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Abstract The emotional valence of animals is challenging to assess, despite being a key component of welfare. In this study, we attempted to assess emotional valence through memory in 1- and 3-week-old piglets. It was hypothesized that piglets would spend less time in a pen where they experienced a negative event (castration) and more time in a pen where they experienced a positive event (enrichment). A testing apparatus was designed with three equally sized pens: two outer sections serving as treatment pens containing unique visual and tactile cues and a center section remaining neutral. Piglets received either negative or positive condition in one outer treatment pen and a sham treatment in the opposite. Various methods were tested (age of piglets, number and length of conditioning sessions, passive vs. active conditioning). Contrary to expectations, piglets did not decrease their time in the pen associated with the negative condition or increase their time in the pen associated with the positive condition. However, when exposed to the positive condition, results indicate older piglets developed an aversion towards the sham treatment. This study provides methodological groundwork for the application of place conditioning in piglets and highlights the nuances important for the use of cognitive tests to assess animal welfare.

Abstract Conventional radiography (CR) is primarily utilized for fracture diagnosis. Artificial intelligence (AI) for CR is a rapidly growing field aimed at enhancing efficiency and increasing diagnostic accuracy. However, the diagnostic performance of commercially available AI fracture detection solutions (CAAI-FDS) for CR in various anatomical regions, their synergy with human assessment, as well as the influence of industry funding on reported accuracy are unknown. Peer-reviewed diagnostic test accuracy (DTA) studies were identified through a systematic review on Pubmed and Embase. Diagnostic performance measures were extracted especially for different subgroups such as product, type of rater (stand-alone AI, human unaided, human aided), funding, and anatomical region. Pooled measures were obtained with a bivariate random effects model. The impact of rater was evaluated with comparative meta-analysis. Seventeen DTA studies of seven CAAI-FDS analyzing 38,978 x-rays with 8,150 fractures were included. Stand-alone AI studies (n = 15) evaluated five CAAI-FDS; four with good sensitivities (> 90%) and moderate specificities (80–90%) and one with very poor sensitivity ( 95%). Pooled sensitivities were good to excellent, and specificities were moderate to good in all anatomical regions (n = 7) apart from ribs (n = 4; poor sensitivity / moderate specificity) and spine (n = 4; excellent sensitivity / poor specificity). Funded studies (n = 4) had higher sensitivity (+ 5%) and lower specificity (-4%) than non-funded studies (n = 11). Sensitivity did not differ significantly between stand-alone AI and human AI aided ratings (p = 0.316) but specificity was significantly higher the latter group (p

Abstract Plants can survive in soils of low micromolar potassium (K+) concentrations. Root K+ intake is accomplished by the K+ channel AKT1 and KUP/HAK/KT type high-affinity K+ transporters. Arabidopsis HAK5 mutants impaired in low K+ acquisition have been identified already more than two decades ago, the molecular mechanism, however, is still a matter of debate also because of lack of direct measurements of HAK5-mediated K+ currents. When we expressed AtHAK5 in Xenopus oocytes together with CBL1/CIPK23, no inward currents were elicited in sufficient K+ media. Under low K+ and inward-directed proton motive force (PMF), the inward K+ current increased indicating that HAK5 energetically couples the uphill transport of K+ to the downhill flux of H+. At extracellular K+ concentrations above 25 μM, the initial rise in current was followed by a concentration-graded inactivation. When we replaced Tyr450 in AtHAK5 to Ala the K+ affinity strongly decreased, indicating that AtHAK5 position Y450 holds a key for K+ sensing and transport. When the soil K+ concentration drops toward the range that thermodynamically cannot be covered by AKT1, the AtHAK5 K+/H+ symporter progressively takes over K+ nutrition. Therefore, optimizing K+ use efficiency of crops, HAK5 could be key for low K+ tolerant agriculture.

Abstract Cancer‐associated cachexia is a multifactorial wasting disorder characterized by anorexia, unintentional weight loss (skeletal muscle mass with or without loss of fat mass), progressive functional impairment, and poor prognosis. This systematic literature review (SLR) examined the relationship between cachexia and survival in patients with colorectal or pancreatic cancer in recent literature. The SLR was conducted following PRISMA guidelines. Embase® and PubMed were searched to identify articles published in English between 1 January 2016 and 10 October 2021 reporting survival in adults with cancer and cachexia or at risk of cachexia, defined by international consensus (IC) diagnostic criteria or a broader definition of any weight loss. Included publications were studies in ≥100 patients with colorectal or pancreatic cancer. Thirteen publications in patients with colorectal cancer and 13 with pancreatic cancer met eligibility criteria. Included studies were observational and primarily from Europe and the United States. Eleven studies (42%) reported cachexia using IC criteria and 15 (58%) reported any weight loss. An association between survival and cachexia or weight loss was assessed across studies using multivariate (n = 23) or univariate (n = 3) analyses and within each study across multiple weight loss categories. Cachexia/weight loss was associated with a statistically significantly poorer survival in at least one weight loss category in 16 of 23 studies that used multivariate analyses and in 1 of 3 studies (33%) that used univariate analyses. Of the 17 studies demonstrating a significant association, 9 were in patients with colorectal cancer and 8 were in patients with pancreatic cancer. Cachexia or weight loss was associated with significantly poorer survival in patients with colorectal or pancreatic cancer in nearly two‐thirds of the studies. The classification of weight loss varied across and within studies (multiple categories were evaluated) and may have contributed to variability. Nonetheless, awareness of cachexia and routine assessment of weight change in clinical practice in patients with colorectal or pancreatic cancer could help inform prognosis and influence early disease management strategies.

Abstract Progressive supranuclear palsy (PSP), a rare Parkinsonian disorder, is characterized by problems with movement, balance, and cognition. PSP differs from Alzheimer’s disease (AD) and other diseases, displaying abnormal microtubule-associated protein tau by both neuronal and glial cell pathologies. Genetic contributors may mediate these differences; however, the genetics of PSP remain underexplored. Here we conduct the largest genome-wide association study (GWAS) of PSP which includes 2779 cases (2595 neuropathologically-confirmed) and 5584 controls and identify six independent PSP susceptibility loci with genome-wide significant (P

