bluuebunny/crossref_metadata_2025 huggingface dataset:
Drivers of inappropriate use of antimicrobials in South Asia: A systematic review of qualitative literature
Authors: Murray Jennifer L. , Leung Daniel T. , Hanson Olivia R. , Ahmed Sharia M. , Pavia Andrew T. , Khan Ashraful I. , Szymczak Julia E. , Vaughn Valerie M. , Patel Payal K. , Biswas Debashish , Watt Melissa H.
Published: September 2023

Antimicrobial resistance is a global public health crisis. Effective antimicrobial stewardship requires an understanding of the factors and context that contribute to inappropriate use of antimicrobials. The goal of this qualitative systematic review was to synthesize themes across levels of the social ecological framework that drive inappropriate use of antimicrobials in South Asia. In September 2023, we conducted a systematic search using the electronic databases PubMed and Embase. Search terms, identifieda priori, were related to research methods, topic, and geographic location. We identified 165 articles from the initial search and 8 upon reference review (n=173); after removing duplicates and preprints (n=12) and excluding those that did not meet eligibility criteria (n=115), 46 articles were included in the review. We assessed methodological quality using the qualitative Critical Appraisal Skills Program checklist. The studies represented 6 countries in South Asia, and included data from patients, health care providers, community members, and policy makers. For each manuscript, we wrote a summary memo to extract the factors that impede antimicrobial stewardship. We coded memos using NVivo software; codes were organized by levels of the social ecological framework. Barriers were identified at multiple levels including the patient (self-treatment with antimicrobials; perceived value of antimicrobials), the provider (antimicrobials as a universal therapy; gaps in knowledge and skills; financial or reputational incentives), the clinical setting (lack of resources; poor regulation of the facility), the community (access to formal health care; informal drug vendors; social norms), and policy (absence of a regulatory framework; poor implementation of existing policies). The findings highlight the importance of working across multiple sectors to design and implement approaches to antimicrobial stewardship in South Asia.
Regulation of Toxic RNA Foci and MutantDMPKTranscripts: Role of MBNL Proteins and RNA Decay Pathways
Authors: Xing Xiaomeng , Markus Robert , Ghosh Tushar , Buxton Sarah , Nieves Daniel J. , Wojciechowska Marzena , Brook J. David
Published: September 2023

Myotonic dystrophy type 1 (DM1) is a progressive, multisystemic disorder caused by an expansion of CTG repeats in the 3’ untranslated region of theDMPKgene. When transcribed the mutant RNAs accumulate in affected tissues appearing as distinct foci when visualised byin situhybridisation. The RNA foci are aggregates of CUG repeat-containing RNAs that sequester RNA-binding proteins, particularly muscleblind-like (MBNL) proteins, leading to their dysfunction and causing downstream molecular and cellular defects. Here we show the double knock-out ofMBNL1and2prevents RNA foci formation and nuclear retention of mutantDMPKmRNA in DM1 cells as well as promoting their degradation and nuclear export. Using stochastic optical reconstruction microscopy (STORM), we find the presence of both large foci and micro foci in DM1 cells. Large foci consist of multiple DMPK transcripts, while many micro foci are (CUG)n fragments. The absence of MBNL proteins not only prevents the aggregation of multipleDMPKtranscripts into large foci, but also promotes their degradation and nuclear processing. However, although a substantial amount of MBNL1 proteins are bound to the mutant transcripts, the pools of free MBNL1 proteins are similar in DM1 nuclei to those in controls. Furthermore, we have identified several factors that are involved in the control of mutantDMPKmRNA turnover, including XRN2, EXOSC10, UPF1 and STAU1. Our study indicates that these factors are implicated in the RNA foci accumulation and the degradation of mutantDMPKmRNA. UPF1 and STAU1 may have additional roles beyond degradation, impacting the nuclear processing of mutantDMPKmRNA. Our study also highlights the critical role of MBNL proteins in regulating mutantDMPKmRNA metabolism: the absence of MBNLs in DM1 appears to expedite the processing of mutantDMPKmRNA mediated by these RNA decay factors.

Significance statement

Our investigations uncovered valuable data on the RNA foci dynamics in DM1, revealing the intricate mechanisms that underlie their formation, stability, and turnover. Our findings also contributed to delineate the complex pathways involved in the transportation and degradation of the mutant mRNA and provided insights into the critical role played by MBNL proteins in these processes. Studying the degradation mechanism of mutantDMPKmRNA in myotonic dystrophy may provide a foundation for comprehending the mechanisms of RNA degradation in other diseases caused by short tandem repeat (STR) mutations, such as Huntington’s disease, Fragile X syndrome, and several types of ataxia. Additionally, the use of cutting-edge STORM technology can provide a valuable tool for investigating RNA foci in other STR expansion disorders.
A taxonomy for human social perception: Data-driven modelling with cinematic stimuli
Authors: Santavirta Severi , Malén Tuulia , Erdemli Asli , Nummenmaa Lauri
Published: September 2023

