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Toxoplasma gondii is an intracellular parasite that establishes a long-term infection in the brain of many warm-blooded hosts, including humans and rodents. Like all obligate intracellular microbes, Toxoplasma uses many effector proteins to manipulate the host cell to ensure parasite survival. While some of these effector proteins are universal to all Toxoplasma strains, some are polymorphic between Toxoplasma strains. One such polymorphic effector is GRA15. The gra15 allele carried by type II strains activates host NF-κB signaling, leading to the release of cytokines such as IL-12, TNF, and IL-1β from immune cells infected with type II parasites. Prior work also suggested that GRA15 promotes early host control of parasites in vivo, but the effect of GRA15 on parasite persistence in the brain and the peripheral immune response has not been well defined. For this reason, we sought to address this gap by generating a new IIΔgra15 strain and comparing outcomes at 3 weeks post infection between WT and IIΔgra15 infected mice. We found that the brain parasite burden and the number of macrophages/microglia and T cells in the brain did not differ between WT and IIΔgra15 infected mice. In addition, while IIΔgra15 infected mice had a lower number and frequency of splenic M1-like macrophages and frequency of PD-1+ CTLA-4+ CD4+ T cells and NK cells compared to WT infected mice, the IFN-γ+ CD4 and CD8 T cell populations were equivalent. In summary, our results suggest that in vivo GRA15 may have a subtle effect on the peripheral immune response, but this effect is not strong enough to alter brain parasite burden or parenchymal immune cell number at 3 weeks post infection.

OBJECTIVE: The unprecedented events of 2020 required a pivot in scientific training to better prepare the biomedical research workforce to address global pandemics, structural racism, and social inequities that devastate human health individually and erode it collectively. Furthermore, this pivot had to be accomplished in the virtual environment given the nation-wide lockdown. METHODS: These needs and context led to leveraging of the San Francisco Building Infrastructure Leading to Diversity (SF BUILD) theories of change to innovate a Virtual BUILD Research Collaboratory (VBRC). The purpose of VBRC was to train Black, Indigenous, and people of color (BIPOC) students to apply their unique perspectives to biomedical research. These training activities were evaluated using a pre-post survey design that included both validated and new psychosocial scales. A new scale was piloted to measure culturally relevant pedagogy. RESULTS: VBRC scholars increased science identity on two items: thinking of myself as a scientist (+1point, p = 0.006) and belonging to a community of scientists (+1point, p = 0.069). Overall, scholars perceived stress also decreased over VBRC (-2.35 points, p = 0.02). Post VBRC, scholars had high agency scores (μ = 11.02, Md = 12, range = 6-12, σ = 1.62) and cultural humility scores (μ = 22.11, Md = 23, range = 12-24, σ = 2.71). No notable race/ethnic differences were found in any measures. CONCLUSIONS: Taken together, our innovative approach to data science training for BIPOC in unprecedented times shows promise for better preparing the workforce critically needed to address the fundamental gaps in knowledge at the intersection of public health, structural racism, and biomedical sciences.

Current models of early human subsistence economies suggest a focus on large mammal hunting. To evaluate this hypothesis, we examine human bone stable isotope chemistry of 24 individuals from the early Holocene sites of Wilamaya Patjxa (9.0-8.7 cal. ka) and Soro Mikaya Patjxa (8.0-6.5 cal. ka) located at 3800 meters above sea level on the Andean Altiplano, Peru. Contrary to expectation, Bayesian mixing models based on the isotope chemistry reveal that plants dominated the diet, comprising 70-95% of the average diet. Paleoethnobotanical data further show that tubers may have been the most prominent subsistence resource. These findings update our understanding of earliest forager economies and the pathway to agricultural economies in the Andean highlands. The findings furthermore suggest that the initial subsistence economies of early human populations adapting to new landscapes may have been more plant oriented than current models suggest.

Campylobacter spp. is often underreported and underrated bacteria that present real health risks to both humans and animals, including non-human primates. It is a commensal microorganism of gastrointestinal tract known to cause gastroenteritis in humans. Commonly found in many wild animals including non-human primates (monkeys- Rhesus macaques) these pathogens are known to be a common cause of diarrhea in humans in many parts of developing and under developed countries. Rhesus macaques from the two holy sites in Kathmandu (Pashupati and Swoyambhu) were included in this cross-sectional study. Diarrheal samples of monkeys were analyzed to detect and characterize the pathogen using 16S rRNA-based PCR screening, followed by DNA sequencing and phylogenetic analysis. Out of a total 67 collected diarrheal samples, Campylobacter spp. were detected in the majority of the samples (n = 64; 96%). DNA sequences of the amplified PCR products were successfully obtained from 13 samples. Phylogenetic analysis identified Candidatus Campylobacter infans (n = 10, Kimura-2 parameter (K2P) pairwise distance values of 0.002287). Remaining three sequences might potentially belong to a novel Campylobacter species/sub-species- closely relating to known species of C. helviticus (K2P pairwise distance of 0.0267). Both Candidatus Campylobacter infans and C. helvitucus are known to infect humans and animals. Additionally, we also detected the bacteria in water and soil samples from the sites. Campylobacter spp. caused the 2018 diarrhea outbreak in Rhesus macaques in the Kathmandu valley. Campylobacter might be one of the important contributing pathogens in diarrheal outbreaks-both in humans and animals (monkeys) in Nepal. Due to close interactions of these animals with humans and other animals, One Health approach might be the most effective way to prevent and mitigate the threat posed by this pathogen.

Background As diagnostic tests for COVID-19 were broadly deployed under Emergency Use Authorization, there emerged a need to understand the real-world utilization and performance of serological testing across the United States. Methods Six health systems contributed electronic health records and/or claims data, jointly developed a master protocol, and used it to execute the analysis in parallel. We used descriptive statistics to examine demographic, clinical, and geographic characteristics of serology testing among patients with RNA positive for SARS-CoV-2. Results Across datasets, we observed 930,669 individuals with positive RNA for SARS-CoV-2. Of these, 35,806 (4%) were serotested within 90 days; 15% of which occurred 30%) in some datasets—limiting our ability to examine differences in serological testing by race. In datasets where race/ethnicity information was available, we observed a greater distribution of White individuals among those serotested; however, the time between RNA and serology tests appeared shorter in Black compared to White individuals. Test manufacturer data was available in half of the datasets contributing to the analysis. Conclusion Our results inform the underlying context of serotesting during the first year of the COVID-19 pandemic and differences observed between claims and EHR data sources–a critical first step to understanding the real-world accuracy of serological tests. Incomplete reporting of race/ethnicity data and a limited ability to link test manufacturer data, lab results, and clinical data challenge the ability to assess the real-world performance of SARS-CoV-2 tests in different contexts and the overall U.S. response to current and future disease pandemics.

Background Real-world performance of COVID-19 diagnostic tests under Emergency Use Authorization (EUA) must be assessed. We describe overall trends in the performance of serology tests in the context of real-world implementation. Methods Six health systems estimated the odds of seropositivity and positive percent agreement (PPA) of serology test among people with confirmed SARS-CoV-2 infection by molecular test. In each dataset, we present the odds ratio and PPA, overall and by key clinical, demographic, and practice parameters. Results A total of 15,615 people were observed to have at least one serology test 14–90 days after a positive molecular test for SARS-CoV-2. We observed higher PPA in Hispanic (PPA range: 79–96%) compared to non-Hispanic (60–89%) patients; in those presenting with at least one COVID-19 related symptom (69–93%) as compared to no such symptoms (63–91%); and in inpatient (70–97%) and emergency department (93–99%) compared to outpatient (63–92%) settings across datasets. PPA was highest in those with diabetes (75–94%) and kidney disease (83–95%); and lowest in those with auto-immune conditions or who are immunocompromised (56–93%). The odds ratios (OR) for seropositivity were higher in Hispanics compared to non-Hispanics (OR range: 2.59–3.86), patients with diabetes (1.49–1.56), and obesity (1.63–2.23); and lower in those with immunocompromised or autoimmune conditions (0.25–0.70), as compared to those without those comorbidities. In a subset of three datasets with robust information on serology test name, seven tests were used, two of which were used in multiple settings and met the EUA requirement of PPA ≥87%. Tests performed similarly across datasets. Conclusion Although the EUA requirement was not consistently met, more investigation is needed to understand how serology and molecular tests are used, including indication and protocol fidelity. Improved data interoperability of test and clinical/demographic data are needed to enable rapid assessment of the real-world performance of in vitro diagnostic tests.

