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Abstract Background Studies have shown that the coronavirus (COVID-19) pandemic negatively impacted the mental health of young Australians. However, there is limited longitudinal research exploring how individual factors and COVID-19 related public-health restrictions influenced mental health in young people over the acute phase of the COVID-19 pandemic. This study aimed to identify risk and protective factors associated with changes in individual symptoms of anxiety and depression among young Australians during the COVID-19 pandemic. Methods This prospective cohort study collected data on anxiety and depression symptoms of young Australians aged 15–29 years old using the Depression, Anxiety and Stress Scale short form (DASS-21). We delivered four online questionnaires from April 2020 to August 2021 at intervals of 3, 6, and 12 months after the initial survey. We implemented linear mixed-effects regression models to determine the association among demographic, socioeconomic, lifestyle and COVID–19 public health restrictions related factors and the severity of anxiety and depression symptoms over time. Results Analyses included 1936 young Australians eligible at baseline. There was a slight increase in DASS-21 anxiety mean scores from timepoint 3 to timepoint 4. DASS-21 depression scores showed slight fluctuations across timepoints with the highest mean score observed in timepoint 2. Factors associated with increases in anxiety and depression severity symptoms included LGBTQIA + identity, financial insecurity both before and during the pandemic, higher levels of loneliness, withdrawal or deferral of studies, spending more time on social media, and difficulties to sleep. Risk factors for only depression symptoms include unemployment during COVID-19 pandemic and being in lockdown. Living with someone was a protective factor for both anxiety and depression symptoms, pre-COVID-19 unemployment for depression symptoms, and older age and unemployment during the pandemic for anxiety symptoms. Conclusion These findings indicate that during the first year of the pandemic in Australia, there were significant changes in young people’s mental health which were associated with multiple demographic, socioeconomic, lifestyle, and lockdown factors. Hence, in future public health crises, we suggest more inclusive guidelines that involve young people in their development and implementation ensuring that their unique perspectives and needs are adequately considered.

Increased neck strength has been linked to a potential decrease in traumatic brain injuries (TBI). The purpose was to determine the efficacy of a neck-strengthening protocol using a novel neck-strengthening device to increase isometric neck strength and rate of force development (RFD). Utilizing self-generated centripetal force, participants trained for 14 weeks. A linear mixed model was used to analyze the relationship between post-assessment measurements and pre-assessments measurements, while accounting for repeated measure random effect at the individual level, and a regular random error term. RFD values were 4.344 times higher in the clockwise direction and 5.978 times higher in the counterclockwise direction when comparing pre and post assessment measurements. Isometric neck strength increased significantly (p ​

Abstract Background At a global scale, the SARS-CoV-2 virus did not remain in its initial genotype for a long period of time, with the first global reports of variants of concern (VOCs) in late 2020. Subsequently, genome sequencing has become an indispensable tool for characterizing the ongoing pandemic, particularly for typing SARS-CoV-2 samples obtained from patients or environmental surveillance. For such SARS-CoV-2 typing, various in vitro and in silico workflows exist, yet to date, no systematic cross-platform validation has been reported. Results In this work, we present the first comprehensive cross-platform evaluation and validation of in silico SARS-CoV-2 typing workflows. The evaluation relies on a dataset of 54 patient-derived samples sequenced with several different in vitro approaches on all relevant state-of-the-art sequencing platforms. Moreover, we present UnCoVar, a robust, production-grade reproducible SARS-CoV-2 typing workflow that outperforms all other tested approaches in terms of precision and recall. Conclusions In many ways, the SARS-CoV-2 pandemic has accelerated the development of techniques and analytical approaches. We believe that this can serve as a blueprint for dealing with future pandemics. Accordingly, UnCoVar is easily generalizable towards other viral pathogens and future pandemics. The fully automated workflow assembles virus genomes from patient samples, identifies existing lineages, and provides high-resolution insights into individual mutations. UnCoVar includes extensive quality control and automatically generates interactive visual reports. UnCoVar is implemented as a Snakemake workflow. The open-source code is available under a BSD 2-clause license at github.com/IKIM-Essen/uncovar.

Background: Acute myocardial infarction (AMI) remains a common reason for admission to the intensive care unit (ICU). However, there is limited data comparing outcomes for patients with AMI admitted to specific ICUs. Objectives: The purpose of this study was to assess clinical outcomes between patients with AMI requiring invasive mechanical ventilation admitted to the medical ICU (MICU) compared to cardiac (CICU). Methods: We utilized the Vizient Clinical Data Base to identify patients with a primary diagnosis of AMI between October 2015 and December 2019 and requiring invasive mechanical ventilation. Using multivariable logistic regression, we compared clinical outcomes for patients admitted to the MICU vs CICU. Results: We identified 12,639 patients, 25.2% (n = 3,185) of which were admitted to a MICU and 74.8% (n = 9,454) to a CICU. Patients admitted to a CICU were more likely to present with STEMI (57.0% vs 42.8%), cardiogenic shock (46.0% vs 37.4%), and require mechanical circulatory support and vasoactive medications (all, P 750 beds), and teaching hospitals (all, P

Background:. Open fractures, a common consequence of road traffic collisions, are associated with a high risk of complications. The introduction of standard guidelines has been shown to improve patient care and reduce the risk of complications in several countries. In September 2021, the Malawi Orthopaedic Association/Arbeitsgemeinschaft für Osteosynthesefragen Alliance (MOA/AOA) guidelines and standards for open fracture management were introduced in Malawi. This study aimed to assess the management of open fractures in hospitals in Malawi, before and after implementing a training course on the MOA/AOA open fracture guidelines. Methods:. This was a descriptive and quantitative, before-and-after study that reviewed the medical files of patients with open fractures at Zomba Central Hospital and Mulanje, Salima, and Mangochi district hospitals over two 3-month periods. Variables included initial assessment; antibiotic prophylaxis; place of debridement; type of anesthesia; treatment of the open fracture in the emergency department, operating room, and wards; and short-term complications requiring hospital treatment. Results:. A total of 88 open-fracture case files were reviewed; 43 were prior and 45 were subsequent to the implementation of the open fracture guidelines. The overall median patient age was 36 years (interquartile range, 27 to 45 years), and 91% (80) were male. Limb neurovascular status assessment and documentation improved from 26% (11) of the patients before the guidelines to 62% (28) afterward (p = 0.0002). The percentage who underwent debridement in the operating room significantly increased from 19% (8) to 69% (31) (p = 0.01). The percentage who underwent debridement under general or spinal anesthesia significantly increased from 5% (2) to 38% (17) and from 12% (5) to 29% (13), respectively (p= 0.001). The wound infection rate decreased from 21% to 11%, but this was not significant, and there was no change in the overall complication rate (p = 0.152). Conclusions:. This study suggests that training on the MOA/AOA open fracture management guidelines followed by their implementation can lead to at least temporary improvement in the management of open fractures. Nevertheless, additional studies need to be performed to understand the effect on long-term patient outcomes. Levels of Evidence:. Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.

