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Objective Janus kinase family members are essential for signaling by multiple cytokines, including many implicated in systemic lupus erythematosus (SLE) pathogenesis. To test whether inhibition of JAK1 signaling can be efficacious in SLE, we used a JAK1‐selective inhibitor (ABT‐317) and evaluated its ability to ameliorate disease in murine SLE. Methods Efficacy of ABT‐317 was evaluated using NZB/W‐F1 mice treated prophylactically and therapeutically. Primary endpoints were proteinuria, survival, and saliva production. Other endpoints included histological analysis of kidneys and salivary glands, flow cytometric analysis of splenic cell populations, and gene expression analysis by RNA sequencing in the kidneys, salivary glands, and blood. Publicly available human kidney gene transcription data were used to assess the translatability of the mouse findings. Results ABT‐317 was efficacious when dosed prophylactically and prevented disease for up to two months after treatment cessation. When dosed therapeutically, ABT‐317 quickly reversed severe proteinuria and restored saliva production, as well as diminished kidney and salivary gland inflammation. ABT‐317‐induced changes in glomerular morphology coincided with normalization of a human nephrotic gene signature, suggesting translatability to human lupus nephritis (LN). Conclusion JAK1 inhibition prevented and reversed kidney and salivary gland manifestations of murine lupus with long‐lasting effects after treatment cessation. These data, along with the presence of JAK1 and nephrotic gene signatures in human LN glomeruli, suggest that a JAK1‐selective inhibitor may be an effective therapeutic in the treatment of human SLE and LN.

Abstract Background Ureteral stents are placed by urologists to ensure ureteral patency in the postoperative period following ureteroscopy to treat kidney stones, with the goal to reduce complications. However, ureteral stents themselves cause pain and urinary symptoms in many patients that can lead to morbidity. Professional society guidelines support stent omission after uncomplicated ureteroscopy, which represents most cases. Despite this, ureteral stents are utilized in more than 80% of all ureteroscopy procedures. One reason for guideline discordance is the low level of evidence supporting stent omission recommendations. Studies are inconclusive on whether stents increase pain and complications. A recent Cochrane review concluded higher quality and large trials are needed to inform decision-making. Furthermore, there is a lack of studies evaluating health-related quality of life (HRQOL), patient-reported outcomes (PROs), and unplanned healthcare utilization. Another factor is that prospective clinical trials are hindered by patient reluctance to be randomized to either stent placement or omission. The outcomes of patients who decline randomization have been ignored in trials, limiting the generalizability of the evidence. Methods Through collaboration with patient partners, we developed a pragmatic multi-center combined randomized and observational cohort study in a quality improvement collaborative. Patients will be prospectively enrolled into a randomized cohort in which assignment to ureteral stent omission (vs. placement) is determined in the operating room using a web-based randomization platform. Patients who decline randomization are invited to take part in an observational (real-world) cohort in which the determination of stent use is at the discretion of the urologist. Patients in both cohorts will complete preoperative and postoperative assessments of PROs including pain, urinary symptoms, interference with usual activities, time taken off work or school, and treatment satisfaction. Unplanned healthcare utilization within 30 days postoperatively will be assessed by review of the electronic health record. Severe adverse events will be recorded. A subgroup of patients and urologists will also participate in qualitative semi-structured interviews focusing on knowledge, preferences, and practice patterns regarding ureteral stenting. Interview transcripts will be thematically analyzed. Discussion This study is designed to evaluate the HRQOL and 30-day healthcare utilization of patients undergoing ureteral stent omission compared to stent placement following uncomplicated ureteroscopic treatment of upper urinary tract stones. Additionally, patient and urologist opinions and preferences related to ureteral stenting will be explored through qualitative interviews, with the aim of identifying key barriers and facilitators of practice change related to stenting practices. Trial registration ClinicalTrials.gov, NCT05866081. Registered on 19 May 2023.

