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Abstract High-grade serous ovarian cancer (HGSOC) is marked by significant molecular diversity, presenting a major clinical challenge due to its aggressive nature and poor prognosis. This study aims to deepen the understanding of HGSOC by characterizing mRNA subtypes and examining their immune microenvironment (TIME) and its role in disease progression. Using transcriptomic data and an advanced computational pipeline, we investigated four mRNA subtypes: immunoreactive, differentiated, proliferative, and mesenchymal, each associated with distinct gene expression profiles and clinical behaviors. We performed differential expression analysis among mRNA subtypes using DESeq2 and conducted Weighted Gene Co-Expression Network Analysis (WGCNA) to identify co-expressed gene modules related to clinical traits, e.g., age, survival, and subtype classification. Gene Ontology (GO) analysis highlighted key pathways involved in tumor progression and immune evasion. Additionally, we utilized TIMER 2.0 to assess immune cell infiltration across different HGSOC subtypes, providing insights into the interplay between tumor immune microenvironment (TIME). Our findings show that the immunoreactive subtype, particularly the M3 module-associated network, was marked by high immune cell infiltration, including M1 (p

ABSTRACT Objective Hypoalbuminemia (HA) is a proxy for malnutrition that has been associated with postoperative complications in several surgical subspecialties. This study investigates the association between pre‐operative HA and complications following major salivary gland (MSG) resection. Methods Patients undergoing outpatient, elective MSG resection were extracted from the 2005 to 2020 National Surgical Quality Improvement Program database. Demographics and comorbidities were compared between HA (preoperative serum albumin

Summary: Background: While people with type 2 diabetes (T2D) are more susceptible to infections, studies potentially underestimate the true burden of infection-related mortality since they rely on clinical coding systems primarily structured by body system, and by only focusing on underlying cause. This study examined cause-specific mortality in people with T2D compared to the general population during 2015–2019, focusing on infections. Methods: 509,403 people aged 41–90 years with T2D alive on 1/1/2015 in Clinical Practice Research Datalink were matched to 976,431 without diabetes on age, sex, and ethnicity. Recorded underlying cause of death was identified through national linked mortality data; infection-related deaths were counted across all ICD-10 (10th revision of the International Classification of Diseases) chapters, not just infection chapters A00-B99. All-cause and cause-specific hazard ratios (HR) for mortality during 2015–2019 compared people with T2D to people without diabetes and were estimated using Cox models adjusting for region. Additional analyses for sepsis related mortality considered the impact of including any mention of sepsis on the death certificate. Findings: 85,367/509,403 (16.8%) people with T2D died during 2015–2019 compared to 106,824/976,431 (10.9%) of people without diabetes of the same sex, age and ethnicity. All infections (11,128/85,367 = 13.0%) represented the third highest underlying cause of death among people with T2D after cardiovascular disease and cancer; a much higher contribution than counting only from specific infection chapters (1046/85,367 = 1.2%). The HR for people with T2D vs non-diabetes for all infection mortality (1.82, 95% CI 1.78–1.86) was higher than that estimated for all-cause (HR = 1.65, 95% CI 1.64–1.66). The estimated mortality rate associated with sepsis among people with T2D was highly dependent on whether any mention was included (2.2 per 1000 person-years) or only underlying cause (0.2 per 1000 person-years); but the HR for people with T2D vs non-diabetes was similar (any mention HR = 2.26, 95% CI 2.19–2.34 vs underlying cause only HR = 2.52, 95% CI 2.27–2.80). Interpretation: People with T2D die from infections at a higher rate than similar people without diabetes, and the overall burden is greater than previously reported. Routine statistics concentrating on underlying cause of death may somewhat under-estimate the importance of infections as causes of death among people with T2D. These findings emphasise the potential importance of awareness, earlier diagnosis and treatment of infections to prevent premature deaths. Funding: National Institute for Health and Care Research.

Abstract Near-infrared (NIR) probes are highly sought after as fluorescent tags for multicolor cellular and in vivo imaging. Here we develop small NIR fluorescent nanobodies, termed NIR-FbLAG16 and NIR-FbLAG30, enabling background-free visualization of various GFP-derived probes and biosensors. We also design a red-shifted variant, NIR-Fb(718), to simultaneously target several antigens within the NIR spectral range. Leveraging the antigen-stabilizing property of the developed NIR-Fbs, we then create two modular systems for precise control of gene expression in GFP-labeled cells. Applying the NIR-Fbs in vivo, we target cells expressing GFP and the calcium biosensor GCaMP6 in the somatosensory cortex of transgenic mice. Simultaneously tracking calcium activity and the reference signal from NIR-FbLAGs bound to GCaMP6 enables ratiometric deep-brain in vivo imaging. Altogether, NIR-FbLAGs present a promising approach for imaging and manipulating various processes in live cells and behaving animals expressing GFP-based probes.

Abstract Head and neck squamous cell carcinomas (HNSCCs) are cancers that arise in the mucosa of the upper aerodigestive tract. The five-year patient survival rate is ~50%. Treatment includes surgery, radiation, and/or chemotherapy and is associated with lasting effects even when successful in irradicating the disease. New molecular targets and therapies must be identified to improve outcomes for HNSCC patients. We recently identified bitter taste receptors (taste family 2 receptors, or T2Rs) as a novel candidate family of receptors that activate apoptosis in HNSCC cells through mitochondrial Ca2+ overload and depolarization. We hypothesized that targeting another component of tumor cell metabolism, namely glycolysis, may increase the efficacy of T2R-directed therapies. GLUT1 (SLC2A1) is a facilitated-diffusion glucose transporter expressed by many cancer cells to fuel their increased rates of glycolysis. GLUT1 is already being investigated as a possible cancer target, but studies in HNSCCs are limited. Examination of immortalized HNSCC cells, patient samples, and The Cancer Genome Atlas revealed high expression of GLUT1 and upregulation in some patient tumor samples. HNSCC cells and tumor tissue express GLUT1 on the plasma membrane and within the cytoplasm (perinuclear, likely co-localized with the Golgi apparatus). We investigated the effects of a recently developed small molecule inhibitor of GLUT1, BAY-876. This compound decreased HNSCC glucose uptake, viability, and metabolism and induced apoptosis. Moreover, BAY-876 had enhanced effects on apoptosis when combined at low concentrations with T2R bitter taste receptor agonists. Notably, BAY-876 also decreased TNFα-induced IL-8 production, indicating an additional mechanism of possible tumor-suppressive effects. Our study demonstrates that targeting GLUT1 via BAY-876 to kill HNSCC cells, particularly in combination with T2R agonists, is a potential novel treatment strategy worth exploring further in future translational studies.

