Single-cell transcriptomic atlas of enteroendocrine cells along the murine gastrointestinal tract.

BACKGROUND: Enteroendocrine cells (EECs) produce over 20 gut hormones which contribute to intestinal physiology, nutrient metabolism and the regulation of food intake. The objective of this study was to generate a comprehensive transcriptomic map of mouse EECs from the stomach to the rectum. METHODS: EECs were purified by flow-cytometry from the stomach, upper small intestine, lower small intestine, caecum and large intestine of NeuroD1-Cre mice, and analysed by single cell RNA sequencing. Regional datasets were analysed bioinformatically and combined into a large cluster map. Findings were validated by L-cell calcium imaging and measurements of CCK secretion in vitro. RESULTS: 20,006 EECs across the full gastrointestinal tract could be subdivided based on their full transcriptome into 10 major clusters, each exhibiting a different pattern of gut hormone expression. EECs from the stomach were largely distinct from those found more distally, even when expressing the same hormone. Cell clustering was also observed when performed only using genes related to GPCR cell signalling, revealing GPCRs predominating in different EEC populations. Mc4r was expressed in 55% of Cck-expressing cells in the upper small intestine, where MC4R agonism was found to stimulate CCK release in primary cultures. Many individual EECs expressed more than one hormone as well as machinery for activation by multiple nutrients, which was supported by the finding that the majority of L-cells exhibited calcium responses to multiple stimuli. CONCLUSIONS: This comprehensive transcriptomic map of mouse EECs reveals patterns of GPCR and hormone co-expression that should be helpful in predicting the effects of nutritional and pharmacological stimuli on EECs from different regions of the gut. The finding that MC4R agonism stimulates CCK secretion adds to our understanding of the melanocortin system.

Vulvar Reconstruction with Freestyle Perforator Flap from the Medial Thigh Using Color Doppler Ultrasound.

Vulvoperineal defects mount significant functional and aesthetic challenges for reconstructive surgeons, often followed by postoperative complications. Ideal flap creation and perforator selection is vital for proper healing, especially in those with comorbid health concerns. This report details an innovative approach to vulvar reconstruction using a freestyle perforator flap from the medial thigh, identified by color flow Doppler ultrasound (CDU). This noninvasive imaging technique made possible the exact identification and confirmation of a well-vascularized pedicle, giving rise to a tailored flap for enhanced postoperative recovery and healing. The flap was designed based on real-time vascular mapping by CDU and validated through indocyanine green angiography, ensuring robust perfusion. The case of this patient demonstrated the successful application of portable CDU to navigate the intricate vessel network of the medial thigh, which would not have been possible with traditional imaging modalities such as computed tomography alone. The lack of postoperative complications following reconstruction demonstrates how this strategy for addressing vulvoperineal defects is advantageous due to meticulous perforator selection based on the patient's unique anatomy. CDU is therefore an important tool for achieving optimal outcomes related to form and function, all while reducing the risks of donor site morbidity.

Dielectric Signatures of Late Carcinoma Immune Cells Using MMTV-PyMT Mammary Carcinoma Models.

Peripheral blood mononuclear cells (PBMCs) are specialized immune cells produced from hematopoietic stem cells (HSC). They actively surveil for any signs of infection, foreign invaders, and abnormal or aberrant cells associated with diseases. Numerous inherent interactions between PBMCs and proliferating cancer cells facilitate cellular communication, inducing alterations in the composition of the PBMCs. These subtle alterations can be detected by using dielectrophoresis (DEP). The ultimate objective is to apply this knowledge in a clinical setting to achieve noninvasive early detection of breast cancer while minimizing the occurrence of false positives and negatives commonly associated with standard screening methods like mammography. To realize our long-term goal, we are probing the dielectric properties of the PBMCs from FVB/N MMTV-PyMT+ (late carcinoma, PyMT+ PBMC) and FVB/N (wild-type, WT-PBMC) age-matched mice at 14+ weeks using dielectrophoresis on a microfluidic platform. The central hypothesis of this research is that the changes triggered in the subcellular components, such as the cytoskeleton, lipid bilayer membrane, cytoplasm, focal adhesion proteins, and extracellular matrix (ECM) at the onset of carcinoma, regulate dielectric properties (conductivity, σ; and permittivity, ε), thus affecting the bioelectric signals that aid in the detection of breast cancer. The ANOVA results suggest a significant difference in PyMT+ PBMCs crossover frequencies at 0.01 and 0.05 S/m medium conductivity levels. Post hoc pairwise analysis of WT-PBMCs showed that the crossover frequencies are distinct across the medium conductivity ranges from 0.01 to 0.05 S/m. This study revealed that on average, PyMT+ PBMCs have increased crossover frequency, polarizability, higher membrane capacitance, and a folding factor compared with the age-matched wild-type PBMCs.

