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Sepsis, an amplified immune response to systemic infection, is characterized by a transient cytokine storm followed by chronic immune dysfunction. Consequently, sepsis survivors are highly susceptible to newly introduced infections, suggesting sepsis can influence the function and composition of the naïve CD8 T cell pool and resulting pathogen-induced primary CD8 T cell responses. Here, we explored the extent to which sepsis induces phenotypic and functional changes within the naïve CD8 T cell pool. To interrogate this, the cecal ligation and puncture (CLP) mouse model of polymicrobial sepsis was used. In normal, non-septic mice, we show type-I interferon (IFN I)-mediated signaling plays an important role in driving the phenotypic and functional heterogeneity in the naïve CD8 T cell compartment leading to increased representation of Ly6C+ naïve CD8 T cells. In response to viral infection after sepsis resolution, naïve Ly6C+ CD8 T cells generated more primary effector and memory CD8 T cells with slower conversion to a central memory CD8 T cell phenotype (Tcm) than Ly6C- naïve CD8 T cells. Importantly, as a potent inducer of cytokine storm and IFN I production, sepsis leads to increased representation of Ly6C+ naïve CD8 T cells that maintained their heightened ability to respond (i.e., effector and memory CD8 T cell accumulation and cytokine production) to primary LCMV infection. Lastly, longitudinal analyses of peripheral blood samples obtained from septic patients revealed profound changes in CD8 T cell subset composition and frequency compared to healthy controls. Thus, sepsis has the capacity to alter the composition of naïve CD8 T cells, directly influencing primary CD8 T cell responses to newly introduced infections. Author summary: Sepsis is an exaggerated host response to systemic infection that can lead to significant mortality and long-lasting immune dysfunction in survivors. Sepsis-induced immune dysfunction contributes to the increased susceptibility to new infections and increased cancer incidence. These findings suggest naïve CD8 T cells are negatively impacted following a septic event, but the underlying mechanism(s) responsible for naïve CD8 T cell impairment and the consequences for CD8 T cell-mediated pathogen control remain understudied. Naïve CD8 T cells are required for the control of newly encountered pathogens and have been recently appreciated as a functionally and phenotypically heterogenous compartment. The present study defines Type I IFN as a signal which shapes naïve CD8 T cell heterogeneity at homeostasis. Following a septic event, the naïve CD8 T cell compartment undergoes lasting phenotypic and functional changes in mice and humans. Type I IFN signaling along with increased cell cycling in the post-septic environment contribute to this compositional change, which may have direct implications for therapeutic intervention to enhance CD8 T cell function in immunocompromised sepsis survivors. [ABSTRACT FROM AUTHOR]

The antigen-specific targeting of autoreactive B cells via their unique B cell receptors (BCRs) is a novel and promising alternative to the systemic suppression of humoral immunity. We generated and characterized cytolytic fusion proteins based on an existing immunotoxin comprising tetanus toxoid fragment C (TTC) as the targeting component and the modified Pseudomonas aeruginosa exotoxin A (ETA') as the cytotoxic component. The immunotoxin was reconfigured to replace ETA' with either the granzyme B mutant R201K or MAPTau as human effector domains. The novel cytolytic fusion proteins were characterized with a recombinant human lymphocytic cell line developed using Transpo-mAb™ technology. Genes encoding a chimeric TTC-reactive immunoglobulin G were successfully integrated into the genome of the precursor B cell line REH so that the cells could present TTC-reactive BCRs on their surface. These cells were used to investigate the specific cytotoxicity of GrB(R201K)-TTC and TTC-MAPTau, revealing that the serpin proteinase inhibitor 9-resistant granzyme B R201K mutant induced apoptosis specifically in the lymphocytic cell line. Our data confirm that antigen-based fusion proteins containing granzyme B (R201K) are suitable candidates for the depletion of autoreactive B cells. [ABSTRACT FROM AUTHOR]

Misfolding of the cellular prion protein (PrPC) into the scrapie prion protein (PrPSc) results in progressive, fatal, transmissible neurodegenerative conditions termed prion diseases. Experimental and epidemiological evidence point toward a protracted, clinically silent phase in prion diseases, yet there is no diagnostic test capable of identifying asymptomatic individuals incubating prions. In an effort to identify early biomarkers of prion diseases, we have compared global transcriptional profiles in brains from pre-symptomatic prion-infected mice and controls. We identified Cst7, which encodes cystatin F, as the most strongly upregulated transcript in this model. Early and robust upregulation of Cst7 mRNA levels and of its cognate protein was validated in additional mouse models of prion disease. Surprisingly, we found no significant increase in cystatin F levels in both cerebrospinal fluid or brain parenchyma of patients with Creutzfeldt-Jakob disease compared to Alzheimer’s disease or non-demented controls. Our results validate cystatin F as a useful biomarker of early pathogenesis in experimental models of prion disease, and point to unexpected species-specific differences in the transcriptional responses to prion infections. [ABSTRACT FROM AUTHOR]

Antibody drug conjugates (ADCs) have recently been proven to be highly potent anti-tumor drugs, typically exceeding the efficacy of conventional monoclonal antibodies (mAbs). ADCs are currently produced by chemical conjugation of a small-molecule toxin to the mAb through lysine or cysteine side chains. This leads to heterogeneous mixtures of ADCs in which variable numbers of drugs are conjugated to individual antibodies and in which the site of conjugation cannot be defined. Consequently, there is currently significant interest in further development of drug conjugation technologies, with a particular focus on site-specific payload conjugation. Here, we present an enzymatic conjugation platform based on the S. aureus sortase A-mediated transpeptidation reaction, allowing the efficient generation of ADCs with toxins conjugated to pre-defined sites at pre-defined drug-to-antibody ratios. For this, two modifications were introduced: first, immunoglobulin heavy (IgH) and light (IgL) chains were modified at their C-termini by addition of the sortase A recognition motif LPETG, and second, the small molecule tubulin polymerization inhibitors monomethylauristatin E (MMAE) and maytansine were modified by addition of a pentaglycine peptide, thus making them suitable substrates for sortase A-mediated transpeptidation. We demonstrate efficient generation and characterization of the anti-CD30 ADC Ac10-vcPAB-MMAE, an enzymatically conjugated counterpart of brentuximab vedotin (Adcetris), as well as several anti-HER-2 ADCs including trastuzumab-maytansine, the counterpart of trastuzumab emtansine (Kadcyla). ADCs generated in this manner were found to display in vitro cell killing activities indistinguishable from the classic conjugates. Further, when tested in vivo in a HER-2-overexpressing ovarian cancer xenograft mouse model, enzymatically generated trastuzumab-maytansine was found to lead to complete regression of established tumors, similar to Kadcyla. [ABSTRACT FROM AUTHOR]

The TERT-CLPTM1L region of chromosome 5p15.33 is a multi-cancer susceptibility locus that encodes the reverse transcriptase subunit, hTERT, of the telomerase enzyme. Numerous cancer-associated single-nucleotide polymorphisms (SNPs), including rs10069690, have been identified within the hTERT gene. The minor allele (A) at rs10069690 creates an additional splice donor site in intron 4 of hTERT, and is associated with an elevated risk of multiple cancers including breast and ovarian carcinomas. We previously demonstrated that the presence of this allele resulted in co-production of full length (FL)-hTERT and an alternatively spliced, INS1b, transcript. INS1b does not encode the reverse transcriptase domain required for telomerase enzyme activity, but we show here that INS1b protein retains its ability to bind to the telomerase RNA subunit, hTR. We also show that INS1b expression results in decreased telomerase activity, telomere shortening, and an increased telomere-specific DNA damage response (DDR). We employed antisense oligonucleotides to manipulate endogenous transcript expression in favor of INS1b, which resulted in a decrease in telomerase activity. These data provide the first detailed mechanistic insights into a cancer risk-associated SNP in the hTERT locus, which causes cell type-specific expression of INS1b transcript from the presence of an additional alternative splice site created in intron 4 by the risk allele. We predict that INS1b expression levels cause subtle inadequacies in telomerase-mediated telomere maintenance, resulting in an increased risk of genetic instability and therefore of tumorigenesis. [ABSTRACT FROM AUTHOR]