Abstract Objective To investigate primary site surgical resection and overall survival (OS) in clinically distantly metastatic (cM1) oral cavity squamous cell carcinoma (OCSCC). Methods The 2006–2018 National Cancer Database was queried for patients presenting with cM1 OCSCC who underwent chemotherapy. Binary logistic, Kaplan–Meier, and multivariable Cox proportional hazards regression models were implemented. Results Of 278 patients satisfying inclusion criteria, 139 (50.0%) underwent chemotherapy alone, 80 (28.8%) underwent chemoradiotherapy, 25 (9.0%) underwent surgical resection + adjuvant chemotherapy, and 34 (12.2%) underwent surgical resection + adjuvant chemoradiotherapy; 5‐year OS was 9.4%, 15.2%, 8.3%, and 23.8%, respectively (p

Abstract Objective Surgical resection is associated with higher overall survival (OS) than definitive radiotherapy (RT) or chemoradiotherapy (CRT) in cT4b sinonasal squamous cell carcinoma (SCC). Our study investigates the survival benefit of surgical resection in cT4b sinonasal non‐SCC. Methods The 2004 to 2019 National Cancer Database was queried for patients with cT4b sinonasal non‐SCC undergoing definitive treatment with (1) surgical resection + additional therapy (RT, chemotherapy, or both), (2) RT alone, or (3) CRT. Surgical resection + additional therapy and definitive RT/CRT were compared with Kaplan–Meier and multivariable Cox regression models. Results Of 629 patients satisfying inclusion criteria, 513 (81.6%) underwent surgical resection + additional therapy and 116 (18.4%) underwent definitive RT/CRT. The most frequent histologic types were undifferentiated carcinoma (23.7%), adenoid cystic carcinoma (22.6%), and adenocarcinoma (20.7%). Few patients presented with clinical nodal metastasis (15.7%). There were 4 (0.8%) mortalities within 90 days of surgical resection. Patients undergoing surgical resection with positive surgical margins had higher 5‐year OS than those undergoing definitive RT/CRT (56.3% vs. 39.4%, p = .039) and similar 5‐year OS as those with negative margins (56.3% vs. 63.9%, p = .059). Patients undergoing neoadjuvant chemotherapy had similar 5‐year OS as those undergoing definitive RT/CRT (60.9% vs. 39.5%, p = .053). Age at diagnosis, tumor diameter, and surgical resection + additional therapy (aHR 0.64, 95% CI 0.45–0.91) were associated with OS (p

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is a human pathogenic virus that encodes for a helicase (SC2Hel) that is essential for viral replication. SC2Hel has the ability to unravel dsRNA or dsDNA in an NTP-dependent manner from the 5′ to 3′ directionality. The standard helicase assay from studies involving SARS-CoV and SARS-CoV-2 have relied on the concept of fluorescence resonance energy transfer. Adding to the collection of helicase assays, herein, we have developed a novel tetrazolium-based colorimetric assay system for the detection of ADP that is produced via SC2Hel activity. This SC2Hel assay combines three enzyme-coupled steps involving the ADP-dependent Thermococcus litoralis glucokinase (TlGlcK), Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase (LmG6PDH), and Clostridium kluyveri diaphorase (CkDIA). Iodonitrotetrazolium chloride (INT), a colorimetric tetrazolium reagent, was used in the final step of the assay that converted into INT-formazan during reduction. INT-formazan in the assay’s buffered solution at pH 7.6 exhibited an intense colorimetric response at a wavelength maximum of 505 nm. The assay exhibited excellent performance characteristics as it revealed a Z’ factor of 0.87 and it has the potential to be further adopted into high-throughput screening studies for therapeutic drug discovery research.

IntroductionThere is a growing interest in characterizing the cognitive-motor processes that underlie superior performance in highly skilled athletes. The aim of this study was to explore neural markers of putting performance in highly skilled golfers by recording mobile EEG (electroencephalogram) during the pre-shot period.MethodsTwenty-eight right-handed participants (20 males) with a mean age of 24.2 years (± 6.4) and an average handicap of +1.7 (± 6.4) completed a testing session. Following the warm-up, participants completed 140 putts from a distance of 8ft (2.4m), with putts taken from 5 different positions. While putting, participants wore an eye tracker and a gel-based EEG system with 32 electrodes. Time and frequency domain features of the EEG signals were extracted to characterize Movement-Related Cortical Potentials (MRCP) and rhythmic modulations of neural activity in theta, alpha, sensorimotor and beta frequency bands associated with putting performance.ResultsEye-tracking data demonstrate that mean Quiet Eye durations are not a reliable marker of expertise as the same duration was found for both successful and unsuccessful putts. Following rigorous data processing data from 12 participants (8 males, mean age 21.6 years ± 5.4, average handicap +1.5 ± 4.4) were included in the EEG analysis. MRCP analysis revealed performance-based differences, with unsuccessful putts having a greater negative amplitude in comparison to successful putts. Time frequency analysis of the EEG data revealed that successful putts exhibit distinct neural activity profiles compared to unsuccessful ones. For successful putts, greater suppression of beta was present in the central region prior to the putt. By contrast, increased frontal theta power was present for unsuccessful putts immediately before the putt (consistent with hesitation and the need for motor plan adjustments prior to execution).DiscussionWe propose that neural activity may provide plausible insights into the mechanisms behind why identical QE durations can lead to both success and failure. From an applied perspective, this study highlights the merits of a multi-measure approach to gain further insights into performance differences within highly skilled golfers. We discuss considerations for future research and solutions to address the challenges related to the complexities of collecting clean EEG signals within naturalistic sporting contexts.

Stroke is a leading cause of long-term disability worldwide. With the advancements in sensor technologies and data availability, artificial intelligence (AI) holds the promise of improving the amount, quality and efficiency of care and enhancing the precision of stroke rehabilitation. We aimed to identify and characterize the existing research on AI applications in stroke recovery and rehabilitation of adults, including categories of application and progression of technologies over time. Data were collected from peer-reviewed articles across various electronic databases up to January 2024. Insights were extracted using AI-enhanced multi-method, data-driven techniques, including clustering of themes and topics. This scoping review summarizes outcomes from 704 studies. Four common themes (impairment, assisted intervention, prediction and imaging, and neuroscience) were identified, in which time-linked patterns emerged. The impairment theme revealed a focus on motor function, gait and mobility, while the assisted intervention theme included applications of robotic and brain–computer interface (BCI) techniques. AI applications progressed over time, starting from conceptualization and then expanding to a broader range of techniques in supervised learning, artificial neural networks (ANN), natural language processing (NLP) and more. Applications focused on upper limb rehabilitation were reviewed in more detail, with machine learning (ML), deep learning techniques and sensors such as inertial measurement units (IMU) used for upper limb and functional movement analysis. AI applications have potential to facilitate tailored therapeutic delivery, thereby contributing to the optimization of rehabilitation outcomes and promoting sustained recovery from rehabilitation to real-world settings.

The incidence of diabetes has been increasing and, in parallel, so has the incidence of females in their childbearing years with diabetes. Preconception care is important in females with diabetes due many factors related to fertility, blood glucose control, and complications. For example, many individuals with Type 2 diabetes are obese, which can affect contraceptive efficacy, fertility, and fetal growth. Additionally, patients with all types of diabetes are at risk for disordered eating, which can be harmful to a developing fetus. Both hypoglycaemia and hyperglycaemia are known to increase the risk of adverse maternal and neonatal outcomes, including during the first trimester, when many females are not aware of pregnancy. Additionally, individuals with diabetes are at risk for complications, such as atherosclerotic cardiovascular disease, nephropathy, retinopathy, and neuropathy, that can lead to complicated pregnancies. Importantly, several of the medications used to control blood glucose, and manage diabetes complications, are not recommended for use during pregnancy due to potential fetal harm. For these reasons, females with diabetes in their childbearing years are encouraged to utilise reliable contraception, so that pregnancies can be planned, or should be treated with medications with low teratogenicity potential. Thus, the preconception care of females with diabetes is complex, and the increasing prevalence of this patient population warrants greater awareness among clinicians. This narrative review summarises the current standard of preconception care for individuals with diabetes, including the management of contraception, weight, blood glucose, hypertension, and dyslipidaemia.