Abstract Background The field of long COVID research is rapidly evolving, however, tools to assess and monitor symptoms and recovery of the disease are limited. The objective of the present study was to develop a new patient-reported outcomes instrument, the Symptoms Evolution of Long COVID‑19 (SE-LC19), and establish its content validity. Methods The 40-item SE-LC19 instrument was developed based on patient-relevant empirical evidence from scientific literature and clinical guidelines that reported symptoms specific to long COVID. A 2-part mixed-method approach was employed. Part 1: Qualitative interviews with a purposive sample of 41 patients with confirmed long COVID were conducted for the content validation of SE-LC19. During cognitive debriefing interviews, patients were asked to describe their understanding of the instrument’s instructions, specific symptoms, response options, and recall period to ensure its relevance and comprehensiveness. Five clinicians of different medical specialties who regularly treated patients with long COVID were also interviewed to obtain their clinical expert opinions on SE-LC19. Part 2: Exploratory Rasch Measurement Theory (RMT) analysis was conducted to evaluate the psychometric properties of the SE-LC19 data collected during the interviews. Results Overall, patients reported that the instructions, questions, recall period, and response options for SE-LC19 were comprehensive and relevant. Minor conceptual gaps reported by patients captured nuances in the experience of some symptoms that could be considered in future studies. Some patients suggested a revision of the recall period from 24 h to 7 days to be able to capture more symptoms given the waxing and waning nature of some symptoms. Clinicians found the instrument comprehensive with minimal suggestions regarding its content. Exploratory RMT analyses provided evidence that the SE-LC19 questionnaire performed as intended. Conclusion The present mixed-methods study in patients with confirmed long COVID supports the content validity and applicability of the SE-LC19 instrument to evaluate the symptoms of patients with long COVID. Further research is warranted to explore the psychometric properties of the instrument and refine a meaningful and robust patient-relevant endpoint for use in different settings such as clinical trials and clinical practice to track the onset, severity, and recovery of long COVID.

Abstract Cachexia, with weight loss (WL) as a major component, is highly prevalent in patients with cancer and indicates a poor prognosis. The primary objective of this study was to conduct a meta‐analysis to estimate the risk of mortality associated with cachexia (using established WL criteria prior to treatment initiation) in patients with non‐small‐cell lung cancer (NSCLC) in studies identified through a systematic literature review. The review was conducted according to PRISMA guidelines. Embase® and PubMed were searched to identify articles on survival outcomes in adult patients with NSCLC (any stage) and cachexia published in English between 1 January 2016 and 10 October 2021. Two independent reviewers screened titles, abstracts and full texts of identified records against predefined inclusion/exclusion criteria. Following a feasibility assessment, a meta‐analysis evaluating the impact of cachexia, defined per the international consensus criteria (ICC), or of pre‐treatment WL ≥ 5% without a specified time interval, on overall survival in patients with NSCLC was conducted using a random‐effects model that included the identified studies as the base case. The impact of heterogeneity was evaluated through sensitivity and subgroup analyses. The standard measures of statistical heterogeneity were calculated. Of the 40 NSCLC publications identified in the review, 20 studies that used the ICC for cachexia or reported WL ≥ 5% and that performed multivariate analyses with hazard ratios (HRs) or Kaplan–Meier curves were included in the feasibility assessment. Of these, 16 studies (80%; n = 6225 patients; published 2016–2021) met the criteria for inclusion in the meta‐analysis: 11 studies (69%) used the ICC and 5 studies (31%) used WL ≥ 5%. Combined criteria (ICC plus WL ≥ 5%) were associated with an 82% higher mortality risk versus no cachexia or WL

Background: As transgender people initiate gender-affirming hormone therapy (GAHT), they are exposed to exogenous sex hormones that have effects that have not yet been fully studied. While exogenous estrogen is associated with a risk of venous thrombosis, the full impact of estrogen on the liver is unknown. Conversely, the erroneous attribution of risks from GAHT presents a barrier to treatment for some patients. We present a case of obliterative portal venopathy (OPV) and possible DILI occurring after the initiation of estrogen in a transgender woman. Case presentation: A 28-year-old transgender woman on GAHT was referred to hepatology for liver enzyme elevations. She did not have any notable comorbid conditions, family history, or psychosocial history. Lab and imaging workup were unremarkable, and the patient underwent liver biopsy. The patient’s biopsy results showed OPV. The patient continued GAHT at a lower dose and liver enzyme elevations resolved. Conclusions: OPV is a vascular disease that falls under the category of porto-sinusoidal vascular disorder. Patients with this condition can present with or without overt clinical signs of portal hypertension. Porto-sinusoidal vascular disorder is rare and given the timing and possible dose dependence, it might be reasonable to consider that the observed OPV was influenced by the exogenous estrogen administered in an association not previously reported. Alternatively, the patient’s continued estrogen treatment without ill effect could suggest that the events were not connected and that the fear of harm could have served as a barrier to the patient receiving indicated care.

Spin models like the Heisenberg Hamiltonian effectively describe the interactions of open-shell transition-metal ions on a lattice and can account for various properties of magnetic solids and molecules. Numerical methods are usually required to find exact or approximate eigenstates, but for small clusters with spatial symmetry, analytical solutions exist, and a few Heisenberg systems have been solved in closed form. This paper presents a simple, generally applicable approach to analytically solve isotropic spin clusters, based on adapting the basis to both total spin and point group symmetry to factor the Hamiltonian matrix into sufficiently small blocks. We demonstrate applications to small rings and polyhedra, some of which are straightforward to solve by successive spin-coupling for Heisenberg terms only; additional interactions, such as biquadratic exchange or multi-center terms necessitate symmetry adaptation.

Abstract High altitude (HA) ascent imposes systemic hypoxia and associated risk of acute mountain sickness. Acute hypoxia elicits a hypoxic ventilatory response (HVR), which is augmented with chronic HA exposure (i.e., ventilatory acclimatization; VA). However, laboratory‐based HVR tests lack portability and feasibility in field studies. As an alternative, we aimed to characterize area under the curve (AUC) calculations on Fenn diagrams, modified by plotting portable measurements of end‐tidal carbon dioxide (PETCO2) against peripheral oxygen saturation (SpO2) to characterize and quantify VA during incremental ascent to HA (n = 46). Secondarily, these participants were compared with a separate group following the identical ascent profile whilst self‐administering a prophylactic oral dose of acetazolamide (Az; 125 mg BID; n = 20) during ascent. First, morning PETCO2 and SpO2 measurements were collected on 46 acetazolamide‐free (NAz) lowland participants during an incremental ascent over 10 days to 5160 m in the Nepal Himalaya. AUC was calculated from individually constructed Fenn diagrams, with a trichotomized split on ranked values characterizing the smallest, medium, and largest magnitudes of AUC, representing high (n = 15), moderate (n = 16), and low (n = 15) degrees of acclimatization. After characterizing the range of response magnitudes, we further demonstrated that AUC magnitudes were significantly smaller in the Az group compared to the NAz group (P = 0.0021), suggesting improved VA. These results suggest that calculating AUC on modified Fenn diagrams has utility in assessing VA in large groups of trekkers during incremental ascent to HA, due to the associated portability and congruency with known physiology, although this novel analytical method requires further validation in controlled experiments. Highlights What is the central question of this study? What are the characteristics of a novel methodological approach to assess ventilatory acclimatization (VA) with incremental ascent to high altitude (HA)? What is the main finding and its importance? Area under the curve (AUC) magnitudes calculated from modified Fenn diagrams were significantly smaller in trekkers taking an oral prophylactic dose of acetazolamide compared to an acetazolamide‐free group, suggesting improved VA. During incremental HA ascent, quantifying AUC using modified Fenn diagrams is feasible to assess VA in large groups of trekkers with ascent, although this novel analytical method requires further validation in controlled experiments.