Understanding how stem cells interact with cardiomyocytes is crucial for cell-based therapies to restore the cardiomyocyte loss that occurs during myocardial infarction and other cardiac diseases. It has been thought that functional myocardial repair and regeneration could be regulated by stem cell-cardiomyocyte contact. However, because various contact modes (junction formation, cell fusion, partial cell fusion, and tunneling nanotube formation) occur randomly in a conventional coculture system, the particular regulation corresponding to a specific contact mode could not be analyzed. In this study, we used laser-patterned biochips to define cell-cell contact modes for systematic study of contact-mediated cellular interactions at the single-cell level. The results showed that the biochip design allows defined stem cell-cardiomyocyte contact-mode formation, which can be used to determine specific cellular interactions, including electrical coupling, mechanical coupling, and mitochondria transfer. The biochips will help us gain knowledge of contact-mediated interactions between stem cells and cardiomyocytes, which are fundamental for formulating a strategy to achieve stem cell-based cardiac tissue regeneration. [ABSTRACT FROM AUTHOR]

Alternative Lengthening of Telomeres (ALT) is a non-telomerase mechanism of telomere lengthening that occurs in about 10% of cancers overall and is particularly common in astrocytic brain tumors and specific types of sarcomas. Somatic cell hybridization analyses have previously shown that normal telomerase-negative fibroblasts and telomerase-positive immortalized cell lines contain repressors of ALT activity, indicating that activation of ALT results from loss of one or more unidentified repressors. More recently, ATRX or DAXX was shown to be mutated both in tumors with telomere lengths suggestive of ALT activity and in ALT cell lines. Here, an ALT cell line was separately fused to each of four telomerase-positive cell lines, and four or five independent hybrid lines from each fusion were examined for expression of ATRX and DAXX and for telomere lengthening mechanism. The hybrid lines expressed either telomerase or ALT, with the other mechanism being repressed. DAXX was expressed normally in all parental cell lines and in all of the hybrids. ATRX was expressed normally in each of the four telomerase-positive parental cell lines and in every telomerase-positive hybrid line, and was abnormal in the ALT parental cells and in all but one of the ALT hybrids. This correlation between ALT activity and loss of ATRX expression is consistent with ATRX being a repressor of ALT. [ABSTRACT FROM AUTHOR]

Telomeres, specialised structures that protect chromosome ends, play a critical role in preserving chromosome integrity. Telomere dynamics in the Tasmanian devil (Sarcophilus harrisii) are of particular interest in light of the emergence of devil facial tumour disease (DFTD), a transmissible malignancy that causes rapid mortality and threatens the species with extinction. We used fluorescent in situ hybridisation to investigate telomere length in DFTD cells, in healthy Tasmanian devils and in four closely related marsupial species. Here we report that animals in the Order Dasyuromorphia have chromosomes characterised by striking telomere length dimorphism between homologues. Findings in sex chromosomes suggest that telomere length dimorphism may be regulated by events in the parental germlines. Long telomeres on the Y chromosome imply that telomere lengthening occurs during spermatogenesis, whereas telomere diminution occurs during oogenesis. Although found in several somatic cell tissue types, telomere length dimorphism was not found in DFTD cancer cells, which are characterised by uniformly short telomeres. This is, to our knowledge, the first report of naturally occurring telomere length dimorphism in any species and suggests a novel strategy of telomere length control. Comparative studies in five distantly related marsupials and a monotreme indicate that telomere dimorphism evolved at least 50 million years ago. [ABSTRACT FROM AUTHOR]

The Alternative Lengthening of Telomeres (ALT) pathway is a telomerase-independent pathway for telomere maintenance that is active in a significant subset of human cancers and in vitro immortalized cell lines. ALT is thought to involve templated extension of telomeres through homologous recombination, but the genetic or epigenetic changes that unleash ALT are not known. Recently, mutations in the ATRX/DAXX chromatin remodeling complex and histone H3.3 were found to correlate with features of ALT in pancreatic neuroendocrine cancers, pediatric glioblastomas, and other tumors of the central nervous system, suggesting that these mutations might contribute to the activation of the ALT pathway in these cancers. We have taken a comprehensive approach to deciphering ALT by applying genomic, molecular biological, and cell biological approaches to a panel of 22 ALT cell lines, including cell lines derived in vitro. Here we show that loss of ATRX protein and mutations in the ATRX gene are hallmarks of ALT--immortalized cell lines. In addition, ALT is associated with extensive genome rearrangements, marked micronucleation, defects in the G2/M checkpoint, and altered double-strand break (DSB) repair. These attributes will facilitate the diagnosis and treatment of ALT positive human cancers. [ABSTRACT FROM AUTHOR]

14-3-3δ is frequently lost in human breast cancers by genetic deletion or promoter methylation. We have now investigated the involvement of 14-3-3δ in the termination of NF-κB signal in mammary cells and its putative role in cancer relapse and metastasis. Our results show that 14-3-3s regulates nuclear export of p65-NF-κB following chronic TNFa stimulation. Restoration of 14-3-3δ in breast cancer cells reduces migration capacity and metastatic abilities in vivo. By microarray analysis, we have identified a genetic signature that responds to TNFa in a 14-3-3δ-dependent manner and significantly associates with different breast and other types of cancer. By interrogating public databases, we have found that overexpression of this signature correlates with poor relapse-free survival in breast cancer patients. Finally, screening of 96 human breast tumors showed that NF-κB activation strictly correlates with the absence of 14-3-3δ and it is significantly associated with worse prognosis in the multivariate analysis. Our findings identify a genetic signature that is important for breast cancer prognosis and for future personalized treatments based on NF-κB targeting [ABSTRACT FROM AUTHOR]

Positron emission tomography (PET) is a non-invasive molecular imaging technique using positron-emitting radioisotopes to study functional processes within the body. High resolution PET scanners designed for imaging rodents and non-human primates are now commonplace in preclinical research. Brain imaging in this context, with motion compensation, can potentially enhance the usefulness of PET by avoiding confounds due to anaesthetic drugs and enabling freely moving animals to be imaged during normal and evoked behaviours. Due to the frequent and rapid motion exhibited by alert, awake animals, optimal motion correction requires frequently sampled pose information and precise synchronisation of these data with events in the PET coincidence data stream. Motion measurements should also be as accurate as possible to avoid degrading the excellent spatial resolution provided by state-of-the-art scanners. Here we describe and validate methods for optimised motion tracking suited to the correction of motion in awake rats. A hardware based synchronisation approach is used to achieve temporal alignment of tracker and scanner data to within 10 ms. We explored the impact of motion tracker synchronisation error, pose sampling rate, rate of motion, and marker size on motion correction accuracy. With accurate synchronisation (,100 ms error), a sampling rate of .20 Hz, and a small head marker suitable for awake animal studies, excellent motion correction results were obtained in phantom studies with a variety of continuous motion patterns, including realistic rat motion (,5% bias in mean concentration). Feasibility of the approach was also demonstrated in an awake rat study. We conclude that motion tracking parameters needed for effective motion correction in preclinical brain imaging of awake rats are achievable in the laboratory setting. This could broaden the scope of animal experiments currently possible with PET. [ABSTRACT FROM AUTHOR]

Approximately 10% of cancers use recombination-mediated Alternative Lengthening of Telomeres (ALT) instead of telomerase to prevent telomere shortening. A characteristic of cells that utilize ALT is the presence of ALT-associated PML nuclear bodies (APBs) containing (TTAGGG)n DNA, telomere binding proteins, DNA recombination proteins, and heterochromatin protein 1 (HP1). The function of APBs is unknown and it is possible that they are functionally heterogeneous. Most ALT cells lack functional p53, and restoration of the p53/p21 pathway in these cells results in growth arrest/senescence and a substantial increase in the number of large APBs that is dependent on two HP1 isoforms, HP1a and HP1c. Here we investigated the mechanism of HP1-mediated APB formation, and found that histone chaperones, HIRA and ASF1a, are present in APBs following activation of the p53/p21 pathway in ALT cells. HIRA and ASF1a were also found to colocalize inside PML bodies in normal fibroblasts approaching senescence, providing evidence for the existence of a senescence-associated ASF1a/HIRA complex inside PML bodies, consistent with a role for these proteins in induction of senescence in both normal and ALT cells. Moreover, knockdown of HIRA but not ASF1a significantly reduced p53-mediated induction of large APBs, with a concomitant reduction of large HP1 foci. We conclude that HIRA, in addition to its physical and functional association with ASF1a, plays a unique, ASF1a-independent role, which is required for the localization of HP1 to PML bodies and thus for APB formation. [ABSTRACT FROM AUTHOR]