Thinning sea ice cover and earlier melt in the Arctic impact primary producer (PP) phenology, causing earlier ice algal bloom termination and phytoplankton bloom commencement. However, logistic constraints limit capturing the complete seasonal evolution of PPs and their physical drivers. Here, we combine spectral irradiance data from subsurface oceanographic moorings with synthetic aperture radar backscatter and meteorological variables to study light in Dease Strait, investigating its relation to timing and magnitude of surface PPs for 2017 and 2019. Ice algal blooms in 2017 and 2019 lasted 66 and 84 days, respectively, peaking within 2 days of snow melt onset. In 2019, lower temperatures and a deeper snowpack before snow melt extended the ice algal bloom. Melt pond formation increased light transmission, enabling a short, 6–7-day under-ice phytoplankton bloom in both years that was likely nutrient-limited. The 2019 phytoplankton bloom was less productive, possibly due to the longer ice algal bloom depleting surface nutrients. After ice break-up in 2019, a 31-day late-summer bloom occurred via wind-driven mixing. Our findings suggest that the combined remote sensing technique has novel applicability in other settings, providing insights into the changing state of PP phenology, and the need for long-term Arctic observations to discern regional climate change effects.

Abstract Understanding neurodegenerative disease pathology requires a close examination of neurons and their processes. However, image-based single-cell analyses of neurons often require laborious and time-consuming manual classification tasks. Here, we present a machine learning (ML) approach leveraging convolutional neural network (CNN) classifiers capable of accurately identifying various classes of neuronal images, including single neurons. We developed the Single Neuron Identification Model 20-Class (SNIM20) which was trained on a dataset of induced pluripotent stem cell (iPSC)-derived motor neurons, containing over 12,000 images from 20 distinct classes. SNIM20 is built in TensorFlow and trained on images of neurons differentiated from iPSC cultures that were stained for nuclei and microtubules. This classifier demonstrated high predictive accuracy (AUC = 0.99) for distinguishing single neurons. Additionally, the 2-stage training framework can be used more broadly for cellular classification tasks. A variation was successfully trained on images of a human osteosarcoma cell line (U2OS) for single-cell classification (AUC = 0.99). While this framework was primarily designed for single-cell microraft-based identification and capture, it also works with cells in standard plate formats. We additionally explore the impact of fluorescent channels and brightfield images, class groupings, and transfer learning on the quality of the classification. This framework can both assist in high throughput neuronal or cellular identification and be used to train a custom classifier for the user’s specific needs.

Abstract Introduction Diagnosing infected joint replacements relies heavily on assessing the neutrophil response to bacteria. Bacteria form biofilms on joint replacements. Biofilms are sessile bacterial communities encased in a protective extracellular matrix, making them notoriously difficult to culture, remarkably tolerant to antibiotics, and able to evade phagocytosis. Phagocytized bacteria dramatically alter cytokine production and compromise macrophage antigen presentation. We hypothesize that a subset of joint replacements have a dormant infection that suppresses the neutrophil response to bacteria but can be distinguished from uninfected joint replacements by the response of the mononuclear phagocyte system (MPS) within periarticular tissue, synovial fluid, and circulating plasma. Methods Single cell RNASeq transcriptomic and OLink proteomic profiling was performed on matched whole blood, synovial fluid, and periarticular tissue samples collected from 4 joint replacements with an active infection and 3 joint replacements without infection as well as 6 joint replacements with a prior infection deemed “infection-free” by the 2018 Musculoskeletal Infection Society criteria (follow-up of 26 ± 3 months). Results The MPS and neutrophil responses differ by infected state; the cellular distribution of the MPS response in the subset of joints with dormant infections resembled actively infected joints (p = 0.843, Chi-square test) but was significantly different from uninfected joints (p

Abstract Transparent conducting oxides (TCOs) are crucial for high-performance displays, solar cells, and wearable sensors. However, their high process temperatures and brittle nature have hindered their use in flexible electronics. In this paper, we overturn these limitations by harnessing Cabrera-Mott oxidation to fabricate large-area, two-dimensional (2D) transparent electrodes via liquid metal printing. Our robotic, vacuum-free process deposits ultrathin (2–10 nm) indium tin oxide (ITO) with exceptional flexibility, transparency (>95%) and conductivity (>1300 S/cm) by utilizing hypoeutectic In-Sn alloys to print at