Eutrophic conditions in estuaries are a globally important stressor to coastal ecosystems and have been suggested as a driver of coastal salt marsh loss. Potential mechanisms in marshes include disturbance caused by macroalgae accumulations, enhanced soil sulfide levels linked to high labile carbon inputs, accelerated decomposition, and declines in belowground biomass that contribute to edge instability, erosion, and slumping. However, results of fertilization studies have been mixed, and it is unclear the extent to which local environmental conditions, such as soil composition and nutrient profiles, help shape the response of salt marshes to nutrient exposure. In this study, we characterized belowground productivity and decomposition, organic matter mineralization rates, soil respiration, microbial biomass, soil humification, carbon and nitrogen inventories, nitrogen isotope ratios, and porewater profiles at high and low marsh elevations across eight marshes in four estuaries in California and New York that have strong contrasts in nutrient inputs. The higher nutrient load marshes were characterized by faster carbon turnover, with higher belowground production and decomposition and greater carbon dioxide efflux than lower nutrient load marshes. These patterns were robust across marshes of the Atlantic and Pacific coasts that varied in plant species composition, soil flooding patterns, and soil texture. Although impacts of eutrophic conditions on carbon cycling appeared clear, it was ambiguous whether high nutrient loads are causing negative effects on long-term marsh sustainability in terms of studied metrics. While high nutrient exposure marshes had high rates of decomposition and soil respiration rates, high nutrient exposure was also associated with increased belowground production, and reduced levels of sulfides, which should lead to greater marsh sustainability. While this study does not resolve the extent to which nutrient loads are negatively affecting these salt marshes, we do highlight functional differences between Atlantic and Pacific wetlands which may be useful for understanding coastal marsh health and integrity.

Current models of early human subsistence economies suggest a focus on large mammal hunting. To evaluate this hypothesis, we examine human bone stable isotope chemistry of 24 individuals from the early Holocene sites of Wilamaya Patjxa (9.0-8.7 cal. ka) and Soro Mik'aya Patjxa (8.0-6.5 cal. ka) located at 3800 meters above sea level on the Andean Altiplano, Peru. Contrary to expectation, Bayesian mixing models based on the isotope chemistry reveal that plants dominated the diet, comprising 70-95% of the average diet. Paleoethnobotanical data further show that tubers may have been the most prominent subsistence resource. These findings update our understanding of earliest forager economies and the pathway to agricultural economies in the Andean highlands. The findings furthermore suggest that the initial subsistence economies of early human populations adapting to new landscapes may have been more plant oriented than current models suggest.

The SARS-CoV-2 pandemic has caused unprecedented worldwide infections from persistent mutant variants with various degrees of infectivity and virulence. The elusiveness of a highly penetrant, worldwide vaccination strategy suggests that the complete eradication of SARS-CoV-2 is unlikely. Even with the advent of new antiviral agents, the disease burden worldwide continues to exceed current preventative and therapeutic strategies. Greater interest has been placed towards the development of affordable,broadly effective antiviral therapeutics. Here, we report that the small branched-chain fatty acid Valproic acid (VPA), approved for maintenance of seizure and bipolar disorder, has a novel anti- coronavirus activity that can be augmented with the addition of a long-chain, polyunsaturated omega-3 fatty acid, Docosahexaenoic acid (DHA). An EMR-based epidemiological study of patients tested for COVID-19 demonstrated a correlation exists between a reduced infection rate in patients treated withVPA of up to 25%, as well as a decreased risk of emergency room visits, hospitalization, ICU admission,and use of mechanical ventilation. In vitro studies have demonstrated that VPA modifies gene expression in MRC5 cells. Interestingly, VPA correlates with the inhibition of several SARS-CoV2 interacting genes and the greater inhibition of alpha-coronavirus HCoV-229E (a "common cold" virus) and SARS-CoV2. The VPA-DHA combination activates pre-existing intracellular antiviral mechanisms normally repressed by coronaviruses. Gene expression profiles demonstrate subtle differences in overall gene expression between VPA-treated and VPA-DHA-treated cells. HCoV-229E infection caused an intensely different response with a marked induction of multiple intracellular inflammatory genes. Changes in gene expression took at least 24 hours to manifest and most likely why prior drug screens failed to identify any antiviral VPA activity despite in silico predictions. This report demonstrates an interaction between HDAC inhibition and the potent activation of cellular antiviral responses. A foundation now exists for a low-cost, highly effective antiviral strategy when supplemented with DHA.

BackgroundThis systematic review aimed to evaluate the performance of machine learning (ML) models in predicting post-treatment survival and disease progression outcomes, including recurrence and metastasis, in head and neck cancer (HNC) using clinicopathological structured data.MethodsA systematic search was conducted across the Medline, Scopus, Embase, Web of Science, and Google Scholar databases. The methodological characteristics and performance metrics of studies that developed and validated ML models were assessed. The risk of bias was evaluated using the Prediction model Risk Of Bias ASsessment Tool (PROBAST).ResultsOut of 5,560 unique records, 34 articles were included. For survival outcome, the ML model outperformed the Cox proportional hazards model in time-to-event analyses for HNC, with a concordance index of 0.70-0.79 vs. 0.66-0.76, and for all sub-sites including oral cavity (0.73-0.89 vs. 0.69-0.77) and larynx (0.71-0.85 vs. 0.57-0.74). In binary classification analysis, the area under the receiver operating characteristics (AUROC) of ML models ranged from 0.75-0.97, with an F1-score of 0.65-0.89 for HNC; AUROC of 0.61-0.91 and F1-score of 0.58-0.86 for the oral cavity; and AUROC of 0.76-0.97 and F1-score of 0.63-0.92 for the larynx. Disease-specific survival outcomes showed higher performance than overall survival outcomes, but the performance of ML models did not differ between three- and five-year follow-up durations. For disease progression outcomes, no time-to-event metrics were reported for ML models. For binary classification of the oral cavity, the only evaluated subsite, the AUROC ranged from 0.67 to 0.97, with F1-scores between 0.53 and 0.89.ConclusionsML models have demonstrated considerable potential in predicting post-treatment survival and disease progression, consistently outperforming traditional linear models and their derived nomograms. Future research should incorporate more comprehensive treatment features, emphasize disease progression outcomes, and establish model generalizability through external validations and the use of multicenter datasets.

Foster care programs at animal shelters have emerged as an important tool for promoting animal welfare and supporting shelter life-saving efforts. Preliminary evidence suggests that foster caregiving may also be health-promoting for humans. The goals of this study were to investigate the experiences of foster caregivers at animal shelters based on measures of positive and negative affect, quality of life, and grief, and to describe human-animal attachment among foster populations. Between March 2022 and 2023, 131 foster caregivers from five shelters in the United States completed three online surveys before, during and after providing foster care to a shelter animal. Positive affect decreased significantly from baseline to post-foster (F = 5.71, p

Fibrotic remodeling is the primary driver of functional loss in chronic kidney disease, with no specific anti-fibrotic agent available for clinical use. Transglutaminase 2 (TG2), a wound response enzyme that irreversibly crosslinks extracellular matrix proteins causing dysregulation of extracellular matrix turnover, is a well-characterized anti-fibrotic target in the kidney. We describe the humanization and characterization of two anti-TG2 monoclonal antibodies (zampilimab [hDC1/UCB7858] and BB7) that inhibit crosslinking by TG2 in human in vitro and rabbit/cynomolgus monkey in vivo models of chronic kidney disease. Determination of zampilimab half-maximal inhibitory concentration (IC50) against recombinant human TG2 was undertaken using the KxD assay and determination of dissociation constant (Kd) by surface plasmon resonance. Efficacy in vitro was established using a primary human renal epithelial cell model of tubulointerstitial fibrosis, to assess mature deposited extracellular matrix proteins. Proof of concept in vivo used a cynomolgus monkey unilateral ureteral obstruction model of chronic kidney disease. Zampilimab inhibited TG2 crosslinking transamidation activity with an IC50 of 0.25 nM and Kd of 50%, with no safety signals. Our data support the clinical investigation of zampilimab for the treatment of kidney fibrosis.