Objectives People living in less urban areas in the UK may have reduced access to mental healthcare compared with people in urban areas. Although this was not reported in the 2000 Adult Psychiatric Morbidity Survey (APMS) data, subsequent changes to mental health provision and economic recession may have impacted care inequalities. We re-examined this, hypothesising that those living in less urban areas of England received less antidepressant medication and psychological interventions, compared with those living in urban areas, after adjusting for covariates including common mental disorders (CMDs) and socioeconomic status.Design National cross-sectional study.Setting Households in England.Participants We analysed data from 7455 participants aged 16 and above from the 2014 English APMS.Exposures Residence in an urban or less urban area.Primary outcome Current receipt of any antidepressant medication or any psychological intervention.Results Participants living in less urban areas were half as likely to be in receipt of any psychological intervention relative to those living in urban areas (adjusted OR (aOR) 0.49; 95% CI 0.30 to 0.79; p=0.004). There was no association between urbanisation and antidepressant receipt (aOR 1.01; 95% CI 0.81 to 1.25; p=0.944).Conclusion People living in less urban areas have a lower likelihood of being in receipt of psychological treatment for CMDs, which may indicate barriers to care access. Our findings differ from the results of a study using a comparable sample conducted 14 years previously. While Improving Access to Psychological Therapies services have increased therapy access nationwide, this may have had more impact in urban areas, potentially widening urban/rural inequalities. Further research is needed to understand how to address such geographical inequalities in access to mental healthcare.

Background: Hospital discharge summaries play an essential role in informing GPs of recent admissions to ensure excellent continuity of care and prevent adverse events; however, they are notoriously poorly written, time-consuming, and can result in delayed discharge. Aim: To evaluate the potential of artificial intelligence (AI) to produce high-quality discharge summaries equivalent to the level of a doctor who has completed the UK Foundation Programme. Design & setting: Feasibility study using 25 mock patient vignettes. Method: Twenty-five mock patient vignettes were written by the authors. Five junior doctors wrote discharge summaries from the case vignettes (five each). The same case vignettes were input into ChatGPT. In total, 50 discharge summaries were generated; 25 by Al and 25 by junior doctors. Quality and suitability were determined through both independent GP evaluators and adherence to a minimum dataset. Results: Of the 25 AI-written discharge summaries 100% were deemed by GPs to be of an acceptable quality compared with 92% of the junior doctor summaries. They both showed a mean compliance of 97% with the minimum dataset. In addition, the ability of GPs to determine if the summary was written by ChatGPT was poor, with only a 60% accuracy of detection. Similarly, when run through an AI-detection tool all were recognised as being very unlikely to be written by AI. Conclusion: AI has proven to produce discharge summaries of equivalent quality to a junior doctor who has completed the UK Foundation Programme; however, larger studies with real-world patient data with NHS-approved AI tools will need to be conducted.

Abstract Background Microsporidia are diverse spore forming, fungal-related obligate intracellular pathogens infecting a wide range of hosts. This diversity is reflected at the genome level with sizes varying by an order of magnitude, ranging from less than 3 Mb in Encephalitozoon species (the smallest known in eukaryotes) to more than 50 Mb in Edhazardia spp. As a paradigm of genome reduction in eukaryotes, the small Encephalitozoon genomes have attracted much attention with investigations revealing gene dense, repeat- and intron-poor genomes characterized by a thorough pruning of molecular functions no longer relevant to their obligate intracellular lifestyle. However, because no Encephalitozoon genome has been sequenced from telomere-to-telomere and since no methylation data is available for these species, our understanding of their overall genetic and epigenetic architectures is incomplete. Methods In this study, we sequenced the complete genomes from telomere-to-telomere of three human-infecting Encephalitozoon spp. —E. intestinalis ATCC 50506, E. hellem ATCC 50604 and E. cuniculi ATCC 50602— using short and long read platforms and leveraged the data generated as part of the sequencing process to investigate the presence of epigenetic markers in these genomes. We also used a mixture of sequence- and structure-based computational approaches, including protein structure prediction, to help identify which Encephalitozoon proteins are involved in telomere maintenance, epigenetic regulation, and heterochromatin formation. Results The Encephalitozoon chromosomes were found capped by TTAGG 5-mer telomeric repeats followed by telomere associated repeat elements (TAREs) flanking hypermethylated ribosomal RNA (rRNA) gene loci featuring 5-methylcytosines (5mC) and 5-hemimethylcytosines (5hmC), themselves followed by lesser methylated subtelomeres and hypomethylated chromosome cores. Strong nucleotide biases were identified between the telomeres/subtelomeres and chromosome cores with significant changes in GC/AT, GT/AC and GA/CT contents. The presence of several genes coding for proteins essential to telomere maintenance, epigenetic regulation, and heterochromatin formation was further confirmed in the Encephalitozoon genomes. Conclusion Altogether, our results strongly support the subtelomeres as sites of heterochromatin formation in Encephalitozoon genomes and further suggest that these species might shutdown their energy-consuming ribosomal machinery while dormant as spores by silencing of the rRNA genes using both 5mC/5hmC methylation and facultative heterochromatin formation at these loci.