Abstract This research focuses on the recent efforts to improve STEM skills among future science educators in Oman. It details the development and application of a questionnaire aimed at evaluating self-efficacy in teaching science through engineering design. A survey was given to 326 aspiring science teachers to gauge their views on incorporating engineering design in science education. A quantitative methodology was used, with Rasch analysis used in the analysis of the survey instrument. The survey revealed two distinct dimensions: Personal Self-efficacy (SE) and Outcome Expectancy (OE). The reliability of person and item measures fell within acceptable limits. The analysis of the category probability curves showed effective rating scale measurements. The Wright map corresponded well with theoretical predictions, offering insightful implications for training science teachers in Oman. This research adds to the existing body of knowledge, particularly in Arab regions, by providing data on science education from Omani preservice science teachers. It further explores how these findings can enhance the quality of science teaching and learning.

Abstract Heart shape captures variation in cardiac structure beyond traditional phenotypes of mass and volume. Although observational studies have demonstrated associations with cardiometabolic risk factors and diseases, its genetic basis is less understood. We utilised cardiovascular magnetic resonance images from 45,683 UK Biobank participants to construct a heart shape atlas from bi-ventricular end-diastolic surface mesh models through principal component (PC) analysis. Genome-wide association studies were performed on the first 11 PCs that captured 83.6% of shape variance. We identified 43 significant loci, 14 were previously unreported for cardiac traits. Genetically predicted PCs were associated with cardiometabolic diseases. In particular two PCs (2 and 3) linked with more spherical ventricles being associated with increased risk of atrial fibrillation. Our study explores the genetic basis of multidimensional bi-ventricular heart shape using PCA, reporting new loci and biology, as well as polygenic risk scores for exploring genetic relationships of heart shape with cardiometabolic diseases.

Abstract Drug-resistant strains of Mycobacterium tuberculosis are a major global health problem. Resistance to the front-line antibiotic isoniazid is often associated with mutations in the katG-encoded bifunctional catalase-peroxidase. We hypothesise that perturbed KatG activity would generate collateral vulnerabilities in isoniazid-resistant katG mutants, providing potential pathway targets to combat isoniazid resistance. Whole genome CRISPRi screens, transcriptomics, and metabolomics were used to generate a genome-wide map of cellular vulnerabilities in an isoniazid-resistant katG mutant strain of M. tuberculosis. Here, we show that metabolic and transcriptional remodelling compensates for the loss of KatG but in doing so generates vulnerabilities in respiration, ribosome biogenesis, and nucleotide and amino acid metabolism. Importantly, these vulnerabilities are more sensitive to inhibition in an isoniazid-resistant katG mutant and translated to clinical isolates. This work highlights how changes in the physiology of drug-resistant strains generates druggable vulnerabilities that can be exploited to improve clinical outcomes.

Abstract Imidazole Propionate (ImP), a gut-derived metabolite from histidine, affects insulin signaling in mice and is elevated in type 2 diabetes (T2D). However, the source of histidine and the role of the gut microbiota remain unclear. We conducted an intervention study in mice and humans, comparing ImP kinetics in mice on a high-fat diet with varying histidine levels and antibiotics, and assessed ImP levels in healthy and T2D subjects with histidine supplementation. Results show that dietary histidine is metabolized to ImP, with antibiotic-induced gut microbiota suppression reducing ImP levels in mice. In contrast, oral histidine supplementation resulted in increases in circulating ImP levels in humans, whereas antibiotic treatment increased ImP levels, which was associated with a bloom of several bacterial genera that have been associated with ImP production, such as Lactobacilli. Our findings highlight the gut microbiota’s crucial role in regulating ImP and the complexity of translating mouse models to humans.