First evolutionary insights into the human otolithic system.

The human otolithic system (utricle and saccule), housed within the bony vestibule of the inner ear, establishes our sense of balance in conjunction with the semicircular canals. Yet, while the morphological evolution of the semicircular canals is actively explored, comparative morphological analyses of the otolithic system are lacking. This is regrettable because functional links with head orientation suggest the otolithic system could be used to track postural change throughout human evolution and across primates more broadly. In this context, we present the first analysis of the evolution of the human otolithic system within an anthropoid primate setting. Using the vestibule as a morphological proxy for the utricle and saccule, we compare humans to 13 other extant anthropoid species, and use phylogenetically-informed methods to find correlations with body size, endocranial flexion, and head-neck posture. Our results, obtained through micro-CT of 136 inner ears, reveal two major evolutionary transitions in hominoids, leading to distinctive vestibular morphology in humans, characterized by otolithic morphology resembling squirrel monkeys (possibly due to reversal), with a pronounced supraovalic fossa. Finally, we find a positional signal embedded in the anthropoid bony vestibule, providing the foundation to further explore the evolution of human head-neck posture using inner ear morphology.

Complementarity and sensitivity of benthic state indicators to bottom-trawl fishing disturbance.

Many indicators have been developed to assess the state of benthic communities and identify seabed habitats most at risk from bottom trawling disturbance. However, the large variety of indicators and their development and application under specific geographic areas and management contexts has made it difficult to evaluate their wider utility. We compared the complementarity/uniqueness, sensitivity, and selectivity of 18 benthic indicators to pressure of bottom trawling. Seventeen common datasets with broad regional representation covering a range of pressure gradients from bottom trawling disturbance (n = 14), eutrophication (n = 1), marine pollution (n = 1), and oxygen depletion (n = 1) were used for the comparison. The outcomes of most indicators were correlated to a certain extent with response to bottom trawling disturbance, and two complementary groups of indicators were identified: diversity-based and biological trait-based indicators. Trait-based indicators that quantify the changes in relative abundance of sensitive taxa were most effective in identifying benthic community change in response to bottom trawling disturbance. None of the indicators responded to the trawling pressure gradient in all datasets, and some showed a response that were opposed to the theoretical expectation for some gradients. Indicators that showed clear responses to bottom trawling disturbance also showed clear responses in at least one other pressure gradient, suggesting those indicators are not pressure specific. These results emphasize the importance of selecting several indicators, at least one from each group (diversity and trait-based), to capture the broader signals of change in benthic communities due to bottom trawling activities. Our systematic approach offers the basis from which scientific advisors and/or managers can select suitable combinations of indicators to arrive at a sensitive and comprehensive benthic status assessment.

Overconfidence in climate overshoot.

Global emission reduction efforts continue to be insufficient to meet the temperature goal of the Paris Agreement1. This makes the systematic exploration of so-called overshoot pathways that temporarily exceed a targeted global warming limit before drawing temperatures back down to safer levels a priority for science and policy2-5. Here we show that global and regional climate change and associated risks after an overshoot are different from a world that avoids it. We find that achieving declining global temperatures can limit long-term climate risks compared with a mere stabilization of global warming, including for sea-level rise and cryosphere changes. However, the possibility that global warming could be reversed many decades into the future might be of limited relevance for adaptation planning today. Temperature reversal could be undercut by strong Earth-system feedbacks resulting in high near-term and continuous long-term warming6,7. To hedge and protect against high-risk outcomes, we identify the geophysical need for a preventive carbon dioxide removal capacity of several hundred gigatonnes. Yet, technical, economic and sustainability considerations may limit the realization of carbon dioxide removal deployment at such scales8,9. Therefore, we cannot be confident that temperature decline after overshoot is achievable within the timescales expected today. Only rapid near-term emission reductions are effective in reducing climate risks.