The role that inhaled particulate matter plays in the development of post-deployment lung disease among US service members deployed to Southwest Asia during the Global War on Terrorism has been difficult to define. There is a persistent gap in data addressing the relationship between relatively short-term (months to a few years) exposures to high levels of particulate matter during deployment and the subsequent development of adverse pulmonary outcomes. Surgical lung biopsies from deployed service members and veterans (DSMs) and non-deployed service members and veterans (NDSMs) who develop lung diseases can be analyzed to potentially identify residual deployment-specific particles and develop associations with pulmonary pathological diagnoses. We examined 52 surgical lung biopsies from 25 DSMs and 27 NDSMs using field emission scanning electron microscopy (FE-SEM) with energy dispersive x-ray spectroscopy (EDS) to identify any between-group differences in the number and composition of retained inorganic particles, then compared the particle analysis results with the original histopathologic diagnoses. We recorded a higher number of total particles in biopsies from DSMs than from NDSMs, and this difference was mainly accounted for by geologic clays (illite, kaolinite), feldspars, quartz/silica, and titanium-rich silicate mixtures. Biopsies from DSMs deployed to other Southwest Asia regions (SWA-Other) had higher particle counts than those from DSMs primarily deployed to Iraq or Afghanistan, due mainly to illite. Distinct deployment-specific particles were not identified. Particles did not qualitatively associate with country of deployment. The individual diagnoses of the DSMs and NDSMs were not associated with elevated levels of total particles, metals, cerium oxide, or titanium dioxide particles. These results support the examination of particle-related lung disease in DSMs in the context of comparison groups, such as NDSMs, to assist in determining the strength of associations between specific pulmonary pathology diagnoses and deployment-specific inorganic particulate matter exposure.

Toxoplasma gondii is an intracellular parasite that establishes a long-term infection in the brain of many warm-blooded hosts, including humans and rodents. Like all obligate intracellular microbes, Toxoplasma uses many effector proteins to manipulate the host cell to ensure parasite survival. While some of these effector proteins are universal to all Toxoplasma strains, some are polymorphic between Toxoplasma strains. One such polymorphic effector is GRA15. The gra15 allele carried by type II strains activates host NF-κB signaling, leading to the release of cytokines such as IL-12, TNF, and IL-1β from immune cells infected with type II parasites. Prior work also suggested that GRA15 promotes early host control of parasites in vivo, but the effect of GRA15 on parasite persistence in the brain and the peripheral immune response has not been well defined. For this reason, we sought to address this gap by generating a new IIΔgra15 strain and comparing outcomes at 3 weeks post infection between WT and IIΔgra15 infected mice. We found that the brain parasite burden and the number of macrophages/microglia and T cells in the brain did not differ between WT and IIΔgra15 infected mice. In addition, while IIΔgra15 infected mice had a lower number and frequency of splenic M1-like macrophages and frequency of PD-1+ CTLA-4+ CD4+ T cells and NK cells compared to WT infected mice, the IFN-γ+ CD4 and CD8 T cell populations were equivalent. In summary, our results suggest that in vivo GRA15 may have a subtle effect on the peripheral immune response, but this effect is not strong enough to alter brain parasite burden or parenchymal immune cell number at 3 weeks post infection.

BackgroundNewborn bloodspot screening is a well-established population health initiative that detects serious, childhood-onset, treatable conditions to improve health outcomes. With genomic technologies advancing rapidly, many countries are actively discussing the introduction of genomic assays into newborn screening programs. While adding genomic testing to Australia's newborn screening program could improve outcomes for infants and families, it must be considered against potential harms, ethical, legal, equity and social implications, and economic and health system impacts. We must ask not only 'can' we use genomics to screen newborns?' but 'should we'?' and 'how much should health systems invest in genomic newborn screening?'.MethodsThis study will use qualitative methods to explore understanding, priorities, concerns and expectations of genomic newborn screening among parents/carers, health professionals/scientists, and health policy makers across Australia. In-depth, semi-structured interviews will be held with 30-40 parents/carers recruited via hospital and community settings, 15-20 health professionals/scientists, and 10-15 health policy makers. Data will be analysed using inductive content analysis. The Sydney Children's Hospital Network Human Research Ethics Committee approved this study protocol [2023/ETH02371]. The Standards for Reporting Qualitative Research will guide study planning, conduct and reporting.DiscussionFew studies have engaged a diverse range of stakeholders to explore the implications of genomics in newborn screening in a culturally and genetically diverse population, nor in a health system underpinned by universal health care. As the first study within a multi-part research program, findings will be used to generate new knowledge on the risks and benefits and importance of ethical, legal, social and equity implications of genomic newborn screening from the perspective of key stakeholders. As such it will be the foundation on which child and family centered criteria can be developed to inform health technology assessments and drive efficient and effective policy decision-making on the implementation of genomics in newborn screening.

ObjectiveThe unprecedented events of 2020 required a pivot in scientific training to better prepare the biomedical research workforce to address global pandemics, structural racism, and social inequities that devastate human health individually and erode it collectively. Furthermore, this pivot had to be accomplished in the virtual environment given the nation-wide lockdown.MethodsThese needs and context led to leveraging of the San Francisco Building Infrastructure Leading to Diversity (SF BUILD) theories of change to innovate a Virtual BUILD Research Collaboratory (VBRC). The purpose of VBRC was to train Black, Indigenous, and people of color (BIPOC) students to apply their unique perspectives to biomedical research. These training activities were evaluated using a pre-post survey design that included both validated and new psychosocial scales. A new scale was piloted to measure culturally relevant pedagogy.ResultsVBRC scholars increased science identity on two items: thinking of myself as a scientist (+1point, p = 0.006) and belonging to a community of scientists (+1point, p = 0.069). Overall, scholars perceived stress also decreased over VBRC (-2.35 points, p = 0.02). Post VBRC, scholars had high agency scores (μ = 11.02, Md = 12, range = 6-12, σ = 1.62) and cultural humility scores (μ = 22.11, Md = 23, range = 12-24, σ = 2.71). No notable race/ethnic differences were found in any measures.ConclusionsTaken together, our innovative approach to data science training for BIPOC in unprecedented times shows promise for better preparing the workforce critically needed to address the fundamental gaps in knowledge at the intersection of public health, structural racism, and biomedical sciences.

The material properties of some bones are known to vary with anatomical location, orientation and position within the bone (e.g., cortical and trabecular bone). Details of the heterogeneity and anisotropy of bone is an important consideration for biomechanical studies that apply techniques such as finite element analysis, as the outcomes will be influenced by the choice of material properties used. Datasets detailing the regional variation of material properties in the bones of the skull are sparse, leaving many finite element analyses of skulls no choice but to employ homogeneous, isotropic material properties, often using data from a different species to the one under investigation. Due to the growing significance of investigating the cranial biomechanics of the rabbit in basic science and clinical research, this study used nanoindentation to measure the elastic modulus of cortical and trabecular bone throughout the skull. The elastic moduli of cortical bone measured in the mediolateral and ventrodorsal direction were found to decrease posteriorly through the skull, while it was evenly distributed when measured in the anteroposterior direction. Furthermore, statistical tests showed that the variation of elastic moduli between separate regions (anterior, middle and posterior) of the skull were significantly different in cortical bone, but was not in trabecular bone. Elastic moduli measured in different orthotropic planes were also significantly different, with the moduli measured in the mediolateral direction consistently lower than that measured in either the anteroposterior or ventrodorsal direction. These findings demonstrate the significance of regional and directional variation in cortical bone elastic modulus, and therefore material properties in finite element models of the skull, particularly those of the rabbit, should consider the heterogeneous and orthotropic properties of skull bone when possible.

BackgroundCommunity-based out-of-hours services are an integral component of end-of-life care. However, there is little understanding of how patients and families utilise these services. This additional analysis of a systematic review aims to understand and identify patterns of out-of-hours service use and produce recommendations for future service design.MethodData on service use was extracted and secondary analysis undertaken, from a systematic review of models of community out-of-hours services. Narrative synthesis was completed, addressing four specific aspects of service use: 1.Times when patients/families/healthcare professionals need to contact out-of-hours services; 2. Who contacts out-of-hours services; 3. Whether a telephone call, centre visit or home visit is provided; 4. Who responds to out-of-hours calls.ResultsCommunity-based out-of-hours palliative care services were most often accessed between 5pm and midnight, especially on weekdays (with reports of 69% of all calls being made out-of-hours). Family members and carers were the most frequent callers to of the services (making between 60% and 80% of all calls). The type of contact (telephone, centre visit or home visit) varied based on what was offered and on patient need. Over half of services were led by a single discipline (nurse).ConclusionsOut-of-hours services are highly used up to midnight, and particularly by patients' family and carers. Recommendations to commissioners and service providers are to: • Increase provision of out-of-hours services between 5pm and midnight to reflect the increased use at these times. • Ensure that family and carers are provided with clear contact details for out-of-hours support. • Ensure patient records can be easily accessed by health professionals responding to calls, making the triage process easier. • Listen to patients, family and carers in the design of out-of-hours services, including telephone services. • Collect data systematically on out-of-hours-service use and on outcomes for patients who use the service.