Abstract The emergence of antimalarial drug resistance is an impediment to malaria control and elimination in Africa. Analysis of temporal trends in molecular markers of resistance is critical to inform policy makers and guide malaria treatment guidelines. In a low and seasonal transmission region of southern Zambia, we successfully genotyped 85.5% (389/455) of Plasmodium falciparum samples collected between 2013 and 2018 from 8 spatially clustered health centres using molecular inversion probes (MIPs) targeting key drug resistance genes. Aside from one sample from 2016 carrying K13 622I, no other World Health Organization-validated or candidate artemisinin partial resistance (ART-R) mutations were observed. However, in the more recent years (2016–2017) five novel K13-propeller-domain mutations, C532S, A578S, Q613E, D680N and G718S were identified at low prevalence. Moreover, 13% (CI, 9.6–17.2) of isolates had the AP2MU 160N mutation, which has been associated with delayed clearance following artemisinin combination therapy in Africa. This mutation increased in prevalence between 2015 and 2018 and bears a genomic signature of selection. During this time period, there was an increase in the MDR1 NFD haplotype that is associated with reduced susceptibility to lumefantrine. Sulfadoxine-pyrimethamine polymorphisms were near fixation. While validated ART-R mutations are rare, a mutation associated with slow parasite clearance in Africa appears to be under selection in southern Zambia.

Abstract Many studies of azobenzene photoswitches are carried out in polar aprotic solvents as a first principles characterization of thermal isomerization. Among the most convenient polar aprotic solvents are chlorinated hydrocarbons, such as DCM. However, studies of azobenzene thermal isomerization in such solvents have led to spurious, inconclusive, and irreproducible results, even when scrupulously cleaned and dried, a phenomenon not well understood. We present the results of a comprehensive investigation into the root cause of this problem. We explain how irradiation of an azopyridine photoswitch with UV in DCM acts not just as a trigger for photoisomerization, but protonation of the pyridine moiety through photodecomposition of the solvent. Protonation markedly accelerates the thermal isomerization rate, and DFT calculations demonstrate that the singlet-triplet rotation mechanism assumed for many azo photoswitches is surprisingly abolished. This study implies exploitative advantages of photolytically-generated protons and finally explains the warning against using chlorinated solvent with UV irradiation in isomerization experiments.

The evaluation of adult patients suspected of ST-segment elevation myocardial infarction (STEMI) includes a focused history and examination, 12-lead electrocardiogram (ECG), and cardiac serum marker analysis. The ECG plays a pivotal role in the early diagnosis and management of STEMI. A number of ECG entities in this patient population will present with ST-segment elevation and other electrocardiographic abnormalities which can mimic STEMI. In this article, we review the most frequent STEMI mimic patterns, highlight their ECG characteristics, and compare these individual ECG entities to the electrocardiographic abnormalities present with STEMI.

Abstract Background Traditionally, ultrasound skills have been taught through a one-on-one approach, where instructors physically guide learners’ hands at the bedside or in the workshop. However, this method is frequently challenged by scheduling and cost limitations. Our objective was to create a tele-education model for point-of-care ultrasound training and evaluate its effectiveness and its impact on learners’ perceived workload compared to conventional education and self-directed learning methods. Methods We conducted a 3-arm randomized trial, comparing tele-education (TE), conventional education (CE) and self-directed learning (SL) methods. All subjects underwent online didactic lectures prior to a hands-on ultrasound workshop. The TE group utilized an ultrasound machine equipped with a speakerphone, a webcam for direct visualization of learner’s hand maneuvers, and an analog-to-video converter for the real-time streaming of ultrasound images. This configuration enabled remote instructors to provide immediate verbal feedback to learners. In contrast, the CE group received in-person coaching, while the SL group had no instructors present. Following the coaching session, subjects completed a scenario-based skill test and a survey on the National Aeronautics and Space Administration task load index (NASA-TLX) to measure their ultrasound competency and perceived workload, respectively. Results Twenty-seven ultrasound novices were randomly allocated into 3 groups. The median skill test score of TE, CE, and SL was 22 [interquartile range (IQR): 18–28], 24 [IQR: 21–31], and 16 [IQR: 15–18], respectively (p

Abstract The nucleus accumbens (NAc) regulates reward-motivated behavior, but the temporal dynamics of NAc neurons that enable “free-willed” animals to obtain rewards remain elusive. Here, we recorded Ca2+ activity from individual NAc neurons when mice performed self-paced lever-presses for sucrose. NAc neurons exhibited three temporally-sequenced clusters, defined by times at which they exhibited increased Ca2+ activity: approximately 0, −2.5 or −5 sec relative to the lever-pressing. Dopamine D1 receptor (D1)-expressing neurons and D2-neurons formed the majority of the −5-sec versus −2.5-sec clusters, respectively, while both neuronal subtypes were represented in the 0-sec cluster. We found that pre-press activity patterns of D1- or D2-neurons could predict subsequent lever-presses. Inhibiting D1-neurons at −5 sec or D2-neurons at −2.5 sec, but not at other timepoints, reduced sucrose-motivated lever-pressing. We propose that the time-specific activity of D1- and D2-neurons mediate key temporal features of the NAc through which reward motivation initiates reward-seeking behavior.