Abstract A recent evidence gaps assessment of the clinical, health‐related quality of life, and economic burden associated with α‐thalassemia is lacking. We conducted a systematic literature review (SLR) following the methodological and reporting requirements of the Preferred Reporting Items for Systematic Reviews and Meta‐Analyses and the Cochrane Handbook for Systematic Reviews, using available literature over the past decade. This SLR identified a considerable evidence gap with regard to understanding the current burden of α‐thalassemia as evident from paucity of studies published in the past 10 years. The limited data available still indicate that patients with α‐thalassemia experience substantial morbidity and quality of life/economic burden that is generally comparable to patients with β‐thalassemia.

ABSTRACT Anaerobic digestion and composting of biowastes are vital pathways to recycle carbon and nutrients for agriculture. However, plastic contamination of soil amendments and fertilizers made from biowastes is a relevant source of (micro‐) plastics in (agricultural) ecosystems. To avoid this contamination, plastic containing biowastes could be pyrolyzed to eliminate the plastic, recycle most of the nutrients, and create carbon sinks when the resulting biochar is applied to soil. Literature suggests plastic elimination mainly by devolatilization at co‐pyrolysis temperatures of > 520°C. However, it is uncertain if the presence of plastic during biomass pyrolysis induces the formation of organic contaminants or has any other adverse effects on biochar properties. Here, we produced biochar from wood residues (WR) obtained from sieving of biowaste derived digestate. The plastic content was artificially enriched to 10%, and this mixture was pyrolyzed at 450°C and 600°C. Beech wood (BW) chips and the purified, that is, (macro‐) plastic‐free WR served as controls. All biochars produced were below limit values of the European Biochar Certificate (EBC) regarding trace element content and organic contaminants. Under study conditions, pyrolysis of biowaste, even when contaminated with plastic, can produce a biochar suitable for agricultural use. However, thermogravimetric and nuclear magnetic resonance spectroscopic analysis of the WR + 10% plastics biochar suggested the presence of plastic residues at pyrolysis temperatures of 450°C. More research is needed to define minimum requirements for the pyrolysis of plastic containing biowaste and to cope with the automated identification and determination of plastic types in biowaste at large scales.

Abstract Wolves are opportunistic generalists that can respond quickly to new and unique food sources. Wolves in some ecosystems will consume berries and other fruits when they are abundant and available; however, many aspects of this behavior remain unknown. In the Greater Voyageurs Ecosystem (GVE), Minnesota, USA, wolves consistently consume berries, particularly blueberries, when they are available. We deployed remote cameras in blueberry patches to record wolves foraging on berries over several years. We captured footage of wolves of all age classes, social statuses, and sex foraging on blueberries alone or with other wolves. Our observations indicate berry consumption by wolves is a widespread behavior in the GVE and likely in similar southern boreal ecosystems. We hope our work spurs researchers across wolf range to examine whether berry consumption by wolves is a widespread and ubiquitous behavior for wolves.

Abstract Effective photoinduced charge transfer makes molecular bimetallic assemblies attractive for applications as active light‐induced proton reduction systems. Developing competitive base metal dyads is mandatory for a more sustainable future. However, the electron transfer mechanisms from the photosensitizer to the proton reduction catalyst in base metal dyads remain so far unexplored. A Fe─Co dyad that exhibits photocatalytic H2 production activity is studied using femtosecond X‐ray emission spectroscopy, complemented by ultrafast optical spectroscopy and theoretical time‐dependent DFT calculations, to understand the electronic and structural dynamics after photoexcitation and during the subsequent charge transfer process from the FeII photosensitizer to the cobaloxime catalyst. This novel approach enables the simultaneous measurement of the transient X‐ray emission at the iron and cobalt K‐edges in a two‐color experiment. With this methodology, the excited state dynamics are correlated to the electron transfer processes, and evidence of the Fe→Co electron transfer as an initial step of proton reduction activity is unraveled.

Objective Pregnant women with COVID-19 are at elevated risk for severe outcomes, but clinical data on management of these patients are limited. Monoclonal antibodies, such as casirivimab plus imdevimab (CAS+IMD), have proven effective in treating non-pregnant adults with COVID-19, prompting further evaluation in pregnant women.Methods A phase 3 portion of an adaptive, multicentre, randomised, double-blind, placebo-controlled trial evaluated the safety, clinical outcomes, pharmacokinetics and immunogenicity of CAS+IMD (1200 mg or 2400 mg) in the treatment of pregnant outpatients with COVID-19 (NCT04425629). Participants were enrolled between December 2020 and November 2021, prior to the emergence of Omicron-lineage variants against which CAS+IMD is not active. Safety was evaluated in randomised participants who received study drug (n=80); clinical outcomes were evaluated in all randomised participants (n=82). Only two pregnant participants received placebo, limiting conclusions regarding treatment effect. Infants born to pregnant participants were followed for developmental outcomes ≤1 year of age.Results In pregnant participants, CAS+IMD was well tolerated, with no grade ≥2 hypersensitivity or infusion-related reactions reported. There were no participant deaths, and only one COVID-19–related medically attended visit. Although two pregnancies (3%) reported issues in the fetus/neonate, they were confounded by maternal history or considered to be due to an alternate aetiology. No adverse developmental outcomes in infants ≤1 year of age were considered related to in utero exposure to the study drug. CAS+IMD 1200 mg and 2400 mg rapidly and similarly reduced viral loads, with a dose-proportional increase in concentrations of CAS+IMD in serum. Pharmacokinetics were consistent with that reported in the general population. Immunogenicity incidence was low.Conclusion CAS+IMD treatment of pregnant outpatients with COVID-19 showed similar safety, clinical outcomes and pharmacokinetic profiles to that observed in non-pregnant adults. There was no evidence of an impact on developmental outcomes in infants ≤1 year of age.Trial registration number NCT04425629.

Abstract Vapor phase infiltration (VPI) has emerged as a promising tool for fabrication of novel hybrid materials. In the field of polymeric gas separation membranes, a beneficial impact on stability and membrane performance is known for several polymers with differing functional groups. This study for the first time investigates VPI of trimethylaluminum (TMA) into poly(1‐trimethylsilyl‐1‐propyne) (PTMSP), featuring a carbon–carbon double bond as functional group. Saturation of the precursor inside the polymer is already attained after 60 s infiltration time leading to significant densification of the material. Depth profiling proves accumulation of aluminum in the polymer itself, but a significantly increased accumulation is visible in the gradient layer between polymer and SiO2 substrate. A reaction pathway is proposed and supplemented by density‐functional theory (DFT) calculations. Infrared spectra derived from both experiments and simulation support the presented reaction pathway. In terms of permeance, a favorable impact on selectivity is observed for infiltration times up to 1 s. Longer infiltration times yield greatly reduced permeance values close or even below the detection limit of the measurement device. The present results of this study set a strong basis for the application of VPI on polymers for gas‐barrier and membrane applications in the future.

