Do high-protein diets have the potential to reduce gut barrier function in a sex-dependent manner?

PURPOSE: Impaired gut barrier function is associated with systemic inflammation and many chronic diseases. Undigested dietary proteins are fermented in the colon by the gut microbiota which produces nitrogenous metabolites shown to reduce barrier function in vitro. With growing evidence of sex-based differences in gut microbiotas, we determined whether there were sex by dietary protein interactions which could differentially impact barrier function via microbiota modification. METHODS: Fermentation systems were inoculated with faeces from healthy males (n = 5) and females (n = 5) and supplemented with 0.9 g of non-hydrolysed proteins sourced from whey, fish, milk, soya, egg, pea, or mycoprotein. Microbial populations were quantified using fluorescence in situ hybridisation with flow cytometry. Metabolite concentrations were analysed using gas chromatography, solid phase microextraction coupled with gas chromatography-mass spectrometry and ELISA. RESULTS: Increased protein availability resulted in increased proteolytic Bacteroides spp (p < 0.01) and Clostridium coccoides (p < 0.01), along with increased phenol (p < 0.01), p-cresol (p < 0.01), indole (p = 0.018) and ammonia (p < 0.01), varying by protein type. Counts of Clostridium cluster IX (p = 0.03) and concentration of p-cresol (p = 0.025) increased in males, while females produced more ammonia (p = 0.02), irrespective of protein type. Further, we observed significant sex-protein interactions affecting bacterial populations and metabolites (p < 0.005). CONCLUSIONS: Our findings suggest that protein fermentation by the gut microbiota in vitro is influenced by both protein source and the donor's sex. Should these results be confirmed through human studies, they could have major implications for developing dietary recommendations tailored by sex to prevent chronic illnesses.

THREE-DECADE changes of reef cover in Pulau Layang-Layang, Malaysia using multitemporal Landsat images.

Over the years, coral reefs in the South China Sea have degraded and faced severe threats from rapid development, coral bleaching, and Crown-of-Thorns Starfish (COTS) outbreak. There is limited knowledge relating to the effects of anthropogenic disturbances and natural events on the coral reefs of Pulau Layang-Layang. This study aims to assess reef cover changes by utilizing Landsat satellite images spanning from 1989 to 2022. Using the object-based image analysis method, this study classified the reef cover into three categories: coral, rock and rubble, and sand. The supervised classification had an overall accuracy of 86.41-87.38 % and Tau's coefficients of 0.80-0.81. The results showed island development and construction of artificial bird sanctuary have led to an increase in coral cover. Furthermore, it was illustrated that the impact of COTS outbreaks in 2010 and 2020 differed significantly, with the latter showing no signs of recovery. Our study underscores the importance of timely intervention to mitigate the spread of COTS. This study provides insights into the resilience and vulnerability of these ecosystems in the face of various stressors.

Achalasia-associated megaoesophagus presenting with dyspnoea and cough.

We present an unusual case of achalasia presenting with dyspnoea and persistent cough. These symptoms persisted for months, leading to the patient undergoing a chest X-ray by her general practitioner which showed right basal consolidation and a density extending along the right mediastinum. CT scan was done which revealed megaoesophagus with a diameter of 7 cm causing tracheal compression, as well as right basal consolidation, consistent with aspiration. Further history revealed 6-month history of progressive swallowing difficulty, retrosternal chest pain and shortness of breath which worsened when eating solid foods. After thorough workup, a diagnosis of idiopathic achalasia (type II) was made. She was treated with laparoscopic Heller cardiomyotomy and Dor fundoplication with significant improvement at follow-up. Dyspnoea and respiratory symptoms are unusual presenting symptoms, suggesting a need to consider achalasia in a wider range of presentations. Successful treatment of achalasia depends on timely diagnosis and intervention prior to oesophageal failure.

Gene drive and genetic sex conversion in the global agricultural pest Ceratitis capitata.