The National Institute of Health Stroke Scale (NIHSS) is used worldwide to classify stroke severity as ‘mild’, ‘moderate’, or ‘severe’ based on neurological impairment. Yet, stroke survivors argue that the classification of ‘mild’ does not represent the holistic experience and impact of stroke on their daily lives. In this observational cohort study, we aimed to identify different types of impairment profiles among stroke survivors classified as ‘mild’. We used survivors of mild stroke’ data from the START longitudinal stroke cohort (n = 73), with measures related to sensorimotor, cognition, depression, functional disability, physical activity, work, and social adjustment over 12 months. Given the multisource, multigranular, and unlabeled nature of the data, we utilized a structure-adapting, unsupervised machine learning approach, the growing self-organizing map (GSOM) algorithm, to generate distinct clinical profiles. These diverse impairment profiles revealed that survivors of mild stroke experience varying degrees of impairment and impact (cognitive, depression, physical activity, work/social adjustment) at different time points, despite the uniformity implied by their NIHSS-classified ‘mild’ stroke. This emphasizes the necessity of creating a holistic and more comprehensive representation of survivors of mild stroke’ needs over the first year after stroke to improve rehabilitation and poststroke care.

Background: The use of anticoagulant medications is complex and prone to inappropriate prescribing that impacts patient safety. Anticoagulation stewardship is a growing field that requires more focus and attention. Aim: This review aimed to retrieve and synthesise the available research studies on the clinical and economic value of Anticoagulation Stewardship Programmes (ASPs) in line with the Anticoagulation Forum Core Elements of Anticoagulation Stewardship. Methods: A comprehensive electronic literature search was conducted using three databases (Cochrane Central Register of Controlled Trials (CENTRAL), Embase and PubMed) from inception up to and including January 2023. Inclusion criteria were primary research studies where the purpose of the study was any hospital ASP intervention applicable to optimising antithrombotic drug use, across all anticoagulation medications and in adult hospital inpatients. Results: A total of 787 records were identified after duplicates were removed. Twenty-eight papers were included in this review. Twenty-four of these studies were single-centre studies; four of these studies were multi-centre studies. Eleven studies took a prospective approach, fifteen took a retrospective approach and two were quasi-experimental studies. Interventions implemented by either multidisciplinary ASP teams or pharmacist-led anticoagulation services included the provision of education to healthcare professionals and patients, undertaking medication reviews and implementing guidelines and protocols, and interventions to facilitate transitions of care. Clinical benefits to patients and cost savings were observed across many studies. Conclusion: Implementing multidisciplinary ASP teams and pharmacist-led anticoagulation services is associated with an overall improvement in the safe use of anticoagulation among hospital patients. Studies identified incorporated some of the core elements of ASP, however further work and research are necessary to standardise the adoption and implementation of ASP and ensure that it is prioritised among healthcare professionals, in the healthcare setting, and among health systems and policy-makers.

Background: Dental unit waterline (DUWL) safety is vital for dental practitioners. Unfortunately, the standard heterotrophic plate count (HPC) methods commonly used to identify bacterial contamination dramatically underestimate planktonic bacteria counts. Flow cytometry (FCM) directly counts bacterial cells using laser-activated fluorescence, providing more reliable and timely results. Methods: Approximately 1,700 DUWL samples were measured by both HPC and FCM methods. HPC samples were plated on R2A agar within 8 hours of receipt, incubated at 25 °C for 7 days, and reported as colony-forming units (CFU)/mL. FCM samples were treated with SYBR Green I fluorescent stain, analyzed and reported as high nucleic acid (HNA) and low nucleic acid (LNA) cell counts. An empirical algorithm was developed using HNA and LNA cell counts compared with the recommended action limit for the Centers for Disease Control and Prevention’s greater than 500 CFU/mL pass or fail. Results: An FCM threshold limit that included both HNA 250 or greater and LNA 750 or greater cell counts was derived by pairing FCM data with R2A samples dichotomized as passing (≤ 500 CFU/mL) or failing (> 500 CFU/mL) to establish corresponding pass or fail criteria that would minimize false pass and false fail compared with R2A results. 1,388 (81.6%) of samples passed the R2A limit of 500 CFU/mL or fewer, whereas 1,427 (83.9%) FCM samples passed the proposed HNA 250 or fewer and LNA 750 or fewer criteria. Conclusions: The data show that FCM analysis and the proposed threshold criteria can be used as a powerful adjunctive tool for monitoring DUWL contamination, which includes greater accuracy and the ability to analyze samples immediately on receipt.

BackgroundMild stroke affects more than half the stroke population, yet there is limited evidence characterizing cognition over time in this population, especially with predictive approaches applicable at the individual-level. We aimed to identify patterns of recovery and the best combination of demographic, clinical, and lifestyle factors predicting individual-level cognitive state at 3- and 12-months after mild stroke.MethodsIn this prospective cohort study, the Montreal Cognitive Assessment (MoCA) was administered at 3-7 days, 3- and 12-months post-stroke. Raw changes in MoCA and impairment rates (defined as MoCAResultsParticipants' (n = 119) MoCA scores improved from baseline to 3-months (pConclusionMore than half of mild-stroke survivors are at risk of cognitive decline one year after stroke, even when preceded by a significantly improving pattern in the first 3-months of recovery. Physical activity was the only modifiable factor independently associated with cognitive recovery. Individual-level prediction methods may inform the timing and personalized application of future interventions to maximize cognitive recovery post-stroke.

Fluorescence imaging of the spinal cord poses challenges, including depth of imaging. Here the authors describe a custom microscope and chronically implanted microprism that enables multicolor translaminar imaging of sensory and motor evoked activity in behaving mice, and show that spinal astrocytes show sensorimotor program-dependent calcium excitation.

BackgroundPeople living in close quarters, such as military trainees, are at increased risk for skin and soft tissue infections (SSTI), especially those caused by methicillin-resistant Staphylococcus aureus (MRSA). The serum immune factors associated with the onset of SSTI are not well understood.MethodsWe conducted a longitudinal study of SSTIs, enrolling US Army trainees before starting military training and following up for 14 weeks. Samples were collected on Day 0, 56, and 90. Serum chemokines and cytokines among 16 SSTI cases and 51 healthy controls were evaluated using an electro-chemiluminescence based multiplex assay platform.ResultsOf 54 tested cytokines, 12 were significantly higher among SSTI cases as compared to controls. Among the cases, there were correlations between factors associated with vascular injury (i.e., VCAM-1, ICAM-1, and Flt1), the angiogenetic factor VEGF, and IL-10. Unsupervised machine learning (Principal Component Analysis) revealed that IL10, IL17A, C-reactive protein, ICAM1, VCAM1, SAA, Flt1, and VGEF were indicative of SSTI.ConclusionThe study demonstrates the power of immunoprofiling for identifying factors predictive of pre-illness state of SSTI thereby identifying early stages of an infection and individuals susceptible to SSTI.

Although both L-glutamate (Glu) and L-glutamine (Gln) have long been considered nutritionally nonessential in ruminants, these two amino acids have enormous nutritional and physiological importance. Results of recent studies revealed that extracellular Gln is extensively degraded by ruminal microbes, but extracellular Glu undergoes little catabolism by these cells due to the near absence of its uptake. Ruminal bacteria hydrolyze Gln to Glu plus ammonia and, intracellularly, use both amino acids for protein synthesis. Microbial proteins and dietary Glu enter the small intestine in ruminants. Both Glu and Gln are the major metabolic fuels and building blocks of proteins, as well as substrates for the syntheses of glutathione and amino acids (alanine, ornithine, citrulline, arginine, proline, and aspartate) in the intestinal mucosa. In addition, Gln and aspartate are essential for purine and pyrimidine syntheses, whereas arginine and proline are necessary for the production of nitric oxide (a major vasodilator) and collagen (the most abundant protein in the body), respectively. Under normal feeding conditions, all diet- and rumen-derived Glu and Gln are extensively utilized by the small intestine and do not enter the portal circulation. Thus, de novo synthesis (e.g., from branched-chain amino acids and α-ketoglutarate) plays a crucial role in the homeostasis of Glu and Gln in the whole body but may be insufficient for maximal growth performance, production (e.g., lactation and pregnancy), and optimal health (particularly intestinal health) in ruminants. This applies to all types of feeding systems used around the world (e.g., rearing on a milk replacer before weaning, pasture-based production, and total mixed rations). Dietary supplementation with the appropriate doses of Glu or Gln [e.g., 0.5 or 1 g/kg body weight (BW)/day, respectively] can safely improve the digestive, endocrine, and reproduction functions of ruminants to enhance their productivity. Both Glu and Gln are truly functional amino acids in the nutrition of ruminants and hold great promise for improving their health and productivity.