Background: Expression profiling, the measurement of all transcripts of a cell or tissue type, is currently the most comprehensive method to describe their physiological states. Given that accurate profiling methods currently available require RNA amounts found in thousands to millions of cells, many fields of biology working with specialized cell types cannot use these techniques because available cell numbers are limited. Currently available alternative methods for expression profiling from nanograms of RNA or from very small cell populations lack a broad validation of results to provide accurate information about the measured transcripts. Methods and Findings: We provide evidence that currently available methods for expression profiling of very small cell populations are prone to technical noise and therefore cannot be used efficiently as discovery tools. Furthermore, we present Pico Profiling, a new expression profiling method from as few as ten cells, and we show that this approach is as informative as standard techniques from thousands to millions of cells. The central component of Pico Profiling is Whole Transcriptome Amplification (WTA), which generates expression profiles that are highly comparable to those produced by others, at different times, by standard protocols or by Real-time PCR. We provide a complete workflow from RNA isolation to analysis of expression profiles. Conclusions: Pico Profiling, as presented here, allows generating an accurate expression profile from cell populations as small as ten cells. [ABSTRACT FROM AUTHOR]

We aimed to describe the attributes and attitudes of Swiss health professionals who treat persons with occupational burnout (POB) in the outpatient sector and explore associated determinants. The study design was descriptive cross-sectional survey, distributed to the 16,883 general practitioners (GP), psychiatrist-psychotherapists (PP), occupational physicians (OP) and psychologists registered in the Swiss Medical Association, the Swiss Federation of Psychologists, and other specialized associations. Using an online questionnaire, we identified professionals who consult and treat POB, their attributes, volume of POB consultations, diagnostics and treatment modalities and outcomes (OB severity, average proportion of POB who returned to work and who relapsed). Multinomial regression analysis was conducted to identify attributes associated with these outcomes. Among 3216 respondents, 2951 reported to consult POB, and 1130 (713 physicians and 410 psychologists) to treat them. POB consultations constitute 5 to 25% of professionals' consultations, which varies across professionals' specialties and specializations and geographic regions. The profile of POB consulted also differs across professionals. Work psychologists reported more often consulting POB at early OB stage, GPs mostly reported having patients with moderate OB, while PPs reported having the largest proportion of patients with severe OB. The treatment practices depend on OB severity but neither latter nor former was associated with the proportion of relapsed POB or POB who return to work. Physicians with waiting time >3 months reported more often having a higher proportion of relapsed patients. Since the study had an exploratory nature using a cross-sectional survey design and aggregated outcomes, these findings should be considered as first descriptive data, motivating further research., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Guseva Canu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Over 9.5 million confirmed cases of COVID-19 infection have been recorded in Africa. The syndrome of post-acute sequelae of SARS-CoV-2 infection (PASC) affects an estimated 32% to 87% of COVID patients globally. Data regarding prevalence and impact of PASC in Botswana are limited. This study used a cross-sectional survey design to query healthcare workers in Kweneng District, Botswana about perceived PASC prevalence, duration, symptoms, impact, and management strategies. The survey was disseminated to participants via pre-existing WhatsApp groups and paper copy. Descriptive statistics were used to analyse quantitative data, including demographic data. 72 respondents consented and completed the survey, from an estimated 650 staff meeting eligibility criteria; 63% were female and 36% were male. The majority (90%) were nurses, with doctors and "other" accounting for 6% and 4% of respondents, respectively; no administrators responded. Over half (72%) worked at primary care facilities and 28% worked in hospitals. Nearly all (93%) indicated seeing patients with PASC on a weekly basis, though the majority (61%) identified these patients as comprising <10% of total patients. The most frequently reported PASC symptom was persistent cough (64%), followed by shortness of breath (54%) and fatigue (49%). A substantial minority of respondents were unsure how to manage common PASC symptoms, with 29% and 36% indicating uncertainty regarding management of persistent cough and fatigue, respectively. Findings indicate that PASC symptoms are frequently encountered in clinical practice in Botswana with significant overlap with acute COVID-19, influenza-like illnesses, and tuberculosis, likely placing increased burden on existing health system processes. Providers reported uncertainty in managing presumed PASC, and current practice patterns may contribute to unintended adverse effects. Clear clinical algorithms for PASC screening, diagnosis, and management should be developed and disseminated in Botswana to mitigate the effects of PASC symptoms and improve the quality of life of COVID-19 survivors., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Mamalelala et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

During a health emergency, there is an urgent need to rapidly develop guidelines that meet minimum quality standards, as exemplified by the development of WHO guidelines on mask use in health care and community settings during the COVID-19 pandemic. Between January 2020 and October 2023, WHO developed 21 guideline updates on the use of masks as part of infection prevention and control (IPC) practices. Guideline developers had to deal with an ever-growing volume of evidence of variable quality. Initially, indirect evidence drawn from other severe respiratory illnesses and established minimum requirements for IPC were used. As direct evidence began to emerge, WHO commissioned a living systematic review on mask use in June 2020, which formed the basis of evidence-to-decision making. As more evidence became available, additional considerations were incorporated into the process of recommendation formulation, including harms, acceptability, feasibility and resource use. Target populations for the mask guidelines expanded to include the general public, including children. A broad range of disciplines supported guideline development, including IPC, epidemiology, infectious diseases, occupational health, engineering, pneumology, paediatrics, and water, sanitation and hygiene, as well as civil society representatives. Additional expertise was engaged in the areas of ventilation and aerobiology to expand the range of perspectives regarding modes of transmission. Despite challenges, the experience of rapidly and regularly updating advice on mask use during an emergency health response has shown that it is possible to apply the minimum standards for ensuring the guideline methodology is trustworthy and transparent, with increasing rigor over time as evidence improves. Overall, the experience of developing guidelines during the COVID-19 pandemic underscores the importance of adapting to evolving evidence, incorporating diverse perspectives, and maintaining transparency to ensure a rigorous methodology and in return guideline trustworthiness and effectiveness., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: RC received funding from the Agency for Healthcare Research and Quality to conduct a review on masks. He also received funding from WHO to conduct a review on risk factors for transmission in HCWs, and consulting fees as the methodologist for the WHO Health Emergencies Programme COVID-19 Infection Prevention and Control Guideline Development Group. NF, HH, VW, KD, JPT, AS, MM, RT, BA and AB work for the World Health Organization. There are no other competing interests., (Copyright: © 2024 Ford et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Crimean-Congo Hemorrhagic Fever virus (CCHFV) is a tick-borne pathogen that causes severe acute fever disease in humans and requires a biosafety level 4 laboratory for handling. Hazara virus (HAZV), belonging to the same virus genus as CCHFV, does not exhibit pathogenesis in humans. To investigate host RNA-binding proteins (RBPs) that regulate CCHFV replication, we generated a series of mutant RAW264.7 cells by CRISPR/Cas9 system and these cells were infected with HAZV. The viral titers in the supernatant of these cells was investigated, and HuR (ELAVL1) was identified. HuR KO RAW264.7 cells reduced HAZV replication. HuR is an RBP that enhances mRNA stability by binding to adenyl-uridine (AU)-rich regions in their 3' non-coding region (NCR). HuR regulates innate immune response by binding to host mRNAs of signaling molecules. The expression of cytokine genes such as Ifnb, Il6, and Tnf was reduced in HuR KO cells after HAZV infection. Although HuR supports the innate immune response during HAZV infection, we found that innate immune activation by HAZV infection did not affect its replication. We then investigated whether HuR regulates HAZV genome RNA stability. HAZV RNA genome was precipitated with an anti-HuR antibody, and HAZV genome RNA stability was lowered in HuR KO cells. We found that HuR associated with HAZV RNA and stabilized it to enhance HAZV replication. Furthermore, HuR-deficiency reduced CCHFV minigenome replication. CCHFV is a negative-strand RNA virus and positive-strand RNA is produced during replication. HuR was associated with positive-strand RNA rather than negative-strand RNA, and AU-rich region in 3'-NCR of S segment was responsible for immunoprecipitation with anti-HuR antibody and minigenome replication. Additionally, HuR inhibitor treatment reduced CCHFV minigenome replication. Our results indicate that HuR aids replication of the CCHFV minigenome by associating with the AU-rich region in the 3'-NCR., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Ikegawa et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Morphology, composition and molecular structure of starch directly affect the functional properties. This study investigated the morphological, compositional, and molecular structure properties of starch from starch branching enzyme gene (SBE) and granule-bound starch synthase gene (GBSS) mutated potato, and their associations with thermal, pasting, and film-making properties. SBE mutations were induced in native variety Desiree while GBSS mutations were herestacked to a selected SBE mutated parental line. Mutations in SBE resulted in smaller starch granules and higher amylose content, while GBSS mutations in the SBE background reduced amylose content. Mutations in SBE, particularly with GBSS mutations, significantly increased total phosphorus content. 31P NMR spectroscopy revealed higher proportions of C6-bound phosphate than of C3-bound phosphate in all studied lines. Amylopectin unit chain and internal chain distributions showed higher proportions of long chains in mutated lines compared with Desiree. These amylopectin long-chains were positively correlated with gelatinizationand, pasting temperatures, and temperature at peak viscosity. Short amylopectin chains showed positive correlations with breakdown viscosity, but negative correlations with the crystal melting temperature of retrograded starch. Total phosphorus content was positively correlated with the crystal melting temperature of retrograded starch. Starch from different lines was used to produce a series of potato starch films that differed in morphology and functional properties. A negative correlation was observed between Young's modulus of films and the long amylopectin-chain fraction. Thermal gravimetric analysis revealed highest thermal stability of Desiree starch films, followed by films from SBE-mutated high-amylose lines. Oxygen transmission rate and oxygen permeability analyses showed that films made with starch from selected GBSS and SBEs mutated line maintained comparable oxygen barrier properties to Desiree film. These insights on the impact of genetic mutations on starch properties indicate potential applications of in-planta starch modification for specific end-uses including packaging., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Jayarathna et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