Homing-based gene drives are recently proposed interventions promising the area-wide, species-specific genetic control of harmful insect populations. Here we characterise a first set of gene drives in a tephritid agricultural pest species, the Mediterranean fruit fly Ceratitis capitata (medfly). Our results show that the medfly is highly amenable to homing-based gene drive strategies. By targeting the medfly transformer gene, we also demonstrate how CRISPR-Cas9 gene drive can be coupled to sex conversion, whereby genetic females are transformed into fertile and harmless XX males. Given this unique malleability of sex determination, we modelled gene drive interventions that couple sex conversion and female sterility and found that such approaches could be effective and tolerant of resistant allele selection in the target population. Our results open the door for developing gene drive strains for the population suppression of the medfly and related tephritid pests by co-targeting female reproduction and shifting the reproductive sex ratio towards males. They demonstrate the untapped potential for gene drives to tackle agricultural pests in an environmentally friendly and economical way.

A multi-reservoir extruder for time-resolved serial protein crystallography and compound screening at X-ray free-electron lasers.

Serial crystallography at X-ray free-electron lasers (XFELs) permits the determination of radiation-damage free static as well as time-resolved protein structures at room temperature. Efficient sample delivery is a key factor for such experiments. Here, we describe a multi-reservoir, high viscosity extruder as a step towards automation of sample delivery at XFELs. Compared to a standard single extruder, sample exchange time was halved and the workload of users was greatly reduced. In-built temperature control of samples facilitated optimal extrusion and supported sample stability. After commissioning the device with lysozyme crystals, we collected time-resolved data using crystals of a membrane-bound, light-driven sodium pump. Static data were also collected from the soluble protein tubulin that was soaked with a series of small molecule drugs. Using these data, we identify low occupancy (as little as 30%) ligands using a minimal amount of data from a serial crystallography experiment, a result that could be exploited for structure-based drug design.

Implementation of the European Society of Cardiology 0/3-hour accelerated diagnostic protocol, using high sensitive troponin T: a clinical practice evaluation of safety and effectiveness involving 3003 patients with suspected acute coronary syndrome.

BACKGROUND: There have been relatively few studies detailing the real-world effectiveness and safety of accelerated diagnostic protocols (ADP), using high sensitivity cardiac troponin (hs-cTn). OBJECTIVE: To analyse the safety and effectiveness of early emergency department (ED) discharge following implementation of the European Society of Cardiology (ESC) 0/3-hour ADP for suspected acute coronary syndromes (ACS). METHOD: We prospectively studied 2 cohorts of consecutive suspected ACS presentations to ED before (n=1642) and after (n=1376, 2 centres) implementation of the ESC 0/3-hour ADP incorporating limit of detection rule out. Safety was defined by MACE (major adverse cardiac events) inclusive of type 1 myocardial infarction (MI) in patients discharged from ED, and clinical effectiveness by percentage ED discharge. Continuous variables and categorical data were evaluated by independent t-test and χ2 test, respectively. Time-to-event data were analysed as survival data and converted to Kaplan-Meier curves for interpretation. RESULTS: In the preimplementation period, there was a higher prevalence of MI. Discharge from ED increased by >100% (from 27.1% to 56.5% of the cohort) with no safety signal (MACE rate 4/444 (0.9%) vs 4/769 (0.52%), p=0.430 for the 2011 and 2018 cohort, respectively). This correlated with a marked reduction in length of stay overall but a more modest reduction for those discharged from ED (6 hours 10 min vs 5 hours 25 min, p<0.001) for the 2011 and 2018 cohort, respectively. There were improvements in presentation to blood draw (163-90 min, p<0.001). Time from presentation to first ECG actually increased (16.2 vs 31.2 min, p<0.001). Analysis of hs-cTn values and ECGs revealed a maximum ED discharge rate of 69%, by applying the 0/3-hour protocol, implying potential for increasing safe ED discharge. CONCLUSIONS: Implementation of an ADP with hs-cTn is safe and effective for early rule-out and discharge of suspected ACS but require considerable resources and education to optimise maximal patient flow.

Body, carcass, and steak dimensions of straightbred Holstein calves and Angus-sired calves from Holstein, Jersey, and crossbred beef dams.