BackgroundPulmonary exacerbations in alpha-1 antitrypsin deficiency (AATD) related lung disease are a significant contributor to disease burden, as with usual COPD. Separating the early stages of an exacerbation from the day-to-day variation in stable COPD is central to the concerns of both clinicians and patients and has been identified as a research priority by NIHR. Clinical tools that distinguish baseline symptoms from those of an exacerbation could allow early and appropriate treatment of AECOPD to reduce the impact and potentially may slow disease progression thereby improving survival and quality of life. Candidate tools include symptom diaries and biomarkers of infection and acute inflammation. Urinary biomarkers of AECOPD have yet to be explored in AATD related COPD.Methods55 patients with AATD related lung disease with a history of 2 or more AECOPD in the preceding year were prospectively followed for 18 months. Each patient recorded symptom scores daily via an electronic symptom diary (eDiary) based on Bronkotest. Urinary biomarkers for AAT, NE, CRP, TIMP1 and desmosine were measured weekly using a home urinary lateral flow device. During self-reported AECOPD patients were asked to perform urine analysis on the first 7 consecutive days.ResultsType I Anthonisen exacerbations and episodes occurring in autumn/winter lasted longer than Type II/III exacerbations and spring/summer episodes respectively. Median urinary CRP concentration across all study participants increased during Type I AECOPD. eDiary adherence was 68% over a median of 17.8 months (IQR 15.7 to 18.5).ConclusionsUse of an eDiary and urinary biomarkers to detect and characterise AECOPD remotely in AATD related lung disease is feasible over a prolonged period and paves the way for precision detection of exacerbations.

African Americans are disproportionately affected by HIV and socio-structural barriers that impact antiretroviral (ART) adherence. Two-way text-messaging interventions have shown promise in supporting adherence in US studies of mostly White people living with HIV (PLWH). However, culturally-appropriate tailoring is necessary to maximize intervention effectiveness among other racial/ethnic groups. Thus, to refine an existing text-messaging intervention, we examined barriers and facilitators to ART adherence among African Americans and perspectives on features to integrate into the extant intervention. Three focus groups, two with African American PLWH (n = 5 and n = 7) and one with providers of care (n = 11) were conducted; transcripts of audio-recordings were thematically analyzed. Adherence supports operated at individual, interpersonal, and structural/environmental levels (e.g., using reminders and pill organizers, wanting to protect partners from HIV, and positive interactions with providers). Adherence barriers also operated at multiple ecological levels (e.g., poor mental health, fear of disclosure of HIV status, and unstable housing). Participant-suggested features for refinement included: i) matching content to participants' comfort with receiving messages referencing HIV or medication-taking, ii) culturally-tailoring content for African Americans, iii) tracking adherence, and iv) encouraging adherence interactions between patients and providers. Feedback from both patients and providers is foundational to designing effective ART interventions among African American PLWH.

Oligosaccharides are important components of milk, serving as substrates for the intestinal microbiota, acting as antimicrobials that prevent pathogen colonization, and supporting the developing gastrointestinal immune system of neonates. Nutrient composition of canine and feline milk samples has been described previously, but little is known about the oligosaccharide content. Therefore, the objective of this study was to characterize canine and feline milk samples using a high-throughput glycomics approach. 23 dogs (9 Labrador retriever and 14 Labrador retriever x golden retriever crossbreed) and 6 domestic shorthair cats were recruited to the study. Milk samples were collected by manual expression at time points after parturition. Samples were collected across 2 phases per species, differentiated by maternal diet. Following extraction, oligosaccharide content was determined by liquid chromatography-mass spectrometry (LC-MS). In canine milk samples, 3 structures accounted for over 90% of all oligosaccharides detected across two diet groups. These were 3'-sialyllactose, 6'-sialyllactose, and 2'-fucosyllactose. In feline samples, a more diverse range of oligosaccharides was detected, with up to 16 structures present at relative abundance >1% of the total. Difucosyllactose-N-hexaose b, 3'-sialyllactose and lacto-N-neohexaose were all detected at abundances >10% in feline milk samples. Statistically significant differences (p

Alternative splicing is a co-transcriptional mechanism that generates protein diversity by including or excluding exons in different combinations, thereby expanding the diversity of protein isoforms of a single gene. Abnormalities in this process can result in deleterious effects to human health, and several xenobiotics are known to interfere with splicing regulation through multiple mechanisms. These changes could lead to human diseases such as cancer, neurological disorders, autoimmune diseases, and developmental disorders. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is an environmental contaminant generated as a byproduct of various industrial activities. Exposure to this dioxin has been linked to a wide range of pathologies through the alteration of multiple cellular processes. However, the effects of TCDD exposure on alternative splicing have not yet been studied. Here, we investigated whether a single po. dose of 5 μg/kg or 500 μg/kg TCDD influence hepatic alternative splicing in adult male C57BL/6Kou mouse. We identified several genes whose alternative splicing of precursor messenger RNAs was modified following TCDD exposure. In particular, we demonstrated that alternative splicing of Cyp1a1, Ahrr, and Actn1 was significantly altered after TCDD treatment. These findings show that the exposure to TCDD has an impact on alternative-splicing, and suggest a new avenue for understanding TCDD-mediated toxicity and pathogenesis.

IntroductionPeople affected by leprosy are at increased risk of impairments and deformities from peripheral nerve damage. This mostly occurs if diagnosis and treatment is delayed and contributes to continued transmission within the community. Champa district of Chhattisgarh state in India is an endemic area with the highest national annual case detection and disability rates for leprosy. The Replicability Model is a system strengthening intervention implemented by the Leprosy Mission Trust India in Champa that aims to promote early diagnosis and treatment of leprosy, improve on-going management of the effects of leprosy and improve welfare for the people affected by leprosy. This protocol presents a plan to describe the overall implementation of the Replicability Model and describe the barriers and facilitators encountered in the process. We will also quantify the effect of the program on one of its key aims- early leprosy diagnosis.MethodsThe replicability model will be implemented over four years, and the work described in this protocol will be conducted in the same timeframe. We have two Work Packages (WPs). In WP1, we will conduct a process evaluation. This will include three methods i) observations of replicability model implementation teams' monthly meetings ii) key informant interviews (n = 10) and interviews with stakeholders (n = 30) iii) observations of key actors (n = 15). Our purpose is to describe the implementation process and identify barriers and facilitators to successful implementation. WP2 will be a quantitative study to track existing and new cases of leprosy using routinely collected data. If the intervention is successful, we expect to see an increase in cases (with a higher proportion detected at an early clinical stage) followed by a decrease in total cases.ConclusionThis study will enable us to improve and disseminate the Replicability Model by identifying factors that promote success. It will also identify its effectiveness in fulfilling one of its aims: reducing the incidence of leprosy by finding and tracking cases at an earlier stage in the disease.

Background"Choosing All Together" (CHAT), is a community engagement tool designed to give the public a voice in how best to allocate limited resources to improve population health. This process evaluation explored the mechanisms through which CHAT generates community engagement.MethodThe CHAT tool was adapted and implemented for use in two rural communities (Nanoro, Burkina Faso, and Navrongo, Ghana) and one urban township (Soweto, South Africa) to prioritize maternal and child nutrition interventions. Community discussions were audio-recorded, transcribed, and translated into English. Twenty-two transcripts, including six each from Navrongo and Soweto and 10 from Nanoro, were analysed thematically to generate data driven codes and themes to explain mechanisms underlying the CHAT process. The process evaluation was based on the UK MRC process evaluation guidance.ResultsSeven themes describing the functions and outcomes of CHAT were identified. Themes described participants deliberating trade-offs, working together, agreeing on priorities, having a shared vision, and increasing their knowledge, also the skills of the facilitator, and a process of power sharing between participants and researchers. Participants came to an agreement of priorities when they had a shared vision. Trained facilitators are important to facilitate meaningful discussion between participants and those with lower levels of literacy to participate fully.ConclusionCHAT has been shown to be adaptable and useful in prioritising maternal and child nutrition interventions in communities in Burkina Faso, Ghana, and South Africa. Conducting CHAT in communities over a longer period and involving policy-makers would increase trust, mutual respect and develop partnerships.

Native ponies are at increased risk of obesity and metabolic perturbations, such as insulin dysregulation (ID), a key risk factor for endocrinopathic laminitis. Management and feeding practices can be adapted to maintain healthy body condition and support metabolic health, but owners may inadvertently provide their ponies with inappropriate management leading to obesity and exacerbating risk of metabolic disease. Adoption of preventative weight management approaches (WMAs), including regular monitoring of body condition, providing appropriate preserved forage, promoting seasonal weight loss, and using exercise accordingly, are key in supporting native ponies' metabolic health. The factors influencing the adoption of WMAs, such as owners' experience and confidence, require exploration. The aim of the current study was to understand factors influencing owners' likelihood to undertake certain WMAs, to develop our understanding of suitable intervention targets. A total of 571 responses to an online cross-sectional questionnaire were analysed. Mediation analysis revealed that whilst long term (≥20 years) experience caring for native ponies was associated with owners increased, self-reported confidence in identifying disease and managing their native ponies, this did not translate to an increased likelihood of implementing WMAs. Conversely, respondents who managed ponies with dietary requirements related to obesity, laminitis, or equine metabolic syndrome were more likely to use WMAs related to feeding, seasonal weight management and exercise. Owner confidence was assessed and rejected as a mediator of the relationship between experience and WMA use. These results highlight the need for further work that elucidates the pathways leading owners to undertake action against obesity without the need for ponies to develop overt disease, as well as suggesting a need for long term managers of native ponies to update management practices with preventative care as the focus.