Every day, humans encounter complex social situations that need to be encoded effectively to allow interaction with others. Yet, principles for organizing the perception of social features from the external world remain poorly characterised. In this large-scale study we investigated the principles of social perception in dynamic scenes. In the primary dataset, we presented 234 movie clips (41 minutes) containing various social situations to 1140 participants and asked them to evaluate the presence of 138 social features in each clip. Analyses of the social feature ratings revealed that some features are perceived categorically (present or absent) and others continuously (intensity) and simple social features requiring immediate response are perceived most consistently across participants. To establish the low-dimensional perceptual organization for social features based on movies, we used principal coordinate analysis and consensus clustering for the feature ratings. These dimension reduction analyses revealed that the social perceptual structure can be modelled with eight main dimensions and that behaviourally relevant perceptual categories emerge from these main dimensions. This social perceptual structure generalized from the perception of unrelated Hollywood movie clips to the perception of a full Finnish movie (70 min), and to the perception of static images (n = 468) and across three independent sets of participants (n = 2254). Based on the results, we proposeeight basic dimensions of social perceptionas a model for rapid social perception where social situations are perceived along eight orthogonal perceptual dimensions (most importantly emotional valence, empathy versus dominance, and cognitive versus physical behaviour).
Open top multi sample dual view light sheet microscope for live imaging of large multicellular systems
Authors: Moos F. , Suppinger S. , de Medeiros G. , Oost K. , Boni A. , Rémy C. , Weevers S. , Tsiairis C. , Strnad P. , Liberali P.
Published: September 2023

Multicellular systems grow over the course of weeks from single cells to tissues or even full organisms composed of several thousands of cells, making live imaging of such samples challenging. To bridge these wide spatiotemporal scales, we present an open top dual view and dual illumination light sheet microscope dedicated for live imaging of large specimens at single cell resolution. The novel configuration of objectives together with a flexible and customizable multi-well mounting system combines dual view with high throughput multi-position imaging. We use the newly developed microscope to image a wide variety of samples and highlight its capabilities to gain quantitative single-cell information in large specimens such as mature intestinal organoids and gastruloids.
Differential utilisation of subcellular skeletal muscle glycogen pools: A comparative analysis between 1 and 15 minutes of maximal exercise
Authors: Schytz Camilla Tvede , Ørtenblad Niels , Gejl Kasper Degn , Nielsen Joachim
Published: September 2023

Distinct subcellular pools of glycogen particles exist within skeletal muscle fibres, distributed both within and between myofibrils and can be found in proximity to, or at a distance from mitochondria. Their precise localisation may influence their degradation rate and role in muscle function. Here, we investigated how exercise at different intensities (1- and 15-min maximal exercise) with known variations in glycogenolytic rate and relative contribution from anaerobic metabolism affects the utilisation of the distinct pools. Further, we investigated how lowered carbohydrate and energy intake affected glycogen volume densities and the storage of glycogen particles (i.e., localisation, size, and number) and their subsequent utilisation during the exercises. Using a randomized, counterbalanced, cross-over design, participants performed two maximal cycle tests of either 1 (n=10) or 15 min (n=10), conducted following consumption of two distinct diets with either high or lowered carbohydrate and energy contents. Muscle biopsies fromm. vastus lateraliswere obtained before and after the exercises. Intermyofibrillar glycogen was preferentially utilised during the 1-min exercise, whereas intramyofibrillar glycogen was preferentially utilised during the 15-min exercise. The lowered carbohydrate and energy intake decreased the particle size across all subcellular pools and reduced the numerical density in the intramyofibrillar and subsarcolemmal pools, with no effects on the glycogen utilisation during the subsequent exercise. In conclusion, the distinct subcellular glycogen pools were differentially utilised during 1-min and 15-min maximal exercise. Additionally, lowered carbohydrate and energy consumption reduces particle size and numerical density, depending on subcellular localisation.
Contributions of hyperactive mutations in Mprofrom SARS-CoV-2 to drug resistance
Authors: Flynn Julia M. , Zvornicanin Sarah N. , Shaqra Ala M. , Yilmaz Nese Kurt , Moquin Stephanie , Dovala Dustin , Schiffer Celia S. , Bolon Daniel N.A.
Published: September 2023

The appearance and spread of mutations that cause drug resistance in rapidly evolving diseases, including infections by SARS-CoV-2 virus, are major concerns for human health. Many drugs target enzymes, and resistance-conferring mutations impact inhibitor binding and/or enzyme activity. Nirmatrelvir, the most widely used inhibitor currently used to treat SARS-CoV-2 infections, targets the main protease (Mpro) preventing it from processing the viral polyprotein into active subunits. Our previous work systematically analyzed resistance mutations in Mprothat reduce binding to inhibitors; here we investigate mutations that affect enzyme function. Hyperactive mutations that increase Mproactivity can contribute to drug resistance but had not been thoroughly studied. To explore how hyperactive mutations contribute to resistance, we comprehensively assessed how all possible individual mutations in Mproaffect enzyme function using a mutational scanning approach with a FRET-based yeast readout. We identified hundreds of mutations that significantly increased Mproactivity. Hyperactive mutations occurred both proximal and distal to the active site, consistent with protein stability and/or dynamics impacting activity. Hyperactive mutations were observed three times more than mutations which reduced apparent binding to nirmatrelvir in recent studies of laboratory grown viruses selected for drug resistance. Hyperactive mutations were also about three times more prevalent than nirmatrelvir-binding mutations in sequenced isolates from circulating SARS-CoV-2. Our findings indicate that hyperactive mutations are likely to contribute to the natural evolution of drug resistance in Mproand provide a comprehensive list for future surveillance efforts.
A Small Multidrug Resistance Transporter inPseudomonas aeruginosaConfers Substrate-Specific Resistance or Susceptibility
Authors: Wegrzynowicz Andrea K. , Heelan William J. , Demas Sydnye P. , McLean Maxwell S. , Peters Jason M. , Henzler-Wildman Katherine A.
Published: September 2023