Early surgical intervention is critical for strangulated Richter's hernia.

Richter's hernia is a rare but serious surgical emergency involving the entrapment or strangulation of part of the bowel's circumference in the hernial orifice, often without causing complete luminal obstruction. This case report describes a man in his 70s presenting with a 3-day history of abdominal pain and vomiting, despite normal bowel movements. Blood results revealed raised inflammatory markers, and computed tomography imaging suggested small bowel obstruction due to an incarcerated left inguinal hernia. Emergency surgery confirmed a Richter's hernia with a strangulated but viable bowel loop. Early surgical intervention led to a positive outcome. This case underscores the importance of high suspicion, early referral, and timely imaging in managing Richter's hernia to prevent severe complications, such as gangrene and perforation. Despite the absence of obstructive symptoms, early surgical intervention is critical when there is clinical concern regarding strangulation.

Molecular mechanisms underlying glucose-dependent insulinotropic polypeptide secretion in human duodenal organoids.

AIMS/HYPOTHESIS: Glucose-dependent insulinotropic polypeptide (GIP) is an incretin hormone secreted by enteroendocrine K cells in the proximal small intestine. This study aimed to explore the function of human K cells at the molecular and cellular levels. METHODS: CRISPR-Cas9 homology-directed repair was used to insert transgenes encoding a yellow fluorescent protein (Venus) or an Epac-based cAMP sensor (Epac-S-H187) in the GIP locus in human duodenal-derived organoids. Fluorescently labelled K cells were purified by FACS for RNA-seq and peptidomic analysis. GIP reporter organoids were employed for GIP secretion assays, live-cell imaging of Ca2+ using Fura-2 and cAMP using Epac-S-H187, and basic electrophysiological characterisation. The G protein-coupled receptor genes GPR142 and CASR were knocked out to evaluate roles in amino acid sensing. RESULTS: RNA-seq of human duodenal K cells revealed enrichment of several G protein-coupled receptors involved in nutrient sensing, including FFAR1, GPBAR1, GPR119, CASR and GPR142. Glucose induced action potential firing and cytosolic Ca2+ elevation and caused a 1.8-fold increase in GIP secretion, which was inhibited by the sodium glucose co-transporter 1/2 (SGLT1/2) blocker sotagliflozin. Activation of the long-chain fatty acid receptor free fatty acid receptor 1 (FFAR1) induced a 2.7-fold increase in GIP secretion, while tryptophan and phenylalanine stimulated secretion by 2.8- and 2.1-fold, respectively. While CASR knockout blunted intracellular Ca2+ responses, a CASR/GPR142 double knockout was needed to reduce GIP secretory responses to aromatic amino acids. CONCLUSIONS/INTERPRETATION: The newly generated human organoid K cell model enables transcriptomic and functional characterisation of nutrient-sensing pathways involved in human GIP secretion. Both calcium-sensing receptor (CASR) and G protein-coupled receptor 142 (GPR142) contribute to protein-stimulated GIP secretion. This model will be further used to identify potential targets for modulation of native GIP secretion in diabetes and obesity.

The impact of locked cabinets for automated external defibrillators (AEDs) on cardiac arrest and AED outcomes: A scoping review.

Background: Rapid public defibrillation with automated external defibrillators (AEDs) is critical to improving out-of-hospital cardiac arrest survival. Concerns about AED theft and vandalism have led to implementing security measures, including locked cabinets. This scoping review, conducted as part of the evidence review for the International Liaison Committee on Resuscitation, explores the impact of securing AEDs in locked cabinets. Methods: Searches of Medline, Embase, Cochrane, CINAHL (from database inception to 25/5/2024) and Google Scholar (first 200 articles). Studies of any type or design, published with an English abstract, examining the impact of locked AED cabinets were included. The included studies were grouped by outcomes, and an iterative narrative synthesis was performed. Results: We screened 2,096 titles and found 10 relevant studies: 8 observational studies (4 published as conference abstracts) and 2 simulation studies. No study reported patient outcomes. Studies reported data on between 36 and 31,938 AEDs. Most studies reported low rates (<2%) of theft/missing/vandalism, including AEDs that were accessible 24/7. The only study comparing unlocked and locked cabinets showed minimal difference in theft and vandalism rates (0.3% vs. 0.1%). Two simulation studies showed significantly slower AED retrieval when additional security measures, included locked cabinets, were used. A survey of first responders reported half (25/50) were injured while accessing an AED that required breaking glass to access. Conclusion: The limited literature suggests that vandalism and the loss of AEDs are rare and occur in locked and unlocked cabinets. Research on this topic is needed that focuses on real-life retrieval and patient outcomes.