SARS-CoV-2 infection during pregnancy was associated with maternal mortality and adverse birth outcomes in the pre-Omicron era, including a stillbirth rate of 5.6% in Botswana. We re-evaluated these outcomes in the Tsepamo Study during the Omicron era. We assessed maternal mortality and adverse birth outcomes for all singleton pregnancies from mid-November 2021 (the start of the Omicron era) to mid-August 2022 at nine Tsepamo sites, among individuals with documented SARS-CoV-2 screening PCR or antigen tests and known HIV status. Of 9,705 women routinely screened for SARS-CoV-2 infection at delivery (64% of deliveries at these sites), 373 (3.8%) tested positive. Women with HIV were as likely to test positive for SARS-CoV-2 (77/1833, 4.2%) as women without HIV (293/6981, 4.2%) (p = 1.0). There were 5 recorded maternal deaths (0.03%), one occurring in a woman with a positive SARS-CoV-2 test result. In contrast, maternal mortality was 3.7% and 0.1% in those with and without SARS-CoV-2, respectively, during the pre-Omicron era. In the Omicron era, there were no differences among infants exposed or unexposed to SARS-CoV-2 in overall adverse birth outcomes (28.1% vs 29.6%; aRR 1.0, 95%CI 0.8-1.1), severe adverse birth outcomes (11.9 vs 10.6%; aRR 1.1, 95%CI 0.8-1.5), preterm delivery (15.1% vs 14.9%; aRR 1.0, 95%CI 0.8-1.3), or stillbirth (1.9% vs 2.3%; aRR 0.8, 95%CI 0.4-1.7). Adverse outcomes among those exposed to both HIV and SARS-CoV-2 were similar to those exposed to HIV alone (31.2% vs. 33.1%; aRR 0.9, 95%CI 0.6-1.3; p = 0.5). Maternal mortality was far lower in Botswana during the Omicron era than in the pre-Omicron era, and adverse birth outcomes were no longer significantly impacted by exposure to SARS-CoV-2 either overall or with HIV co-exposure. Increased population immunity to SARS-CoV-2, less stress on the hospital systems in the Omicron era, and possible differences in viral pathogenicity may combine to explain these findings., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Banga et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

An acute bout of exercise in the moments after learning benefits the retention of new memories. This finding can be explained, at least partly, through a consolidation account: exercise provides a physiological state that is conducive to the early stabilisation of labile new memories, which supports their retention and subsequent retrieval. The modification of consolidation through non-invasive exercise interventions offers great applied potential. However, it remains poorly understood whether effects of exercise translate from the laboratory to naturalistic settings and whether the intensity of exercise determines the effect in memory. To this end, adult endurance runners were recruited as participants and completed two study sessions spaced two weeks apart. In each session, participants were presented with a list of words and asked to recall them on three occasions: (i) immediately following their presentation, (ii) after a 30-minute retention interval, and (iii) after 24 hours. Crucially, the 30-minute retention interval comprised our experimental manipulation: higher intensity exercise (running) in the first session and lower intensity exercise (walking) in the second, both completed in a naturalistic setting around participants' existing physical activity training programmes. Exertion was recorded through heart rate and rate of perceived exertion data. Alertness, mood, and arousal ratings were also collected before and after the 30-minute retention interval. Immediate memory for the two wordlists was matched, but participants retained significantly more words after 30 minutes and 24 hours when encoding was followed by higher than lower intensity exercise. Exertion data revealed that participants experienced vigorous and light exercise in the higher and lower intensity conditions, respectively. Significant improvements in alertness, mood, and arousal were observed following both exercise conditions, but especially in the higher intensity condition. These outcomes reveal that experiencing higher intensity physical activity in the field is conducive to declarative memory retention, possibly because it encourages consolidation., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Makepeace, Craig. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

In sub-Saharan African countries, mass media is critical in disseminating health information, including the need for HIV testing. Yet, in Cameroon, there is a dearth of studies examining how exposure to mass media is effective in the uptake of HIV testing. Using the 2018 Cameroon Demographic and Health Survey, we examined the association between exposure to mass media and HIV testing among sexually active women (n = 12,619) and men (n = 5,607). Our findings revealed a generally low uptake of HIV testing although more women (78%) have ever tested for HIV compared to men (67%). Adjusting for demographic, socioeconomic, and psychosocial factors, we found for both women and men their exposure at least once a week to the Internet (aOR = 1.57, p<0.001 for women; aOR = 1.76, p<0.001 for men), print media (aOR = 1.59, p<0.05 for women; aOR = 2.04, p<0.001 for men), radio (aOR = 1.34, p<0.01 for women; aOR = 1.57, p<0.001 for men), and television (aOR = 1.74, p<0.001 for women; aOR = 1.94, p<0.001 for men) was significantly associated with a higher likelihood of testing for HIV compared to their counterparts with no exposure at all. Our findings underscore the importance of further integrating mass media in HIV messaging in Cameroon as the country aims to achieve UNAIDS target 95-95-95 by 2023., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Antabe et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