Small Multidrug Resistance (SMR) transporters are key players in the defense of multidrug-resistant pathogens to toxins and other homeostasis-perturbing compounds. However, recent evidence demonstrates that EmrE, an SMR fromEscherichia coliand a model for understanding transport, can also induce susceptibility to some compounds by drug-gated proton leak. This runs down the ΔpH component of the Proton Motive Force (PMF), reducing viability of the affected bacteria. Proton leak may provide an unexplored drug target distinct from the targets of most known antibiotics. Activating proton leak requires an SMR to be merely present, rather than be the primary resistance mechanism, and dissipates the energy source for many other efflux pumps. PAsmr, an EmrE homolog fromP. aeruginosa, transports many EmrE substrates in cells and purified systems. We hypothesized that PAsmr, like EmrE, may confer susceptibility to some compounds via drug-gated proton leak. Growth assays ofE. coliexpressing PAsmr displayed substrate-dependent resistance and susceptibility phenotypes, andin vitrosolid-supported membrane electrophysiology experiments revealed that PAsmr performs both antiport and substrate-gated proton uniport, demonstrating the same functional promiscuity observed in EmrE. Growth assays ofP. aeruginosastrain PA14 demonstrated that PAsmr contributes resistance to some antimicrobial compounds, but no growth defect is observed with susceptibility substrates, suggestingP. aeruginosacan compensate for the proton leak occurring through PAsmr. These phenotypic differences betweenP. aeruginosaandE. coliadvance our understanding of underlying resistance mechanisms inP. aeruginosaand prompt further investigation into the role that SMRs play in antibiotic resistance in pathogens.

IMPORTANCE

Small multidrug resistance transporters are a class of efflux pumps found in many pathogens, but whose contributions to antibiotic resistance are not fully understood. We hypothesize that these transporters may confer not only resistance, but also susceptibility, by dissipating the proton-motive force. This means to use an SMR transporter as a target, it merely needs to be present (as opposed to being the primary resistance mechanism). Here, we test this hypothesis with an SMR transporter found inPseudomonas aeruginosaand find that it can perform both antiport (conferring resistance) and substrate-gated proton leak. Proton leak is detrimental to growth inE. colibut notP. aeruginosa, suggesting thatP. aeruginosaresponds differently to or can altogether prevent ΔpH dissipation.
High density aromatic peptides protect retinal ganglion cells in mouse retina following optic nerve crush injury
Authors: MacNeil Margaret A. , Garcia-Marin Virginia , Birk Alexander
Published: September 2023

Purpose

Mitochondrial dysfunction plays a significant role in the death of retinal ganglion cells (RGCs) and their axons in retinal neurodegenerative diseases. We studied the effect of the mitochondrial-targeted HDAP2 (High Density Aromatic Peptide 2) to protect MDBK cells following serum starvation in cultured cells, and in mouse retinal ganglion cells following optic nerve crush.

Methods

Uptake, toxicity, and efficacy of HDAP2 was studied in cultured cells under conditions of serum starvation. Once preliminary safety and efficacy of the peptide was establishedin vitro, HDAP2 was administered to mice following unilateral optic nerve crush and the distribution and survival of RBPMS-labeled RGCs was assessed.

Results

We found that water-soluble HDAP2 localizes to mitochondria and improves the mitochondrial membrane potential, prevents oxidative stress, and reduces death of cells grown in serum-free culture media. In mice treated systemically with biotinylated peptide, we found that HDAP2 labeled all layers of the retina in a pattern similar to the mitochondrial marker Cox IV, and that the peptide was colocalized with retinal ganglion cells and Müller glia. Animals treated with HDAP2 following optic nerve crush showed significantly greater survival of retinal ganglion cells compared to untreated animals at all retinal eccentricities.

Conclusion

Our results demonstrate that HDAP2 protects cells against serum starvation-induced decreases in mitochondrial membrane potential and leads to significant protection of retinal ganglion cells in the mouse retina following optic nerve crushin vivo.These results suggest that therapeutic agents preserving the mitochondrial membrane potential may help protect neurons following trauma or from neurodegenerative disease.
mRNA N1-2’O-methylation by CMTR1 affects NVL2 mRNA splicing
Authors: Wolter Steven , Hennig Thomas , Wallerath Christina , Schlee-Guimaraes Thais M. , Kirchhoff Alexander , Urban Christian , Piras Antonio , Stukalov Alexey , Juranek Stefan , Engelke Michael , Boehm Volker , Gehring Niels H. , Hartmann Gunther , Pichlmair Andreas , Friedel Caroline C. , Dölken Lars , Schlee Martin
Published: September 2023

Cap0-mRNA is characterized by a 5’-5’triphosphate-linked N7-methylated guanosine(m7G). In higher eukaryotes, the methyltransferase CMTR1 additionally methylates the 2’O-position of the penultimate mRNA nucleotide(N1) ribose (cap1-mRNA). While the m7G cap is essential for mRNA export and translation initiation by the eIF4F complex, the N1-2’O-methylation prevents recognition of cap1-mRNA by the antiviral RNA receptors RIG-I and IFIT1, but a function beyond immunotolerance remained elusive. Here, we generatedCMTR1-knockout(CMTR1-/-) cells and found that type-I-interferon(IFN-I) treatment resulted in IFIT1-mediated reduction of cell viability and broad mRNA translation. Consequently, stimulation of the antiviral receptor RIG-I inCMTR1-/-cells revealed an IFIT1 dependent dramatic reduction of IFN-I and chemokine protein induction, demonstrating the importance of N1-2’O-methylation for antiviral responses.