Beef genetics are used with increasing frequency on commercial dairies. Although use of beef genetics improves calf value, variability has been reported in beef × dairy calf phenotype for traits related to muscularity and carcass composition. The objective of this study was to characterize morphometric and compositional differences between beef, beef × dairy, and dairy-fed cattle. Tested treatment groups included Angus-sired straightbred beef steers and heifers (A × B; n = 45), Angus × Holstein crossbreds (A × H; n = 15), Angus × Jersey crossbreds (A × J; n = 16), and straightbred Holsteins (H, n = 16). Cattle were started on trial at mean BW of 302 ±â€…29.9 kg and then fed at 196 ±â€…3.4 d. Morphometric measures were recorded every 28 d during the finishing period, ultrasound measures were recorded every 56 d, and morphometric carcass measures were recorded upon slaughter. Muscle biopsies were collected from the longissimus thoracis of a subset of steers (n = 43) every 56 d. Strip loins were collected from carcasses (n = 78) for further evaluation. Frame size measured as hip height, hip width, and body length was greatest for H cattle (P < 0.05), and A × H cattle had greater hip height than A × J cattle (P < 0.05). Relative to BW as a percentage of mature size, ribeye area of all cattle increased at a decreasing rate (negative quadratic term: P < 0.01), and all ultrasound measures of fat depots increased at an increasing rate (positive quadratic term: P < 0.01). Although no difference was observed in muscle fiber area across the finishing period from the longissimus thoracis (P = 0.80), H cattle had a more oxidative muscle phenotype than A × B cattle (P < 0.05). Additionally, H cattle had the smallest area of longissimus lumborum in the posterior strip loin, greatest length-to-width ratio of longissimus lumborum in the posterior strip loin, and least round circumference relative to round length (P < 0.05). Beef genetics improved muscularity in portions of the carcass distal to the longissimus thoracis.

Divergent selection of beef and dairy breeds has caused differences in skeletal size and muscularity. When calves from dairy systems enter the beef supply chain, variability in mature size and carcass composition are introduced. The objective of this study was to characterize morphometric differences in cattle populations with different proportions of beef and dairy genetics. Body measurements confirmed differences in mature size of beef-type cattle, dairy-type cattle, and beef × dairy cattle; Holstein influence was associated with greater skeletal growth. With advancing maturity, the rate of muscle accretion decreased quadratically while the rate of fat accretion increased quadratically. Although muscularity across all cattle types was similar in the longissimus near the last rib, differences were observed in the posterior end of the strip loin, the forearm, and the round. Differences in mature size, muscularity, and steak dimensions were observed between beef-type cattle, dairy-type cattle, and beef × dairy cattle.

Exploring the Interplay Between Vitamin B12-related Biomarkers, DNA Methylation, and Gene-Nutrition Interaction in Esophageal Precancerous Lesions.

BACKGROUND: Vitamin B12 depletion has been suggested to be associated with esophageal precancerous lesions (EPL). However, the potential mechanisms remain unclear. AIMS: This study aims to evaluate the role of vitamin B12 and its regulated epigenetic modification in EPL and provide preliminary information on the identification of potential molecular biomarkers for the early prediction of EPL. METHODS: We collected information and samples from the Early Diagnosis and Early Treatment Project of Esophageal Cancer database from 200 EPL cases and 200 matched controls. Vitamin B12, one-carbon metabolism biomarkers, genetic polymorphism of TCN2 C776G, and DNA methylation were compared. Preliminarily identified candidate promoters of differentially methylated CpG positions were further verified by targeted bisulfite sequencing. RESULTS: EPL cases had significantly lower serum levels of vitamin B12 and transcobalamin II, and higher serum levels of homocysteine and 5-methyltetrahydrofolate than controls. The TCN2 C776G polymorphism was found to be associated with susceptibility to EPL and may interact with vitamin B12 nutritional status to influence the risk of EPL in male subjects. In addition, global hypomethylation related to vitamin B12 depletion was observed in EPL cases, along with region-specific hypermethylation of UGT2B15 and FGFR2 promoters. CONCLUSIONS: This study suggests that vitamin B12 depletion may be associated with aberrant DNA methylation and increased risk of EPL through the one-carbon metabolism pathway, presents that the TCN2 C776G polymorphism may interact with vitamin B12 nutritional status to affect EPL risk in males, and also identifies specific locations in the UGT2B15 and FGFR2 promoters with potential as promising molecular biomarkers.