New methodologies have enabled viable sap yields from juvenile sugar maple trees. To further improve yields, a better understanding of sap exudation is required. To achieve this, the anatomy of the xylem must first be fully characterised. We examine juvenile maple saplings using light optical microscopy (LOM) and scanning electron microscopy (SEM), looking at sections cut along differing orientations as well as macerations. From this we measure various cell parameters. We find diameter and length of vessel elements to be 28 ± 8 μm and 200 ± 50 μm, for fibre cells 8 ± 3 μm and 400 ± 100 μm, and for ray parenchyma cells 8 ± 2 μm and 50 ± 20 μm. We also examine pitting present on different cell types. On vessel elements we observe elliptical bordered pits connecting to other vessel elements (with major axis of 2.1 ± 0.7 μm and minor 1.3 ± 0.3 μm) and pits connecting to ray parenchyma (with major axis of 4 ± 2 μm and minor 2.0 ± 0.7 μm). We observe two distinct pit sizes on fibres with circular pits 0.7 ± 0.2 μm in diameter and ellipsoidal pits 1.6 ± 0.4 μm by 1.0 ± 0.3 μm. We do not observe distinct pitting patterns on different fibre types. The various cell and pit measurements obtained generally agree with the limited data available for mature trees, with the exception of vessel element and fibre length, both of which were significantly smaller than reported values.

Seasonal peaks in infectious disease incidence put pressures on health services. Therefore, early warning of the timing and magnitude of peak activity during seasonal epidemics can provide information for public health practitioners to take appropriate action. Whilst many infectious diseases have predictable seasonality, newly emerging diseases and the impact of public health interventions can result in unprecedented seasonal activity. We propose a Machine Learning process for generating short-term forecasts, where models are selected based on their ability to correctly forecast peaks in activity, and can be useful during atypical seasons. We have validated our forecasts using typical and atypical seasonal activity, using respiratory syncytial virus (RSV) activity during 2019-2021 as an example. During the winter of 2020/21 the usual winter peak in RSV activity in England did not occur but was 'deferred' until the Spring of 2021. We compare a range of Machine Learning regression models, with alternate models including different independent variables, e.g. with or without seasonality or trend variables. We show that the best-fitting model which minimises daily forecast errors is not the best model for forecasting peaks when the selection criterion is based on peak timing and magnitude. Furthermore, we show that best-fitting models for typical seasons contain different variables to those for atypical seasons. Specifically, including seasonality in models improves performance during typical seasons but worsens it for the atypical seasons.

Many publications lack sufficient background information (e.g. location) to be interpreted, replicated, or reused for synthesis. This impedes scientific progress and the application of science to practice. Reporting guidelines (e.g. checklists) improve reporting standards. They have been widely taken up in the medical sciences, but not in ecological and agricultural research. Here, we use a community-centred approach to develop a reporting checklist (AgroEcoList 1.0) through surveys and workshops with 23 experts and the wider agroecological community. To put AgroEcoList in context, we also assessed the agroecological community's perception of reporting standards in agroecology. A total of 345 researchers, reviewers, and editors, responded to our survey. Although only 32% of respondents had prior knowledge of reporting guidelines, 76% of those that had said guidelines improved reporting standards. Overall, respondents agreed on the need of AgroEcolist 1.0; only 24% of respondents had used reporting guidelines before, but 78% indicated they would use AgroEcoList 1.0. We updated AgroecoList 1.0 based on respondents' feedback and user-testing. AgroecoList 1.0 consists of 42 variables in seven groups: experimental/sampling set-up, study site, soil, livestock management, crop and grassland management, outputs, and finances. It is presented here, and is also available on github (https://github.com/AgroecoList/Agroecolist). AgroEcoList 1.0 can serve as a guide for authors, reviewers, and editors to improve reporting standards in agricultural ecology. Our community-centred approach is a replicable method that could be adapted to develop reporting checklists in other fields. Reporting guidelines such as AgroEcoList can improve reporting standards and therefore the application of research to practice, and we recommend that they are adopted more widely in agriculture and ecology.

IntroductionThis study aimed to determine whether the COVID-19 pandemic had an impact on essential primary healthcare services at public primary healthcare facilities.MethodsThe number of weekly consultations for antenatal care (ANC), outpatient (OPD), immunisations (EPI), family planning (FP) and HIV services, between January 2018 and December 2020, were collected from 25 facilities in Masaka district, Uganda, 21 in Goma, and 29 in Kambia district, Sierra Leone. Negative binomial regression models accounting for clustering and season were used to analyse changes in activity levels between 2018, 2019 and 2020.ResultsIn Goma, we found no change in OPD, EPI or ANC consultations, FP was 17% lower in March-July 2020 compared to 2019, but this recovered by December 2020. New diagnoses of HIV were 34% lower throughout 2020 compared to 2019. In Sierra Leone, compared to the same periods in 2019, facilities had 18-29% fewer OPD consultations throughout 2020, and 27% fewer DTP3 doses in March-July 2020. There was no evidence of differences in other services. In Uganda there were 20-35% fewer under-5 OPD consultations, 21-66% fewer MCV1 doses, and 48-51% fewer new diagnoses of HIV throughout 2020, compared to 2019. There was no difference in the number of HPV doses delivered.ConclusionsThe level of disruption varied across the different settings and qualitatively appeared to correlate with the strength of lockdown measures and reported attitudes towards the risk posed by COVID-19. Mitigation strategies such as health communications campaigns and outreach services may be important to limit the impact of lockdowns on primary healthcare services.

BackgroundExertional dyspnea and exercise intolerance are frequently endorsed in Veterans of post 9/11 conflicts in Southwest Asia (SWA). Studying the dynamic behavior of ventilation during exercise may provide mechanistic insight into these symptoms. Using maximal cardiopulmonary exercise testing (CPET) to experimentally induce exertional symptoms, we aimed to identify potential physiological differences between deployed Veterans and non-deployed controls.Materials and methodsDeployed (n = 31) and non-deployed (n = 17) participants performed a maximal effort CPET via the Bruce treadmill protocol. Indirect calorimetry and perceptual rating scales were used to measure rate of oxygen consumption ([Formula: see text]), rate of carbon dioxide production ([Formula: see text]), respiratory frequency (f R), tidal volume (VT), minute ventilation ([Formula: see text]), heart rate (HR), perceived exertion (RPE; 6-20 scale), and dyspnea (Borg Breathlessness Scale; 0-10 scale). A repeated measures analysis of variance (RM-ANOVA) model (2 groups: deployed vs non-deployed X 6 timepoints: 0%, 20%, 40%, 60%, 80%, and 100% [Formula: see text]) was conducted for participants meeting valid effort criteria (deployed = 25; non-deployed = 11).ResultsSignificant group (η2partial = 0.26) and interaction (η2partial = 0.10) effects were observed such that deployed Veterans exhibited reduced f R and a greater change over time relative to non-deployed controls. There was also a significant group effect for dyspnea ratings (η2partial = 0.18) showing higher values in deployed participants. Exploratory correlational analyses revealed significant associations between dyspnea ratings and fR at 80% (R2 = 0.34) and 100% (R2 = 0.17) of [Formula: see text], but only in deployed Veterans.ConclusionRelative to non-deployed controls, Veterans deployed to SWA exhibited reduced fR and greater dyspnea during maximal exercise. Further, associations between these parameters occurred only in deployed Veterans. These findings support an association between SWA deployment and affected respiratory health, and also highlight the utility of CPET in the clinical evaluation of deployment-related dyspnea in Veterans.