Additionally, IFN-I- and IFIT1-independent effects were observed:CMTR1-/-cells were smaller, divided slower, and exhibited a reduced transcription of mRNAs coding ribosomal proteins (RP), 5’TOP-RNA and snoRNA host genes(SNHG). Additionally, proteome and transcriptome analysis revealed that expression of NVL2, an essential factor in ribosome biogenesis, is strongly suppressed by an alternative-splicing event ofNVL2 mRNAin CMTR1-/-cells. This reduction could only be rescued by catalytically active CMTR1. Altogether, besides antiviral immunity N1-2’O-methylation by CMTR1 has broad effects on cellular physiology and controls splicing of NVL2.
Statistics to prioritize rare variants in family-based sequencing studies with disease subtypes
Authors: Nieuwoudt Christina , Farooq Fabiha Binte , Brooks-Wilson Angela , Bureau Alexandre , Graham Jinko
Published: September 2023

Family-based sequencing studies are increasingly used to find rare genetic variants of high risk for disease traits with familial clustering. In some studies, families with multiple disease subtypes are collected and the exomes of affected relatives are sequenced for shared rare variants. Since different families can harbor different causal variants and each family harbors many rare variants, tests to detect causal variants can have low power in this study design. Our goal is rather to prioritize shared variants for further investigation by, e.g., pathway analyses or functional studies. The transmission-disequilibrium test prioritizes variants based on departures from Mendelian transmission in parent-child trios. Extending this idea to families, we propose methods to prioritize rare variants shared in affected relatives with two disease subtypes, with one subtype more heritable than the other. Global approaches condition on a variant being observed in the study and assume a known probability of carrying a causal variant. In contrast, local approaches condition on a variant being observed in specific families to eliminate the carrier probability. Our simulation results indicate that global approaches are robust to mis-specification of the carrier probability and prioritize more effectively than local approaches even when the carrier probability is misspecified.
Standard methods for marking caudate amphibians do not impair animal welfare over the short term: an experimental approach
Authors: Daversa DR , Baxter E , Rosa GM , Sergeant C , Garner TWJ
Published: September 2023

Major advancements in ecology and biodiversity conservation have been made thanks to methods for marking and individually tracking animals. Marking animals is both widely used and controversial due to the potential consequences to animal welfare, which are often incompletely evaluated before implementation. Two outstanding knowledge gaps concerning the welfare consequences of individual marking concerns their short-term behavioural impacts and the relative impacts from marking versus the handling of animals while carrying out procedures. We addressed these knowledge gaps through an experimental study of alpine newts (Ichthyosaura alpestris) in which we varied handling and marking procedures. Examining individual responses to handling, toe-clipping and visible implant elastomer (VIE) injection over 21 days showed that handling and marking elicited increased newt activity and hesitancy to feed compared to animals that did not get handled or marked. These effects were apparent even when animals were handled only (not marked), and marking did not further increase the magnitude of responses. Increases in newt activity and feeding hesitancy were transient; they were not observed in the weeks following handling and marking. Whereas previous studies emphasize the welfare impacts of marking procedures themselves, these findings highlight that handling alone can elicit behavioural changes with possible costs to welfare. Yet, the transient nature of behavioural responses observed here suggests that immediate costs of handling may be subsequently compensated for.
Resource conflicts leading to moral distress: A longitudinal study among physicians in Norway
Authors: Miljeteig Ingrid , Førde Reidun , Rø Karin Isaksson , Bååthe Fredrik , Bringedal Berit
Published: September 2023

Background

The scarcity of resources represents ethical challenges and involves moral distress for health professionals. There are no longitudinal studies of moral distress among representative samples of physicians.

Method

Surveys of the Norwegian Physician Panel (NPP) compared the extent of moral distress in 2004 and 2021. Descriptive statistics and regression analysis were used in the study.

Results

Response rates were 67% (1004/1499) in 2004 and 70% (1639/2316) in 2021. That patient care is deprived due to time constraints is the most severe dimension of moral distress among physicians, and it has increased comparing 2021 with 2004 (68.3% in 2004 to 75.1% in 2021 reported “somewhat” or “very morally distressing”). Moral distress also increased concerning patients who “cry the loudest” get better and faster treatment than others. Moral distress was reduced on statements about long waiting times, treatment not provided due to economic limitations, deprioritisation of older patients, and acting against one’s conscience. Women reported higher moral distress than men in both years, and there were significant gender differences for six statements in 2021 and one in 2004. Though not consistently, the physicians’ age and workplace influenced the reported moral distress.