The alternative splicing and genetic variations across the whole MHC-E locus of Rhesus Macaques (Mamu-E) are analysed, which provides a foundation for more comprehensive research of Mamu-E and informs translational research of CMV-based vaccines.

BackgroundBreakthrough infections with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants are increasingly observed in vaccinated individuals. Immune responses towards SARS-CoV-2 variants, particularly Omicron-BA.5, are poorly understood. We investigated the humoral and cellular immune responses of hospitalized COVID-19 patients during Delta and Omicron infection waves.MethodsThe corresponding SARS-CoV-2 variant of the respective patients were identified by whole genome sequencing. Humoral immune responses were analyzed by ELISA and a cell culture-based neutralization assay against SARS-CoV-2 D614G isolate (wildtype), Alpha, Delta (AY.43) and Omicron (BA.1 and BA.5). Cellular immunity was evaluated with an IFN-γ ELISpot assay.ResultsOn a cellular level, patients showed a minor IFN-γ response after stimulating PBMCs with mutated regions of SARS-CoV-2 variants. Neutralizing antibody titers against Omicron-BA.1 and especially BA.5 were strongly reduced. Double-vaccinated patients with Delta breakthrough infection showed a significantly increased neutralizing antibody response against Delta compared to double-vaccinated uninfected controls (median complete neutralization titer (NT100) 640 versus 80, p

Motor impairment has a profound impact on a significant number of individuals, leading to a substantial demand for rehabilitation services. Through brain–computer interfaces (BCIs), people with severe motor disabilities could have improved communication with others and control appropriately designed robotic prosthetics, so as to (at least partially) restore their motor abilities. BCI plays a pivotal role in promoting smoother communication and interactions between individuals with motor impairments and others. Moreover, they enable the direct control of assistive devices through brain signals. In particular, their most significant potential lies in the realm of motor rehabilitation, where BCIs can offer real-time feedback to assist users in their training and continuously monitor the brain’s state throughout the entire rehabilitation process. Hybridization of different brain-sensing modalities, especially functional near-infrared spectroscopy (fNIRS) and electroencephalography (EEG), has shown great potential in the creation of BCIs for rehabilitating the motor-impaired populations. EEG, as a well-established methodology, can be combined with fNIRS to compensate for the inherent disadvantages and achieve higher temporal and spatial resolution. This paper reviews the recent works in hybrid fNIRS-EEG BCIs for motor rehabilitation, emphasizing the methodologies that utilized motor imagery. An overview of the BCI system and its key components was introduced, followed by an introduction to various devices, strengths and weaknesses of different signal processing techniques, and applications in neuroscience and clinical contexts. The review concludes by discussing the possible challenges and opportunities for future development.

Abstract Basic information on the ecology of species is key for their conservation. Here we study the ecology of the little‐known yellow‐throated bunting Emberiza elegans based on a multi‐year study on its breeding grounds in the Russian Far East. For the first time in this species, we quantified breeding habitat parameters, calculated sex‐specific apparent survival, and determined individual nonbreeding locations using light‐level geolocation. We found that the habitat around song posts of male yellow‐throated buntings is characterized by tree and shrub layers on richly littered moist ground. Habitat use overlaps with co‐occurring Tristram's Buntings Emberiza tristrami and Black‐faced Buntings E. spodocephala, but territories differ especially in tree cover and litter cover. Based on 4 years of color‐ringing data of 72 individuals, we calculated an apparent survival rate of 36%, with higher survival estimates for male than for female yellow‐throated buntings. We found no effect of carrying a geolocator on survival. We retrieved six geolocators from males. All birds migrated south‐westward during autumn and spent the nonbreeding season at locations in China 700–1700 km away from their breeding sites. At least two individuals spent the boreal winter outside of the known range in northern or central China. Birds left the breeding area between early October and early November and returned between mid‐March and mid‐April. Our data on habitat use, survival rate, and migratory connectivity will help to assess threats to the populations of this enigmatic species, which might include habitat loss due to forest fires on the breeding grounds, and unsustainable harvest for consumption during the nonbreeding season.

Abstract SARS-CoV-2 is spreading worldwide with continuously evolving variants, some of which occur in the Spike protein and appear to increase viral transmissibility. However, variants that cause severe COVID-19 or lead to other breakthroughs have not been well characterized. To discover such viral variants, we assembled a cohort of 683 COVID-19 patients; 388 inpatients (“cases”) and 295 outpatients (“controls”) from April to August 2020 using electronically captured COVID test request forms and sequenced their viral genomes. To improve the analytical power, we accessed 7137 viral sequences in Washington State to filter out viral single nucleotide variants (SNVs) that did not have significant expansions over the collection period. Applying this filter led to the identification of 53 SNVs that were statistically significant, of which 13 SNVs each had 3 or more variant copies in the discovery cohort. Correlating these selected SNVs with case/control status, eight SNVs were found to significantly associate with inpatient status (q-values

Introduction: With the introduction of portable handheld ultrasounds, higher levels of technology are more easily available for patients in rural and underserved communities. Point-of-care ultrasound (POCUS) increases accessibility for patients with limited resources, thus reducing costs and decreasing the risk of non-compliance or subsequent loss to follow-up. Despite the increasing utility of ultrasonography, literature demonstrates a lack of sufficient training in POCUS and ultrasound-guided techniques for Family Medicine residents. Integrating unfixed cadavers into the preclinical curriculum may be an ideal adjunct to simulating pathologies and screening sensitive regions.Methods: In total, 27 unfixed, de-identified cadavers were scanned with a handheld portable ultrasound. Sixteen body systems were screened: ocular, thyroid, carotid artery/internal jugular vein, brachial plexus, heart, kidneys, pancreas, gallbladder, liver, aorta and inferior vena cava, femoral artery and vein, knee, popliteal vessels, uterus, scrotum, and shoulder.Results: Eight of the sixteen body systems, including the ocular, thyroid, carotid artery/internal jugular vein, brachial plexus, liver, knee, scrotum, and shoulder, consistently showed accurate anatomy and pathology. A physician skilled in ultrasound reviewed images obtained from the cadavers and concluded that differences in anatomy and common pathologies of unfixed cadavers were indiscernible compared with live patient ultrasound images.Discussion: Using unfixed cadavers in POCUS training can be a valuable educational tool in preparing Family Medicine Physicians for rural or remote practices because the cadavers display accurate anatomy and pathology under ultrasound evaluation in multiple body systems. Further studies should explore creating artificial pathologies in cadaveric models to broaden the scope of application.