The alteration of environmental conditions has two major outcomes on the demographics of living organisms: population decline of the common species and extinction of the rarest ones. Halting the decline of abundant species as well as the erosion of biodiversity require solutions that may be mismatched, despite being rooted in similar causes. In this study, we demonstrate how rank abundance distribution (RAD) models are mathematical representations of a dominance-diversity dilemma. Across 4,375 animal communities from a range of taxonomic groups, we found that a reversed RAD model correctly predicts species richness, based solely on the relative dominance of the most abundant species in a community and the total number of individuals. Overall, predictions from this RAD model explained 69% of the variance in species richness, compared to 20% explained by simply regressing species richness on the relative dominance of the most abundant species. Using the reversed RAD model, we illustrate how species richness is co-limited by the total abundance of a community and the relative dominance of the most common species. Our results highlight an intrinsic trade-off between species richness and dominance that is present in the structure of RAD models and real-world animal community data. This dominance-diversity dilemma suggests that withdrawing individuals from abundant populations might contribute to the conservation of species richness. However, we posit that the positive effect of harvesting on biodiversity is often offset by exploitation practices with negative collateral consequences, such as habitat destruction or species bycatches.

Campylobacter spp. is often underreported and underrated bacteria that present real health risks to both humans and animals, including non-human primates. It is a commensal microorganism of gastrointestinal tract known to cause gastroenteritis in humans. Commonly found in many wild animals including non-human primates (monkeys- Rhesus macaques) these pathogens are known to be a common cause of diarrhea in humans in many parts of developing and under developed countries. Rhesus macaques from the two holy sites in Kathmandu (Pashupati and Swoyambhu) were included in this cross-sectional study. Diarrheal samples of monkeys were analyzed to detect and characterize the pathogen using 16S rRNA-based PCR screening, followed by DNA sequencing and phylogenetic analysis. Out of a total 67 collected diarrheal samples, Campylobacter spp. were detected in the majority of the samples (n = 64; 96%). DNA sequences of the amplified PCR products were successfully obtained from 13 samples. Phylogenetic analysis identified Candidatus Campylobacter infans (n = 10, Kimura-2 parameter (K2P) pairwise distance values of 0.002287). Remaining three sequences might potentially belong to a novel Campylobacter species/sub-species- closely relating to known species of C. helviticus (K2P pairwise distance of 0.0267). Both Candidatus Campylobacter infans and C. helvitucus are known to infect humans and animals. Additionally, we also detected the bacteria in water and soil samples from the sites. Campylobacter spp. caused the 2018 diarrhea outbreak in Rhesus macaques in the Kathmandu valley. Campylobacter might be one of the important contributing pathogens in diarrheal outbreaks-both in humans and animals (monkeys) in Nepal. Due to close interactions of these animals with humans and other animals, One Health approach might be the most effective way to prevent and mitigate the threat posed by this pathogen.

Craniofacial defects require a treatment approach that provides both robust tissues to withstand the forces of mastication and high geometric fidelity that allows restoration of facial architecture. When the surrounding soft tissue is compromised either through lack of quantity (insufficient soft tissue to enclose a graft) or quality (insufficient vascularity or inducible cells), a vascularized construct is needed for reconstruction. Tissue engineering using customized 3D printed bioreactors enables the generation of mechanically robust, vascularized bony tissues of the desired geometry. While this approach has been shown to be effective when utilized for reconstruction of non-load bearing ovine angular defects and partial segmental defects, the two-stage approach to mandibular reconstruction requires testing in a large, load-bearing defect. In this study, 5 sheep underwent bioreactor implantation and the creation of a load-bearing mandibular defect. Two bioreactor geometries were tested: a larger complex bioreactor with a central groove, and a smaller rectangular bioreactor that were filled with a mix of xenograft and autograft (initial bone volume/total volume BV/TV of 31.8 ± 1.6%). At transfer, the tissues generated within large and small bioreactors were composed of a mix of lamellar and woven bone and had BV/TV of 55.3 ± 2.6% and 59.2 ± 6.3%, respectively. After transfer of the large bioreactors to the mandibular defect, the bioreactor tissues continued to remodel, reaching a final BV/TV of 64.5 ± 6.2%. Despite recalcitrant infections, viable osteoblasts were seen within the transferred tissues to the mandibular site at the end of the study, suggesting that a vascularized customized bony flap is a potentially effective reconstructive strategy when combined with an optimal stabilization strategy and local antibiotic delivery prior to development of a deep-seated infection.

BackgroundAs diagnostic tests for COVID-19 were broadly deployed under Emergency Use Authorization, there emerged a need to understand the real-world utilization and performance of serological testing across the United States.MethodsSix health systems contributed electronic health records and/or claims data, jointly developed a master protocol, and used it to execute the analysis in parallel. We used descriptive statistics to examine demographic, clinical, and geographic characteristics of serology testing among patients with RNA positive for SARS-CoV-2.ResultsAcross datasets, we observed 930,669 individuals with positive RNA for SARS-CoV-2. Of these, 35,806 (4%) were serotested within 90 days; 15% of which occurred 30%) in some datasets-limiting our ability to examine differences in serological testing by race. In datasets where race/ethnicity information was available, we observed a greater distribution of White individuals among those serotested; however, the time between RNA and serology tests appeared shorter in Black compared to White individuals. Test manufacturer data was available in half of the datasets contributing to the analysis.ConclusionOur results inform the underlying context of serotesting during the first year of the COVID-19 pandemic and differences observed between claims and EHR data sources-a critical first step to understanding the real-world accuracy of serological tests. Incomplete reporting of race/ethnicity data and a limited ability to link test manufacturer data, lab results, and clinical data challenge the ability to assess the real-world performance of SARS-CoV-2 tests in different contexts and the overall U.S. response to current and future disease pandemics.

The Directed Acyclic Graph (DAG) is a graph representing causal pathways for informing the conduct of an observational study. The use of DAGs allows transparent communication of a causal model between researchers and can prevent over-adjustment biases when conducting causal inference, permitting greater confidence and transparency in reported causal estimates. In the era of 'big data' and increasing number of observational studies, the role of the DAG is becoming more important. Recent best-practice guidance for constructing a DAG with reference to the literature has been published in the 'Evidence synthesis for constructing DAGs' (ESC-DAG) protocol. We aimed to assess adherence to these principles for DAGs constructed within perioperative literature. Following registration on the International Prospective Register of Systematic Reviews (PROSPERO) and with adherence to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) reporting framework for systematic reviews, we searched the Excerpta Medica dataBASE (Embase), the Medical Literature Analysis and Retrieval System Online (MEDLINE) and Cochrane databases for perioperative observational research incorporating a DAG. Nineteen studies were included in the final synthesis. No studies demonstrated any evidence of following the mapping stage of the protocol. Fifteen (79%) fulfilled over half of the translation and integration one stages of the protocol. Adherence with one stage did not guarantee fulfilment of the other. Two studies (11%) undertook the integration two stage. Unmeasured variables were handled inconsistently between studies. Only three (16%) studies included unmeasured variables within their DAG and acknowledged their implication within the main text. Overall, DAGs that were constructed for use in perioperative observational literature did not consistently adhere to best practice, potentially limiting the benefits of subsequent causal inference. Further work should focus on exploring reasons for this deviation and increasing methodological transparency around DAG construction.

BackgroundReal-world performance of COVID-19 diagnostic tests under Emergency Use Authorization (EUA) must be assessed. We describe overall trends in the performance of serology tests in the context of real-world implementation.MethodsSix health systems estimated the odds of seropositivity and positive percent agreement (PPA) of serology test among people with confirmed SARS-CoV-2 infection by molecular test. In each dataset, we present the odds ratio and PPA, overall and by key clinical, demographic, and practice parameters.ResultsA total of 15,615 people were observed to have at least one serology test 14-90 days after a positive molecular test for SARS-CoV-2. We observed higher PPA in Hispanic (PPA range: 79-96%) compared to non-Hispanic (60-89%) patients; in those presenting with at least one COVID-19 related symptom (69-93%) as compared to no such symptoms (63-91%); and in inpatient (70-97%) and emergency department (93-99%) compared to outpatient (63-92%) settings across datasets. PPA was highest in those with diabetes (75-94%) and kidney disease (83-95%); and lowest in those with auto-immune conditions or who are immunocompromised (56-93%). The odds ratios (OR) for seropositivity were higher in Hispanics compared to non-Hispanics (OR range: 2.59-3.86), patients with diabetes (1.49-1.56), and obesity (1.63-2.23); and lower in those with immunocompromised or autoimmune conditions (0.25-0.70), as compared to those without those comorbidities. In a subset of three datasets with robust information on serology test name, seven tests were used, two of which were used in multiple settings and met the EUA requirement of PPA ≥87%. Tests performed similarly across datasets.ConclusionAlthough the EUA requirement was not consistently met, more investigation is needed to understand how serology and molecular tests are used, including indication and protocol fidelity. Improved data interoperability of test and clinical/demographic data are needed to enable rapid assessment of the real-world performance of in vitro diagnostic tests.