Conclusion

In both years, moral distress among physicians related to scarcity of or unfair distribution of resources was high. Moral distress associated with resource scarcity and acting against one’s conscience decreased, which might indicate improvements in the healthcare system. On the other hand, it might suggest that physicians have reduced their ideals or expectations or are morally fatigued.
PROxAb Shuttle: A non-covalent plug-and-play platform for the rapid generation of tumor-targeting antibody-PROTAC conjugates
Authors: Schneider Hendrik , Jäger Sebastian , Könning Doreen , Rasche Nicolas , Schröter Christian , Elter Desislava , Evers Andreas , Lecomte Marc , Sirtori Federico Riccardi , Schwarz Daniel , Wegener Ansgar , Hartung Ingo , Rieker Marcel
Published: September 2023

Proteolysis-targeting chimeras (PROTACs) have evolved in recent years from an academic idea to a therapeutic modality with more than 25 active clinical programs. However, achieving oral bioavailability and cell-type specificity remains a challenge, especially for PROTACs recruiting the von Hippel-Lindau (VHL) E3 ligase. Herein, we present an unprecedented, plug- and-play platform for VHL-recruiting PROTACs to overcome these limitations. Our platform allows for the generation of non-covalent antibody-PROTAC complexes within minutes and obviates the need for prior PROTAC modification, antibody-drug linker chemistry optimization or bioconjugation. Our technology relies on camelid-derived antibody domains (VHHs) which can easily be engineered into existing therapeutic antibody scaffolds. The resulting targeted, bispecific fusion proteins can be complexed with PROTACs at defined PROTAC-to-antibody ratios and have been termed PROxAb Shuttles. PROxAb Shuttles can prolong the half-life of PROTACs from hours to days, demonstrate anti-tumor efficacyin vivoand have the potential for reloadingin vivoto further boost efficacy.
Voluntary wheel running mitigates disease in an Orai1 gain-of-function mouse model of tubular aggregate myopathy
Authors: O’Connor Thomas N. , Zhao Nan , Orciuoli Haley M. , Brasile Alice , Pietrangelo Laura , He Miao , Groom Linda , Leigh Jennifer , Mahamed Zahra , Liang Chen , Malik Sundeep , Protasi Feliciano , Dirksen Robert T.
Published: September 2023

Tubular aggregate myopathy (TAM) is an inherited skeletal muscle disease associated with progressive muscle weakness, cramps, and myalgia. Tubular aggregates (TAs) are regular arrays of highly ordered and densely packed SR straight-tubes in muscle biopsies; the extensive presence of TAs represent a key histopathological hallmark of this disease in TAM patients. TAM is caused by gain-of-function mutations in proteins that coordinate store-operated Ca2+entry (SOCE): STIM1 Ca2+sensor proteins in the sarcoplasmic reticulum (SR) and Ca2+-permeable ORAI1 channels in the surface membrane. We have previously shown that voluntary wheel running (VWR) prevents formation of TAs in aging mice. Here, we assessed the therapeutic potential of endurance exercise (in the form of VWR) in mitigating the functional and structural alterations in a knock-in mouse model of TAM (Orai1G100S/+or GS mice) based on a gain-of-function mutation in the ORAI1 pore. WT and GS mice were singly-housed for six months (from two to eight months of age) with either free-spinning or locked low profile wheels. Six months of VWR exercise significantly increased soleus peak tetanic specific force production, normalized FDB fiber Ca2+store content, and markedly reduced TAs in EDL muscle from GS mice. Six months of VWR exercise normalized the expression of mitochondrial proteins found to be altered in soleus muscle of sedentary GS mice in conjunction with a signature of increased protein translation and biosynthetic processes. Parallel proteomic analyses of EDL muscles from sedentary WT and GS mice revealed changes in a tight network of pathways involved in formation of supramolecular complexes, which were also normalized following six months of VWR. In summary, sustained voluntary endurance exercise improved slow twitch muscle function, reduced the presence of TAs in fast twitch muscle, and normalized the muscle proteome of GS mice consistent with protective adaptions in proteostasis, mitochondrial structure/function, and formation of supramolecular complexes.
Genetic diversity of domestic cat hepadnavirus in Taiwan
Authors: Ilagan Silva Benji Brayan , Chen Jin-Yang , Avanceña Villanueva Brian Harvey , Lu Zi-Ying , Zhen Hsing-Hua , Montecillo Andrew D. , Shofa Maya , Minh Hoang , Chuang Jen-Pin , Huang Huai-Ying , Saito Akatsuki , Chuang Kuo-Pin
Published: September 2023