Abstract The Kunming‐Montreal Global Biodiversity Framework lays out an ambitious set of goals and targets aimed at halting and reversing biodiversity loss. The extent of natural ecosystems has been selected as one of a small set of headline indicators against which countries will report progress under this framework. We evaluate the effectiveness with which this indicator is expected to capture the overall scope of the ecosystem‐focused component of Goal A, and interlinkages with the species‐focused component of this same goal, using extensive global data on the integrity, connectivity, and plant species composition of forests. Results generated for all forest‐supporting countries demonstrate that consideration of these additional factors can profoundly alter understanding of the state of forest biodiversity relative to that based on extent alone. Employment of ecosystem extent as a headline indicator must therefore be augmented by appropriate use of component and complementary indicators addressing other key dimensions of ecosystem change.

Abstract Introduction Pea is one of the most important pulse legumes globally due to its high protein, which is due to its ability to fix atmospheric nitrogen through a symbiotic relationship with nitrogen‐fixing rhizobia. Symbiotic nitrogen fixation (SNF) should be optimized to maximize nitrogen fixation and achieve higher yields with more grain protein. The use of humic‐based soil amendments in crop production has garnered considerable attention in recent years due to their biostimulatory effect in improving plant growth, yield, nutritional quality and soil health. This study examines the impact of a humic‐based soil amendment (Humalite) on root nodulation, SNF, plant growth, yield and grain protein of pea. Materials and Methods Chemical characterization was performed using Fourier transform infrared spectroscopy (FTIR). Pea plants inoculated with Rhizobia were grown in pots under greenhouse conditions with five different Humalite rates (0, 200, 400, 800 and 1600 kg ha−1). SNF capacity was assessed using the 15N‐isotope dilution method. Results FTIR analysis revealed the abundant presence of hydroxyl (‐OH) and carboxyl (‐CO) functional groups in Humalite. Plants treated with Humalite displayed augmented root traits [root length (21%–50%), root surface area (24%–51%), volume (26%–53%), average nodule weight (11%–91%)], plant biomass [shoots (13%–29%) and roots (29%–54%)], shoot nitrogen concentration (12%–33%), shoot total nitrogen content (38%–53%), percentage nitrogen derived from the atmosphere (8%–14%) and total shoot nitrogen fixed (48%–80%) compared to the control plants at the flowering stage. Furthermore, at seed maturity stage, plants treated with Humalite at 400 and 1600 kg ha−1 exhibited a significant increase in plant biomass (4%–14%), number of seeds (8%–16%), seed weight (3%–11%), seed nitrogen content (8%–20%) and total seed nitrogen fixed (7%–22%) compared to the control plants. Conclusions These findings demonstrate that humic‐based soil amendment can effectively enhance plant growth, root nodulation, SNF and seed yield, thereby supporting sustainable agricultural practices.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for nearly 7 million deaths worldwide since its outbreak in late 2019. Even with the rapid development and production of vaccines and intensive research, there is still a huge need for specific anti-viral drugs that address the rapidly arising new variants. To address this concern, the National Institute of Allergy and Infectious Diseases (NIAID) established nine Antiviral Drug Discovery (AViDD) Centers, tasked with exploring approaches to target pathogens with pandemic potential, including SARS-CoV-2. In this study, we sought inhibitors of SARS-CoV2 non-structural protein 13 (nsP13) as potential antivirals, first developing a HTS-compatible assay to measure SARS-CoV2 nsP13 helicase activity. Here we present our effort in implementing the assay in a 1,536 well-plate format and in identifying nsP13 inhibitor hit compounds from a ∼650,000 compound library. The primary screen was robust (average Z’ = 0.86 ± 0.05) and resulted in 7,009 primary hits. 1,763 of these compounds upon repeated retests were further confirmed, showing consistent inhibition. Following in-silico analysis, an additional orthogonal assay and titration assays, we identified 674 compounds with IC50

Emerging highly pathogenic viruses can pose profound impacts on global health, the economy, and society. To meet that challenge, the National Institute of Allergy and Infectious Diseases (NIAID) established nine Antiviral Drug Discovery (AViDD) centers for early-stage identification and validation of novel antiviral drug candidates against viruses with pandemic potential. As part of this initiative, we established paired entry assays that simultaneously screen for inhibitors specifically targeting SARS-CoV-2 (SARS2), Lassa virus (LASV) and Machupo virus (MACV) entry. To do so we employed a dual pseudotyped virus (PV) infection system allowing us to screen ∼650,000 compounds efficiently and cost-effectively. Adaptation of these paired assays into 1536 well-plate format for ultra-high throughput screening (uHTS) resulted in the largest screening ever conducted in our facility, with over 2.4 million wells completed. The paired infection system allowed us to detect two PV infections simultaneously: LASV + MACV, MACV + SARS2, and SARS2 + LASV. Each PV contains a different luciferase reporter gene which enabled us to measure the infection of each PV exclusively, albeit in the same well. Each PV was screened at least twice utilizing different reporters, which allowed us to select the inhibitors specific to a particular PV and to exclude those that hit off targets, including cellular components or the reporter proteins. All assays were robust with an average Z’ value ranging from 0.5 to 0.8. The primary screening of ∼650,000 compounds resulted in 1812, 1506, and 2586 unique hits for LASV, MACV, and SARS2, respectively. The confirmation screening narrowed this list further to 60, 40, and 90 compounds that are unique to LASV, MACV, and SARS2, respectively. Of these compounds, 8, 35, and 50 compounds showed IC50 value < 10 μM, some of which have much greater potency and excellent antiviral activity profiles specific to LASV, MACV, and SARS2, and none are cytotoxic. These selected compounds are currently being studied for their mechanism of action and to improve their specificity and potency through chemical modification.

Introduction: Streptococcal meningoencephalitis (SME) is a rare, and frequently lethal, acute infection, and inflammation of the central nervous system parenchyma, with associated meningeal involvement. Bacterial meningoencephalitis is generally associated with high rates of morbidity and mortality, despite available antimicrobial and corticosteroid treatments. While Streptococcus pneumoniae is well recognised to cause bacterial meningitis, direct extension into the central nervous system parenchyma is rare. Case Presentation: A previously well 49-year-old man presented with sudden onset severe headache, fevers, neck stiffness, and reduced consciousness. The manifestations of SME in this patient were bilateral pupil-involving third-nerve palsies, wall-eyed bilateral internuclear ophthalmoplegia (WEBINO), bilateral blindness, bilateral deafness, a right lower motor neuron facial palsy, and upper motor neuron signs in his limbs. Initially, a partial response to high dose intravenous antibiotics occurred, but with administration of intravenous corticosteroids, further substantial resolution of the patient’s neurological and neuro-ophthalmological deficits occurred. Conclusion: This case highlights the benefit of multidisciplinary diagnostic and therapeutic interventions in a case of SME complicated by bilateral pupil-involving third-nerve palsies, WEBINO, bilateral blindness, bilateral deafness, a right lower motor neuron facial palsy, and upper motor neuron signs. It appears to be the first reported case of SME with this rare collection of neuro-ophthalmological abnormalities.