The primary aims of this study were to examine the application of maturity status bio-banding within professional soccer academy programmes and understand the methods employed, the intended objectives, and the potential barriers to bio-banding. Using a mixed method design, twenty-five professional soccer academy practitioners completed an online survey designed to examine their perceptions of the influence of maturation on practice, their perceptions and application of bio-banding, and the perceived barriers to the implementation of this method. Frequency and percentages of responses for individual items were calculated. In the next phase of the study, seven participants who had experience with, or knowledge of, the bio-banding process within an academy youth soccer setting were recruited to complete a semi-structured interview. Interview data was transcribed and analysed using a combination of deductive and inductive approaches to identify key themes. The main findings across the two phases of the study were that [1] there is consensus among the practitioners that the individual effect of maturation impacts their ability to accurately assess the soccer competencies, [2] the majority (80%) of the sample had implemented bio-banding, with practitioners showing a clear preference for using the Khamis and Roche method to bio-band players, with the greatest perceived benefit being during maturity-matched formats, specifically for late or post-PHV players, [3] Practitioners perceived that bio-banding enhances their ability to assess academy soccer players, and [4] practitioners who have used bio-banding believe that the method is an effective way of enhancing the perception of challenge thereby providing a number of psycho-social benefits. Findings suggest that a collaborative and multi-disciplinary approach is required to enhance the likelihood of bio-banding being successfully implemented within the typical training schedules across the adolescent phase of the player development pathway.

Determining the most likely source of an invasive pest species might help to improve their management by establishing efficient quarantine measures and heading the search of efficient biological control agents. Planococcus ficus is an invasive mealybug pest of vineyards in Argentina, California, Mexico, Peru and South Africa. This mealybug pest had a previously known geographic distribution spanning southern Europe, the Middle East, and parts of northern Africa. In North America, Pl. ficus was first discovered in the early 1990s and soon thereafter in Mexico. To determine the origin of invasive populations in North America, Pl. ficus from California and Mexico were compared with material throughout its presumptive native range in the Mediterranean region, as well as material collected from an older invasion in South Africa and recently invaded Argentina. From each sample location, genomic DNA was sequenced for the nuclear internal transcribed spacer one (ITS1) and the mitochondrial cytochrome c. oxidase one (CO1). Phylogenetic analyses of CO1, ITS1 and concatenated CO1 and ITS1 data-sets using Bayesian and neighbor-joining analysis support two major divisions: a European grouping (Europe, Tunisia, Turkey) and a Middle Eastern grouping (Israel and Egypt). The invasive populations in Argentina and South Africa align with the European group and the invasive populations in North America align with the Middle Eastern group, with one Israel sample aligning closely with the North American clade, suggesting that Israel was the origin of those populations.

BackgroundEvaluation of impaired left ventricular (LV) filling has focused on intrinsic causes of LV dysfunction; however, pulmonary vascular changes may contribute to reduced LV filling and dyspnea. We hypothesized that lower total pulmonary vascular volume (TPVV) on computed tomography (CT) would be associated with dyspnea and decrements in LV end-diastolic volume, particularly among ever-smokers.MethodsThe Multi-Ethnic Study of Atherosclerosis recruited adults without clinical cardiovascular disease in 2000-02. In 2010-12, TPVV was ascertained as the volume of arteries and veins in the lungs detectable on non-contrast chest CT (vessels ≥1 mm diameter). Cardiac measures were assessed by magnetic resonance imaging (MRI). Dyspnea was self-reported.ResultsOf 2303 participants, 53% had ever smoked cigarettes. Among ever-smokers, a lower TPVV was associated with a lower LV end-diastolic volume (6.9 mL per SD TPVV), stroke volume, and cardiac output and with dyspnea (all P-values

The prevalence of lipids devoid of phosphorus suggests that the availability of phosphorus limits microbial growth and activity in many anoxic, stratified environments. To better understand the response of anaerobic bacteria to phosphate limitation and starvation, this study combines microscopic and lipid analyses with the measurements of fitness of pooled barcoded transposon mutants of the model sulfate reducing bacterium Desulfovibrio alaskensis G20. Phosphate-limited G20 has lower growth rates and replaces more than 90% of its membrane phospholipids by a mixture of monoglycosyl diacylglycerol (MGDG), glycuronic acid diacylglycerol (GADG) and ornithine lipids, lacks polyphosphate granules, and synthesizes other cellular inclusions. Analyses of pooled and individual mutants reveal the importance of the high-affinity phosphate transport system (the Pst system), PhoR, and glycolipid and ornithine lipid synthases during phosphate limitation. The phosphate-dependent synthesis of MGDG in G20 and the widespread occurrence of the MGDG/GADG synthase among sulfate reducing ∂-Proteobacteria implicate these microbes in the production of abundant MGDG in anaerobic environments where the concentrations of phosphate are lower than 10 μM. Numerous predicted changes in the composition of the cell envelope and systems involved in transport, maintenance of cytoplasmic redox potential, central metabolism and regulatory pathways also suggest an impact of phosphate limitation on the susceptibility of sulfate reducing bacteria to other anthropogenic or environmental stresses.

The COVID-19 pandemic has strained healthcare systems in many parts of the United States. During the early months of the pandemic, there was substantial uncertainty about whether the large number of COVID-19 patients requiring hospitalization would exceed healthcare system capacity. This uncertainty created an urgent need to accurately predict the number of COVID-19 patients that would require inpatient and ventilator care at the local level. As the pandemic progressed, many healthcare systems relied on such predictions to prepare for COVID-19 surges and to make decisions regarding staffing, the discontinuation of elective procedures, and the amount of personal protective equipment (PPE) to purchase. In this work, we develop a Bayesian Susceptible-Infectious-Hospitalized-Ventilated-Recovered (SIHVR) model to predict the burden of COVID-19 at the healthcare system level. The Bayesian SIHVR model provides daily estimates of the number of new COVID-19 patients admitted to inpatient care, the total number of non-ventilated COVID-19 inpatients, and the total number of ventilated COVID-19 patients at the healthcare system level. The model also incorporates county-level data on the number of reported COVID-19 cases, and county-level social distancing metrics, making it locally customizable. The uncertainty in model predictions is quantified with 95% credible intervals. The Bayesian SIHVR model is validated with an extensive simulation study, and then applied to data from two regional healthcare systems in South Carolina. This model can be adapted for other healthcare systems to estimate local resource needs.

BackgroundRacial/ethnic disparities during the first six months of the COVID-19 pandemic led to differences in COVID-19 testing and adverse outcomes. We examine differences in testing and adverse outcomes by race/ethnicity and sex across a geographically diverse and system-based COVID-19 cohort collaboration.MethodsObservational study among adults (≥18 years) within six US cohorts from March 1, 2020 to August 31, 2020 using data from electronic health record and patient reporting. Race/ethnicity and sex as risk factors were primary exposures, with health system type (integrated health system, academic health system, or interval cohort) as secondary. Proportions measured SARS-CoV-2 testing and positivity; attributed hospitalization and death related to COVID-19. Relative risk ratios (RR) with 95% confidence intervals quantified associations between exposures and main outcomes.Results5,958,908 patients were included. Hispanic patients had the highest proportions of SARS-CoV-2 testing (16%) and positivity (18%), while Asian/Pacific Islander patients had the lowest portions tested (11%) and White patients had the lowest positivity rates (5%). Men had a lower likelihood of testing (RR = 0.90 [0.89-0.90]) and a higher positivity risk (RR = 1.16 [1.14-1.18]) compared to women. Black patients were more likely to have COVID-19-related hospitalizations (RR = 1.36 [1.28-1.44]) and death (RR = 1.17 [1.03-1.32]) compared with White patients. Men were more likely to be hospitalized (RR = 1.30 [1.16-1.22]) or die (RR = 1.70 [1.53-1.89]) compared to women. These racial/ethnic and sex differences were reflected in both health system types.ConclusionsThis study supports evidence of disparities by race/ethnicity and sex during the COVID-19 pandemic that persisted even in healthcare settings with reduced barriers to accessing care. Further research is needed to understand and prevent the drivers that resulted in higher burdens of morbidity among certain Black patients and men.