While most individuals suffer progressive disease following HIV infection, a small fraction spontaneously controls the infection. Although CD8 T-cells have been implicated in this natural control, their mechanistic roles are yet to be established. Here, we combined mathematical modeling and analysis of previously published data from 16 SIV-infected macaques, of which 12 were natural controllers, to elucidate the role of CD8 T-cells in natural control. For each macaque, we considered, in addition to the canonical in vivo plasma viral load and SIV DNA data, longitudinal ex vivo measurements of the virus suppressive capacity of CD8 T-cells. Available mathematical models do not allow analysis of such combined in vivo-ex vivo datasets. We explicitly modeled the ex vivo assay, derived analytical approximations that link the ex vivo measurements with the in vivo effector function of CD8-T cells, and integrated them with an in vivo model of virus dynamics, thus developing a new learning framework that enabled the analysis. Our model fit the data well and estimated the recruitment rate and/or maximal killing rate of CD8 T-cells to be up to 2-fold higher in controllers than non-controllers (p = 0.013). Importantly, the cumulative suppressive capacity of CD8 T-cells over the first 4-6 weeks of infection was associated with virus control (Spearman's ρ = -0.51; p = 0.05). Thus, our analysis identified the early cumulative suppressive capacity of CD8 T-cells as a predictor of natural control. Furthermore, simulating a large virtual population, our model quantified the minimum capacity of this early CD8 T-cell response necessary for long-term control. Our study presents new, quantitative insights into the role of CD8 T-cells in the natural control of HIV infection and has implications for remission strategies., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Vemparala et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Loss of function mutations in the checkpoint kinase gene CHEK2 are associated with increased risk of breast and other cancers. Most of the 3,188 unique amino acid changes that can result from non-synonymous single nucleotide variants (SNVs) of CHEK2, however, have not been tested for their impact on the function of the CHEK2-enocded protein (CHK2). One successful approach to testing the function of variants has been to test for their ability to complement mutations in the yeast ortholog of CHEK2, RAD53. This approach has been used to provide functional information on over 100 CHEK2 SNVs and the results align with functional assays in human cells and known pathogenicity. Here we tested all but two of the 4,887 possible SNVs in the CHEK2 open reading frame for their ability to complement RAD53 mutants using a high throughput technique of deep mutational scanning (DMS). Among the non-synonymous changes, 770 were damaging to protein function while 2,417 were tolerated. The results correlate well with previous structure and function data and provide a first or additional functional assay for all the variants of uncertain significance identified in clinical databases. Combined, this approach can be used to help predict the pathogenicity of CHEK2 variants of uncertain significance that are found in susceptibility screening and could be applied to other cancer risk genes., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 McCarthy-Leo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: The costs associated with healthcare are of critical importance to both decision-makers and users, given the limited resources allocated to the health sector. However, the available scientific evidence on healthcare costs in low- and middle-income countries, such as Peru, is scarce. In the Peruvian context, the health system is fragmented, and the private health insurance and its financing models have received less research attention. We aimed to analyse user cost-sharing and associated factors within the private healthcare system., Methods: Our study was cross-sectional, using open data from the Electronic Transaction Model of Standardized Billing Data-TEDEF-SUSALUD, between 2021-2022. Our unit of analysis is the user's medical bills. We considered the total amount of cost-sharing, proportion of total payments as cost-sharing, and cost-sharing as a proportion of minimum salaries. We use a multiple regression model to perform the analyses., Results: Our study included 5,286,556 health services provided to users of the private health insurance in Peru. We found a significant difference was observed in the cost-sharing for hospitalization-related services, with an average of 419.64 soles per day (95% CI: 413.44 to 425.85). Also, we identified that for hospitalization-related services per day is, on average, 0.41 (95% CI: 0.41 to 0.41) minimum salaries more expensive than outpatient care, although cost-sharing per day of hospitalization represent on average only 14% of the total amount submitted., Conclusions: Our study provides a detailed overview of cost-sharing in the private healthcare system in Peru and the factors associated with them. Policymakers can use the study's finding that higher cost-sharing for inpatient hospitalization compared to outpatient care in private insurance can create inequities in access to healthcare to design policies aimed at reducing these costs and promoting a more equitable and accessible healthcare system in Peru., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Bellido-Boza et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Trachoma is a leading cause of infection-related blindness worldwide. This disease is caused by recurrent Chlamydia trachomatis (Ct) infections of the conjunctiva and develops in two phases: i) active (acute trachoma, characterized by follicular conjunctivitis), then long-term: ii) scarring (chronic trachoma, characterized by conjunctival fibrosis, corneal opacification and eyelid malposition). Scarring trachoma is driven by the number and severity of reinfections. The immune system plays a pivotal role in trachoma including exacerbation of the disease. Hence the immune system may also be key to developing a trachoma vaccine. Therefore, we characterized clinical and local immune response kinetics in a non-human primate model of acute conjunctival Ct infection and disease., Methodology/principal Findings: The conjunctiva of non-human primate (NHP, Cynomolgus monkeys-Macaca fascicularis-) were inoculated with Ct (B/Tunis-864 strain, B serovar). Clinical ocular monitoring was performed using a standardized photographic grading system, and local immune responses were assessed using multi-parameter flow cytometry of conjunctival cells, tear fluid cytokines, immunoglobulins, and Ct quantification. Clinical findings were similar to those observed during acute trachoma in humans, with the development of typical follicular conjunctivitis from the 4th week post-exposure to the 11th week. Immunologic analysis indicated an early phase influx of T cells in the conjunctiva and elevated interleukins 4, 8, and 5, followed by a late phase monocytic influx accompanied with a decrease in other immune cells, and tear fluid cytokines returning to initial levels., Conclusion/significance: Our NHP model accurately reproduces the clinical signs of acute trachoma, allowing for an accurate assessment of the local immune responses in infected eyes. A progressive immune response occurred for weeks after exposure to Ct, which subsided into a persistent innate immune response. An understanding of these local responses is the first step towards using the model to assess new vaccine and therapeutic strategies for disease prevention., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Paulet et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Infections with the pathogenic free-living amoebae Naegleria fowleri can lead to life-threatening illnesses including catastrophic primary amoebic meningoencephalitis (PAM). Efficacious treatment options for these infections are lacking and the mortality rate remains >95% in the US. Glycolysis is very important for the infectious trophozoite lifecycle stage and inhibitors of glucose metabolism have been found to be toxic to the pathogen. Recently, human enolase 2 (ENO2) phosphonate inhibitors have been developed as lead agents to treat glioblastoma multiforme (GBM). These compounds, which cure GBM in a rodent model, are well-tolerated in mammals because enolase 1 (ENO1) is the predominant isoform used systemically. Here, we describe findings that demonstrate these agents are potent inhibitors of N. fowleri ENO (NfENO) and are lethal to amoebae. In particular, (1-hydroxy-2-oxopiperidin-3-yl) phosphonic acid (HEX) was a potent enzyme inhibitor (IC50 = 0.14 ± 0.04 μM) that was toxic to trophozoites (EC50 = 0.21 ± 0.02 μM) while the reported CC50 was >300 μM. Molecular docking simulation revealed that HEX binds strongly to the active site of NfENO with a binding affinity of -8.6 kcal/mol. Metabolomic studies of parasites treated with HEX revealed a 4.5 to 78-fold accumulation of glycolytic intermediates upstream of NfENO. Last, nasal instillation of HEX increased longevity of amoebae-infected rodents. Two days after infection, animals were treated for 10 days with 3 mg/kg HEX, followed by one week of observation. At the end of the one-week observation, eight of 12 HEX-treated animals remained alive (resulting in an indeterminable median survival time) while one of 12 vehicle-treated rodents remained, yielding a median survival time of 10.9 days. However, intranasal HEX delivery was not curative as brains of six of the eight survivors were positive for amoebae. These findings suggest that HEX requires further evaluation to develop as a lead for treatment of PAM., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Milanes et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Traumatic spinal injury (TSI) is a disease of significant global health burden, particularly in low and middle-income countries where road traffic-related trauma is increasing. This study compared the demographics, injury patterns, and outcomes of TSI caused by road traffic accidents (RTAs) to non-traffic related TSI., Methods: A retrospective analysis was conducted using a neurotrauma registry from the Muhimbili Orthopaedic Institute (MOI) in Tanzania, a national referral center for spinal injuries. Patient sociodemographic characteristics, injury level, and severity were compared across mechanisms of injury. Neurological improvement, neurological deterioration, and mortality were compared between those sustaining TSI through an RTA versus non-RTA, using univariable and multivariable analyses., Results: A total of 626 patients were included, of which 302 (48%) were RTA-related. The median age was 34 years, and 532 (85%) were male. RTAs had a lower male preponderance compared to non-RTA causes (238/302, 79% vs. 294/324, 91%, p<0.001) and a higher proportion of cervical injuries (144/302, 48% vs. 122/324, 38%, p<0.001). No significant differences between RTA and non-RTA mechanisms were found in injury severity, time to admission, length of hospital stay, surgical intervention, neurological outcomes, or in-hospital mortality. Improved neurological outcomes were associated with incomplete injuries (AIS B-D), while higher mortality rates were linked to cervical injuries and complete (AIS A) injuries., Conclusion: Our study in urban Tanzania finds no significant differences in outcomes between spinal injuries from road traffic accidents (RTAs) and non-RTA causes, suggesting the need for equitable resource allocation in spine trauma programs. Highlighting the critical link between cervical injuries and increased mortality, our findings call for targeted interventions across all causes of traumatic spinal injuries (TSI). We advocate for a comprehensive trauma care system that merges efficient pre-hospital care, specialized treatment, and prevention measures, aiming to enhance outcomes and ensure equity in trauma care in low- and middle-income countries., Competing Interests: The authors report no conflict of interest concerning the materials or methods used in this study or the findings specified in this paper. Scott L. Zuckerman is a consultant for the National Football League and Medtronic. Roger Hartl is a DePuy Synthes and Brainlab consultant and has royalties from Zimmer Biomet. He is also an advisor for 3D Bio and Real Spine. No other author declares any financial interests or personal relationships., (Copyright: © 2024 Ikwuegbuenyi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