Summary: Advances in cellular engineering, as well as gene, and cell therapy, may be used to produce human tissues with programmable genetically enhanced functions designed to model and/or treat specific diseases. Fabrication of synthetic human liver tissue with these programmable functions has not been described. By generating human iPSCs with target gene expression controlled by a guide RNA-directed CRISPR-Cas9 synergistic-activation-mediator, we produced synthetic human liver tissues with programmable functions. Such iPSCs were guide-RNA-treated to enhance expression of the clinically relevant CYP3A4 and UGT1A1 genes, and after hepatocyte-directed differentiation, cells demonstrated enhanced functions compared to those found in primary human hepatocytes. We then generated human liver tissue with these synthetic human iPSC-derived hepatocytes (iHeps) and other non-parenchymal cells demonstrating advanced programmable functions. Fabrication of synthetic human liver tissue with modifiable functional genetic programs may be a useful tool for drug discovery, investigating biology, and potentially creating bioengineered organs with specialized functions.

Abstract Radiotherapy of head and neck squamous cell carcinoma can lead to long-term complications like osteoradionecrosis, resulting in severe impairment of the jawbone. Current standard procedures require a 6-month wait after irradiation before dental reconstruction can begin. A comprehensive characterization of the irradiation-induced molecular and functional changes in bone cells could allow the development of novel strategies for an earlier successful dental reconstruction in patients treated by radiotherapy. The impact of ionizing radiation on the bone-forming alveolar osteoblasts remains however elusive, as previous studies have relied on animal-based models and fetal or animal-derived cell lines. This study presents the first in vitro data obtained from primary human alveolar osteoblasts. Primary human alveolar osteoblasts were isolated from healthy donors and expanded. After X-ray irradiation with 2, 6 and 10 Gy, cells were cultivated under osteogenic conditions and analyzed regarding their proliferation, mineralization, and expression of marker genes and proteins. Proliferation of osteoblasts decreased in a dose-dependent manner. While cells recovered from irradiation with 2 Gy, application of 6 and 10 Gy doses not only led to a permanent impairment of proliferation, but also resulted in altered cell morphology and a disturbed structure of the extracellular matrix as demonstrated by immunostaining of collagen I and fibronectin. Following irradiation with any of the examined doses, a decrease of marker gene expression levels was observed for most of the investigated genes, revealing interindividual differences. Primary human alveolar osteoblasts presented a considerably changed phenotype after irradiation, depending on the dose administered. Mechanisms for these findings need to be further investigated. This could facilitate improved patient care by re-evaluating current standard procedures and investigating faster and safer reconstruction concepts, thus improving quality of life and social integrity.

Abstract The population of the Yellow‐breasted Bunting Emberiza aureola, a formerly widely distributed and abundant songbird of northern Eurasia, suffered a catastrophic decline and a strong range contraction between 1980 and 2013. There is evidence that the decline was driven by illegal trapping during migration, but potential contributions of other factors to the decline, such as land‐use change, have not yet been evaluated. Before the effects of land‐use change can be evaluated, a basic understanding of the ecological requirements of the species is needed. We therefore compared habitat use in ten remaining breeding regions across the range, from European Russia to Japan and the Russian Far East. We also assessed large‐scale variation in habitat parameters across the breeding range. We found large variation in habitat use, within and between populations. Differences were related to the cover and height of trees and shrubs at Yellow‐breasted Bunting territories. In many regions, Yellow‐breasted Buntings occupied early successional stages, including anthropogenic habitats characterized by mowing, grazing, or fire regimes. We found that the probability of presence can be best predicted with the cover of shrubs, herbs, and grasses. Highest probabilities were found at shrub cover values of 40%–70%. Differences in habitat use along a longitudinal gradient were small, but we found strong differences across latitudes, possibly related to habitat availability. We conclude that the remaining Yellow‐breasted Bunting populations are not limited to specific habitat types. Our results provide important baseline information to model the range‐wide distribution of this critically endangered species and to guide targeted conservation measures.

Background: Open tibia fractures are a common injury following road traffic accidents in Malawi and can lead to long term disability. Very little is known about patients’ experiences of the healthcare system and the disability in low-income countries following this injury. The aim of the study was to explore patient experiences of treatment and disability following an open tibia fracture in Malawi. Methods: A qualitative study was conducted using semi-structured interviews with ten patients with open tibia fractures at a central hospital in Blantyre, Malawi. A mixed deductive-inductive thematic analysis was used to identify broad themes of treatment and disability. Written informed consent was obtained from all participants. Results: Patient characteristics included an average age of 39.1 years old (22-63) and 80% were male. Broad themes found were delays in receiving treatment, change in individuals’ societal role and delayed recovery associated with pain and immobility. Conclusions: Open tibia fractures in Malawi have a devastating impact on patients and their families. Further studies are required to explore the reasons for the delays in open fracture emergency treatment.

COVID-19 vaccine hesitancy is context specific and highly dynamic in the United States. The objective of this study was to examine the underlying causes of COVID-19 vaccine use and hesitancy among students at two colleges in Central New York (USA) in order to better understand how to promote vaccination in this demographic. A Knowledge, Attitudes, Beliefs & Behaviors survey was designed to measure the relationship between gender, political ideology, media consumption, beliefs, mental health and anxiety, and pandemic-related behaviors. The survey was distributed by email to students in April 2021. Of 8,894 eligible respondents, 627 students completed the survey. Structural Equation Modeling was utilized to explore both direct and indirect relationships. Modeling suggests that the effect of political ideology (being more conservative) is to promote the consumption of right wing news, which in turn tends to increase vaccine hesitancy. Conservative political ideology does not directly lead to vaccine hesitancy, but it does encourage the consumption of news from sources that promote fear and hesitation about the vaccines. News sources significantly contribute to vaccine attitudes among this demographic.