Stroke recovery is multifaceted and complex. Machine learning approaches have potential to identify patterns of brain activity associated with clinical outcomes, providing new insights into recovery. We aim to use machine learning to characterise the contribution of and potential interaction between resting state functional connectivity networks in predicting touch discrimination outcomes in a well-phenotyped, but small, stroke cohort. We interrogated and compared a suite of automated machine learning approaches to identify patterns of brain activity associated with clinical outcomes. Using feature reduction, the identification of combined ‘golden features’, and five-fold cross-validation, two golden features patterns emerged. These golden features identified patterns of resting state connectivity involving interactive relationships: 1. The difference between right insula and right superior temporal lobe correlation and left cerebellum and vermis correlation; 2. The ratio between right inferior temporal lobe and left cerebellum correlation and left frontal inferior operculum and left supplementary motor area correlation. Our findings demonstrate evidence of the potential for automated machine learning to provide new insights into brain network patterns and their interactions associated with the prediction of quantitative touch discrimination outcomes, through the automated identification of robust associations and golden feature brain patterns, even in a small cohort of stroke survivors.

Abstract Objective The reconstruction of large scalp defects poses both functional and cosmetic challenges. While free tissue transfer remains the standard for defects larger than 30 cm2, prolonged anesthesia and postoperative complications remain significant limitations. The purpose of this study is to evaluate the use of O‐Z flaps for the reconstruction of large scalp defects and to describe the techniques employed. Methods This is a retrospective analysis of ten patients who underwent reconstructive surgery using an O‐Z flap approach for large scalp defects between July 2017 and June 2019. The parameters included in this study were patient demographics, tumor characteristics, and postoperative management, collected for at least a year after surgery. Results In this cohort, the mean age was 76.1 years and 90% were male. All patients were treated for neoplastic skin lesions, with 70% located on the vertex and 30% located on the temporoparietal region. The mean size of defect was 52.0 cm2 (range: 38.6 to 63.8 cm2). The maximum hospital stay was two days, and no patients were readmitted within 30 days of surgery. There were no cases of wound infection or flap necrosis. All patients reported pain control with acetaminophen and ibuprofen. Four patients received adjuvant radiation, and there was no delay to receiving treatment following surgery. Conclusions The O‐Z flap is a reliable alternative for the reconstruction of non‐irradiated scalp defects in the vertex and temporoparietal regions up to 63 cm2. This technique provides advantages for patients, including hairline preservation, shorter hospital stays, and decreased postoperative complications.

Background The renin‐angiotensin system plays a crucial role in human physiology, and its main hormone, angiotensin, activates 2 G‐protein–coupled receptors, the angiotensin type‐1 and type‐2 receptors, in almost every organ. However, controversy exists about the location, distribution, and expression levels of these receptors. Concerns have been raised over the low sensitivity, low specificity, and large variability between lots of commercially available antibodies for angiotensin type‐1 and type‐2 receptors, which makes it difficult to reconciliate results of different studies. Here, we describe the first non–antibody‐based sensitive and specific targeted quantitative mass spectrometry assay for angiotensin receptors. Methods and Results Using a technique that allows targeted analysis of multiple peptides across multiple samples in a single mass spectrometry analysis, known as TOMAHAQ (triggered by offset, multiplexed, accurate mass, high resolution, and absolute quantification), we have identified and validated specific human tryptic peptides that permit identification and quantification of angiotensin type‐1 and type‐2 receptors in biological samples. Several peptide sequences are conserved in rodents, making these mass spectrometry assays amenable to both preclinical and clinical studies. We have used this method to quantify angiotensin type‐1 and type‐2 receptors in postmortem frontal cortex samples of older adults (n=28) with Alzheimer dementia. We correlated levels of angiotensin receptors to biomarkers classically linked to renin‐angiotensin system activation, including oxidative stress, inflammation, amyloid‐β load, and paired helical filament‐tau tangle burden. Conclusions These robust high‐throughput assays will not only catalyze novel mechanistic studies in the angiotensin research field but may also help to identify patients with an unbalanced angiotensin receptor distribution who would benefit from angiotensin receptor blocker treatment.

Glacial lake outburst floods (GLOFs) are among the most concerning consequences of retreating glaciers in mountain ranges worldwide. GLOFs have attracted significant attention amongst scientists and practitioners in the past 2 decades, with particular interest in the physical drivers and mechanisms of GLOF hazard and in socioeconomic and other human-related developments that affect vulnerabilities to GLOF events. This increased research focus on GLOFs is reflected in the gradually increasing number of papers published annually. This study offers an overview of recent GLOF research by analysing 594 peer-reviewed GLOF studies published between 2017 and 2021 (Web of Science and Scopus databases), reviewing the content and geographical focus as well as other characteristics of GLOF studies. This review is complemented with perspectives from the first GLOF conference (7–9 July 2021, online) where a global GLOF research community of major mountain regions gathered to discuss the current state of the art of integrated GLOF research. Therefore, representatives from 17 countries identified and elaborated trends and challenges and proposed possible ways forward to navigate future GLOF research, in four thematic areas: (i) understanding GLOFs – timing and processes; (ii) modelling GLOFs and GLOF process chains; (iii) GLOF risk management, prevention and warning; and (iv) human dimensions of GLOFs and GLOF attribution to climate change.

Room-temperature macromolecular crystallography allows protein structures to be determined under close-to-physiological conditions, permits dynamic freedom in protein motions and enables time-resolved studies. In the case of metalloenzymes that are highly sensitive to radiation damage, such room-temperature experiments can present challenges, including increased rates of X-ray reduction of metal centres and site-specific radiation-damage artefacts, as well as in devising appropriate sample-delivery and data-collection methods. It can also be problematic to compare structures measured using different crystal sizes and light sources. In this study, structures of a multifunctional globin, dehaloperoxidase B (DHP-B), obtained using several methods of room-temperature crystallographic structure determination are described and compared. Here, data were measured from large single crystals and multiple microcrystals using neutrons, X-ray free-electron laser pulses, monochromatic synchrotron radiation and polychromatic (Laue) radiation light sources. These approaches span a range of 18 orders of magnitude in measurement time per diffraction pattern and four orders of magnitude in crystal volume. The first room-temperature neutron structures of DHP-B are also presented, allowing the explicit identification of the hydrogen positions. The neutron data proved to be complementary to the serial femtosecond crystallography data, with both methods providing structures free of the effects of X-ray radiation damage when compared with standard cryo-crystallography. Comparison of these room-temperature methods demonstrated the large differences in sample requirements, data-collection time and the potential for radiation damage between them. With regard to the structure and function of DHP-B, despite the results being partly limited by differences in the underlying structures, new information was gained on the protonation states of active-site residues which may guide future studies of DHP-B.

