COVID-19 vaccine, compared to definitely getting the vaccine. We used Poisson regression models to evaluate factors influencing vaccine hesitancy among HCP. Approximately 65.6% were nurses and the mean age (±SD) of participants was 35.8 (±9.7) years. At least 67% of the HCP reported being vaccinated. Reasons for low COVID-19 vaccine uptake included concern about vaccine effectiveness, side effects and fear of receiving unsafe and experimental vaccines. COVID-19 vaccine hesitancy affected 45.7% of the HCP in Burkina Faso, 25.7% in Tanzania, 9.8% in Ethiopia, 9% in Ghana and 8.1% in Nigeria. Respondents reporting that COVID-19 vaccines are very effective (RR:0.21, 95% CI:0.08, 0.55), and older HCP (45 or older vs.20-29 years, RR:0.65, 95% CI: 0.44,0.95) were less likely to be vaccine-hesitant. Nurses were more likely to be vaccine-hesitant (RR 1.38, 95% CI: 1.00,1.89) compared to doctors. We found higher vaccine hesitancy among HCP in Burkina Faso and Tanzania. Information asymmetry among HCP, beliefs about vaccine effectiveness and the endorsement of vaccines by the public health institutions may be important. Efforts to address hesitancy should address information and knowledge gaps among different cadres of HCP and should be coupled with efforts to increase vaccine supply.
Bacillus velezensisSQR9 sways the rhizosphere community and its sociality toward cooperation and plant growth promotion
Authors: Liu Yan , Xu Zhihui , Xun Weibing , Stefanic Polonca , Yang Tianjie , Miao Youzhi , Zhang Nan , Zhang Ruifu , Shen Qirong , Mandic-Mulec Ines 
Published: October 2022

ABSTRACTAccording to the Lotka-Volterra competition model, intraspecific competition in plant-associated microbial communities should be stronger than interspecific competition. However, there is limited information on whether microbial communities follow this pattern and how disturbance by a newcomer affects them. Given the increasing popularity of probiotics, filling this knowledge gap could help guide future coexistence research. Here, we show that inoculation with a known probiotic,B. velezensisSQR9 shifts species co-occurrence patterns by decreasing the diversity of more distant species and promoting the growth of more closely related species, especially within theBacilluscommunity. By testing the sociality ofBacillusrhizosphere isolates, we then demonstrated that SQR9 increases the frequency of cooperative interactions in theBacilluscommunity, which may contribute to the promotion of plant growth. Finally, we provide an ecosystem framework comprising the strain’s genetic relatedness, metabolic niche space and social compatibility for the efficient and reliable assembly ofBacillusconsortia. These findings shed new light on the ecological mechanisms underlying the beneficial effects of probiotics on host fitness.
Strain and serovar variants ofSalmonella entericaexhibit diverse tolerance to food chain-related stress
Authors: Pye Hannah V. , Thilliez Gaëtan , Acton Luke , Kolenda Rafał , Al-Khanaq Haider , Grove Stephen , Kingsley Robert A. 
Published: October 2022

AbstractNon-TyphoidalSalmonella(NTS) continues to be a leading cause of foodborne illness worldwide. Food manufacturers implement hurdle technology by combining more than one approach to control food safety and quality, including preservatives such as organic acids, refrigeration, and heating. We assessed the variation in survival in stresses of genotypically diverse isolates ofSalmonella entericato identify genotypes with potential elevated risk to sub-optimal processing or cooking. Sub-lethal heat treatment, survival in desiccated conditions and growth in the presence of NaCl or organic acids were investigated.S. Gallinarum strain 287/91 was most sensitive to all stress conditions. While none of the strains replicated in a food matrix at 4°C,S. Infantis strain S1326/28 retained the greatest viability, and six strains exhibited a significantly reduced viability. AS. Kedougou strain exhibited the greatest resistance to incubation at 60°C in a food matrix that was significantly greater thanS. Typhimurium U288, S Heidelberg,S. Kentucky,S. Schwarzengrund andS. Gallinarum strains. Two isolates of monophasicS. Typhimurium, S04698-09 and B54 Col9 exhibited the greatest tolerance to desiccation that was significantly more than for theS. Kentucky andS. Typhimurium U288 strains. In general, the presence of 12mM acetic acid or 14mM citric acid resulted in a similar pattern of decreased growth in broth, but this was not observed forS. Enteritidis, andS. Typhimurium strains ST4/74 and U288 S01960-05. Acetic acid had a moderately greater effect on growth despite the lower concentration tested. A similar pattern of decreased growth was observed in the presence of 6% NaCl, with the notable exception thatS. Typhimurium strain U288 S01960-05 exhibited enhanced growth in elevated NaCl concentrations. An understanding of the molecular basis of phenotypic variation in response to stress has the potential to improve process validation during food challenge tests, improve processing, and result in more reliable risk assessments in the food industry.
Depletion Assisted Hemin Affinity (DAsHA) Proteomics Reveals an Expanded Landscape of Heme Binding Proteins
Authors: Kim Hyojung , Moore Courtney M. , Mestre-Fos Santi , Hanna David A. , Williams Loren Dean , Reddi Amit R. , Torres Matthew P. 
Published: October 2022