Managing and Preventing Migraine in the Emergency Department: A Review.

Migraine is a leading cause of disability worldwide, and acute migraine attacks are a common reason for patients to seek care in the emergency department (ED). There have been recent advancements in the care of patients with migraine, specifically emerging evidence for nerve blocks and new pharmacological classes of medications like gepants and ditans. This article serves as a comprehensive review of migraine in the ED, including diagnosis and management of acute complications of migraine (eg, status migrainosus, migrainous infarct, persistent aura without infarction, and aura-triggered seizure) and use of evidence-based migraine-specific treatments in the ED. It highlights the role of migraine preventive medications and provides a framework for emergency physicians to prescribe them to eligible patients. Finally, it evaluates the evidence for nerve blocks in the treatment of migraine and introduces the possible role of gepants and ditans in the care of patients with migraine in the ED.

Factors associated with retention of mother-baby pairs in the elimination of mother-to-child transmission of HIV program in Kaberamaido district: A longitudinal analysis.

BACKGROUND: Retention along the elimination of Mother to Child Transmission (eMTCT) cascade in Uganda remains poor as only 62.7%-69.5% are followed up to 18months. The objective of this study was to determine the rates of retention of mother-baby pairs at two levels of the eMTCT cascade (12 and 18 months) and associated factors. METHODS: This was a longitudinal analysis of 368 mother-baby pairs who were enrolled into the eMTCT program in Kaberamaido district from January 2013 to December 2018. Data was extracted from early infant diagnosis (EID) and mothers' ART registers, entered into Microsoft Excel and then exported to Stata statistical software package version 14.0 for management and analysis. Descriptive statistics such as mean and frequencies were computed at univariate level. At the bivariate level, Cox proportional hazard regression was performed to assess the level of association between the primary outcome and each independent variable, while Cox proportional hazard regression model was built at multivariate level to determine the factors independently associated with retention of mother-baby pairs in the eMTCT program. RESULTS: Of the 368 mothers enrolled into the study, their average age was 29.7years (SD = 6.6). Nearly two-thirds of the mothers were married/cohabiting, (n = 232, 63.0%). The 368 mother baby pairs were observed for a total time of 6340 person months, with majority, 349 (94.8%, 95%CI = 92.0-96.7) still active in eMTCT care, while 19(5.2%, 95%CI = 3.3-8.0) were lost to follow up at 12months. At 18 months, 323 (87.8%, 95%CI = 84.0-90.8) were active in eMTCT program while 45(12.2%, 95 CI = 9.2-16.0) were lost to follow up. At bivariate level, marital status, health facility level of enrolment, mothers' ART treatment supporter, and mothers' ART enrolment time were significantly associated with survival/lost to follow up (LTFU) of mother-baby pairs along the eMTCT cascade. At multivariable level, the mothers' time of ART initiation was significantly associated with survival/lost to follow up (LTFU) of mother-baby pairs at along the eMTCT cascade, with mothers-baby pairs who were initiated during the antenatal/post-natal periods having higher hazards of LTFU compared to those who initiated ART before Antenatal period (before pregnancy), aHR = 4.37(95%CI, 1.62-11.76, P = 0.003). Mother-baby pairs who were enrolled into the eMTCT program after the implementation of HIV test and treat policy (year 2017 and 2018) had higher hazards of LTFU as compared to those enrolled before the implementation of test and treat policy in Uganda (year 2013-2016), aHR = 2.22(95% CI, 1.15-4.30, P = 0.017). All the other factors had no significant association with lost to follow up and cascade completion at multivariate level. CONCLUSION: There was high level of retention of mother-baby pairs in the eMTCT program in Kaberamaido at 12 months, but it was suboptimal at 18months. ART initiation during the antenatal and/or post-natal period was significantly associated with suboptimal retention of mother-baby pairs along the eMTCT cascade.