Abstract The human microbiome is comprised of a complex and diverse community of organisms that is subject to dynamic changes over time. As such, cross-sectional studies of the microbiome provide a multitude of information for a specific body site at a particular time, but they fail to account for temporal changes in microbial constituents resulting from various factors. To address this shortcoming, longitudinal research studies of the human microbiome investigate the influence of various factors on the microbiome of individuals within a group or community setting. These studies are vital to address the effects of host and/or environmental factors on microbiome composition as well as the potential contribution of microbiome members during the course of an infection. The relationship between microbial constituents and disease development has been previously explored for skin and soft tissue infections (SSTIs) within congregate military trainees. Accordingly, approximately 25% of the population carries Staphylococcus aureus within their nasal cavity, and these colonized individuals are known to be at increased risk for SSTIs. To examine the evolution of the nasal microbiota of U.S. Army Infantry trainees, individuals were sampled longitudinally from their arrival at Fort Benning, Georgia, until completion of their training 90 days later. These samples were then processed to determine S. aureus colonization status and to profile the nasal microbiota using 16S rRNA gene-based methods. Microbiota stability differed dramatically among the individual trainees; some subjects exhibited great stability, some subjects showed gradual temporal changes and some subjects displayed a dramatic shift in nasal microbiota composition. Further analysis utilizing the available trainee metadata suggests that the major drivers of nasal microbiota stability may be S. aureus colonization status and geographic origin of the trainees. Nasal microbiota evolution within the congregate setting imposed by military training is a complex process that appears to be affected by numerous factors. This finding may indicate that future campaigns to prevent S. aureus colonization and future SSTIs among high-risk military trainees may require a ‘personalized’ approach.

The Kieffer model is a widely accepted explanation for seasonal modification of the Martian surface by CO _2 ice sublimation and the formation of a “zoo” of intriguing surface features. However, the lack of in situ observations and empirical laboratory measurements of Martian winter conditions hampers model validation and refinement. We present the first experiments to investigate all three main stages of the Kieffer model within a single experiment: (i) CO _2 condensation on a thick layer of Mars regolith simulant; (ii) sublimation of CO _2 ice and plume, spot, and halo formation; and (iii) the resultant formation of surface features. We find that the full Kieffer model is supported on the laboratory scale as (i) CO _2 diffuses into the regolith pore spaces and forms a thin overlying conformal layer of translucent ice. When a buried heater is activated, (ii) a plume and dark spot develop as dust is ejected with pressurized gas, and the falling dust creates a bright halo. During plume activity, (iii) thermal stress cracks form in a network similar in morphology to certain types of spiders, dendritic troughs, furrows, and patterned ground in the Martian high south polar latitudes. These cracks appear to form owing to sublimation of CO _2 within the substrate, instead of surface scouring. We discuss the potential for this process to be an alternative formation mechanism for “cracked” spider-like morphologies on Mars. Leveraging our laboratory observations, we also provide guidance for future laboratory or in situ investigations of the three stages of the Kieffer model.

The role played by transient impact-induced endogenous brines in the formation of geomorphic features has been proposed on airless worlds such as Europa, Vesta, and Ceres, as well as on worlds with thin atmospheres such as Mars. After liquefaction, the hypothesized brines flow in a debris-flow-like process, incising curvilinear gullies and constructing lobate deposits within newly formed craters. Here we investigate what parameters (if any) could enable liquid to be transiently present for a sufficient time (∼tens of minutes) under postimpact transient atmospheric pressures (10 ^−4 –10 ^−5 torr) to form curvilinear gullies and lobate deposits, as have been seen on Vesta and more tentatively Ceres. We report that water likely vacuum-freezes too quickly, while NaCl brine enables flow longevity. We also find that frozen lid formation facilitates longer liquid lifetimes, as with lava in terrestrial lava tubes and model predictions for cryovolcanic flows on Europa. This work provides additional contributions to the growing body of literature that investigates the role of transient brines in sculpting the surfaces of airless worlds.

The authors demonstrate a compact multi-modal electro-optical coaxial microprobe design that offers small cross-sectional dimensions and tunable lengths for minimally invasive interfacing with intrinsic neural circuits.

The factors that determine the distinct profiles of NRAS mutants across different tumor types remain unclear. Here, the authors use an allelic series of conditional mouse models to investigate the molecular mechanisms underlying the enrichment of specific NRAS mutants in human melanoma

Abstract Originally identified on the tongue for their chemosensory role, the receptors for sweet, umami, and bitter taste are expressed in some cancers where they regulate important cellular processes including apoptosis and proliferation. We examined DNA mutations (n = 5103), structural variation (n = 7545), and expression (n = 6224) of genes encoding sweet or umami receptors (TAS1Rs) and bitter receptors (TAS2Rs) in 45 solid tumors subtypes compared to corresponding normal tissue using The Cancer Genome Atlas and the Genotype Tissue Expression Project databases. Expression of TAS1R and TAS2R genes differed between normal and cancer tissue, and nonsilent mutations occurred in many solid tumor taste receptor genes (~ 1–7%). Expression levels of certain TAS1Rs/TAS2Rs were associated with survival differences in 12 solid tumor subtypes. Increased TAS1R1 expression was associated with improved survival in lung adenocarcinoma (mean survival difference + 1185 days, p = 0.0191). Increased TAS2R14 expression was associated with worse survival in adrenocortical carcinoma (−1757 days, p

Abstract Coronavirus disease 2019 (COVID‐19) patients have increased thrombosis risk. With increasing age, there is an increase in COVID‐19 severity. Additionally, adults with a history of vasculopathy have the highest thrombotic risk in COVID‐19. The mechanisms of these clinical differences in risk remain unclear. Human umbilical vein endothelial cells (HUVECs) were infected with SARS‐CoV‐2, influenza A/Singapore/6/86 (H1N1) or mock‐infected prior to incubation with plasma from healthy children, healthy adults or vasculopathic adults. Fibrin on surface of cells was observed using scanning electron microscopy, and fibrin characteristics were quantified. This experiment was repeated in the presence of bivalirudin, defibrotide, low‐molecular‐weight‐heparin (LMWH) and unfractionated heparin (UFH). Fibrin formed on SARS‐CoV‐2 infected HUVECs was densely packed and contained more fibrin compared to mock‐infected cells. Fibrin generated from child plasma was the thicker than fibrin generated in vasculopathic adult plasma (p = 0.0165). Clot formation was inhibited by LMWH (0.5 U/ml) and UFH (0.1–0.7 U/ml). We show that in the context of the SARS‐CoV‐2 infection on an endothelial culture, plasma from vasculopathic adults produces fibrin clots with thinner fibrin, indicating that the plasma coagulation system may play a role in determining the thrombotic outcome of SARS‐CoV‐2 infection. Heparinoid anticoagulants were most effective at preventing clot formation.

Better management of head and neck squamous cell carcinomas (HNSCCs) requires a clearer understanding of tumor biology and disease risk. Bitter taste receptors (T2Rs) have been studied in several cancers, including thyroid, salivary, and GI, but their role in HNSCC has not been explored. We found that HNSCC patient samples and cell lines expressed functional T2Rs on both the cell and nuclear membranes. Bitter compounds, including bacterial metabolites, activated T2R‐mediated nuclear Ca2+ responses leading to mitochondrial depolarization, caspase activation, and ultimately apoptosis. Buffering nuclear Ca2+ elevation blocked caspase activation. Furthermore, increased expression of T2Rs in HNSCCs from The Cancer Genome Atlas is associated with improved overall survival. This work suggests that T2Rs are potential biomarkers to predict outcomes and guide treatment selection, may be leveraged as therapeutic targets to stimulate tumor apoptosis, and may mediate tumor‐microbiome crosstalk in HNSCC.