Abstract Background Diet is considered a first-line treatment option for prediabetes, a condition that affects 96 million United States (U.S.) adults. Diet patterns that prioritize whole foods (e.g., Mediterranean) are currently recommended to treat prediabetes. However, no studies have tested whether a U.S.-style diet pattern that prioritizes whole foods can be used to treat prediabetes. The purpose of this study was to assess the feasibility and acceptability of a whole foods diet for adults with prediabetes and their offspring prior to conducting a larger clinical trial. Methods A 2-week single-arm pre-post experimental controlled-feeding intervention of a 2020–2025 Dietary Guidelines for Americans adapted whole foods diet (e.g., primarily focused on foods that have undergone limited processing or refinement) was conducted in adults (25–59 years) with prediabetes and their biological offspring (6–17 years). Families received 2 weeks of menus and grocery delivery and weekly counseling by a registered dietitian. Families were invited to attend an optional focus group session. Feasibility was based on having a ≥ 50% family completion rate with ≥ 80% completion of study outcomes. Measures included adult–child anthropometrics (weight [kg], body mass index [BMI] including BMI% and Z-scores for offspring, waist circumference [cm]) and child diet quality estimated using the 2015 Healthy Eating Index (HEI) from a single random food record. Wilcoxon signed rank was used to compare differences between baseline and 2-week anthropometrics measures and offspring total HEI scores. Qualitative data were analyzed using thematic analysis to understand factors attributed to diet adherence and acceptability. Results Eight families enrolled (n = 8 adults, n = 12 offspring), with 7 families completing the study (12% attrition) and completing 100% of study outcome measures. Adults experienced a median weight loss of − 1.0 kg from baseline to 2 weeks (79.5 kg to 78.5 kg). Offspring had a 24-point increase in total 2015 HEI scores (median difference 50 to 74). Focus group participants (n = 4 adults) reported being satisfied with the program and expressed a willingness to continue the diet despite identified barriers. Conclusions A whole foods diet that provides dietary support was found to be feasible and acceptable for families at risk for T2D. Future studies are needed to test the effects of the diet on prediabetes outcomes, diet quality, and diet adherence in adults and families. Trial registration NCT05483972 at ClinicalTrials.gov. Registered July 25, 2022. https://clinicaltrials.gov/study/NCT05483972?cond=prediabetes&term=whole%20foods%20&rank=1

Abstract Parkinson’s Disease (PD) is a prevalent, progressive neurodegenerative disease with motor and non-motor symptoms. Vagus Nerve Stimulation (VNS) has emerged as a potential therapeutic approach for PD, but published research on this topic varies widely. This scoping review maps existing literature on VNS for PD, highlighting stimulation methods, operational parameters, safety profiles, neurophysiological mechanisms, and clinical outcomes in human and animal models. Online databases were used to identify 788 papers published between 2013 and 2023, from which 17 publications on invasive and non-invasive VNS in PD were selected. Studies showed high variability in VNS parameters and study design. Evidence in animal models and human subjects suggests potential neurophysiological effects on PD-related pathology and motor function improvements. However, significant gaps in the literature remain. Future research should include rigorous reporting of study design, standardization of stimulation parameters, and larger sample sizes to ultimately facilitate translation of VNS into clinical practice.