Report on progress in physics: observation of the Breit-Wheeler process and vacuum birefringence in heavy-ion collisions.

This report reviews the effort over several decades to observe the linear Breit-Wheeler process (γγ→e+e-) and vacuum birefringence (VB) in high-energy particle and heavy-ion collider experiment. This report, motivated by the STAR collaboration's recent observations, attempts to summarize the key issues related to the interpretation of polarizedγγ→l+l-measurements in high-energy experiments. To that end, we start by reviewing the historical context and essential theoretical developments, before focusing on the decades of progress made in high-energy collider experiments. Special attention is given to the evolution in experimental approaches in response to various challenges, to the demanding detector capabilities required to unambiguously identify the linear Breit-Wheeler process, and to the connections with VB. We close the report with a discussion, followed by a look at near-future opportunities for utilizing these discoveries and for testing quantum electrodynamics in previously unexplored regimes.

Ultrafast structural changes direct the first molecular events of vision.

Vision is initiated by the rhodopsin family of light-sensitive G protein-coupled receptors (GPCRs)1. A photon is absorbed by the 11-cis retinal chromophore of rhodopsin, which isomerizes within 200 femtoseconds to the all-trans conformation2, thereby initiating the cellular signal transduction processes that ultimately lead to vision. However, the intramolecular mechanism by which the photoactivated retinal induces the activation events inside rhodopsin remains experimentally unclear. Here we use ultrafast time-resolved crystallography at room temperature3 to determine how an isomerized twisted all-trans retinal stores the photon energy that is required to initiate the protein conformational changes associated with the formation of the G protein-binding signalling state. The distorted retinal at a 1-ps time delay after photoactivation has pulled away from half of its numerous interactions with its binding pocket, and the excess of the photon energy is released through an anisotropic protein breathing motion in the direction of the extracellular space. Notably, the very early structural motions in the protein side chains of rhodopsin appear in regions that are involved in later stages of the conserved class A GPCR activation mechanism. Our study sheds light on the earliest stages of vision in vertebrates and points to fundamental aspects of the molecular mechanisms of agonist-mediated GPCR activation.

Effects of Angled Dynamic Compression Holes in a Tibial Plateau Levelling Osteotomy Plate on Cranially Directed Fragment Displacement.

OBJECTIVE: To compare angled dynamic compression holes in a tibial plateau levelling osteotomy (TPLO) plate to a commercially available TPLO plate in an ovine cadaveric model. STUDY DESIGN: Ovine tibias (40 bones) were mounted on a custom-made securement device and radiopaque markers were placed to aid radiographic measurements. A standard TPLO procedure was performed on each tibia with either a custom-made six-hole 3.5 mm angled compression hole plate (APlate) or a six-hole 3.5 mm standard commercial plate (SPlate). Radiographs were obtained before and after tightening of the cortical screws and evaluated by an observer blinded to the plate. Measurements of cranio-caudal displacement (CDisplacement), proximo-distal displacement (PDisplacement) in relation to the long axis of the tibia, and change in tibial plateau angle (TPA) were determined. RESULTS: CDisplacement was significantly greater in APlate (median 0.85 mm, Q1-Q3: 0.575-1.325 mm) compared to SPlate (median 0.00 mm, Q1-Q3: -0.35-0.50 mm, p < 0.0001). There were no significant differences in the PDisplacement (median 0.55 mm, Q1-Q3: 0.075-1.00 mm, p = 0.5066) or TPA change (median -0.50°, Q1-Q3: -1.225-0.25°, p = 0.1846) between the two plate types. CONCLUSION: APlate increases cranially directed displacement of the osteotomy in a TPLO procedure without causing TPA change. The reduced interfragmentary distance across the whole osteotomy could improve osteotomy healing compared to standard commercial TPLO plates.