Background Cardiac metabolic abnormalities are present in heart failure. Few studies have followed metabolic changes accompanying diastolic and systolic heart failure in the same model. We examined metabolic changes during the development of diastolic and severe systolic dysfunction in spontaneously hypertensive rats (SHR). Methods and Results We serially measured myocardial glucose uptake rates with dynamic 2‐[18F] fluoro‐2‐deoxy‐d‐glucose positron emission tomography in vivo in 9‐, 12‐, and 18‐month‐old SHR and Wistar Kyoto rats. Cardiac magnetic resonance imaging determined systolic function (ejection fraction) and diastolic function (isovolumetric relaxation time) and left ventricular mass in the same rats. Cardiac metabolomics was performed at 12 and 18 months in separate rats. At 12 months, SHR hearts, compared with Wistar Kyoto hearts, demonstrated increased isovolumetric relaxation time and slightly reduced ejection fraction indicating diastolic and mild systolic dysfunction, respectively, and higher (versus 9‐month‐old SHR decreasing) 2‐[18F] fluoro‐2‐deoxy‐d‐glucose uptake rates (Ki). At 18 months, only few SHR hearts maintained similar abnormalities as 12‐month‐old SHR, while most exhibited severe systolic dysfunction, worsening diastolic function, and markedly reduced 2‐[18F] fluoro‐2‐deoxy‐d‐glucose uptake rates. Left ventricular mass normalized to body weight was elevated in SHR, more pronounced with severe systolic dysfunction. Cardiac metabolite changes differed between SHR hearts at 12 and 18 months, indicating progressive defects in fatty acid, glucose, branched chain amino acid, and ketone body metabolism. Conclusions Diastolic and severe systolic dysfunction in SHR are associated with decreasing cardiac glucose uptake, and progressive abnormalities in metabolite profiles. Whether and which metabolic changes trigger progressive heart failure needs to be established.

The authors estimate the associations between community socioeconomic composition and changes in coronavirus disease 2019 (COVID-19) vaccination levels in eight large cities at three time points. In March, communities with high socioeconomic status (SES) had significantly higher vaccination rates than low-SES communities. Between March and April, low-SES communities had significantly lower changes in percentage vaccinated than high-SES communities. Between April and May, this difference was not significant. Thus, the large vaccination gap between communities during restricted vaccine eligibility did not narrow when eligibility opened up. The link between COVID-19 vaccination and community disadvantage may lead to a bifurcated recovery whereby advantaged communities move on from the pandemic more quickly while disadvantaged communities continue to suffer.

IntroductionBitter taste receptors (T2Rs) are G protein-coupled receptors identified on the tongue but expressed all over the body, including in airway cilia and macrophages, where T2Rs serve an immune role. T2R isoforms detect bitter metabolites (quinolones and acyl-homoserine lactones) secreted by gram negative bacteria, including Pseudomonas aeruginosa, a major pathogen in cystic fibrosis (CF). T2R activation by bitter bacterial products triggers calcium-dependent nitric oxide (NO) production. In airway cells, the NO increases mucociliary clearance and has direct antibacterial properties. In macrophages, the same pathway enhances phagocytosis. Because prior studies linked CF with reduced NO, we hypothesized that CF cells may have reduced T2R/NO responses, possibly contributing to reduced innate immunity in CF.MethodsImmunofluorescence, qPCR, and live cell imaging were used to measure T2R localization, calcium and NO signaling, ciliary beating, and antimicrobial responses in air-liquid interface cultures of primary human nasal epithelial cells and immortalized bronchial cell lines. Immunofluorescence and live cell imaging was used to measure T2R signaling and phagocytosis in primary human monocyte-derived macrophages.ResultsPrimary nasal epithelial cells from both CF and non-CF patients exhibited similar T2R expression, localization, and calcium signals. However, CF cells exhibited reduced NO production also observed in immortalized CFBE41o- CF cells and non-CF 16HBE cells CRISPR modified with CF-causing mutations in the CF transmembrane conductance regulator (CFTR). NO was restored by VX-770/VX-809 corrector/potentiator pre-treatment, suggesting reduced NO in CF cells is due to loss of CFTR function. In nasal cells, reduced NO correlated with reduced ciliary and antibacterial responses. In primary human macrophages, inhibition of CFTR reduced NO production and phagocytosis during T2R stimulation.ConclusionsTogether, these data suggest an intrinsic deficiency in T2R/NO signaling caused by loss of CFTR function that may contribute to intrinsic susceptibilities of CF patients to P. aeruginosa and other gram-negative bacteria that activate T2Rs.

Background People with neuromuscular disease (NMD) experience a broader range of chronic diseases and health symptoms compared to the general population. However, no comprehensive analysis has directly quantified this to our knowledge. Methods We used a large UK primary care database (Clinical Practice Research Datalink) to compare the prevalence of chronic diseases and other health conditions, including recent infections between 23,876 patients with NMD ever recorded by 2019 compared to 95,295 age-sex-practice matched patients without NMD. Modified Poisson regression estimated Prevalence Ratios (PR) to summarise the presence of the disease/condition ever (or for infections in 2018) in NMD patients versus non-NMD patients. Results Patients with NMD had significantly higher rates for 16 of the 18 conditions routinely recorded in the primary care Quality and Outcomes Framework (QOF). Approximately 1-in-10 adults with NMD had ≥4 conditions recorded (PR = 1.39, 95%CI 1.33–1.45). Disparities were more pronounced at younger ages (18–49). For other (non-QOF) health conditions, significantly higher recorded levels were observed for rarer events (pulmonary embolism PR = 1.96 95%CI 1.76–2.18, hip fractures PR = 1.65 95%CI 1.47–1.85) as well as for more common primary care conditions (constipation PR = 1.52 95%CI 1.46–1.57, incontinence PR = 1.52 95%CI 1.44–1.60). The greatest co-morbidity burden was in patients with a myotonic disorder. Approximately 1-in-6 (17.1%) NMD patients had an infection recorded in the preceding year, with the risk of being hospitalised with an infection nearly double (PR = 1.92, 95%CI 1.79–2.07) compared to non-NMD patients. Conclusion The burden of chronic co-morbidity among patients with NMD is extremely high compared to the general population, and they are also more likely to present in primary and secondary care for acute events such as infections.

Objective: Visual impairment restricts performance in activities of daily living. The aim of this study was to classify types of visual loss associated with function in older adults based on clinical data and World Health Organization (WHO) acuity and visual field criteria. Methods: Seven hundred retrospective medical records of older adults seen at the outpatient ophthalmology clinics of a Singapore hospital were reviewed. Extracted data was mapped to the WHO low vision criteria. A flow chart was developed to classify the main types of visual loss aligned with function. Results: The flow chart developed describes four major types of visual loss: (1) full visual field with decreased visual acuity, (2) any visual field loss with greater than ten degrees of available field, (3) peripheral field loss with less than 10° of available field, and (4) any visual field loss due to a cortical event. Within each major type, sub-categories were identified reflecting the complexity of the visual impact of the eye conditions. Conclusion: The flow chart can be applied to outpatient records to identify older adults with different types of visual loss to inform targeted rehabilitation linked with function.