Abstract Background Artificial intelligence (AI) uses in healthcare changed dramatically with large language models. While work is underway to integrate generative AI safely and effectively into medical practice, little is known about how resident physicians are already using these tools in their work. Objectives To characterize the current usage patterns and attitudes towards large language models among internal medicine residents in North Carolina. Methods Cross-sectional survey-based study of Internal medicine residents in North Carolina. Results 152 of 523 total residents completed the survey (29% response rate). Many identified a role for LLMs in both clinical (56%) and non-clinical medicine (72%) today, although only 26% reported current use. More perceived a future role in clinical (71%) and non-clinical (80%) medicine. None had received formal training, but 94% expressed interest in learning more and 87% thought residencies maybe or definitely should have formal curriculum. Top clinical uses include forming a differential diagnosis, researching treatments, and creating educational materials. Top non-clinical uses included self-directed learning, scientific writing, and summarizing literature. Conclusions Medical educators and administrators should be aware that residents are using generative AI in professional settings. Residents believe in a current and future role for AI in medicine and desire curriculum and learning opportunities during training.

Abstract Background The western province of Anbar, and the district of Hadeetha, have suffered direct impacts from the second United States led invasion (2003) through the ISIS invasion (2014–2017). With the primary health care centers being closed or inaccessible, the remaining population experienced changes in health seeking. The area of Anbar, Iraq remains largely remote from the discourse of health system recovery post-conflict. The objective of this study was to describe changes in health seeking behaviors of caregivers of children ages 12–72 months of age in Hadeetha, Anbar, Iraq from the conflict period of ISIS (Islamic State of Syria and Iraq), 2014–2017, to the post-conflict period, 2021. Methods This was a mixed-methods study composed of a cross-sectional 415 household survey and focus groups in Hadeetha, Anbar, Iraq. Caretakers of children were interviewed from February to April 2021. Children were sampled from a list of children who missed at least one vaccination appointment without a return for follow-up from the birth cohorts of 2014 to 2020. Healthcare workers focus groups and key informant interviews occurred from 2019 to 2021 centered around experiences from the 2014 to 2021 period. Results In the post-conflict period, there were no differences in health seeking based upon provider type between respondents. Physicians were primary healthcare providers in the post-conflict period for 79% of respondents versus only 47% in the conflict period. Healthcare workers described major barriers to delivering services in Hadeetha during the ISIS occupation due to infrastructure damage, threats of violence, decreased medical personnel, lack of compensation and disruptions in medical supplies from 2014 to 2017. Conclusion This study provides insight into health seeking challenges among the many individuals who remained in Hadeetha during the ISIS occupation. Health use patterns by provider type mirror the concerns the healthcare providers shared: limited availability, efflux of professional workers, limited resources and security challenges to providing care. Positive trends toward increased access to physicians during periods of peace are an encouraging marker for continued population resilience during the post-conflict period. Recovery efforts continue to be hampered by internal sectarian discord within Iraq as well as insufficient resources to maintain health services as well as provide catch-up health services, such as immunizations.

Abstract Recurrent breast cancers often develop resistance to standard-of-care therapies. Identifying targetable factors contributing to cancer recurrence remains the rate-limiting step in improving long-term outcomes. In this study, we identify tumor cell-derived osteopontin as an autocrine and paracrine driver of tumor recurrence. Osteopontin promotes tumor cell proliferation, recruits macrophages, and synergizes with IL-4 to further polarize them into a pro-tumorigenic state. Macrophage depletion and osteopontin inhibition decrease recurrent tumor growth. Furthermore, targeting osteopontin in primary tumor-bearing female mice prevents metastasis, permits T cell infiltration and activation, and improves anti-PD-1 immunotherapy response. Clinically, osteopontin expression is higher in recurrent metastatic tumors versus female patient-matched primary breast tumors. Osteopontin positively correlates with macrophage infiltration, increases with higher tumor grade, and its elevated pathway activity is associated with poor prognosis and long-term recurrence. Our findings suggest clinical implications and an alternative therapeutic strategy based on osteopontin’s multiaxial role in breast cancer progression and recurrence.

Abstract While the critical role of NKX2-1 and its transcriptional targets in lung morphogenesis and pulmonary epithelial cell differentiation is increasingly known, mechanisms by which chromatin accessibility alters the epigenetic landscape and how NKX2-1 interacts with other co-activators required for alveolar epithelial cell differentiation and function are not well understood. Combined deletion of the histone methyl transferases Prdm3 and Prdm16 in early lung endoderm causes perinatal lethality due to respiratory failure from loss of AT2 cells and the accumulation of partially differentiated AT1 cells. Combination of single-cell RNA-seq, bulk ATAC-seq, and CUT&RUN data demonstrate that PRDM3 and PRDM16 regulate chromatin accessibility at NKX2-1 transcriptional targets critical for perinatal AT2 cell differentiation and surfactant homeostasis. Lineage specific deletion of PRDM3/16 in AT2 cells leads to lineage infidelity, with PRDM3/16 null cells acquiring partial AT1 fate. Together, these data demonstrate that NKX2-1-dependent regulation of alveolar epithelial cell differentiation is mediated by epigenomic modulation via PRDM3/16.