Watching the release of a photopharmacological drug from tubulin using time-resolved serial crystallography.

The binding and release of ligands from their protein targets is central to fundamental biological processes as well as to drug discovery. Photopharmacology introduces chemical triggers that allow the changing of ligand affinities and thus biological activity by light. Insight into the molecular mechanisms of photopharmacology is largely missing because the relevant transitions during the light-triggered reaction cannot be resolved by conventional structural biology. Using time-resolved serial crystallography at a synchrotron and X-ray free-electron laser, we capture the release of the anti-cancer compound azo-combretastatin A4 and the resulting conformational changes in tubulin. Nine structural snapshots from 1 ns to 100 ms complemented by simulations show how cis-to-trans isomerization of the azobenzene bond leads to a switch in ligand affinity, opening of an exit channel, and collapse of the binding pocket upon ligand release. The resulting global backbone rearrangements are related to the action mechanism of microtubule-destabilizing drugs.

A spatio-temporal framework for modelling wastewater concentration during the COVID-19 pandemic.

The potential utility of wastewater-based epidemiology as an early warning tool has been explored widely across the globe during the current COVID-19 pandemic. Methods to detect the presence of SARS-CoV-2 RNA in wastewater were developed early in the pandemic, and extensive work has been conducted to evaluate the relationship between viral concentration and COVID-19 case numbers at the catchment areas of sewage treatment works (STWs) over time. However, no attempt has been made to develop a model that predicts wastewater concentration at fine spatio-temporal resolutions covering an entire country, a necessary step towards using wastewater monitoring for the early detection of local outbreaks. We consider weekly averages of flow-normalised viral concentration, reported as the number of SARS-CoV-2N1 gene copies per litre (gc/L) of wastewater available at 303 STWs over the period between 1 June 2021 and 30 March 2022. We specify a spatially continuous statistical model that quantifies the relationship between weekly viral concentration and a collection of covariates covering socio-demographics, land cover and virus associated genomic characteristics at STW catchment areas while accounting for spatial and temporal correlation. We evaluate the model's predictive performance at the catchment level through 10-fold cross-validation. We predict the weekly viral concentration at the population-weighted centroid of the 32,844 lower super output areas (LSOAs) in England, then aggregate these LSOA predictions to the Lower Tier Local Authority level (LTLA), a geography that is more relevant to public health policy-making. We also use the model outputs to quantify the probability of local changes of direction (increases or decreases) in viral concentration over short periods (e.g. two consecutive weeks). The proposed statistical framework can predict SARS-CoV-2 viral concentration in wastewater at high spatio-temporal resolution across England. Additionally, the probabilistic quantification of local changes can be used as an early warning tool for public health surveillance.

Systematic literature review and network meta-analysis of lurasidone, brexpiprazole and cariprazine for schizophrenia.

A systematic review was undertaken to identify randomized controlled trials (RCTs) comparing the efficacy and safety of lurasidone, brexpiprazole and cariprazine (selected because of a shared safety profile) with each other or placebo in adult patients with schizophrenia. Key outcomes included: Positive and Negative Syndrome Scales (PANSS), Clinical Global Impression-Severity (CGI-S) scores and cardiovascular and metabolic parameters. A feasibility assessment evaluated the trials' suitability for inclusion in a Bayesian network meta-analysis (NMA). Random effects models were used. In total, 1138 records were identified and 19 RCTs contributed to the NMA. Lurasidone doses of 160 mg performed best in terms of change in PANSS and CGI-S scores at 6 weeks, with stronger evidence when compared with brexpiprazole than cariprazine. The safety outcomes were variable; for all treatments, the 95% credible intervals usually contained 'no difference'. Active treatments were associated with lower odds of discontinuation due to any cause, and higher odds of experiencing any adverse event. Lurasidone was comparable to brexpiprazole and cariprazine for efficacy and safety outcomes assessed at 6 weeks, with the 160 mg dose being superior for the change in PANSS and CGI-S outcomes. The lurasidone results were relatively consistent across doses compared with brexpiprazole and cariprazine.