Altered somatosensory function is common among stroke survivors, yet is often poorly characterized. Methods of profiling somatosensation that illustrate the variability in impairment within and across different modalities remain limited. We aimed to characterize post-stroke somatosensation profiles (“fingerprints”) of the upper limb using an unsupervised machine learning cluster analysis to capture hidden relationships between measures of touch, proprioception, and haptic object recognition. Raw data were pooled from six studies where multiple quantitative measures of upper limb somatosensation were collected from stroke survivors (n = 207) using the Tactile Discrimination Test (TDT), Wrist Position Sense Test (WPST) and functional Tactile Object Recognition Test (fTORT) on the contralesional and ipsilesional upper limbs. The Growing Self Organizing Map (GSOM) unsupervised machine learning algorithm was used to generate a topology-preserving two-dimensional mapping of the pooled data and then separate it into clusters. Signature profiles of somatosensory impairment across two modalities (TDT and WPST; n = 203) and three modalities (TDT, WPST, and fTORT; n = 141) were characterized for both hands. Distinct impairment subgroups were identified. The influence of background and clinical variables was also modelled. The study provided evidence of the utility of unsupervised cluster analysis that can profile stroke survivor signatures of somatosensory impairment, which may inform improved diagnosis and characterization of impairment patterns.

Objectives: This study sought to evaluate the remineralization of ex vivo human teeth using commercially available artificial saliva, SalivaMAX®, a supersaturated calcium phosphate rinse (SSCPR). Methods: early enamel lesions were artificially induced on ex vivo human teeth by chemical means. The teeth were exposed to the SSCPR for two minutes (experimental) or dH2O (control) four times per day for a total of 35 days. At time points of 0, 2.5, 21, and 35 days, micro-CT was utilized to determine the mineral density profile across the lesion and evaluate lesion depth. The relative percent remineralization was calculated from the initial lesion depth (Time 0) at each evaluation time. Student’s t-test was used to compare the extent of remineralization between the SSCPR and control groups for statistical significance at each time. To evaluate the changes in percent remineralization over time, a two-way ANOVA was used. Results: At Time 0 and 2.5 days, there was no difference in the percent remineralization between the SSCPR and control groups (p > 0.05). After 21 days, the teeth exposed to the SSCPR remineralized 56.7 ± 3.7%, while the control only remineralized 10.7 ± 11.0% (p < 0.0001). At day 35, the remineralization was 73.7 ± 5.4% and 18.2 ± 10.8% (p < 0.0001) for the SSCPR and control groups, respectively. Conclusions: A marked increase in remineralization occurred with the use of the SSCPR. Notably, the remineralization of the SSCPR occurred deep within the tooth and progressed toward the surface over time.

Abstract Background Chronic lymphocytic leukemia (CLL) is an indolent heme malignancy characterized by the accumulation of CD5+ CD19+ B cells and episodes of relapse. The biological signaling that influence episodes of relapse in CLL are not fully described. Here, we identify gene networks associated with CLL relapse and survival risk. Methods Networks were investigated by using a novel weighted gene network co-expression analysis method and examining overrepresentation of upstream regulators and signaling pathways within co-expressed transcriptome modules across clinically annotated transcriptomes from CLL patients (N = 203). Gene Ontology analysis was used to identify biological functions overrepresented in each module. Differential Expression of modules and individual genes was assessed using an ANOVA (Binet Stage A and B relapsed patients) or T-test (SF3B1 mutations). The clinical relevance of biomarker candidates was evaluated using log-rank Kaplan Meier (survival and relapse interval) and ROC tests. Results Eight distinct modules (M2, M3, M4, M7, M9, M10, M11, M13) were significantly correlated with relapse and differentially expressed between relapsed and non-relapsed Binet Stage A CLL patients. The biological functions of modules positively correlated with relapse were carbohydrate and mRNA metabolism, whereas negatively correlated modules to relapse were protein translation associated. Additionally, M1, M3, M7, and M13 modules negatively correlated with overall survival. CLL biomarkers BTK, BCL2, and TP53 were co-expressed, while unmutated IGHV biomarker ZAP70 and cell survival-associated NOTCH1 were co-expressed in modules positively correlated with relapse and negatively correlated with survival days. Conclusions This study provides novel insights into CLL relapse biology and pathways associated with known and novel biomarkers for relapse and overall survival. The modules associated with relapse and overall survival represented both known and novel pathways associated with CLL pathogenesis and can be a resource for the CLL research community. The hub genes of these modules, e.g., ARHGAP27P2, C1S, CASC2, CLEC3B, CRY1, CXCR5, FUT5, MID1IP1, and URAHP, can be studied further as new therapeutic targets or clinical markers to predict CLL patient outcomes.

Abstract Background Dementia is currently the leading certified underlying cause of death in England. We assess how dementia recording on Office for National Statistics death certificates (ONS) corresponded to recording in general practice records (GP) and Hospital Episode Statistics (HES). Methods Retrospective study of deaths (2001-15) in 153 English General Practices contributing to the Clinical Practice Research Datalink, with linked ONS and HES records. Results Of 207,068 total deaths from any cause, 19,627 mentioned dementia on the death certificate with 10,253 as underlying cause; steady increases occurred from 2001 to 2015 (any mention 5.3 to 15.4 %, underlying cause 2.7 to 10 %). Including all data sources, recording of any dementia increased from 13.2 to 28.6 %. In 2015, only 53.8 % of people dying with dementia had dementia recorded on their death certificates. Among deaths mentioning dementia on the death certificate, the recording of a prior diagnosis of dementia in GP and HES rose markedly over the same period. In 2001, only 76.3 % had a prior diagnosis in GP and/or HES records; by 2015 this had risen to 95.7 %. However, over the same period the percentage of all deaths with dementia recorded in GP or HES but not mentioned on the death certificate rose from 7.9 to 13.3 %. Conclusions Dementia recording in all data sources increased between 2001 and 2015. By 2015 the vast majority of deaths mentioning dementia had supporting evidence in primary and/or secondary care. However, death certificates were still providing an inadequate picture of the number of people dying with dementia.

Abstract Background Administration of progesterone (P4) to ewes during the first 9 to 12 days of pregnancy accelerates blastocyst development by day 12 of pregnancy, likely due to P4-induced up-regulation of key genes in uterine epithelia responsible for secretion and transport of components of histotroph into the uterine lumen. This study determined if acceleration of blastocyst development induced by exogenous P4 during the pre-implantation period affects fetal-placental development on day 125 of pregnancy. Suffolk ewes (n = 35) were mated to fertile rams and assigned randomly to receive daily intramuscular injections of either corn oil vehicle (CO, n = 18) or 25 mg progesterone in CO (P4, n = 17) for the first 8 days of pregnancy. All ewes were hysterectomized on day 125 of pregnancy and: 1) fetal and placental weights and measurements were recorded; 2) endometrial and placental tissues were analyzed for the expression of candidate mRNAs involved in nutrient transport and arginine metabolism; and 3) maternal plasma, fetal plasma, allantoic fluid, and amniotic fluid were analyzed for amino acids, agmatine, polyamines, glucose, and fructose. Results Treatment of ewes with exogenous P4 did not alter fetal or placental growth, but increased amounts of aspartate and arginine in allantoic fluid and amniotic fluid, respectively. Ewes that received exogenous P4 had greater expression of mRNAs for SLC7A1, SLC7A2, SLC2A1, AGMAT, and ODC1 in endometria, as well as SLC1A4, SLC2A5, SLC2A8 and ODC1 in placentomes. In addition, AZIN2 protein was immunolocalized to uterine luminal and glandular epithelia in P4-treated ewes, whereas AZIN2 localized only to uterine luminal epithelia in CO-treated ewes. Conclusions This study revealed that exogenous P4 administered in early pregnancy influenced expression of selected genes for nutrient transporters and the expression of a protein involved in polyamine synthesis on day 125 of pregnancy, suggesting a ‘programming’ effect of P4 on gene expression that affected the composition of nutrients in fetal-placental fluids.