Abstract Jawbone differs from other bones in many aspects, including its developmental origin and the occurrence of jawbone-specific diseases like MRONJ (medication-related osteonecrosis of the jaw). Although there is a strong need, adequate in vitro models of this unique environment are sparse to date. While previous approaches are reliant e.g. on scaffolds or spheroid culture, 3D bioprinting enables free-form fabrication of complex living tissue structures. In the present work, production of human jawbone models was realised via projection-based stereolithography. Constructs were bioprinted containing primary jawbone-derived osteoblasts and vasculature-like channel structures optionally harbouring primary endothelial cells. After 28 days of cultivation in growth medium or osteogenic medium, expression of cell type-specific markers was confirmed on both the RNA and protein level, while prints maintained their overall structure. Survival of endothelial cells in the printed channels, co-cultured with osteoblasts in medium without supplementation of endothelial growth factors, was demonstrated. Constructs showed not only mineralisation, being one of the characteristics of osteoblasts, but also hinted at differentiation to an osteocyte phenotype. These results indicate the successful biofabrication of an in vitro model of the human jawbone, which presents key features of this special bone entity and hence appears promising for application in jawbone-specific research.

The assumption that conservation of sequence implies the action of purifying selection is central to diverse methodologies to infer functional importance. GC-biased gene conversion (gBGC), a meiotic mismatch repair bias strongly favouring GC over AT, can in principle mimic the action of selection, this being thought to be especially important in mammals. As mutation is GC→AT biased, to demonstrate that gBGC does indeed cause false signals requires evidence that an AT-rich residue is selectively optimal compared to its more GC-rich allele, while showing also that the GC-rich alternative is conserved. We propose that mammalian stop codon evolution provides a robust test case. Although in most taxa TAA is the optimal stop codon, TGA is both abundant and conserved in mammalian genomes. We show that this mammalian exceptionalism is well explained by gBGC mimicking purifying selection and that TAA is the selectively optimal codon. Supportive of gBGC, we observe (i) TGA usage trends are consistent at the focal stop codon and elsewhere (in UTR sequences); (ii) that higher TGA usage and higher TAA→TGA substitution rates are predicted by a high recombination rate; and (iii) across species the difference in TAA TGA substitution rates between GC-rich and GC-poor genes is largest in genomes that possess higher between-gene GC variation. TAA optimality is supported both by enrichment in highly expressed genes and trends associated with effective population size. High TGA usage and high TAA→TGA rates in mammals are thus consistent with gBGC's predicted ability to "drive" deleterious mutations and supports the hypothesis that sequence conservation need not be indicative of purifying selection. A general trend for GC-rich trinucleotides to reside at frequencies far above their mutational equilibrium in high recombining domains supports the generality of these results.

Background: Patients with diabetes mellitus (DM) have worse outcomes following percutaneous coronary intervention than nondiabetic patients. The novel Supreme DES is a biodegradable polymer sirolimus-eluting stent designed to synchronize early drug delivery, limiting the potential for long-term inflammatory response. The purpose of this study was to evaluate the safety and efficacy of the Supreme DES in patients with DM. Methods: This is a prespecified analysis of the diabetic subgroup from the PIONEER III randomized (2:1), controlled trial, comparing the Supreme DES with a durable polymer everolimus-eluting stent (DP-EES). The primary safety and efficacy composite endpoint was target lesion failure at 1 year, a composite of cardiac death, target vessel myocardial infarction, or clinically driven target lesion revascularization. Results: The PIONEER III trial randomized 1629 patients, of which 494 (30.3%) had DM with 331 (398 lesions) randomly assigned to Supreme DES and 163 (208 lesions) to DP-EES. Among patients with DM, target lesion failure at 1 year was 6.1% (20/331) with Supreme DES vs 3.7% (6/163) with DP-EES (hazard ratio = 1.65; 95% confidence interval = 0.66-4.10, P = .28). The composite of cardiac death or target vessel myocardial infarction was 3.3% (11/331) with Supreme DES and 3.7% (6/163) with DP-EES (hazard ratio = 0.90; 95% confidence interval = 0.33-2.44, P = .83). There were no significant differences in other secondary endpoints. Conclusions: This prespecified substudy of the PIONEER III trial demonstrated the relative safety and efficacy of the novel Supreme DES when compared with commercially available DP-EES in diabetics at 1 year. Longer term follow-up will be required to ensure continued safety and efficacy of the Supreme DES.

Ageing of the haematopoietic system is accompanied by declining erythropoiesis and lymphopoiesis. Here the authors uncover upregulated IL-6 and TGFβ signalling in aged bone marrow stroma; inhibition of these signals reverses age-related haematopoietic defects, re-balancing haematopoietic stem cell lineage output.

Abstract Background The socioeconomic status of applicants to Canadian medical schools has been understudied in the past two decades. Institutional efforts have been made to address the lack of socioeconomic diversity across Canada during this time. We investigated the income characteristics of medical school applicants, as well as the relationship between applicant income and offer of admission, to characterize the current state of socioeconomic diversity in medical admissions. Methods We conducted a retrospective cohort study on 26,120 applicants at one Ontario medical school from 2013 to 2018. Characteristics of applicants who were offered admission were compared to the general population and applicants not offered admission. Regression analyses were used to investigate the association between median total neighborhood income and successful admission. Results The median total neighborhood income for medical school applicants was $98,816, which was approximately $28,480 higher than the Canadian general population. Those not admitted to the medical school had a median total neighborhood income of $98,304 compared to $105,984 for those offered admission (p 75th percentile income group had 54% increased odds of being offered admission when compared to applicants in the

National cardiac life support training program initiated in October 2016, it is a systematic approach to management of a victim of cardiac arrest. The program aimed to address the training needs of a junior physician such as the one undergoing postgraduate medical education and other health-care professionals. It is a 2-day “hands-on” course that recommends an instructor–candidate ratio of 1:6 and limits the number of candidates to 30 per session to maintain adequate mentee–mentor interaction, monitoring, and evaluation. This program is developed in collaboration with Society for Emergency Medicine in India, Indian College of Anesthesiologists, Indian College of Cardiology, and Association of Healthcare Providers India. In addition, this program has been endorsed by West Bengal University and National Board of Examinations, New Delhi. Currently, The Simulation Society which deals with online teaching simulation has lent its support by endorsing this program.