BackgroundThe COVID-19 pandemic has had broad impacts on individuals, families and communities which will continue to require multidimensional responses from service providers, program developers, and policy makers.ObjectivesThe purpose of this study was to use Life Course theory to understand and imagine public health and policy responses to the multiple and varied impacts of the COVID-19 pandemic on different groups.Methods"The Cost of COVID-19" was a research study carried out in Kingston, Frontenac, Lennox and Addington counties in South Eastern Ontario, Canada, between June and December 2020. Data included 210 micronarrative stories collected from community members, and 31 in-depth interviews with health and social service providers. Data were analyzed using directed content analysis to explore the fit between data and the constructs of Life Course theory.ResultsSocial pathways were significantly disrupted by changes to education and employment, as well as changes to roles which further altered anticipated pathways. Transitions were by and large missed, creating a sense of loss. While some respondents articulated positive turning points, most of the turning points reported were negative, including fundamental changes to relationships, family structure, education, and employment with lifelong implications. Participants' trajectories varied based on principles including when they occurred in their lifespan, the amount of agency they felt or did not feel over circumstances, where they lived (rural versus urban), what else was going on in their lives at the time the pandemic struck, how their lives were connected with others, as well as how the pandemic impacted the lives of those dear to them. An additional principle, that of Culture, was felt to be missing from the Life Course theory as currently outlined.ConclusionsA Life Course analysis may improve our understanding of the multidimensional long-term impacts of the COVID-19 pandemic and associated public health countermeasures. This analysis could help us to anticipate services that will require development, training, and funding to support the recovery of those who have been particularly affected. Resources needed will include education, mental health and job creation supports, as well as programs that support the development of individual and community agency.

IntroductionThe composition of the nasal microbiota in surgical patients in the context of general anesthesia and nasal povidone-iodine decolonization is unknown. The purpose of this quality improvement study was to determine: (i) if general anesthesia is associated with changes in the nasal microbiota of surgery patients and (ii) if preoperative intranasal povidone-iodine decolonization is associated with changes in the nasal microbiota of surgery patients.Materials and methodsOne hundred and fifty-one ambulatory patients presenting for surgery were enrolled in a quality improvement study by convenience sampling. Pre- and post-surgery nasal samples were collected from patients in the no intranasal decolonization group (control group, n = 54). Pre-decolonization nasal samples were collected from the preoperative intranasal povidone-iodine decolonization group (povidone-iodine group, n = 97). Intranasal povidone-iodine was administered immediately prior to surgery and continued for 20 minutes before patients proceeded for surgery. Post-nasal samples were then collected. General anesthesia was administered to both groups. DNA from the samples was extracted for 16S rRNA sequencing on an Illumina MiSeq.ResultsIn the control group, there was no evidence of change in bacterial diversity between pre- and post-surgery samples. In the povidone-iodine group, nasal bacterial diversity was greater in post-surgery, relative to pre-surgery (Shannon's Diversity Index (P = 0.038), Chao's richness estimate (P = 0.02) and Inverse Simpson index (P = 0.027). Among all the genera, only the relative abundance of the genus Staphylococcus trended towards a decrease in patients after application (FDR adjusted P = 0.06). Abundant genera common to both povidone-iodine and control groups included Staphylococcus, Bradyrhizobium, Corynebacterium, Dolosigranulum, Lactobacillus, and Moraxella.ConclusionsWe found general anesthesia was not associated with changes in the nasal microbiota. Povidone-iodine treatment was associated with nasal microbial diversity and decreased abundance of Staphylococcus. Future studies should examine the nasal microbiota structure and function longitudinally in surgical patients receiving intranasal povidone-iodine.

This study quantifies the association between patient reported measures (PRMs) and readmission to inform efforts to improve hospital care. A retrospective, cross-sectional study was conducted with adults who had chronic obstructive pulmonary disease (COPD) or congestive heart failure (CHF) and were admitted for acute care in a public hospital in New South Wales, Australia for any reason (n = 2394 COPD and 2476 CHF patients in 2018-2020). Patient- level survey data were linked with inpatient data for one year prior to risk-adjust outcomes and after discharge to detect all cause unplanned readmission to a public or private hospital. Ninety-day readmission rates for respondents with COPD or CHF were 17% and 19%. Crude rates for adults with COPD were highest among those who reported that hospital care and treatment helped "not at all" (28%), compared to those who responded, "to some extent" (20%) or "definitely" (15%). After accounting for patient characteristics, adults with COPD or CHF who said care and treatment didn't help at all were at twice the risk of readmission compared to those who responded that care and treatment helped "definitely" (Hazard ratio for COPD 1.97, CI: 1.17-3.32; CHF 2.07, CI 1.25-3.42). Patients who offered the most unfavourable ratings of overall care, understandable explanations, organised care, or preparedness for discharge were at a 1.5 to more than two times higher risk of readmission. Respect and dignity, effective and clear communications, and timely and coordinated care also matter. PRMs are strong predictors of readmission even after accounting for risk related to age and co-morbidities. More moderate ratings were associated with attenuation of risk, and the most positive ratings were associated with the lowest readmission rate. These results suggest that increasing each patient's positive experiences progressively reduces the risk of adults with chronic conditions returning to acute care.

IntroductionMaternal weight management services have been recognised as a good opportunity to influence lifestyle and dietary behaviour of mothers and families. Exploring women's views of maternal weight management services is paramount to understand what constitutes the most suitable service. This study therefore explored experiences among women with a raised body mass index (BMI) of maternal weight management service provision and the barriers and facilitators to weight management during pregnancy.MethodThirteen women with a BMI≥40kg/m² undertook semi-structured interviews around weight management experiences during pregnancy. Interviews were audio recorded and transcribed verbatim. Inductive thematic analysis was undertaken.ResultsFour themes emerged. 1). "Understanding where I am at" showed current readiness and motivation of women varied, from being avoidant to being motivated to make changes. 2). "Getting information" revealed inconsistent information provision during pregnancy. Women particularly wanted practical advice. Some attempted to find this for themselves from friends or the internet, however this left some women feeling confused when different sources provided inconsistent advice. 3). "Difficulties I face" identified physical, emotional and financial barriers and the strategies some women used to overcome these. 4). "Encountering professionals-a mixed experience" demonstrated women wanted to be treated with respect and sensitivity and that how weight management information was addressed was more important than who provided it. The fine line professionals tread was demonstrated by women thinking that they had received inadequate information and yet too much focus was placed on their weight and the associated risks during pregnancy without practical solutions to their weight management challenges.DiscussionWomen were empowered when practical advice was provided, not just the continual repetition of the risks of being obese during pregnancy. Antenatal weight management services need to be clear, sensitive and respectful. Services centred on individual women's needs and on their current and previous experiences are required. The psychological and social contexts of weight management also need to be addressed.

This study investigated whether hot water immersion (HWI) could heat acclimate athletes and improve intermittent running performance and perception of in-game running ability, during a competitive Australian Rules Football (ARF) season. Fifteen male semi-professional ARF athletes (Mean (SD); age: 22 (3) years, height: 182.3 (6.5) cm, mass: 80.5 (5.1) kg) completed either HWI (HEAT, N = 8, 13 (2) sessions, 322 (69) min exposure, 39.5 (0.3) °C) or acted as a control (CON, N = 7, no water immersion) over 6-weeks. Athletes completed a 30-15 Intermittent Fitness Test pre and post-intervention to assess intermittent running performance (VIFT), with perception of in-game running ability measured. Heat acclimation was determined via change in resting plasma volume, as well as physiological and perceptual responses during HWI. HEAT elicited large PV expansion (mean ± 90% CI: d = 1.03 ± 0.73), large decreases in heart rate (d = -0.89 ± 0.70), thermal sensation (d = -2.30 ± 1.15) and tympanic temperature (d = -1.18 ± 0.77). Large improvements in VIFT were seen in HEAT (d = 1.67 ± 0.93), with HEAT showing a greater improvement in VIFT when compared to CON (d = 0.81 ± 0.88). HEAT also showed greater belief that in-game running ability improved post-intervention (d = 2.15 ± 1.09) compared to CON. A 6-week HWI intervention can elicit heat acclimation, improve perception of in-game running ability, and potentially improve VIFT in semi-professional ARF athletes.

The Questionnaire of Cognitive and Affective Empathy (QCAE) is a multiple dimensional measure of cognitive empathy [comprising primary factors for perspective taking (PT), online simulation (OS)], and affective empathy [comprising primary factors for emotion contagion (EC), proximal responsivity (PRO), and peripheral responsivity (PER)]. This study used independent clusters confirmatory factor analysis (ICM-CFA) and exploratory structural equation modeling (ESEM) to examine the scale's factor structure. A general community sample of 203 (men = 43, women = 160) between 17 and 63 years completed the QCAE. Although both the five-factor oblique and second order factor models showed good model fit, and clarity in the pattern of factor loadings, in the second-order factor model, none of the primary factors loaded significantly on their respective secondary factors, thereby favoring the five-factor oblique model. The factors in this model were supported in terms of external validity. Despite this, the factor for PRO in this model showed low reliability for meaning interpretation. A revised four-factor oblique model without the PRO factor showed good fit, clarity in the pattern of factor loadings, and reliability and validity for the factors in this model, thereby suggesting this to be the best model to represent ratings on the QCAE.