ABSTRACT The coronavirus SARS-CoV-2 infects host cells by binding to the angiotensin-converting enzyme 2 (ACE2) receptor, which belongs to an anti-inflammatory, anti-thrombotic counter-regulatory arm of the renin-angiotensin system (RAS). ACE2 dysfunction and RAS dysregulation has been explored as a driving force in acute respiratory distress syndrome (ARDS), but data from COVID-19 patients has been inconsistent and inconclusive. We sought to identify disruptions of the classical (ACE)/angiotensin (Ang) II/Ang II type-1 receptor (AT1R) and the counter-regulatory ACE2/Ang 1-7/Mas Receptor (MasR) pathways in patients with COVID-19 and correlate these with severity of infection and markers of inflammation and coagulation. Ang II and Ang 1–7 levels in plasma were measured by enzyme-linked immunosorbent assay (ELISA) for 230 patients, 166 of whom were SARS-CoV-2+. Ang 1–7 was repressed in COVID-19 patients compared to that in SARS-CoV-2 negative outpatient controls. Since the control cohort was less sick than the SARS-CoV-2+ group, this association between decreased Ang 1–7 and COVID-19 cannot be attributed to COVID-19 specifically as opposed to critical illness more generally. Multivariable logistic regression analyses demonstrated that every 10-pg/mL increase in plasma Ang 1–7 was associated with a 3% reduction in the odds of hospitalization (adjusted odds ratio [AOR] 0.97, confidence interval [CI] 0.95 to 0.99) and a 3% reduction in odds of requiring oxygen supplementation (AOR 0.97, CI 0.95 to 0.99) and/or ventilation (AOR 0.97, CI 0.94 to 0.99). Ang 1–7 was also inversely associated with pro-inflammatory cytokines and d-dimer in this patient cohort, suggesting that reduced activity in this protective counter-regulatory arm of the RAS contributes to the hyper-immune response and diffuse coagulation activation documented in COVID-19. IMPORTANCE Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes a unique disease, COVID-19, which ranges in severity from asymptomatic to causing severe respiratory failure and death. Viral transmission throughout the world continues at a high rate despite the development and widespread use of effective vaccines. For those patients who contract COVID-19 and become severely ill, few therapeutic options have been shown to provide benefits and mortality rates are high. Additionally, the pathophysiology underlying COVID-19 disease presentation, progression, and severity is incompletely understood. The significance of our research is in confirming the role of renin-angiotensin system dysfunction in COVID-19 pathogenesis in a large cohort of patients with diverse disease severity and outcomes. Additionally, to our knowledge, this is the first study to pair angiotensin peptide levels with inflammatory and thrombotic markers. These data support the role of ongoing clinical trials examining renin-angiotensin system-targeted therapeutics for the treatment of COVID-19.

Abstract Background Transposable element (TE) sequences are classified into families based on the reconstructed history of replication, and into subfamilies based on more fine-grained features that are often intended to capture family history. We evaluate the reliability of annotation with common subfamilies by assessing the extent to which subfamily annotation is reproducible in replicate copies created by segmental duplications in the human genome, and in homologous copies shared by human and chimpanzee. Results We find that standard methods annotate over 10% of replicates as belonging to different subfamilies, despite the fact that they are expected to be annotated as belonging to the same subfamily. Point mutations and homologous recombination appear to be responsible for some of this discordant annotation (particularly in the young Alu family), but are unlikely to fully explain the annotation unreliability. Conclusions The surprisingly high level of disagreement in subfamily annotation of homologous sequences highlights a need for further research into definition of TE subfamilies, methods for representing subfamily annotation confidence of TE instances, and approaches to better utilizing such nuanced annotation data in downstream analysis.

Evidence of observed negative impacts on natural and human systems from anthropogenic climate change is increasing. However, human systems in particular are dynamic and influenced by multiple drivers and hence identifying an anthropogenic climate signal is challenging. Here we analyze the case of lake Palcacocha in the Andes of Peru, which offers a representative model for other glacier lakes and related risks around the world because it features a dynamic evolution of flood risk driven by physical and socioeconomic factors and processes. Furthermore, it is the object of a prominent climate litigation case, wherein a local Peruvian citizen sued a large German energy producer over risk of flooding from lake Palcacocha. Adopting a conceptual model of cascading impacts and multiple drivers of risk, we first study climatic and other geophysical drivers of flood risk. We find that an anthropogenic signal from flood risk to greenhouse gas emissions is traceable. In parallel, flood risk has been strongly shaped (and increased) by interacting socioeconomic, institutional and cultural processes over the past few decades. The case raises important questions about the differentiation of responsibilities relating to flood risk of both global and local agents, which are, however, difficult to address in cases like Palcacocha, where we reveal a complex network of interlinked global, national and local drivers. Following from this, we outline a normative framework with a differentiated perspective on responsibility, implying that global emitters commit to support strengthening capacities in affected regions and localities and that local institutions and societies engage in local risk reduction measures and policies in collaboration with and driven by local communities.

Abstract Background In recent decades, there has been a growing interest in the impact of insults during pregnancy on postnatal health and disease. It is known that changes in placental development can impact fetal growth and subsequent susceptibility to adult onset diseases; however, a method to collect sufficient placental tissues for both histological and gene expression analyses during gestation without compromising the pregnancy has not been described. The ewe is an established biomedical model for the study of fetal development. Due to its cotyledonary placental type, the sheep has potential for surgical removal of materno-fetal exchange tissues, i.e., placentomes. A novel surgical procedure was developed in well-fed control ewes to excise a single placentome at mid-gestation. Results A follow-up study was performed in a cohort of nutrient-restricted ewes to investigate rapid placental changes in response to undernutrition. The surgery averaged 19 min, and there were no viability differences between control and sham ewes. Nutrient restricted fetuses were smaller than controls (4.7 ± 0.1 kg vs. 5.6 ± 0.2 kg; P

Introduction The target of a class of antiplatelet medicines, P2Y12R inhibitors, exists both on platelets and on brain immune cells (microglia). This protocol aims to describe a causal (based on a counterfactual model) approach for analysing whether P2Y12R inhibitors prescribed for secondary prevention poststroke may increase the risk of cognitive disorder or dementia via their actions on microglia, using real-world evidence.Methods and analysis This will be a cohort study nested within the Swedish National Health and Medical Registers, including all people with incident stroke from 2006 to 2016. We developed directed acyclic graphs to operationalise the causal research question considering potential time-independent and time-dependent confounding, using input from several experts. We developed a study protocol following the components of the target trial approach described by Hernan et al and describe the data structure that would be required in order to make a causal inference. We also describe the statistical approach required to derive the causal estimand associated with this important clinical question; that is, a time-to-event analysis for the development of cognitive disorder or dementia at 1, 2 and 5-year follow-up, based on approaches for competing events to account for the risk of all-cause mortality. Causal effect estimates and the precision in these estimates will be quantified.Ethics and dissemination This study has been approved by the Ethics Committee of the University of Gothenburg and Confidentiality Clearance at Statistics Sweden with Dnr 937-18, and an approved addendum with Dnr 2019-0157. The analysis and interpretation of the results will be heavily reliant on the structure, quality and potential for bias of the databases used. When we implement the protocol, we will consider and document any biases specific to the dataset and conduct appropriate sensitivity analyses. Findings will be disseminated to local stakeholders via conferences, and published in appropriate scientific journals.