HIV is a major risk factor for active Tuberculosis (TB.) This raises patients' risk of original infection, reinfection, and TB reactivation. Providing healthcare to KPLHIV in developing countries requires TB prevalence research. This study aims to determine the prevalence of TB and HIV co-infection and associated factors among KPLHIV. This is a retrospective cross-sectional study among KP's living with HIV enrolled on care in One Stop Shop (OSS) of Heartland Alliance Ltd/GTE across six states in Nigeria. Data were analysed using IBM SPSS version 25.0. Secondary data analysis of client's records from the RADET files of the KPCARE 1 project from 6 states was conducted. Means with standard deviations were computed for continuous variables like age, and frequency tables were generated for categorical variables. Chi-square tests and t-tests were used for the bivariate analysis of variables. All tests were done at a 5% level of statistical significance (p = 0.05).TB prevalence was 19.1% among KP's living with HIV, with variations observed in age groups, geographic locations, target populations, marital status, educational backgrounds, clinical characteristics, and antiretroviral therapy (ART) history. KPs aged 51 and above exhibited the highest TB prevalence (21.0%), while those aged below 20 years had the lowest (18.2%). Jigawa KPs recorded the highest TB prevalence (38.4%), and Niger had the least (13.3%). TB was more prevalent among People who inject drugs (20.3%), divorced (32.3%), and those who attained Qur'anic education (29.7%). KPs who had to restart ART exhibited the highest TB prevalence (22.0%), whereas those who experienced Interruption in treatment (IIT) reported the lowest at 10.0%. Immune-suppressed KPs (CD4 counts < 200 cells/m3) had a higher TB prevalence of 26.6%. TB prevalence among KPs living with HIV varies greatly, underlining the need for targeted treatments, especially for high-risk categories, to improve HIV treatment outcomes and reduce TB prevalence., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Ochonye et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: Stigma and discrimination against people living with HIV (PLHIV) remain a major barrier to effective HIV prevention. Despite the understanding that the creation of a socially inclusive environment for PLHIV is crucial for the promotion of testing, status disclosure, and treatment uptake, HIV stigma persists. Additionally, evidence suggests the endorsement of HIV stigma may be gender specific. Nonetheless, very little is known about the factors influencing men's discrimination against PLHIV in the Ghanaian context. Guided by the theory of planned behavior, our study fills this void by exploring the factors associated with the endorsement of HIV stigma in Ghana., Methods: Utilizing a nationally representative data from the 2022 Ghana Demographic and Health Survey (DHS) (N = 7044 men with ages ranging from 15-49 years), and applying logistic regression models, this study examined the factors associated with the endorsement of HIV-related stigma in Ghana., Results: The notion that HIV can be transmitted through the sharing of food with PLHIV was significantly associated with increased odds of stigma endorsement against children with HIV (OR = 3.381; P<0.001) and vendors with HIV (OR = 3.00; P<0.001). On the contrary, knowing that a healthy-looking person can have HIV was significantly associated with decreased odds of endorsement of stigma against children living with HIV (OR = 0.505; P<0.001), and vendors living with HIV (OR = 0.573; P<0.001). Likewise, having knowledge of drugs that help PLHIV to live longer, was significantly associated with decreased odds of stigma endorsement against children living with HIV (OR = 0.768; P<0.001), and vendors living with HIV (OR = 0.719; P<0.001). Moreover, participants with higher educational attainment reported lower odds of stigma endorsement against children living with HIV (OR = 0.255; P<0.01), and vendors living with HIV (OR = 0.327; P<0.01). Furthermore, age was significant and inversely associated with the endorsement of HIV stigma against children living with HIV (OR = 0.951; P<0.05), and vendors living with HIV (OR = 0.961; P<0.05). Also, wealth, ethnicity, and the region of residence significantly predicted endorsement of HIV stigma., Conclusion: For Ghana to achieve UNAIDS target 95-95-95 by 2030, targeted educational campaigns are necessary to dispel misconceptions about HIV and to promote social inclusion for reducing HIV-related stigma and discrimination in the country., Competing Interests: The author declares that there is no competing interest., (Copyright: © 2024 Saaka, Antabe. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Echinococcosis is a natural focal, highly prevalent disease in China. Factors influencing the spread of echinococcosis are not only related to personal exposure but also closely related to the environment itself. The purpose of this study was to explore the influence of environmental factors on the prevalence of human echinococcosis and to provide a reference for prevention and control of echinococcosis in the future., Methods: Data were collected from 370 endemic counties in China in 2018. By downloading Modis, DEM and other remote-sensing images in 2018. Data on environmental factors, i.e., elevation, land surface temperature (LST) and normalized difference vegetation index (NDVI) were collected. Rank correlation analysis was conducted between each environmental factor and the prevalence of echinococcosis at the county level. Negative binomial regression was used to analyze the impact of environmental factors on the prevalence of human echinococcosis at the county level., Results: According to rank correlation analysis, the prevalence of human echinococcosis in each county was positively correlated with elevation, negatively correlated with LST, and negatively correlated with NDVI in May, June and July. Negative binomial regression showed that the prevalence of human echinococcosis was negatively correlated with annual LST and summer NDVI, and positively correlated with average elevation and dog infection rate. The prevalence of human cystic echinococcosis was inversely correlated with the annual average LST, and positively correlated with both the average elevation and the prevalence rate of domestic animals. The prevalence of human alveolar echinococcosis was positively correlated with both NDVI in autumn and average elevation, and negatively correlated with NDVI in winter., Conclusion: The prevalence of echinococcosis in the population is affected by environmental factors. Environmental risk assessment and prediction can be conducted in order to rationally allocate health resources and improve both prevention and control efficiency of echinococcosis., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Wang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: The aim of this study was to use cluster analysis based on the trajectory of five cognitive-emotional processes (worry, rumination, metacognition, cognitive reappraisal and expressive suppression) over time to explore differences in clinical and performance variables in primary care patients with emotional symptoms., Methods: We compared the effect of adding transdiagnostic cognitive-behavioural therapy (TD-CBT) to treatment as usual (TAU) according to cluster membership and sought to determine the variables that predicted cluster membership. 732 participants completed scales about cognitive-emotional processes, anxiety and depressive symptoms, functioning, and quality of life (QoL) at baseline, posttreatment, and at 12 months. Longitudinal cluster analysis and logistic regression analyses were carried out., Results: A two-cluster solution was chosen as the best fit, named as "less" or "more" improvement in cognitive-emotional processes. Individuals who achieved more improvement in cognitive-emotional processes showed lower emotional symptoms and better QoL and functioning at all three time points. TAU+TD-CBT, income level, QoL and anxiety symptoms were significant predictors of cluster membership., Conclusions: These results underscore the value of adding TD-CBT to reduce maladaptive cognitive-emotional regulation strategies. These findings highlight the importance of the processes of change in therapy and demonstrate the relevance of the patient's cognitive-emotional profile in improving treatment outcomes., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Barrio-Martínez et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Resilience-based management is essential to protect ecosystems in the Anthropocene. Unlike large-scale climate threats to Great Barrier Reef (GBR) corals, outbreaks of coral-eating crown-of-thorns starfish (COTS; Acanthaster cf. solaris) can be directly managed through targeted culling. Here, we evaluate the outcomes of a decade of strategic COTS management in suppressing outbreaks and protecting corals during the 4th COTS outbreak wave at reef and regional scales (sectors). We compare COTS density and coral cover dynamics during the 3rd and 4th outbreak waves. During the 4th outbreak wave, sectors that received limited to no culling had sustained COTS outbreaks causing significant coral losses. In contrast, in sectors that received timely and sufficient cull effort, coral cover increased substantially, and outbreaks were suppressed with COTS densities up to six-fold lower than in the 3rd outbreak wave. In the Townsville sector for example, despite exposure to comparable disturbance regimes during the 4th outbreak wave, effective outbreak suppression coincided with relative increases in sector-wide coral cover (44%), versus significant coral cover declines (37%) during the 3rd outbreak wave. Importantly, these estimated increases span entire sectors, not just reefs with active COTS control. Outbreaking reefs with higher levels of culling