This viewpoint summarizes the discussion that occurred during the “Translating Policy to Practice in Telehealth–Lessons from Global Implementation Experiences” panel that was held virtually at Telemedicon2020, December 18-20, 2020. This panel brought together policy and implementation experts from some countries of South Asia, Kuwait, and the European Union to share their experiences in the development and implementation of telehealth standards and of the scale up of telehealth interventions within health systems. Several common themes arose from the discussion, including the significant role of people; encouragement by respective government policymakers; addressing concerns, particularly related to privacy, confidentiality, and security; and capacity building of human resources. These are discussed in turn, along with the future directions identified by the panelists, which emphasized the need for active encouragement toward the adoption and diffusion of digital health in general and of telehealth in particular. All stakeholders, ranging from governmental policymakers to common citizens, need to come together to build trusting partnerships to realize the advantages offered by telehealth.

In search of more detailed explanations for body-mind interactions in physical activity, neural and physiological effects, especially regarding more strenuous sports activities, increasingly attract interest. Little is known about the underlying manifold (neuro-)physiological impacts induced by different motor learning approaches. The various influences on brain or cardiac function are usually studied separately and modeled linearly. Limitations of these models have recently led to a rapidly growing application of nonlinear models. This study aimed to investigate the acute effects of various sequences of rope skipping on irregularity of the electrocardiography (ECG) and electroencephalography (EEG) signals as well as their interaction and whether these depend on different levels of active movement noise, within the framework of differential learning theory. Thirty-two males were randomly and equally distributed to one of four rope skipping conditions with similar cardiovascular but varying coordinative demand. ECG and EEG were measured simultaneously at rest before and immediately after rope skipping for 25 mins. Signal irregularity of ECG and EEG was calculated via the multiscale fuzzy measure entropy (MSFME). Statistically significant ECG and EEG brain area specific changes in MSFME were found with different pace of occurrence depending on the level of active movement noise of the particular rope skipping condition. Interaction analysis of ECG and EEG MSFME specifically revealed an involvement of the frontal, central, and parietal lobe in the interplay with the heart. In addition, the number of interaction effects indicated an inverted U-shaped trend presenting the interaction level of ECG and EEG MSFME dependent on the level of active movement noise. In summary, conducting rope skipping with varying degrees of movement variation appears to affect the irregularity of cardiac and brain signals and their interaction during the recovery phase differently. These findings provide enough incentives to foster further constructive nonlinear research in exercise-recovery relationship and to reconsider the philosophy of classical endurance training.

Cement augmentation of pedicle screws is one of the most promising approaches to enhance the anchoring of screws in the osteoporotic spine. To date, there is no ideal cement for pedicle screw augmentation. The purpose of this study was to investigate whether an injectable, bioactive, and degradable calcium sulfate/hydroxyapatite (CaS/HA) cement could increase the maximum pull-out force of pedicle screws in osteoporotic vertebrae. Herein, 17 osteoporotic thoracic and lumbar vertebrae were obtained from a single fresh-frozen human cadaver and instrumented with fenestrated pedicle screws. The right screw in each vertebra was augmented with CaS/HA cement and the un-augmented left side served as a paired control. The cement distribution, interdigitation ability, and cement leakage were evaluated using radiographs. Furthermore, pull-out testing was used to evaluate the immediate mechanical effect of CaS/HA augmentation on the pedicle screws. The CaS/HA cement presented good distribution and interdigitation ability without leakage into the spinal canal. Augmentation significantly enhanced the maximum pull-out force of the pedicle screw in which the augmented side was 39.0% higher than the pedicle-screw-alone side. Therefore, the novel biodegradable biphasic CaS/HA cement could be a promising material for pedicle screw augmentation in the osteoporotic spine.

Background: Road traffic injury (RTI) is the largest cause of death amongst 15–39-year-old people worldwide, and the burden of injuries such as open tibia fractures are rapidly increasing in Malawi. This study aims to investigate disability and economic outcomes of people with open tibia fractures in Malawi and improve these with locally delivered implementation of open fracture guidelines. Methods: This is a prospective cohort study describing function, quality of life and economic burden of open tibia fractures in Malawi. In total, 160 participants will be recruited across six centres and will be followed-up with face-to-face interviews at six weeks, three months, six months and one year following injury. The primary outcome will be function at one year measured by the short musculoskeletal functional assessment (SMFA) score. Secondary outcomes will include quality of life measured by EuroQol EQ-5D-3L, catastrophic loss of income and implementation outcomes (acceptability, adoption, appropriateness, costs, feasibility, fidelity, penetration, and sustainability) at one year. A nested pilot pre-post implementation study of an interventional bundle for all open fractures will be developed based on other implementation studies from low- and middle-income countries (LMICs). Regression analysis will be used to model and investigate associations between SMFA score and fracture severity, infection and the pre- and post-training course period. Outcome: This prospective cohort study will report patient reported outcomes from open tibia fractures in low-resource settings. Subsequent detailed evaluation of both the clinical and implementation components of the study will promote sustainability of improved open fractures management in the study sites and further scale-up of open fracture management guidelines. Ethics: Ethics approval has been obtained from the Liverpool School of Tropical Medicine and College of Medicine Research and Ethics committee.

The critically endangered Yellow-breasted Bunting has undergone population collapse globally because of illegal hunting and habitat deterioration. It was listed as critically endangered (CR) by the International Union for Conservation of Nature (IUCN) in 2017 and designated a Class I (highest level) national conservation bird species in China in 2021. Birdsong in the breeding season is the main communicative signal under sexual selection, and song variations have long been considered critical evidence of divergence among subspecies or populations. We compared the songs of 89 males from 18 populations to test subspecies taxonomy. We found that songs of the Yellow-breasted Bunting Emberiza aureola are subspecies specific and that three subspecies can be clearly discriminated by song divergences. Moreover, an analysis of multiple vocal traits supports the claim that insulana is distinct from aureola and ornata. Finally, at the geographic population level, populations can be clearly classified in accordance with the three subspecies, although the aureola population in Xinjiang, China is differentiated from other populations of the same subspecies. The results of this study demonstrate that all populations and subspecies are unique and should be protected to maintain intraspecies song diversity. In addition, several specific populations, such as insulana populations in Japan and the Xinjiang, China population of aureola, need to be paid special attention to prevent the extinction of unique or local taxa.