Domestic cat hepadnavirus (DCH) is an infectious disease associated with chronic hepatitis in cats, suggesting a similarity with hepatitis B virus infection in humans. Since its first identification in Australia in 2018, DCH has been reported in several countries with varying prevalence rates, but its prevalence in Taiwan has not yet been investigated. Here, we aimed to identify the presence and prevalence of DCH infections in Taiwan. Among 71 samples tested, eight (11.27%) were positive for DCH. Of these positive cases, three cats had elevated levels of alanine transaminase (ALT) and aspartate transaminase (AST), suggesting an association between DCH infection and chronic hepatitis. Four DCH-positive samples were also tested for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) co-infection, one (25%) was positive for FIV while none for FeLV (0%). In addition, we performed whole genome sequencing of six samples to determine the viral genome sequences. Phylogenetic analyses identified a distinct lineage compared with previously reported sequences. Considering the recent findings suggesting the potential risk of DCH for interspecies or zoonotic transmission, this study suggests the importance of continuous surveillance of DCH and further research to elucidate the pathophysiology and transmission route of DCH.
Temperature elevation synergises with and enhances the type-I IFN-mediated restriction of MPXV
Authors: Davis Chris , Epifano Ilaria , Dee Kieran , McFarlane Steven , Wojtus Joanna K. , Brennan Benjamin , Gu Quan , Sreenu Vattipally B. , Nomikou Kyriaki , Tong Lily , Orr Lauren , Filipe Ana Da Silva , Barr David A. , Ho Antonia , Thomson Emma C. , Boutell Chris
Published: September 2023

Fever is an evolutionary conserved host pro-inflammatory immune response that governs the regulation of multiple biological processes to control the outcome of infection. In January 2022, the World Health Organization (WHO) reported a global outbreak in mpox cases with a high incidence of human-to-human transmission. A frequent prodromal symptom of monkeypox virus (MPXV) infection is fever, with a febrile temperature range of 38.3 to 40.5 °C. However, the outcome of temperature elevation on MPXV infection remains poorly defined. Here, we isolated a circulating strain of MPXV from a patient who presented with fever (38.5 °C) and rash from the 2022 outbreak. Genomic sequencing identified this isolate to belong to the epidemic Clade IIb.B1. Transcriptomic analysis of infected cells demonstrated this virus to induce a strong IL6 pro-inflammatory immune response, consistent with a role for this pyrogen in the regulation of fever. We identify host-cell temperature at both physiological skin (33 °C) and clinical febrile temperatures (38.5 and 40 °C) to be a key determinant in the outcome of infection through the differential regulation of MPXV transcription and associated amplitude of host cytokine response to infection. Incubation of infected cells at 38.5 or 40 °C led to a restriction or ablation in MPXV replication, respectively. Importantly, this thermal inhibition was reversible upon temperature downshift to 37 °C without detriment to viral replication fitness. Co-stimulation of the type-I interferon (IFN) response led to a dose- and temperature-dependent inhibition in MPXV replication that restricted the re-establishment of infection upon temperature downshift and withdrawal of IFN as an immune stimulus. Our data identify febrile temperatures associated with mpox disease to be a critical component of the host pro-inflammatory immune response to infection which can synergise with the type-I IFN response to enhance the host-cell mediated restriction of MPXV.
Reversible conjugation of a CBASS nucleotide cyclase regulates immune response to phage infection
Authors: Krüger Larissa , Gaskell-Mew Laura , Graham Shirley , Shirran Sally , Hertel Robert , White Malcolm F
Published: September 2023

Antiviral defence systems build the prokaryotic immune system and their proper regulation is vital for survival and fitness. While it is important that they are readily available in case of infection, they need to be tightly controlled to prevent activation of an unnecessary cellular response. Here we describe how the bacterial cyclic oligonucleotide-based antiphage signalling system (CBASS) uses its intrinsic protein modification system to regulate the nucleotide cyclase. By integrating a Type II CBASS system fromBacillus cereusinto the cognate hostBacillus subtilis, we show that the Cap2 protein conjugates the cyclase exclusively to the conserved phage shock protein A (PspA) in the absence of phage. This cyclase-PspA conjugation is reversed after infection by the isopeptidase Cap3. Finally, we propose a model in which the cyclase is held in an inactive state by conjugation to PspA in the absence of phage and that this conjugation is released upon infection, priming the cyclase for activation.
Discovery of First-in-Class PROTAC Degraders of SARS-CoV-2 Main Protease
Authors: Alugubelli Yugendar R. , Xiao Jing , Khatua Kaustav , Kumar Sathish , Ma Yuying , Ma Xinyu R. , Vulupala Veerabhadra R. , Atla Sandeep R. , Blankenship Lauren , Coleman Demonta , Neuman Benjamin W. , Liu Wenshe Ray , Xu Shiqing
Published: September 2023

We have witnessed three coronavirus (CoV) outbreaks in the past two decades, including the COVID-19 pandemic caused by SARS-CoV-2. Main protease (MPro) is a highly conserved and essential protease that plays key roles in viral replication and pathogenesis among various CoVs, representing one of the most attractive drug targets for antiviral drug development. Traditional antiviral drug development strategies focus on the pursuit of high-affinity binding inhibitors against MPro. However, this approach often suffers from issues such as toxicity, drug resistance, and a lack of broad-spectrum efficacy. Targeted protein degradation represents a promising strategy for developing next-generation antiviral drugs to combat infectious diseases. Here we leverage the proteolysis targeting chimera (PROTAC) technology to develop a new class of small-molecule antivirals that induce the degradation of SARS-CoV-2 MPro. Our previously developed MProinhibitors MPI8 and MPI29 were used as MProligands to conjugate a CRBN E3 ligand, leading to compounds that can both inhibit and degrade SARS-CoV-2 MPro. Among them, MDP2 was demonstrated to effectively reduce MProprotein levels in 293T cells (DC50= 296 nM), relying on a time-dependent, CRBN-mediated, and proteasome-driven mechanism. Furthermore, MPD2 exhibited remarkable efficacy in diminishing MProprotein levels in SARS-CoV-2-infected A549-ACE2 cells, concurrently demonstrating potent anti-SARS-CoV-2 activity (EC50= 492 nM). This proof-of-concept study highlights the potential of PROTAC-mediated targeted protein degradation of MProas an innovative and promising approach for COVID-19 drug discovery.
Organoid-based in vitro system and reporter for the study of Cryptosporidium parvum sexual reproduction
Authors: Korwin-Mihavics Bethany R , Dews Emmett A , Miller Peter Jonathon , Cameron Alexandra , Martorelli di Genova Bruno , Huston Christopher Dwight
Published: September 2023