Abstract Stalls or crates are a very common type of housing used on pig farms that restrict an animal’s movement. How this confinement impacts the animal’s affective states is seldom investigated. We conducted a preference test over 7 days where trios of gilts (n = 10 trios, 27.4 ± 1.5 weeks old) had free access between individual self-locking stalls (~ 1.2 m2) and a shared open area allowing 2.8 m2/animal (71% of total area). Gilts had access to ad libitum feed and water both inside the crates and in the open area. After 7 days, personality traits of the animals were assessed with open field (OF) and novel object (NO) tests. Principal Component Analysis (PCA) yielded two main components, which we defined as Passivity and Engagement. The median time spent outside the crate was 95.2% as 21/29 of the gilts exhibited a significant preference for pen over crate during the 7-day trial (p

Abstract The development of carbon dioxide removal methods, coupled with decreased CO2 emissions, is fundamental to achieving the targets outlined in the Paris Agreement limiting global warming to 1.5 °C. Here we are investigating the importance of the organic carbon feedstock to support silicate mineral weathering in small-scale flow through bioreactors and subsequent CO2 sequestration. Here, we combine two bacteria and two fungi, widely reported for their weathering potential, in simple flow through bioreactors (columns) consisting of forsterite and widely available, cheap organic carbon sources (wheat straw, bio-waste digestate of pig manure and biowaste, and manure compost), over six weeks. Compared to their corresponding abiotic controls, the inoculated straw and digestate columns release more total alkalinity (~2 times more) and produce greater dissolved and solid inorganic carbon (29% for straw and 13% for digestate), suggesting an increase in CO2 sequestration because of bio-enhanced silicate weathering. Microbial biomass is higher in the straw columns compared to the digestate and manure compost columns, with a phospholipid fatty acid derived total microbial biomass 10 x greater than the other biotic columns. Scanning Electron Microscopy imaging shows the most extensive colonisation and biofilm formation on the mineral surfaces in the straw columns. The biotic straw and digestate columns sequester 50 and 14 mg C more than their abiotic controls respectively, while there is no difference in the manure columns. The selection of organic carbon sources to support microbial communities in the flow through bioreactors controlls the silicate weathering rates and CO2 sequestration.

Abstract Background The primary treatment of ulcerative colitis (UC) is medical therapy using a standard step-up approach. An appendectomy might modulate the clinical course of UC, decreasing the incidence of relapses and reducing need for medication. The objective of the ACCURE trial is to assess the efficacy of laparoscopic appendectomy in addition to standard medical treatment in maintaining remission in UC patients. This article presents the statistical analysis plan to evaluate the outcomes of the ACCURE trial. Design and methods The ACCURE trial was designed as a multicentre, randomised controlled trial. UC patients with a new diagnosis or a disease relapse within the past 12 months, treated with 5-ASA, corticosteroids, or immunomodulators until complete clinical and endoscopic remission (defined as total Mayo score 150 or intensified medical therapy other than 5-ASA therapy). Secondary outcomes include number of relapses per patient, time to first relapse, disease activity, number of colectomies, medication usage, and health-related quality of life. Discussion The ACCURE trial will provide comprehensive evidence whether adding an appendectomy to maintenance treatment is superior to maintenance treatment only in maintaining remission in UC patients. Trial registration Dutch Trial Register (NTR) NTR2883 . Registered May 3, 2011. ISRCTN, ISRCTN60945764 . Registered August 12, 2019.

Abstract In this work, we use a simple multi-agent-based-model (MABM) of a social network, implementing selfish algorithm (SA) agents, to create an adaptive environment and show, using a modified diffusion entropy analysis (DEA), that the mutual-adaptive interaction between the parts of such a network manifests complexity synchronization (CS). CS has been shown to exist by processing simultaneously measured time series from among organ-networks (ONs) of the brain (neurophysiology), lungs (respiration), and heart (cardiovascular reactivity) and to be explained theoretically as a synchronization of the multifractal dimension (MFD) scaling parameters characterizing each time series. Herein, we find the same kind of CS in the emergent intelligence of groups formed in a self-organized social interaction without macroscopic control but with biased self-interest between two groups of agents playing an anti-coordination game. This computational result strongly suggests the existence of the same CS in real-world social phenomena and in human–machine interactions as that found empirically in ONs.

Abstract Arp2/3 complex nucleates branched actin filaments for cell and organelle movements. Here we report a 2.7 Å resolution cryo-EM structure of the mature branch junction formed by S. pombe Arp2/3 complex that provides details about interactions with both mother and daughter filaments. We determine a second structure at 3.2 Å resolution with the phosphate analog BeFx bound with ADP to Arp3 and ATP bound to Arp2. In this ADP-BeFx transition state the outer domain of Arp3 is rotated 2° toward the mother filament compared with the ADP state and makes slightly broader contacts with actin in both the mother and daughter filaments. Thus, dissociation of Pi from the ADP-Pi transition state reduces the interactions of Arp2/3 complex with the actin filaments and may contribute to the lower mechanical stability of mature branch junctions with ADP bound to the Arps. Our structures also reveal that the mother filament in contact with Arp2/3 complex is slightly bent and twisted, consistent with the preference of Arp2/3 complex binding curved actin filaments. The small degree of twisting constrains models of actin filament mechanics.

Abstract Caspase-2, one of the most evolutionarily conserved members of the caspase family, is an important regulator of the cellular response to oxidative stress. Given that ferroptosis is suppressed by antioxidant defense pathways, such as that involving selenoenzyme glutathione peroxidase 4 (GPX4), we hypothesized that caspase-2 may play a role in regulating ferroptosis. This study provides the first demonstration of an important and unprecedented function of caspase-2 in protecting cancer cells from undergoing ferroptotic cell death. Specifically, we show that depletion of caspase-2 leads to the downregulation of stress response genes including SESN2, HMOX1, SLC7A11, and sensitizes mutant-p53 cancer cells to cell death induced by various ferroptosis-inducing compounds. Importantly, the canonical catalytic activity of caspase-2 is not required for its role and suggests that caspase-2 regulates ferroptosis via non-proteolytic interaction with other proteins. Using an unbiased BioID proteomics screen, we identified novel caspase-2 interacting proteins (including heat shock proteins and co-chaperones) that regulate cellular responses to stress. Finally, we demonstrate that caspase-2 limits chaperone-mediated autophagic degradation of GPX4 to promote the survival of mutant-p53 cancer cells. In conclusion, we document a novel role for caspase-2 as a negative regulator of ferroptosis in cells with mutant p53. Our results provide evidence for a novel function of caspase-2 in cell death regulation and open potential new avenues to exploit ferroptosis in cancer therapy.