RBPMS promotes contractile phenotype splicing in human embryonic stem cell derived vascular smooth muscle cells.

AIMS: Differentiated Vascular Smooth Muscle Cells (VSMCs) express a unique network of mRNA isoforms via smooth muscle specific alternative splicing (SM-AS) in functionally critical genes, including those comprising the contractile machinery. We previously described RNA Binding Protein Multiple Splicing (RBPMS) as a potent driver of differentiated SM-AS in the rat PAC1 VSMC cell line. What is unknown is how RBPMS affects VSMC phenotype and behaviour. Here, we aimed to dissect the role of RBPMS in SM-AS in human cells and determine the impact on VSMC phenotypic properties. METHODS AND RESULTS: We used human embryonic stem cell-derived VSMCs (hESC-VSMCs) as our platform. hESC-VSMCs are inherently immature and we found that they display only partially differentiated SM-AS patterns while RBPMS protein levels are low. We found that RBPMS overexpression induces SM-AS patterns in hESC-VSMCs akin to the contractile tissue VSMC splicing patterns. We present in silico and experimental findings that support RBPMS' splicing activity as mediated through direct binding and via functional cooperativity with splicing factor RBFOX2 on a significant subset of targets. We also demonstrate that RBPMS can alter the motility and the proliferative properties of hESC-VSMCs to mimic a more differentiated state. CONCLUSIONS: Overall, this study emphasizes a critical role for RBPMS in establishing the contractile phenotype splicing program of human VSMCs.

Can Repetition-based Training in a High-fidelity Model Enhance Critical Trauma Surgical Skills Among Trainees and Attending Surgeons Equally?