AbstractHemeb(iron protoporphyrin IX) plays important roles in biology as a metallocofactor and signaling molecule. However, the targets of heme signaling and the network of proteins that mediate the exchange of heme from sites of synthesis or uptake to heme dependent or regulated proteins are poorly understood. Herein, we describe a quantitative mass spectrometry-based chemoproteomics strategy to identify exchange labile hemoproteins in human embryonic kidney HEK293 cells that may be relevant to heme signaling and trafficking. The strategy involves depleting endogenous heme with the heme biosynthetic inhibitor succinylacetone (SA), leaving putative heme binding proteins in theirapo-state, followed by the capture of those proteins using hemin-agarose resin and finally elution and identification by mass spectrometry. By identifying only those proteins that interact with high specificity to hemin-agarose relative to control beaded agarose in a SA-dependent manner, we have expanded the number of proteins and ontologies that may be involved in binding and buffering labile heme or are targets of heme signaling. Notably, these include proteins involved in chromatin remodeling, DNA damage response, RNA splicing, cytoskeletal organization and vesicular trafficking, many of which have been associated with heme through complimentary studies published recently. Taken together, these results provide support for the emerging role for heme in an expanded set of cellular processes from genome integrity to protein trafficking and beyond.
Therapeutic trial of anle138b in mouse models of genetic prion disease
Authors: Vallabh Sonia M , Zou Dan , Pitstick Rose , O’Moore Jill , Peters Janet , Silvius Derek , Kriz Jasna , Jackson Walker S , Carlson George A , Minikel Eric Vallabh , Cabin Deborah E 
Published: October 2022

AbstractPhenotypic screening has yielded small molecule inhibitors of prion replication that are effective in vivo against certain prion strains but not others. Here we sought to test the small molecule anle138b in multiple mouse models of prion disease. In mice inoculated with the RML strain of prions, anle138b doubled survival and durably suppressed astrogliosis measured by live animal bioluminescence imaging. In knock-in mouse models of the D178N and E200K mutations that cause genetic prion disease, however, we were unable to identify a clear, quantifiable disease endpoint against which to measure therapeutic efficacy. Among untreated animals, the mutations did not impact overall survival, and bioluminescence remained low out to >20 months of age. Vacuolization and PrP deposition were observed in some brain regions in a subset of mutant animals, but appeared unable to carry the weight of a primary endpoint in a therapeutic study. We conclude that not all animal models of prion disease are suited to well-powered therapeutic efficacy studies, and care should be taken in choosing the models that will support drug development programs.
Differential roles of beta-cell IP3R and RyR ER Ca2+channels in ER stress-induced alterations of beta-cell Ca2+homeostasis
Authors: Zhang Irina X. , Hermann Andrea , Leon Juan , Arunagiri Anoop , Arvan Peter , Satin Leslie S. 
Published: October 2022

AbstractPancreatic beta cells maintain glucose homeostasis by secreting pulses of insulin in response to a rise in glucose. Pulsatile secretion occurs due to glucose-induced oscillations in beta-cell cytosolic Ca2+. The endoplasmic reticulum (ER) helps regulate beta-cell cytosolic Ca2+, and ER stress can lead to ER Ca2+depletion, beta-cell dysfunction and an increased risk of type 2 diabetes. To determine the effects of tunicamycin-induced ER stress on ER inositol 1,4,5-triphosphate receptors (IP3Rs) and ryanodine receptors (RyRs) and their involvement in subsequent Ca2+dysregulation, INS-1 832/13 cells and primary mouse islets were treated with tunicamycin. This increased RyR1 mRNA and potentiated RyR-mediated Ca2+signaling without affecting RyR2 mRNA. TM treatment also enhanced IP3R function, while it decreased IP3R1 and IP3R3 mRNA. Stress reduced ER Ca2+, triggered oscillations in cytosolic Ca2+under subthreshold glucose conditions, and increased apoptosis; these changes were prevented by cotreatment with the RyR1 inhibitor dantrolene. In contrast, inhibiting IP3Rs with xestospongin-C failed to suppress the cytosolic Ca2+oscillations due to tunicamycin treatment and did not protect beta cells from tunicamycin-induced apoptosis, although xestospongin-C inclusion prevented ER Ca2+depletion. Taken together, changes in RyR1 function were shown to play a critical role in ER stress induced Ca2+dysfunction and beta-cell apoptosis.
Brand love and brand addiction and their effects on consumers' negative behaviors
Authors: Junaid Muhammad , Fetscherin Marc , Hussain Khalid , Hou Fujun 
Published: October 2022

Purpose
This study aims to investigate the relationship between brand love and brand addiction and their effects on consumers' negative behaviors with respect to excessive spending, trash-talking and the feeling of anxiety.

Design/methodology/approach
A sample of 352 young fashion brand consumers responded to a structured questionnaire. The resulting data were analyzed with structural equation modeling in MPlus.

Findings
While brand love and brand addiction are related concepts, their effects on negative consumer behaviors differ. In the presence of brand addiction as a mediator of brand love, brand addiction has a significant effect on the three negative behaviors, and the authors observe a suppression effect of brand love on the outcome variables, with total effects (direct and indirect) being insignificant.

Research limitations/implications
The main limitation of this study was its single-country cross-sectional convenience sample.

Practical implications
While brand addiction could aid brands by leading consumers to spend excessively on them and trash-talk rival brands, it may also lead to increased consumer anxiety.

Originality/value
To the best of the authors’ knowledge, this is the first study to empirically assess the relationship between brand love and brand addiction and their effects on three distinctive negative consumer behaviors. This shows that brand love is an important antecedent of brand addiction.