Abstract Determining the genetic architecture of Alzheimer’s disease pathologies can enhance mechanistic understanding and inform precision medicine strategies. Here, we perform a genome-wide association study of cortical tau quantified by positron emission tomography in 3046 participants from 12 independent studies. The CYP1B1-RMDN2 locus is associated with tau deposition. The most significant signal is at rs2113389, explaining 4.3% of the variation in cortical tau, while APOE4 rs429358 accounts for 3.6%. rs2113389 is associated with higher tau and faster cognitive decline. Additive effects, but no interactions, are observed between rs2113389 and diagnosis, APOE4, and amyloid beta positivity. CYP1B1 expression is upregulated in AD. rs2113389 is associated with higher CYP1B1 expression and methylation levels. Mouse model studies provide additional functional evidence for a relationship between CYP1B1 and tau deposition but not amyloid beta. These results provide insight into the genetic basis of cerebral tau deposition and support novel pathways for therapeutic development in AD.

Abstract Background Giving information to trial participants who stop taking part could support them through what can be a difficult process. We previously developed guidance around the ethical acceptability of such information provision, and about how trialists can develop suitable communication materials. There is limited evidence about what research ethics committees think of this issue, and limited guidance about what level of oversight they should have over the proposed communications, or post-consent participant communications generally. We conducted a survey of UK ethics committee members to address these points. Methods The survey was co-developed by public contributors and trialists who had previously worked together on the communications guidance. We asked respondents if they agreed with the general idea of informing participants who stop taking part, if they had ever been requested to review similar communications, and what level of ethics committee review they might recommend. The survey was primarily conducted online. It was reviewed by three ethics committee members before finalisation and shared directly with all UK ethics committee members. We analysed quantitative questions descriptively and used inductive analysis for open questions to identify common themes. Results Ninety-one ethics committee members participated (nearly 10% of all UK members). The sample was similar to reported data about all members in terms of several personal characteristics. Most respondents (83%) agreed with our project’s rationale. Only 23% of respondents reported having been asked to review an end-of-participation information sheet before. Respondents gave various answers about the level of ethics committee review required, but most supported a relatively proportionate review process. Common concerns were about the risk of coercion or making participants feel pressured. Conclusions Our survey suggests that ethics committee members generally support providing information to trial participants who stop taking part, if risks to participants are mitigated. We believe our guidance already addresses the main concerns raised. Our respondents’ lack of prior experience with end-of-participation information sheets suggests that participants are not getting information they want or need when they stop participating. Our results help clarify how ethics committee should oversee post-consent participant communications, but further guidance from research regulators could be helpful.

Abstract Climate warming disproportionately impacts countries in the Global South by increasing extreme heat exposure. However, geographic disparities in adaptation capacity are unclear. Here, we assess global inequality in green spaces, which urban residents critically rely on to mitigate outdoor heat stress. We use remote sensing data to quantify daytime cooling by urban greenery in the warm seasons across the ~500 largest cities globally. We show a striking contrast, with Global South cities having ~70% of the cooling capacity of cities in the Global North (2.5 ± 1.0 °C vs. 3.6 ± 1.7 °C). A similar gap occurs for the cooling adaptation benefits received by an average resident in these cities (2.2 ± 0.9 °C vs. 3.4 ± 1.7 °C). This cooling adaptation inequality is due to discrepancies in green space quantity and quality between cities in the Global North and South, shaped by socioeconomic and natural factors. Our analyses further suggest a vast potential for enhancing cooling adaptation while reducing global inequality.