had net increases in coral cover, while the rate of coral loss was more than halved on reefs with lower levels of cull effort. Our results also indicate that outbreak wave progression to adjoining sectors has been delayed, probably via suppression of COTS larval supply. Our findings provide compelling evidence that proactive, targeted, and sustained COTS management can effectively suppress COTS outbreaks and deliver coral growth and recovery benefits at reef and sector-wide scales. The clear coral protection outcomes demonstrate the value of targeted manual culling as both a scalable intervention to mitigate COTS outbreaks, and a potent resilience-based management tool to "buy time" for coral reefs, protecting reef ecosystem functions and biodiversity as the climate changes., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Matthews et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Women are biologically more susceptible to the Human Immunodeficiency Virus (HIV) and other sexually transmitted Infections (STIs) because receptive sex is riskier than insertive. Despite condom use being the staple preventive method for HIV infection (over 80% efficacy), in Sub-Saharan African countries like Burkina Faso, a high burden of HIV and the unmet need for condom use coexist. Moreover, even though women in SSA are disproportionately HIV positive, they are reportedly less capable of negotiating condom use for HIV risk reduction. Thus, using the Health Believe Model (HBM), this study explored the factors that influence condom use among women within the context of HIV prevention, with a key interest in condom use negotiation. Using the women's dataset of the 2021 Burkina Faso Demographic and Health Survey and applying logistic regression models, this study examined the factors associated with condom use for HIV risk reduction. Women who had confidence to negotiate condom use with their partners (OR = 1.57, P<0.001, 95%CI: 1.29, 1.91), those with secondary education (OR = 1.38, P<0.05, 95%CI: 1.07 1.77), from richest households (OR = 1.64, P<0.05, 95%CI: 1.08, 2.47), the employed (OR = 1.23, P<0.05, 95%CI: 1.02, 1.49), women with knowledge of sexually transmitted infections (OR = 1.58, P<0.001, 95%CI: 1.26, 1.97), those who have ever been tested for HIV (OR = 1.85, P<0.001, 95%CI: 1.52 2.24), as well as those who knew that a healthy-looking person can have HIV (OR = 2.64, P<0.001, 95%CI: 2.15, 3.24) were all significantly more likely to practice condom use for HIV risk reduction. Also, religion and geographical location of participants significantly predicted condom use for HIV risk reduction in the study context. The ability to negotiate condom use, knowledge of HIV and STIs, the socioeconomic status of women, as well as their geographical location, influence their practice of safer sex for HIV risk reduction in Burkina Faso., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Saaka et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Robust prognostic and predictive factors for hepatocellular carcinoma, a leading cause of cancer-related deaths worldwide, have not yet been identified. Previous studies have identified potential HCC determinants such as genetic mutations, epigenetic alterations, and pathway dysregulation. However, the clinical significance of these molecular alterations remains elusive. MicroRNAs are major regulators of protein expression. MiRNA functions are frequently altered in cancer. In this study, we aimed to explore the prognostic value of differentially expressed miRNAs in HCC, to elucidate their associated pathways and their impact on treatment response. To this aim, bioinformatics techniques and clinical dataset analyses were employed to identify differentially expressed miRNAs in HCC compared to normal hepatic tissue. We validated known associations and identified a novel miRNA signature with potential prognostic significance. Our comprehensive analysis identified new miRNA-targeted pathways and showed that some of these protein coding genes predict HCC patients' response to the tyrosine kinase inhibitor sorafenib., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Smith et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: The use of routinely collected health data for secondary research purposes is increasingly recognised as a methodology that advances medical research, improves patient outcomes, and guides policy. This secondary data, as found in electronic medical records (EMRs), can be optimised through conversion into a uniform data structure to enable analysis alongside other comparable health metric datasets. This can be achieved with the Observational Medical Outcomes Partnership Common Data Model (OMOP-CDM), which employs a standardised vocabulary to facilitate systematic analysis across various observational databases. The concept behind the OMOP-CDM is the conversion of data into a common format through the harmonisation of terminologies, vocabularies, and coding schemes within a unique repository. The OMOP model enhances research capacity through the development of shared analytic and prediction techniques; pharmacovigilance for the active surveillance of drug safety; and 'validation' analyses across multiple institutions across Australia, the United States, Europe, and the Asia Pacific. In this research, we aim to investigate the use of the open-source OMOP-CDM in the PATRON primary care data repository., Methods: We used standard structured query language (SQL) to construct, extract, transform, and load scripts to convert the data to the OMOP-CDM. The process of mapping distinct free-text terms extracted from various EMRs presented a substantial challenge, as many terms could not be automatically matched to standard vocabularies through direct text comparison. This resulted in a number of terms that required manual assignment. To address this issue, we implemented a strategy where our clinical mappers were instructed to focus only on terms that appeared with sufficient frequency. We established a specific threshold value for each domain, ensuring that more than 95% of all records were linked to an approved vocabulary like SNOMED once appropriate mapping was completed. To assess the data quality of the resultant OMOP dataset we utilised the OHDSI Data Quality Dashboard (DQD) to evaluate the plausibility, conformity, and comprehensiveness of the data in the PATRON repository according to the Kahn framework., Results: Across three primary care EMR systems we converted data on 2.03 million active patients to version 5.4 of the OMOP common data model. The DQD assessment involved a total of 3,570 individual evaluations. Each evaluation compared the outcome against a predefined threshold. A 'FAIL' occurred when the percentage of non-compliant rows exceeded the specified threshold value. In this assessment of the primary care OMOP database described here, we achieved an overall pass rate of 97%., Conclusion: The OMOP CDM's widespread international use, support, and training provides a well-established pathway for data standardisation in collaborative research. Its compatibility allows the sharing of analysis packages across local and international research groups, which facilitates rapid and reproducible data comparisons. A suite of open-source tools, including the OHDSI Data Quality Dashboard (Version 1.4.1), supports the model. Its simplicity and standards-based approach facilitates adoption and integration into existing data processes., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Ward et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Stigmatization and discrimination within healthcare settings deter key populations (KPs) from seeking mental health and psychosocial support (MHPS). Consequently, understanding the prevalence, associated factors, and impact of the MHPSS intervention on psychological distress among Nigeria's KPs is crucial., Method: This is a cross-sectional study focused on KPs, including Female Sex Workers (FSW), Men who have Sex with Men (MSM), and People Who Inject Drugs (PWID) enrolled in Heartland Alliance LTD/GTE across 17 One-Stop Shops (OSS) in six states of Nigeria. Data were extracted from the databases of the OSS. PD was assessed using the Mental Health Screening Form III (MHSF-III). Descriptive statistics and univariable and multivariable binary logistic regression models were done using IBM-SPSS version 28., Results: The prevalence of PD among the KPs was 9.7%. Higher rates were observed among FSWs (12.0%). Of the 22310 KPs, the prevalence of PD was 9.7%. The major dependants of PD include being a PWID with PD prevalence of 8.5% and AOR of 1.95 (95% CI: 0.60-0.98, p = 0.015), alcohol intake with PD prevalence of 97.7% and AOR of 21.83 (95% CI: 15.13-56.83, p<0.001), and having experienced gender-based violence with PD prevalence of 99.0% and AOR of 25.70(95% CI: 17.10-38.73, p<0.001). All Participants (100%) were given brief intervention, and 1595 of 2159 (73.8%) were referred for further psychological intervention. The services with the highest proportion were psychoeducation (21.20%), followed by coping skills training (17.70%) and motivational enhancement (12.90%)., Conclusion: The study highlights the critical need for targeted mental health interventions among KPs in Nigeria, primarily focusing on those with a history of substance abuse and gender-based violence. Despite universal brief interventions, the proportion enrolled in further Psychosocial support indicates a need to improve mental health service utilization among the KPs in Nigeria., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Ochonye et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The circadian clock controls behavior and metabolism in various organisms. However, the exact timing and strength of rhythmic phenotypes can vary significantly between individuals of the same species. This is highly relevant for rhythmically complex marine