Many advances have been made recently in our understanding of Cryptosporidium asexual cycle and sexual differentiation. However, the process of fertilization, which is required for transmission of infectious oocysts, is not well understood. Typical cancer cell-based culture only allows robust exploration of asexual cycle and sexual differentiation of Cryptosporidium. To facilitate exploration of sexual reproduction in C. parvum we developed an organoid-based culture system that supports Cryptosporidium full life cycle and a novel fertilization reporter. Organoid derived monolayers (ODMs) supported fertilization and oocyst production and maintained the infection for up to 3 weeks. ODM derived oocysts were infectious in vivo. Fertilization was confirmed by successfully mating two strains of C. parvum and with a novel fertilization switch reporter. The fertilization switch reporter utilizes a DiCre system in which cre fragments are expressed under the control of sexual stage promoters resulting in a rapamycin inducible switch in fluorescent protein expression from mCherry to mNeonGreen after fertilization that is spatially and temporally controlled. This results in mCherry positive parasites in the first generation and offspring that express mNeonGreen. In vivo validation of the fertilization switch reporter demonstrated the precision and efficiency of the fertilization switch reporter and confirmed excision of the mCherry gene sequence only after rapamycin treatment. The start of a second generation of parasites was also shown in the ODMs and rarely in HCT8s. Use of this reporter in ODMs can help investigate the Cryptosporidium lifecycle post sexual differentiation in a physiologically relevant in vitro system.
Disease-specific selective vulnerability and neuroimmune pathways in dementia revealed by single cell genomics
Authors: Rexach Jessica E. , Cheng Yuyan , Chen Lawrence , Polioudakis Damon , Lin Li-Chun , Mitri Vivianne , Elkins Andrew , Yin Anna , Calini Daniela , Kawaguchi Riki , Ou Jing , Huang Jerry , Williams Christopher , Robinson John , Gaus Stephanie E. , Spina Salvatore , Lee Edward B. , Grinberg Lea T. , Vinters Harry , Trojanowski John Q. , Seeley William W. , Malhotra Dheeraj , Geschwind Daniel H.
Published: September 2023

Summary/Abstract

The development of successful therapeutics for dementias requires an understanding of their shared and distinct molecular features in the human brain. We performed single-nuclear RNAseq and ATACseq in Alzheimer disease (AD), Frontotemporal degeneration (FTD), and Progressive Supranuclear Palsy (PSP), analyzing 40 participants, yielding over 1.4M cells from three brain regions ranging in vulnerability and pathological burden. We identify 35 shared disease-associated cell types and 14 that are disease-specific, replicating those previously identified in AD. Disease-specific cell states represent molecular features of disease-specific glial-immune mechanisms and neuronal vulnerability in each disorder, layer 4/5 intra-telencephalic neurons in AD, layer 2/3 intra-telencephalic neurons in FTD, and layer 5/6 near-projection neurons in PSP. We infer intrinsic disease-associated gene regulatory networks, which we empirically validate by chromatin footprinting. We find that causal genetic risk acts in specific neuronal and glial cells that differ across disorders, primarily non-neuronal cells in AD and specific neuronal subtypes in FTD and PSP. These data illustrate the heterogeneous spectrum of glial and neuronal composition and gene expression alterations in different dementias and identify new therapeutic targets by revealing shared and disease-specific cell states.
Parameter estimation of Wiener-Hammerstein system based on multi-population self-adaptive differential evolution algorithm
Authors: Chu Jie , Li Junhong , Jiang Yizhe , Song Weicheng , Zong Tiancheng
Published: September 2023

Purpose

The Wiener-Hammerstein nonlinear system is made up of two dynamic linear subsystems in series with a static nonlinear subsystem, and it is widely used in electrical, mechanical, aerospace and other fields. This paper considers the parameter estimation of the Wiener-Hammerstein output error moving average (OEMA) system.

Design/methodology/approach

The idea of multi-population and parameter self-adaptive identification is introduced, and a multi-population self-adaptive differential evolution (MPSADE) algorithm is proposed. In order to confirm the feasibility of the above method, the differential evolution (DE), the self-adaptive differential evolution (SADE), the MPSADE and the gradient iterative (GI) algorithms are derived to identify the Wiener-Hammerstein OEMA system, respectively.

Findings

From the simulation results, the authors find that the estimation errors under the four algorithms stabilize after 120, 30, 20 and 300 iterations, respectively, and the estimation errors of the four algorithms converge to 5.0%, 3.6%, 2.7% and 7.3%, which show that all four algorithms can identify the Wiener-Hammerstein OEMA system.