BACKGROUND: The evolution of warfare has resulted in a surge of high-energy blast injuries predominantly involving the lower extremities. Once thought to impact only forward-deployed military, such mechanisms of injury are becoming a harsh reality even in civilian territory. Proficiency in surgical techniques for extremity damage control is vital for surgeons, regardless of specialty. To evaluate and train surgical residents and attending surgeons in critical limb-salvage techniques, Theater Hospital Operations Replication (THOR) has been proposed as a high-fidelity training platform to enhance extremity surgery teaching practices and assess proficiency in fasciotomy, vascular shunting, and knee-spanning external fixation in a simulated, resource-limited environment. QUESTIONS/PURPOSES: Given the importance of proficiency and efficiency in surgeon ability to perform fasciotomies, vascular shunting, and placement of external fixators, in this study, we proposed two research questions: (1) Does repetition-based training within the THOR model improve surgeon knowledge and technical skill? (2) Are there differences in skill outcome when utilizing the THOR model based on surgical specialty (orthopaedic versus general) and/or level in training (attending versus resident)? METHODS: This was an observational learning outcome study performed in the bioskills lab at a single institution, the Naval Medical Center Portsmouth, in which 26 surgeons completed a same-day course consisting of pretraining assessment, cadaver training on three damage-control surgery techniques in a high-fidelity THOR environment, and post-training assessment. The surgeons were either general surgeons or orthopaedic surgeons and consisted of both residents and attending surgeons. Subjects underwent a pretraining knowledge assessment, followed by two rounds of performing simulated surgery. The first round of surgery included guidance and instruction from board-certified orthopaedic surgeons. After a short break for the subjects to review the material, the second round was completed without instruction. To answer our first question of how repetition-based training impacts surgical skill, our metrics included: Objective Structured Assessment of Technical Skills (OSATS) scores (range 1 to 5, scored lowest to highest, where higher scores represent optimal skill performance), procedural accuracy, a 10-question knowledge assessment administered before and after training (scored 0 to 10, where higher scores represent competency of the anatomy and procedure steps), and total procedure time. To address our second question, data on demographic characteristics were collected on all participants, which included surgical specialty, year in training, and gender. RESULTS: The general surgery residents' cohort demonstrated improvement in both mean ± SD OSATS scores (2.4 ± 0.7 before training versus 3.6 ± 0.6 after training, mean difference 1.2 [95% CI 0.4 to 2.0]; p = 0.01) and procedure duration (23 ± 7 minutes before training versus 16 ± 5 minutes after training, mean difference 6 minutes [95% CI 4 to 9]; p = 0.001) for external fixator procedures after THOR. Similar results were seen in fasciotomies after THOR. The orthopaedic surgery residents' cohort showed similar improvements in reducing procedure duration when performing knee-spanning external fixator surgery, as well as exhibited improvements during vascular shunting procedures in both mean ± SD OSATS scores (previously 3.3 ± 0.9 versus 4.0 ± 0.9, mean difference 0.7 [95% CI 0.2 to 1.2]; p = 0.01) and procedure duration (23 ± 5 minutes versus 14 ± 8 minutes, mean difference 10 minutes [95% CI 4 to 15]; p = 0.003) after THOR. When we compared the cohorts, general surgery attending surgeons demonstrated an improvement in vascular shunting procedure duration after instruction with the THOR model (19 ± 3 minutes versus 12 ± 4 minutes, mean difference 7 minutes [95% CI 1 to 13]; p = 0.03) and in overall knowledge scores (4.3 + 0.5 versus 8.0 ± 1.6, mean difference 3.8 [95% CI 0.5 to 7.0]; p = 0.04). The orthopaedic surgery attending surgeons yielded no improvement for any metric, pretraining and post-training. Using the prescores as a covariate, for almost all metrics analyzed, there were no differences between surgeon groups based on specialty after completing the repeated training. CONCLUSION: Use of the high-fidelity THOR training model improved proficiency in the examined sample of extremity damage-control procedures, thus demonstrating its efficacy in enhancing surgical skills and short-interval knowledge retention while simultaneously highlighting the importance of cadaver simulation training in enhancing surgical preparedness for complex trauma. CLINICAL RELEVANCE: The high-fidelity THOR training model represents a promising modality for preparing surgeons for extremity combat trauma management in resource-limited, high-stress environments. Whether in a wartime environment or in the civilian sector, efficient and skilled intervention are crucial in managing patient morbidity. Differences of a few minutes per procedure can mean all the difference in damage control, especially when compounded in situations where vascular shunting, knee-spanning external fixation, and lower leg four-compartment fasciotomy have to be performed together by both orthopaedic and general surgeons. The observational period in this study spanned 1 day and thus does not provide information on retention of long-term skills or knowledge. A follow-on study can assess how time from initial training to final evaluation impacts overall skill competency and knowledge.

Controlling and predicting alkyl-onium electronic structure.

X-ray photoelectron spectroscopy (XPS) and ab initio calculations show that fully alkylated onium cation electronic structure can be tuned using both the alkyl chains and the central onium atom. The key for tuning the central onium atom is methyl versus longer alkyl chains, allowing selection of the optimum cation for a wide range of applications, including catalysis and biocides.

High radiative forcing climate scenario relevance analyzed with a ten-million-member ensemble.

Developing future climate projections begins with choosing future emissions scenarios. While scenarios are often based on storylines, here instead we produce a probabilistic multi-million-member ensemble of radiative forcing trajectories to assess the relevance of future forcing thresholds. We coupled a probabilistic database of future greenhouse gas emission scenarios with a probabilistically calibrated reduced complexity climate model. In 2100, we project median forcings of 5.1 watt per square meters (5th to 95th percentiles of 3.3 to 7.1), with roughly 0.5% probability of exceeding 8.5 watt per square meters, and a 1% probability of being lower than 2.6 watt per square meters. Although the probability of 8.5 watt per square meters scenarios is low, our results support their continued utility for calibrating damage functions, characterizing climate in the 22nd century (the probability of exceeding 8.5 watt per square meters increases to about 7% by 2150), and assessing low-probability/high-impact futures.

LAIR-1 and PECAM-1 function via the same signaling pathway to inhibit GPVI-mediated platelet activation.