environments where organismal rhythmic diversity likely permits the occupation of different microenvironments. When investigating circadian locomotor behavior of Platynereis dumerilii, a model system for marine molecular chronobiology, we found strain-specific, high variability between individual worms. The individual patterns were maintained for several weeks. A diel head transcriptome comparison of behaviorally rhythmic versus arrhythmic wild-type worms showed that 24-h cycling of core circadian clock transcripts is identical between both behavioral phenotypes. While behaviorally arrhythmic worms showed a similar total number of cycling transcripts compared to their behaviorally rhythmic counterparts, the annotation categories of their transcripts, however, differed substantially. Consistent with their locomotor phenotype, behaviorally rhythmic worms exhibit an enrichment of cycling transcripts related to neuronal/behavioral processes. In contrast, behaviorally arrhythmic worms showed significantly increased diel cycling for metabolism- and physiology-related transcripts. The prominent role of the neuropeptide pigment-dispersing factor (PDF) in Drosophila circadian behavior prompted us to test for a possible functional involvement of Platynereis pdf. Differing from its role in Drosophila, loss of pdf impacts overall activity levels but shows only indirect effects on rhythmicity. Our results show that individuals arrhythmic in a given process can show increased rhythmicity in others. Across the Platynereis population, rhythmic phenotypes exist as a continuum, with no distinct "boundaries" between rhythmicity and arrhythmicity. We suggest that such diel rhythm breadth is an important biodiversity resource enabling the species to quickly adapt to heterogeneous or changing marine environments. In times of massive sequencing, our work also emphasizes the importance of time series and functional tests., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Häfker et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: To assess the cost-effectiveness and budget impact of olaparib as a maintenance therapy in platinum-responsive, metastatic pancreatic cancer patients harboring a germline BRCA1/2 mutation, using the Swiss context as a model., Methods: Based on data from the POLO trial, published literature and local cost data, we developed a partitioned survival model of olaparib maintenance including full costs for BRCA1/2 germline testing compared to FOLFIRI maintenance chemotherapy and watch-and-wait. We calculated the incremental cost-effectiveness ratio (ICER) for the base case and several scenario analyses and estimated 5-year budget impact., Results: Comparing olaparib with watch-and wait and maintenance chemotherapy resulted in incremental cost-effectiveness ratios of CHF 2,711,716 and CHF 2,217,083 per QALY gained, respectively. The 5-year costs for the olaparib strategy in Switzerland would be CHF 22.4 million, of which CHF 11.4 million would be accounted for by germline BRCA1/2 screening of the potentially eligible population. This would amount to a budget impact of CHF 15.4 million (USD 16.9 million) versus watch-and-wait., Conclusions: Olaparib is not a cost-effective maintenance treatment option. Companion diagnostics are an equally important cost driver as the drug itself., Competing Interests: All authors have read the journal’s policy and the authors of this manuscript have the following competing interests. AW: no conflicts of interest for this article AS: no conflicts of interest for this article KD: no conflicts of interest for this article TK: no conflicts of interest for this article JL: no conflicts of interest for this article MS: unrelated to the present work, research funding from AbbVie, Biogen, Bristol Myers Squibb, Merck Sharpe & Dohme, Mundipharma, Novartis, and Roche via employment institution; personal fees from Sandoz RvM: consultancy fees from Astra Zeneca, Eili Lilly, Gilead Science, GlaxoSmithKline, Merck, MSD, Novartis, Pierre Fabre, Pharmamar, Sanofi and Vifor. Travel grants from Pierre Fabre, Takeda TM: no conflicts of interest for this article This study was partially funded by Swiss Study Group for Clinical Oncological Research (SAKK, Schweizerische Arbeitsgemeinschaft für Klinische Krebsforschung). This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2024 Mehra et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The feeding strategies of the first domesticated herds had to manage the risks arising from the novelty of livestock practices in territories often distant from the animals' primary habitats. The Iberian Peninsula is characterised by a great diversity of environments, which undoubtedly influenced these dynamics. At the beginning of the Neolithic period these led the possibility to combine diverse livestock farming practices based on different animal feeding habits. This variability is also consistent with the rythms of adoption of domesticated animals, being later on the northern area. In order to address this issue, this work focuses on the dietary regimes of early sheep herds from southern Iberia, an area for which information is currently scarce. This study utilises high-resolution radiocarbon dating and stable isotope data on teeth to investigate sheep husbandry management strategies in Cueva de El Toro (Antequera, Málaga). The radiocarbon dates on the analysed remains evidenced they were deposited at the site over a short period, supporting the recurrent use of the cave. The sequential analysis of oxygen and carbon isotopes in tooth enamel reveals distinct livestock management strategies, reproduction patterns, feeding habits, and mobility during this short period. This variability demonstrates that livestock management practices in the western Mediterranean are more diverse than previously considered. Furthermore, these findings support the hypothesis that early Neolithic communities in the southern Iberian Peninsula were able to adopt different feeding strategies within the same herd, depending on their ecological and productive needs., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Sierra et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Transdiagnostic group cognitive behavioural therapy (TD-GCBT) is more effective in improving symptoms and severity of emotional disorders (EDs) than treatment as usual (TAU; usually pharmacological treatment). However, there is little research that has examined the effects of these treatments on specific symptoms. This study used Network Intervention Analysis (NIA) to investigate the direct and differential effects of TD-GCBT + TAU and TAU on specific symptoms of anxiety and depression. Data are from a multicentre randomised clinical trial (N = 1061) comparing TD-GCBT + TAU versus TAU alone for EDs. The networks included items from the PHQ-9 (depression) and GAD-7 (anxiety) questionnaire and mixed graphical models were estimated at pre-treatment, post-treatment and 3-, 6- and 12-month follow-up. Results revealed that TD-GCBT + TAU was associated with direct effects, mainly on several anxiety symptoms and depressed mood after treatment. New direct effects on other depressive symptoms emerged during the follow-up period promoted by TD-GCBT compared to TAU. Our results suggest that the improvement of anxiety symptoms after treatment might precipitate a wave of changes that favour a decrease in depressive symptomatology. NIA is a methodology that can provide fine-grained insight into the likely pathways through which treatments exert their effects., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Jurado-González et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: Monthly intravenous infusion of broadly neutralizing monoclonal antibodies may be an attractive alternative to daily oral antiretroviral treatment for children living with HIV. However, acceptability among caregivers remains unknown., Methods: We evaluated monthly infusion of dual bNAbs (VRCO1LS and 10-1074) as a treatment alternative to ART among children participating in the Tatelo Study in Botswana. Eligible children aged 2-5 years received 8-32 weeks of bNAbs overlapping with ART, and up to 24 weeks of bNAbs alone as monthly intravenous infusion. Using closed-ended questionnaires, we evaluated caregiver acceptability of each treatment strategy prior to the first bNAb administration visit (pre-intervention) and after the completion of the final bNAb administration visit (post-intervention)., Results: Twenty-five children completed the intervention phase of the study, and acceptability data were available from 24 caregivers at both time points. Responses were provided by the child's mother at both visits (60%), an extended family member at both visits (28%), or a combination of mother and an extended family member (12%). Caregiver acceptance of monthly bNAb infusions was extremely high both pre-and post-intervention, with 21/24 (87.5%) preferring bNAbs to ART pre-intervention, and 21/25 (84%) preferring bNAbs post-intervention. While no caregiver preferred ART pre-intervention, 2/25 preferred it post-intervention. Pre-intervention, 3 (13%) caregivers had no preference between monthly bNAbs or daily ART, and 2 (8%) had no preference post-intervention. Pre-intervention, the most common reasons for preferring bNAbs over ART were the perception that bNAbs were better at suppressing the virus than ART (n = 10) and the fact that infusions were dosed once monthly compared to daily ART (n = 9). Post-intervention, no dominant reason for preferring bNAbs over ART emerged from caregivers., Conclusions: Monthly intravenous bNAb infusions were highly acceptable to caregivers of children with HIV in Botswana and preferred over standard ART by the majority of caregivers., Clinical Trial Number: NCT03707977., Competing Interests: The authors have declared that no competing interest exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)