Advances in genome sequencing have greatly accelerated the identification of sex chromosomes in a variety of species. Many of these species have experienced structural rearrangements that reduce recombination between the sex chromosomes, allowing the accumulation of sequence differences over many megabases. Identification of the genes that are responsible for sex determination within these sometimes large regions has proved difficult. Here, we identify an XY sex chromosome system on LG19 in the West African cichlid fish Chromidotilapia guntheri in which the region of differentiation extends over less than 400 kb. We develop high-quality male and female genome assemblies for this species, which confirm the absence of structural variants, and which facilitate the annotation of genes in the region. The peak of differentiation lies within rin3, which has experienced several debilitating mutations on the Y chromosome. We suggest two hypotheses about how these mutations might disrupt endocytosis, leading to Mendelian effects on sexual development.

A number of post-mortem studies conducted in transplanted Huntington’s disease (HD) patients from various trials have reported the presence of pathological and misfolded proteins, in particular mutant huntingtin (mHtt) and phosphorylated tau neuropil threads, in the healthy grafted tissue. Here, we extended these observations with histological analysis of post-mortem tissue from three additional HD patients who had received similar striatal allografts from the fetal tissue transplantation trial conducted in Los Angeles in 1998. Immunohistochemical staining was performed using anti-mHtt antibodies, EM48 and MW7, as well as anti-hyperphosphorylated tau antibodies, AT8 and CP13. Immunofluorescence was used to assess the colocalization of EM48+ mHtt aggregates with the neuronal marker MAP2 and/or the extracellular matrix protein phosphacan in both the host and grafts. We confirmed the presence of mHtt aggregates within grafts of all three cases as well as tau neuropil threads in the grafts of two of the three transplanted HD patients. Phosphorylated tau was also variably expressed in the host cerebral cortex of all three subjects. While mHtt inclusions were present within neurons (immunofluorescence co-localization of MAP2 and EM48) as well as within the extracellular matrix of the host (immunofluorescence co-localization of phosphacan and EM48), their localization was limited to the extracellular matrix in the grafted tissue. This study corroborates previous findings that both mHtt and tau pathology can be found in the host and grafts of HD patients years post-grafting.

Abstract In a warming climate, an intensifying fire regime and higher likelihood of extreme rain are expected to increase watershed sediment yield in many regions. Understanding regional variability in landscape response to fire and post‐fire rainfall is essential for managing water resources and infrastructure. We measured sediment yield resulting from sequential wildfire and extreme rain and flooding in the upper Carmel River watershed (116 km2), on the central California coast, USA, using changes in sediment volume mapped in a reservoir. We determined that the sediment yield after fire and post‐fire flooding was 854–1,100 t/km2/yr, a factor of 3.5–4.6 greater than the long‐term yield from this watershed and more than an order of magnitude greater than during severe drought conditions. In this first large‐scale field validation test of the WEPPcloud/wepppy framework for the Water Erosion Prediction Project (WEPP) model on a burned landscape, WEPP predicted 81%–106% of the measured sediment yield. These findings will facilitate assessing and predicting future fire effects in steep watersheds with a Mediterranean climate and indicate that the increasingly widespread use of WEPP is appropriate for evaluating post‐fire hillslope erosion even across 100‐km2 scales under conditions without debris flows.

IntroductionSoil organic carbon (SOC) content can vary significantly across a given plot. Therefore, a representative sampling is a prerequisite to obtain meaningful results from analysis and of utter importance when SOC quantification is used to quantify (temporary) carbon dioxide removal (CDR). However, certain management practices aiming to increase SOC further increase the level of heterogeneity and may challenge representative sampling schemes. This includes concentrated root-zone application of biochar, which immediately increases SOC with the input of biochar-C and may promote the local enrichment of non-biochar SOC over time.MethodsHere, we used numerical modelling to quantify the number of single sampling points (soil cores) needed to achieve a representative sample of biochar-C and total SOC on a plot after application of biochar in rows, e.g., for growing vegetables, or in the circumference of trees in agroforestry systems.ResultsAfter row application of 5 t ha−1 biochar in soil with rather low SOC content (26 t ha−1), 140 soil cores (per 0.25 ha plot) where necessary to achieve representative sampling of C-stocks (±5% error) in 90% of the repeated sampling simulation cases. Compared to realistic and cost-effective soil sampling scenarios in agronomic practice, we conclude that concentrated root zone application of biochar makes representative sampling for quantification of SOC in soils with low baseline C-stocks virtually impossible.DiscussionThis finding calls into question the soil-sampling and SOC-analysis-based (“result-based”) monitoring of SOC as a (temporary) CDR when biochar might have been applied. Considering the rapid scaling of biochar production and use in agriculture, this is a considerable challenge for SOC certification. Instead, action-based incentives, rewarding farmers for carrying out specific practices, could be applied to promote carbon farming practices.

Abstract Isoprene, a volatile hydrocarbon, is typically emitted from the leaves of many plant species. Given its well‐known function in plant growth and defense aboveground, we examined its effects on root physiology. We used isoprene‐emitting (IE) lines and a non‐emitting (NE) line of Arabidopsis and investigated their performance by analyzing root phenotype, hormone levels, transcriptome, and metabolite profiles under both normal and salt stress conditions. We show that IE lines emitted tiny amounts of isoprene from roots and showed an increased root/shoot ratio compared with NE line. Isoprene emission exerted a noteworthy influence on hormone profiles related to plant growth and stress response, promoting root development and salt‐stress resistance. Methyl erythritol 4‐phosphate pathway metabolites, precursors of isoprene and hormones, were higher in the roots of IE lines than in the NE line. Transcriptome data indicated that the presence of isoprene increased the expression of key genes involved in hormone metabolism/signaling. Our findings reveal that constitutive root isoprene emission sustains root growth under saline conditions by regulating and/or priming hormone biosynthesis and signaling mechanisms and expression of key genes relevant to salt stress defense.