Background: Inhibition of platelet responsiveness is important for controlling thrombosis. It is well established that platelet endothelial cell adhesion molecule-1 (PECAM-1) serves as a physiological negative regulator of platelet-collagen interactions. We recently demonstrated that leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1) is a negative regulator of platelet production and reactivity. It is however not known if LAIR-1 and PECAM-1 function in the same or different inhibitory pathways. Objectives: In this study, we investigated the role of LAIR-1 alongside PECAM-1 in megakaryocyte development and platelet production and determined the functional redundancy through characterization of a LAIR-1/PECAM-1 double knockout (DKO) mouse model. Methods: LAIR-1 and PECAM-1 expression in megakaryocytes were evaluated by western blotting. Megakaryocyte ploidy and proplatelet formation were evaluated by flow cytometry and fluorescent microscopy. Platelet function and signalling were compared in wild-type, LAIR-1 -/- , PECAM-1 -/-  and DKO mice using aggregometry, flow cytometry and western blotting. Thrombosis was evaluated using the FeCl 3  carotid artery model. Results: We show that LAIR-1/PECAM-1 DKO mice exhibit a 17% increase in platelet count. Bone marrow-derived megakaryocytes from all 3 mouse models had normal ploidy in vitro, suggesting that neither LAIR-1 nor PECAM-1 regulates megakaryocyte development. Furthermore, relative to wild-type platelets, platelets derived from LAIR-1, PECAM-1, and DKO mice were equally hyperresponsive to collagen in vitro, indicating that LAIR-1 and PECAM-1 participate in the same inhibitory pathway. Interestingly, DKO mice exhibited normal thrombus formation in vivo due to DKO mouse platelets lacking the enhanced Src family kinase activation previously shown in platelets from LAIR-1-deficient mice. Conclusion: Findings from this study reveal that LAIR-1 and PECAM-1 act to inhibit GPVI-mediated platelet activation via the same signaling pathway. Mice lacking LAIR-1 and PECAM-1 do not however exhibit an increase in thrombus formation despite minor increase in platelet count and reactivity to collagen. This study adds to the growing evidence that immunoreceptor tyrosine-based inhibition motif-containing receptors are important regulators of platelet count and function.

Reconstructing 120 years of climate change impacts on Joshua tree flowering.

Quantifying how global change impacts wild populations remains challenging, especially for species poorly represented by systematic datasets. Here, we infer climate change effects on masting by Joshua trees (Yucca brevifolia and Y. jaegeriana), keystone perennials of the Mojave Desert, from 15 years of crowdsourced observations. We annotated phenophase in 10,212 geo-referenced images of Joshua trees on the iNaturalist crowdsourcing platform, and used them to train machine learning models predicting flowering from annual weather records. Hindcasting to 1900 with a trained model successfully recovers flowering events in independent historical records and reveals a slightly rising frequency of conditions supporting flowering since the early 20th Century. This reflects increased variation in annual precipitation, which drives masting events in wet years-but also increasing temperatures and drought stress, which may have net negative impacts on recruitment. Our findings reaffirm the value of crowdsourcing for understanding climate change impacts on biodiversity.

Insurance-Related Disparities in Withdrawal of Life Support and Mortality After Spinal Cord Injury.

Importance: Identifying disparities in health outcomes related to modifiable patient factors can improve patient care. Objective: To compare likelihood of withdrawal of life-supporting treatment (WLST) and mortality in patients with complete cervical spinal cord injury (SCI) with different types of insurance. Design, Setting, and Participants: This retrospective cohort study collected data between 2013 and 2020 from 498 trauma centers participating in the Trauma Quality Improvement Program. Participants included adult patients (older than 16 years) with complete cervical SCI. Data were analyzed from November 1, 2023, through May 18, 2024. Exposure: Uninsured or public insurance compared with private insurance. Main Outcomes and Measures: Coprimary outcomes were WLST and mortality. The adjusted odds ratio (aOR) of each outcome was estimated using hierarchical logistic regression. Propensity score matching was used as an alternative analysis to compare public and privately insured patients. Process of care outcomes, including the occurrence of a hospital complication and length of stay, were compared between matched patients. Results: The study included 8421 patients with complete cervical SCI treated across 498 trauma centers (mean [SD] age, 49.1 [20.2] years; 6742 male [80.1%]). Among the 3524 patients with private insurance, 503 had WLST (14.3%) and 756 died (21.5%). Among the 3957 patients with public insurance, 906 had WLST (22.2%) and 1209 died (30.6%). Among the 940 uninsured patients, 156 had WLST (16.6%) and 318 died (33.8%). A significant difference was found between uninsured and privately insured patients in the adjusted odds of WLST (aOR, 1.49; 95% CI, 1.11-2.01) and mortality (aOR, 1.98; 95% CI, 1.50-2.60). Similar results were found in subgroup analyses. Matched public compared with private insurance patients were found to have significantly greater odds of hospital complications (odds ratio, 1.27; 95% CI, 1.14-1.42) and longer hospital stay (mean difference 5.90 days; 95% CI, 4.64-7.20), which was redemonstrated on subgroup analyses. Conclusions and Relevance: Health insurance type was associated with significant differences in the odds of WLST, mortality, hospital complications, and days in hospital among patients with complete cervical SCI in this study. Future work is needed to incorporate patient perspectives and identify strategies to close the quality gap for the large number of patients without private insurance.