Originality/value

Compared with DE, SADE and GI algorithm, the MPSADE algorithm not only has higher parameter estimation accuracy but also has a faster convergence speed. Finally, the input–output relationship of laser welding system is described and identified by the MPSADE algorithm. The simulation results show that the MPSADE algorithm can effectively identify parameters of the laser welding system.
Key policy mechanisms supporting the University–Industry collaboration in the Danube region: case study of academic HPC centres and SMEs
Authors: Besednjak Valič Tamara , Kolar Janez , Lamut Urša , Pandiloska Jurak Alenka
Published: September 2023

Purpose

This paper aims to explore the key anchors of the National Innovation System shaping the nature of collaboration between academic high-performance computing centres (academic HPC centres) and small- to medium-sized enterprises (SMEs) working in the automotive and electronics sectors of the Danube region. With two main research questions, it discusses the importance of knowledge transfer and technology transfer for collaboration between University and Industry (U-I collaboration) in three groups of developmentally distinct countries: competitively advanced, competitively intermediate and competitively lagging. As main anchors of the innovation system, stable legal environment, exciting innovation policies and strong R&D funding are recognised.

Design/methodology/approach

A qualitative empirical study in 14 Danube region countries included 92 focus group participants, expert representatives of academic HPC centres and SMEs. The data were audio recorded, transcribed and analysed.

Findings

The findings show the main prerequisites of the framework conditions for efficient U-I collaboration evolve through a goal-oriented National Innovation Policy and developed and functioning legal environment supporting labour market and intellectual property (IP) protection and enforcement. Additionally, skilled people are needed to be able to operate with HPC, where it seems all the countries lack such skilled workforce. In competitively lagging countries, the high levels of brain drain exhibit strong impact to U-I collaboration.

Research limitations/implications

Research into relationships between academic HPC centres and SMEs conducted was qualitative; therefore, limitations in terms of generalisation arise from it. On the other hand, the research is promising in terms of offering the guidance for policy makers who can use the findings when delivering innovation policy mix, adjusted to developmental level of own innovation ecosystem.

Originality/value

The study is among the pioneering work in U-I collaboration between academic HPC centres and SMEs from automotive and electronics industries in the Danube region. The research addresses the dynamics of collaboration and offers policy implications to strengthen the particular U-I collaboration.
Moderating effect of Nigerian government policy support on the relationship between project management framework and emerging construction contractors' sustainability
Authors: Ebekozien Andrew , Aigbavboa Clinton , Samsurijan Mohamad Shaharudin , Amadi Godspower C. , Duru Okechukwu Dominic Saviour
Published: September 2023

Purpose

In most developing countries, indigenous emerging construction contractors (ECCs) face severe problems of not adopting a project management framework (PMF) in their business activities. It has increased their business risk and threatened their sustainability. Studies showed that government policy support (GPS) helps mitigate business risks. Thus, there is a paucity of literature concerning GPS on emerging Nigerian construction contractors' business sustainability. Therefore, the paper aims to investigate the moderating effect of GPS on the relationship between PMF and ECCs in Nigeria.

Design/methodology/approach

SmartPLS was used to analyse the collected data from the useable 310 questionnaires retrieved from respondents in Abuja and Lagos, Nigeria. Systems Theory was used to support the developed framework.

Findings

Findings show that government policy support significantly moderates the relationships between PMF and ECCs in the Nigerian construction sector. It implies that the study's results offer more understanding regarding issues affecting construction entrepreneurs' sustainable business cycle via applying PMF to mitigate business sustainable associated risks.

Practical implications

The study will stir Nigeria's ECCs and policymakers to promote construction business sustainability for a new entrepreneur, emphasising business risk management via PMF and GPS to enhance the sustainable business cycle.

Originality/value

The research (PMF and GPS) is strategies to enhance ECCs business sustainability in the Nigerian construction sector and other developing countries with similar political and economic attributes. Besides the study guiding old and intending ECCs and policymakers in the developing countries industries, it would contribute to bridge the theoretical gap regarding PMF and ECC, especially ECCs in developing countries with similar business sustainability issues.
Job embeddedness and turnover intention in the public sector: the role of life satisfaction and ethical leadership
Authors: Nguyen Tuan Duong , Nguyen Thuy Thi , Nguyen Phuong Cam
Published: September 2023

Purpose

This study aimed to investigate the impact of job embeddedness (JE) on the turnover intention (TI) of the public sector with the mediating effect of the individual factor (i.e. life satisfaction [LS]) and the moderating effect of the leadership style (i.e. ethical leadership [EL]).

Design/methodology/approach

This study adopted a quantitative design to collect data from 236 employees working in the public sector in Vietnam through field research using structured questionnaires. Data were analysed using partial least squares structural equation modelling (PLS-SEM) to test the hypotheses.

Findings

The results show that JE negatively affects the TI of public sector employees through the mediation effect of LS. Additionally, this study indicates that EL moderates the relationship between JE and TI.

Practical implications

This study implies that public sector stakeholders should consider both individual and contextual factors to manage and retain employees. In addition to addressing employees' embeddedness with the organisation and community and their LS, public organisations need to focus on hiring, training and promoting ethical leaders.

Originality/value

This study highlights the role of embeddedness within both the organisation and the community, along with the role of EL in the LS and TI of public sector employees.