Abstract Metabolites are essential molecules involved in various metabolic processes, and their deficiencies and excessive concentrations can trigger significant physiological consequences. The detection of multiple metabolites within a non‐invasively collected biofluid could facilitate early prognosis and diagnosis of severe diseases. Here, a metal oxide heterojunction transistor (HJ‐TFT) sensor is developed for the label‐free, rapid detection of uric acid (UA) and 25(OH)Vitamin‐D3 (Vit‐D3) in human saliva. The HJ‐TFTs utilize a solution‐processed In2O3/ZnO channel functionalized with uricase enzyme and Vit‐D3 antibody for the selective detection of UA and Vit‐D3, respectively. The ultra‐thin tri‐channel architecture facilitates strong coupling between the electrons transported along the buried In2O3/ZnO heterointerface and the electrostatic perturbations caused by the interactions between the surface‐immobilized bioreceptors and target analytes. The biosensors can detect a wide range of concentrations of UA (from 500 nm to 1000 µM) and Vit‐D3 (from 100 pM to 120 nm) in human saliva within 60 s. Moreover, the biosensors exhibit good linearity with the physiological concentration of metabolites and limit of detections of ≈152 nm for UA and ≈7 pM for Vit‐D3 in real saliva. The specificity is demonstrated against various interfering species, including other metabolites and proteins found in saliva, further showcasing its capabilities.

Summary: PAX3/7 fusion-negative rhabdomyosarcoma (FN-RMS) is a childhood mesodermal lineage malignancy with a poor prognosis for metastatic or relapsed cases. Limited understanding of advanced FN-RMS is partially attributed to the absence of sequential invasion and dissemination events and the challenge in studying cell behavior, using, for example, non-invasive intravital microscopy (IVM), in currently used xenograft models. Here, we developed an orthotopic tongue xenograft model of FN-RMS to study cell behavior and the molecular basis of invasion and metastasis using IVM. FN-RMS cells are retained in the tongue and invade locally into muscle mysial spaces and vascular lumen, with evidence of hematogenous dissemination to the lungs and lymphatic dissemination to lymph nodes. Using IVM of tongue xenografts reveals shifts in cellular phenotype, migration to blood and lymphatic vessels, and lymphatic intravasation. Insight from this model into tumor invasion and metastasis at the tissue, cellular, and subcellular level can guide new therapeutic avenues for advanced FN-RMS. Motivation: Current models for studying fusion-negative rhabdomyosarcoma (FN-RMS) are limited in terms of their physiological relevance and ability to inform on invasion and metastasis. As a step toward advancing our understanding of molecular mechanisms of advanced FN-RMS, we developed an orthotopic tongue xenograft model of FN-RMS for intravital multi-photon microscopy. Longitudinal non-invasive intravital microscopy of cell line-based xenografts in live animals allows the dissection of the cellular behavior and related molecular basis in vivo. Combined with the tongue xenograft model, the system facilitates the examination of cellular behavior in FN-RMS local invasion, dissemination, and metastasis.

This study aimed to determine whether obese men with nonalcoholic fatty liver disease (NAFLD) display differences between those with simple steatosis versus steatohepatitis (NASH) in splanchnic and hepatic FFA and VLDL-triglycerides (VLDL-TG) balances. The study involved 17 obese men with biopsy-proven NAFLD (9 with NASH and 8 with simple steatosis). We used hepatic vein catheterization in combination with [3H]palmitate and [14C]VLDL-TG tracers to measure splanchnic palmitate and VLDL-TG uptake and release rates during basal and hyperinsulinemic conditions. Indocyanine green was used to measure splanchnic plasma flow. Splanchnic palmitate uptake was similar in the two groups and significantly reduced during hyperinsulinemia (NASH: 62 (48–77) versus 38 (18–58) μmol/min; simple steatosis: 62 (46–78) versus 45 (25–65) μmol/min, mean (95% CI), basal versus clamp periods, respectively, P = 0.02 time-effect). Splanchnic palmitate release was also comparable between groups and nonsignificantly diminished during hyperinsulinemia. The percent palmitate delivered to the liver originating from visceral adipose tissue lipolysis was similar and unchanged by hyperinsulinemia. Splanchnic uptake and release of VLDL-TG were similar between groups. Hyperinsulinemia suppressed VLDL-TG release (P

Inflammation, reduced gamma-aminobutyric acidergic (GABAergic) function and altered neuroplasticity are co-occurring pathophysiologies in major depressive disorder (MDD). However, the link between these biological changes remains unclear. We hypothesized that inflammation induces deficits in GABAergic interneuron markers and that this effect is mediated by brain-derived neurotrophic factor (BDNF). We report here that systemic inflammation induced by intraperitoneal injection of lipopolysaccharide (LPS) (0.125, 0.25, 0.5, 1, 2 mg/kg) in the first cohort of C57BL/6 mice (n = 72; 10–11 weeks; 50% female) resulted in increased interleukin 1-beta and interleukin-6 in prefrontal cortex (PFC) and hippocampus (HPC), as measured using enzyme-linked immunosorbent assay (ELISA). Quantitative real-time polymerase reaction (qPCR) was used to explore the effect of LPS on the expression of GABAergic interneuron markers. In the PFC of the second cohort (n = 39; 10–11 weeks; 50% female), 2 mg/kg of LPS decreased the expression of somatostatin (Sst) (p = 0.0014), parvalbumin (Pv) (p = 0.0257), cortistatin (Cort) (p = 0.0003), neuropeptide Y (Npy) (p = 0.0033) and cholecystokinin (Cck) (p = 0.0041), and did not affect corticotropin-releasing hormone (Crh) and vasoactive intestinal peptide (Vip) expression. In the HPC, 2 mg/kg of LPS decreased the expression of Sst (p = 0.0543), Cort (p = 0.0011), Npy (p = 0.0001), and Cck (p

In higher education, generative chatbots have infiltrated teaching and learning. Concerns about how and if to utilize chatbots in the classroom are at the forefront of scholarly discussion. This quick-hit article presents a plan to teach learners about generative AI writing tools and their ethical use for writing purposes. As generative AI tools continue to emerge, this guide can support instructors from all disciplines to engage learners in getting the most accurate information.

ABSTRACT Pea production across the world is significantly limited by root rot disease, which is caused by many fungal and oomycetes pathogens. In Canada, Fusarium avenaceum is the most devastating pathogen of the Fusarium root rot complex of pea. Host genetic resistance is the most effective control method for this disease. Evaluation of global pea accessions and Canadian varieties for F. avenaceum root rot resistance has not been reported to date. This study evaluated 20 pea accessions of different market classes with pigmented or nonpigmented seed coats and flowers for F. avenaceum resistance under controlled conditions. The pea accessions CDC Acer, CDC Vienna, PBA OURA, Morgan, CDC Blazer, CDC Dakota, and PI 280609, which have pigmented flowers and seed coats, were identified as resistant or partially resistant to F. avenaceum. This was based on their root rot severity scores and ability to tolerate F. avenaceum infection without significant (p > 0.05) reductions in plant height, shoot dry weight, and root dry weight. Among the varieties with nonpigmented flowers and seed coats, only Cameor showed partial resistance to F. avenaceum when challenged with reduced conidial concentration. Root dry weight (R = −0.86), plant height (R = −0.82), and shoot dry weight (R = −0.78) had a strong negative correlation (p