Resuscitation on the field of play: a best-practice guideline from Resuscitation Council UK.

Sudden cardiac arrest (SCA) is the leading cause of sudden death in athletes during high-level, organised sport. Patient-related and event-related factors provide an opportunity for rapid intervention and the potential for high survival rates. The aim of this consensus was to develop a best-practice guideline for dedicated field-of-play medical teams responding to SCA during an organised sporting event. A task-and-finish group from Resuscitation Council UK identified a stakeholder group of relevant experts and cardiac arrest survivors in March and April 2022. Together, they developed a best-practice guideline using the best available evidence. A public consultation period further refined the guideline before it was finalised in December 2023. Any sudden collapse, without rapid recovery during sporting activity, should be considered an SCA until proven otherwise. Field-of-play medical teams should be empowered to access the collapsed athlete as soon as possible and perform initial essential interventions in situ. This includes a suggested minimum of three cycles of cardiopulmonary resuscitation and defibrillation in persistent shockable rhythms while other aspects of advanced life support are initiated. There should be careful organisation and practice of the medical response, including plans to transport athletes to dedicated facilities for definitive medical care. This best-practice guideline complements, rather than supersedes, existing resuscitation guidelines. It provides a clear approach to how to best treat an athlete with SCA and how to organise the medical response so treatments are delivered effectively and optimise outcomes.

Barcoded overexpression screens in gut Bacteroidales identify genes with roles in carbon utilization and stress resistance.

A mechanistic understanding of host-microbe interactions in the gut microbiome is hindered by poorly annotated bacterial genomes. While functional genomics can generate large gene-to-phenotype datasets to accelerate functional discovery, their applications to study gut anaerobes have been limited. For instance, most gain-of-function screens of gut-derived genes have been performed in Escherichia coli and assayed in a small number of conditions. To address these challenges, we develop Barcoded Overexpression BActerial shotgun library sequencing (Boba-seq). We demonstrate the power of this approach by assaying genes from diverse gut Bacteroidales overexpressed in Bacteroides thetaiotaomicron. From hundreds of experiments, we identify new functions and phenotypes for 29 genes important for carbohydrate metabolism or tolerance to antibiotics or bile salts. Highlights include the discovery of a D-glucosamine kinase, a raffinose transporter, and several routes that increase tolerance to ceftriaxone and bile salts through lipid biosynthesis. This approach can be readily applied to develop screens in other strains and additional phenotypic assays.

A morphometric comparison of the ductus reuniens in humans and guinea pigs, with a note on its evolutionary importance.

The mammalian inner ear contains the sensory organs responsible for balance (semicircular canals, utricle, and saccule) and hearing (cochlea). While these organs are functionally distinct, there exists a critical structural connection between the two: the ductus reuniens (DR). Despite its functional importance, comparative descriptions of DR morphology are limited, hindering our understanding of the evolutionary diversification of hearing and balance systems among mammals. Using virtual 3D models derived from micro-CT, we examine the morphology of the DR and its relationship to the bony labyrinth in humans compared to that in a commonly used animal model, the guinea pig. Anatomical reconstructions and univariate measurements were carried out in the software 3D Slicer. Data indicate similarities in DR morphology between humans and guinea pigs in terms of overall shape. However, there are considerable differences in relative DR length and width between humans and guinea pigs. Humans possess a relatively shorter and narrower DR but with wider openings to the saccule and cochlear duct. This results in a relatively more constricted DR lumen in humans which may differentially limit fluid transfer between the saccule and cochlea. Our results reveal previously hidden morphological diversity in the communication between the hearing and balance systems of the mammalian inner ear which may indicate alternative strategies for isolating the Organ of Corti from the peripheral vestibular system throughout mammalian evolution.