Chimeric antigen receptor (CAR) T cell performance against solid tumors in mouse models and clinical trials is often less effective than predicted by CAR construct selection in two-dimensional (2D) cocultures. Three-dimensional (3D) solid tumor architecture is likely to be crucial for CAR T cell efficacy. We used a three-dimensional (3D) bioprinting approach for large-scale generation of highly reproducible 3D human tumor models for the test case, neuroblastoma, and compared these to 2D cocultures for evaluation of CAR T cells targeting the L1 cell adhesion molecule, L1CAM. CAR T cells infiltrated the model, and both CAR T and tumor cells were viable for long-term experiments and could be isolated as single-cell suspensions for whole-cell assays quantifying CAR T cell activation, effector function and tumor cell cytotoxicity. L1CAM-specific CAR T cell activation by neuroblastoma cells was stronger in the 3D model than in 2D cocultures, but neuroblastoma cell lysis was lower. The bioprinted 3D neuroblastoma model is highly reproducible and allows detection and quantification of CAR T cell tumor infiltration, representing a superior in vitro analysis tool for preclinical CAR T cell characterization likely to better select CAR T cells for in vivo performance than 2D cocultures.

Canada is warming at double the rate of the global average caused in part to a fast-growing population and large land transformations, where urban surfaces contribute significantly to the urban heat island (UHI) phenomenon. The federal government released the strengthened climate plan in 2020, which emphasizes using nature-based solutions (NBSs) to combat the effects of UHI phenomenon. Here, the effects of two NBSs techniques are reviewed and analysed: increasing surface greenery/vegetation (ISG) and increasing surface reflectivity (ISR). Policymakers have the challenge of selecting appropriate NBSs to meet a wide range of objectives within the urban environment and Canadian-specific knowledge of how NBSs can perform at various scales is lacking. As such, this state-of-the-art review intends to provide a snapshot of the current understanding of the benefits and risks associated with the implantation of NBSs in urban spaces as well as a review of the current techniques used to model, and evaluate the potential effectiveness of UHI under evolving climate conditions. Thus, if NBSs are to be adopted to mitigate UHI effects and extreme summertime temperatures in Canadian municipalities, an integrated, comprehensive analysis of their contributions is needed. As such, developing methods to quantify and evaluate NBSs’ performance and tools for the effective implementation of NBSs are required.

Background Several studies have shown improved outcomes in closed compared with open medical and surgical intensive care units. However, very little is known about the ideal organizational structure in the modern cardiac intensive care unit (CICU). Methods and Results We retrospectively reviewed consecutive unique admissions (n=3996) to our tertiary care CICU from September 2013 to October 2017. The aim of our study was to assess for differences in clinical outcomes between an open compared with a closed CICU. We used multivariable logistic regression adjusting for demographics, comorbidities, and severity of illness. The primary outcome was in‐hospital mortality. We identified 2226 patients in the open unit and 1770 in the closed CICU. The unadjusted in‐hospital mortality in the open compared with closed unit was 9.6% and 8.9%, respectively (P=0.42). After multivariable adjustment, admission to the closed unit was associated with a lower in‐hospital mortality (odds ratio [OR], 0.69; 95% CI: 0.53–0.90, P=0.007) and CICU mortality (OR, 0.70; 95% CI, 0.52–0.94, P=0.02). In subgroup analysis, admissions for cardiac arrest (OR, 0.42; 95% CI, 0.20–0.88, P=0.02) and respiratory insufficiency (OR, 0.43; 95% CI, 0.22–0.82, P=0.01) were also associated with a lower in‐hospital mortality in the closed unit. We did not find a difference in CICU length of stay or total hospital charges (P>0.05). Conclusions We found an association between lower in‐hospital and CICU mortality after the transition to a closed CICU. These results may help guide the ongoing redesign in other tertiary care CICUs.

Importance Injuries cause 30% more deaths than HIV, TB and malaria combined, and a prospective fracture care registry was established to investigate the fracture burden and treatment in Malawi to inform evidence-based improvements. Objective To use the analysis of prospectively-collected fracture data to develop evidence-based strategies to improve fracture care in Malawi and other similar settings. Design Multicentre prospective registry study. Setting Two large referral centres and two district hospitals in Malawi. Participants All patients with a fracture (confirmed by radiographs)—including patients with multiple fractures—were eligible to be included in the registry. Exposure All fractures that presented to two urban central and two rural district hospitals in Malawi over a 3.5-year period (September 2016 to March 2020). Main outcome(s) and measure(s) Demographics, characteristics of injuries, and treatment outcomes were collected on all eligible participants. Results Between September 2016 and March 2020, 23,734 patients were enrolled with a median age of 15 years (interquartile range: 10–35 years); 68.7% were male. The most common injuries were radius/ulna fractures (n = 8,682, 36.8%), tibia/fibula fractures (n = 4,036, 17.0%), humerus fractures (n = 3,527, 14.9%) and femoral fractures (n = 2,355, 9.9%). The majority of fractures (n = 21,729, 91.6%) were treated by orthopaedic clinical officers; 88% (20,885/2,849) of fractures were treated non-operatively, and 62.7% were treated and sent home on the same day. Open fractures (OR:53.19, CI:39.68–72.09), distal femoral fractures (OR:2.59, CI:1.78–3.78), patella (OR:10.31, CI:7.04–15.07), supracondylar humeral fractures (OR:3.10, CI:2.38–4.05), ankle fractures (OR:2.97, CI:2.26–3.92) and tibial plateau fractures (OR:2.08, CI:1.47–2.95) were more likely to be treated operatively compared to distal radius fractures. Conclusions and relevance The current model of fracture care in Malawi is such that trained orthopaedic surgeons manage fractures operatively in urban referral centres whereas orthopaedic clinical officers mainly manage fractures non-operatively in both district and referral centres. We recommend that orthopaedic surgeons should supervise orthopaedic clinical officers to manage non operative injuries in central and district hospitals. There is need for further studies to assess the clinical and patient reported outcomes of these fracture cases, managed both operatively and non-operatively.