Background: Retrospective studies and large-scale reviews suggest multiple properties of effective checklist implementation, such as active leadership, ongoing feedback, and local modification. We hypothesized that the use of these effective checklist implementation strategies would yield better results as compared to a more fixed implementation strategy of the Pre-anesthesia Induction Patient Safety (PIPS) checklist. Methods: We conducted a prospective, pre-post, pragmatic cohort trial of a checklist implementation using a prospective difference-in-difference design observing simultaneous implementation of the same PIPS checklist in two community hospitals. At one hospital, we created a multidisciplinary committee of stakeholders to oversee implementation and provide active leadership, ongoing feedback, and local modification of the checklist (“Dynamic Implementation”). At the second hospital, a single clinical champion implemented the checklist with no local modification (“Static Implementation”). We examined checklist compliance and proximal outcomes three months before and nine months after implementation. We conducted surveys and interviews to examine attitudes and implementation barriers. Results: We observed 477 anesthesia inductions. The “Dynamic” cohort demonstrated significantly greater checklist compliance post-implementation compared to the “Static” cohort (B = 0.26, p

Abstract Mutations in the α-Synuclein (αS) gene promote αS monomer aggregation that causes neurodegeneration in familial Parkinson’s disease (fPD). However, most mouse models expressing single-mutant αS transgenes develop neuronal aggregates very slowly, and few have dopaminergic cell loss, both key characteristics of PD. To accelerate neurotoxic aggregation, we previously generated fPD αS E46K mutant mice with rationally designed triple mutations based on the α-helical repeat motif structure of αS (fPD E46K→3 K). The 3 K variant increased αS membrane association and decreased the physiological tetramer:monomer ratio, causing lipid- and vesicle-rich inclusions and robust tremor-predominant, L-DOPA responsive PD-like phenotypes. Here, we applied an analogous approach to the G51D fPD mutation and its rational amplification (G51D → 3D) to generate mutant mice. In contrast to 3 K mice, G51D and 3D mice accumulate monomers almost exclusively in the cytosol while also showing decreased αS tetramer:monomer ratios. Both 1D and 3D mutant mice gradually accumulate insoluble, higher-molecular weight αS oligomers. Round αS neuronal deposits at 12 mos immunolabel for ubiquitin and pSer129 αS, with limited proteinase K resistance. Both 1D and 3D mice undergo loss of striatal TH+ fibers and midbrain dopaminergic neurons by 12 mos and a bradykinesia responsive to L-DOPA. The 3D αS mice have decreased tetramer:monomer equilibria and recapitulate major features of PD. These fPD G51D and 3D mutant mice should be useful models to study neuronal αS-toxicity associated with bradykinetic motor phenotypes.

Abstract Reduced inhibition by somatostatin-expressing interneurons is associated with depression. Administration of positive allosteric modulators of α5 subunit-containing GABAA receptor (α5-PAM) that selectively target this lost inhibition exhibit antidepressant and pro-cognitive effects in rodent models of chronic stress. However, the functional effects of α5-PAM on the human brain in vivo are unknown, and currently cannot be assessed experimentally. We modeled the effects of α5-PAM on tonic inhibition as measured in human neurons, and tested in silico α5-PAM effects on detailed models of human cortical microcircuits in health and depression. We found that α5-PAM effectively recovered impaired cortical processing as quantified by stimulus detection metrics, and also recovered the power spectral density profile of the microcircuit EEG signals. We performed an α5-PAM dose-response and identified simulated EEG biomarker candidates. Our results serve to de-risk and facilitate α5-PAM translation and provide biomarkers in non-invasive brain signals for monitoring target engagement and drug efficacy.

Abstract Background More than 650 million people are obese (BMI > 30) worldwide, which increases their risk for several metabolic diseases and cancer. While cachexia and obesity are at opposite ends of the weight spectrum, leading many to suggest a protective effect of obesity against cachexia, mechanistic support for obesity's benefit is lacking. Given that obesity and cachexia are both accompanied by metabolic dysregulation, we sought to investigate the impact of obesity on skeletal muscle mass loss and mitochondrial dysfunction in murine cancer cachexia. Methods Male C57BL/6 mice were given a purified high fat or standard diet for 16 weeks before being implanted with 106 Lewis lung carcinoma (LLC) cells. Mice were monitored for 25 days, and hindlimb muscles were collected for cachexia indices and mitochondrial assessment via western blotting, high‐resolution respirometry and transmission electron microscopy (TEM). Results Obese LLC mice experienced significant tumour‐free body weight loss similar to lean (−12.8% vs. −11.8%, P = 0.0001) but had reduced survival (33.3% vs. 6.67%, χ2 = 10.04, P = 0.0182). Obese LLC mice had reduced muscle weights (−24%, P

Abstract Sex chromosome replacement is frequent in many vertebrate clades, including fish, frogs, and lizards. In order to understand the mechanisms responsible for sex chromosome turnover and the early stages of sex chromosome divergence, it is necessary to study lineages with recently evolved sex chromosomes. Here we examine sex chromosome evolution in a group of African cichlid fishes (tribe Tropheini) which began to diverge from one another less than 4 MYA. We have evidence for a previously unknown sex chromosome system, and preliminary indications of several additional systems not previously reported in this group. We find a high frequency of sex chromosome turnover and estimate a minimum of 14 turnovers in this tribe. We date the origin of the most common sex determining system in this tribe (XY-LG5/19) near the base of one of two major sub-clades of this tribe, about 3.4 MY ago. Finally, we observe variation in the size of one sex-determining region that suggests independent evolution of evolutionary strata in species with a shared sex-determination system. Our results illuminate the rapid rate of sex chromosome turnover in the tribe Tropheini and set the stage for further studies of the dynamics of sex chromosome evolution in this group.

Abstract A prevalent side-reaction of succinate dehydrogenase oxidizes malate to enol-oxaloacetate (OAA), a metabolically inactive form of OAA that is a strong inhibitor of succinate dehydrogenase. We purified from cow heart mitochondria an enzyme (OAT1) with OAA tautomerase (OAT) activity that converts enol-OAA to the physiological keto-OAA form, and determined that it belongs to the highly conserved and previously uncharacterized Fumarylacetoacetate_hydrolase_domain-containing protein family. From all three domains of life, heterologously expressed proteins were shown to have strong OAT activity, and ablating the OAT1 homolog caused significant growth defects. In Escherichia coli, expression of succinate dehydrogenase was necessary for OAT1-associated growth defects to occur, and ablating OAT1 caused a significant increase in acetate and other metabolites associated with anaerobic respiration. OAT1 increased the succinate dehydrogenase reaction rate by 35% in in vitro assays with physiological concentrations of both succinate and malate. Our results suggest that OAT1 is a universal metabolite repair enzyme that is required to maximize aerobic respiration efficiency by preventing succinate dehydrogenase inhibition.