Unstructured citizen-science data are increasingly used for analysing the abundance and distribution of species. Here we test the usefulness of such data to predict the seasonal distribution of migratory songbirds, and to analyse patterns of migratory connectivity.We used bird occurrence data from eBird, one of the largest global citizen science databases, to predict the year-round distribution of eight songbird taxa (Agropsar philippensis, Calliope calliope, Cecropis daurica, Emberiza aureola, Hirundo rustica, Locustella certhiola, Oriolus chinensis, Saxicola torquatus stejnegeri) that migrate through East Asia, a region especially poor in data but globally important for the conservation of migratory land birds. Maximum entropy models were built to predict spring stopover, autumn stopover and wintering areas. Ring recovery and geolocator tracking data were then used to evaluate, how well the predicted occurrence at a given period of the annual cycle matched sites where the species were known to be present from ringing and tracking data.Predicted winter ranges were generally smaller than those on published extent-of-occurrence maps (the hitherto only available source of distribution information). There was little overlap in stopover regions. The overlap between areas predicted as suitable from the eBird data and areas that had records from geolocator tracking was high in winter, and lower for spring and autumn migration. Less than 50% of the ringing recoveries came from locations within the seasonal predicted areas, with the highest overlap in autumn. The seasonal range size of a species affected the matching of tracking/ringing data with the predictions. Strong migratory connectivity was evident in Siberian Rubythroats and Barn Swallows. We identified two migration corridors, one over the eastern mainland of China, and one along a chain of islands in the Pacific.We show that the combination of disparate data sources has great potential to gain a better understanding of the non-breeding distribution and migratory connectivity of Eastern Palearctic songbirds. Citizen-science observation data are useful even in remote areas to predict the seasonal distribution of migratory species, especially in periods when birds are sedentary and when supplemented with tracking data.

Background Hypertrophic cardiomyopathy (HCM) is the most common inherited cardiomyopathy. Current guidelines endorse management in expert centers, but patient socioeconomic status can affect access to specialty care. The effect of socioeconomic status and specialty care access on HCM outcomes has not been examined. Methods and Results We conducted a retrospective cohort study that examined outcomes among HCM patients receiving care in the Yale New Haven Health System between June 2011 and December 2017. Patients were assigned to lower or higher socioeconomic status groups (LSES/HSES) based on medical insurance provider and to receivers of specialty care (SC) at Yale's Inherited Cardiomyopathy clinic or general cardiology care (GC). The primary outcome was all‐cause death, and the secondary outcome was all‐cause hospitalization. We identified 953 HCM patients; 820 (86%) were HSES and 133 (14%) were LSES. Forty‐three (4.5%) patients died from cardiac and noncardiac causes. LSES patients within the general cardiology care cohort had significantly higher all‐cause mortality compared with HSES patients (adjusted hazard ratio, [95% CI]=10.06 [4.38–23.09]; P

The novel coronavirus SARS-CoV-2 is the seventh identified human coronavirus. Understanding the extent of pre-existing immunity induced by seropositivity to endemic seasonal coronaviruses and the impact of cross-reactivity on COVID-19 disease progression remains a key research question in immunity to SARS-CoV-2 and the immunopathology of COVID-2019 disease. This paper describes a panel of lentiviral pseudotypes bearing the spike (S) proteins for each of the seven human coronaviruses (HCoVs), generated under similar conditions optimized for high titre production allowing a high-throughput investigation of antibody neutralization breadth. Optimal production conditions and most readily available permissive target cell lines were determined for spike-mediated entry by each HCoV pseudotype: SARS-CoV-1, SARS-CoV-2 and HCoV-NL63 best transduced HEK293T/17 cells transfected with ACE2 and TMPRSS2, HCoV-229E and MERS-CoV preferentially entered HUH7 cells, and CHO cells were most permissive for the seasonal betacoronavirus HCoV-HKU1. Entry of ACE2 using pseudotypes was enhanced by ACE2 and TMPRSS2 expression in target cells, whilst TMPRSS2 transfection rendered HEK293T/17 cells permissive for HCoV-HKU1 and HCoV-OC43 entry. Additionally, pseudotype viruses were produced bearing additional coronavirus surface proteins, including the SARS-CoV-2 Envelope (E) and Membrane (M) proteins and HCoV-OC43/HCoV-HKU1 Haemagglutinin-Esterase (HE) proteins. This panel of lentiviral pseudotypes provides a safe, rapidly quantifiable and high-throughput tool for serological comparison of pan-coronavirus neutralizing responses; this can be used to elucidate antibody dynamics against individual coronaviruses and the effects of antibody cross-reactivity on clinical outcome following natural infection or vaccination.

A microfluidic protein aggregation device (microPAD) that allows the user to perform a series of protein incubations with various concentrations of two reagents is demonstrated. The microfluidic device consists of 64 incubation chambers to perform individual incubations of the protein at 64 specific conditions. Parallel processes of metering reagents, stepwise concentration gradient generation, and mixing are achieved simultaneously by pneumatic valves. Fibrillation of bovine insulin was selected to test the device. The effect of insulin and sodium chloride (NaCl) concentration on the formation of fibrillar structures was studied by observing the growth rate of partially folded protein, using the fluorescent marker Thioflavin-T. Moreover, dual gradients of different NaCl and hydrochloric acid (HCl) concentrations were formed, to investigate their interactive roles in the formation of insulin fibrils and spherulites. The chip-system provides a bird’s eye view on protein aggregation, including an overview of the factors that affect the process and their interactions. This microfluidic platform is potentially useful for rapid analysis of the fibrillation of proteins associated with many misfolding-based diseases, such as quantitative and qualitative studies on amyloid growth.