Abstract Background The foodborne bacterium Listeria monocytogenes (Lm) causes a range of diseases, from mild gastroenteritis to invasive infections that have high fatality rate in vulnerable individuals. Understanding the population genomic structure of invasive Lm is critical to informing public health interventions and infection control policies that will be most effective especially in local and regional communities. Methods We sequenced the whole draft genomes of 936 Lm isolates from human clinical samples obtained in a two-decade active surveillance program across 58 counties in New York State, USA. Samples came mostly from blood and cerebrospinal fluid. We characterized the phylogenetic relationships, population structure, antimicrobial resistance genes, virulence genes, and mobile genetic elements. Results The population is genetically heterogenous, consisting of lineages I–IV, 89 clonal complexes, 200 sequence types, and six known serogroups. In addition to intrinsic antimicrobial resistance genes (fosX, lin, norB, and sul), other resistance genes tetM, tetS, ermG, msrD, and mefA were sparsely distributed in the population. Within each lineage, we identified clusters of isolates with ≤ 20 single nucleotide polymorphisms in the core genome alignment. These clusters may represent isolates that share a most recent common ancestor, e.g., they are derived from the same contamination source or demonstrate evidence of transmission or outbreak. We identified 38 epidemiologically linked clusters of isolates, confirming eight previously reported disease outbreaks and the discovery of cryptic outbreaks and undetected chains of transmission, even in the rarely reported Lm lineage III (ST3171). The presence of animal-associated lineages III and IV may suggest a possible spillover of animal-restricted strains to humans. Many transmissible clones persisted over several years and traversed distant sites across the state. Conclusions Our findings revealed the bacterial determinants of invasive listeriosis, driven mainly by the diversity of locally circulating lineages, intrinsic and mobile antimicrobial resistance and virulence genes, and persistence across geographical and temporal scales. Our findings will inform public health efforts to reduce the burden of invasive listeriosis, including the design of food safety measures, source traceback, and outbreak detection.

The geologic framework of the Seattle fault zone (SFZ) has been extensively studied, but the structure and fault strand locations in the central portion of the fault zone through the city of Seattle have remained controversial. Much of what is known about the SFZ has come from light detection and ranging (lidar)-topographic surveys and paleoseismic investigations of fault scarps primarily west of Puget Sound, regional gravity and aeromagnetic modeling, and multiscale marine seismic imaging in waters both west and east of Seattle. We analyze ∼24 km of land-based P-wave seismic-reflection data that fill in a critical gap in our understanding of the SFZ beneath the urban areas of West Seattle, south-central Seattle, and Mercer Island. These data image deformed strata in the upper 1 km, including upwarped Tertiary rock and younger sediments. Collectively, these data provide evidence for multiple Quaternary-active thrust faults, back thrusts, and sub-basins within the SFZ beneath the city of Seattle. The results indicate that multiple and potentially active back thrusts in the upper ∼500 m extend across the length of the SFZ and the entire urban corridor that may be analogous to those on Bainbridge Island west of Puget Sound.

Abstract At the start of the Zika virus (ZIKV) epidemic in 2015, ZIKV spread across South and Central America, and reached parts of the southern United States placing pregnant women at risk for fetal microcephaly, fetal loss, and other adverse pregnancy outcomes associated with congenital ZIKA syndrome (CZS). For this reason, testing of a safe and efficacious ZIKV vaccine remains a global health priority. Here we report that a single immunization with Ad26.M.Env ZIKV vaccine, when administered prior to conception, fully protects pregnant rhesus macaques from ZIKV viral RNA in blood and tissues with no adverse effects in dams and fetuses. Furthermore, vaccination prevents ZIKV distribution to fetal tissues including the brain. ZIKV associated neuropathology was absent in offspring of Ad26.M.Env vaccinated dams, although pathology was limited in fetuses from non-immunized, challenged dams. Vaccine efficacy is associated with induction of ZIKV neutralizing antibodies in pregnant rhesus macaques. These data suggest the feasibility of vaccine prevention of CZS in humans.

Abstract The retina is light-sensitive neuronal tissue in the back of the eye. The phospholipid composition of the retina is unique and highly enriched in polyunsaturated fatty acids, including docosahexaenoic fatty acid (DHA). While it is generally accepted that a high DHA content is important for vision, surprisingly little is known about the mechanisms of DHA enrichment in the retina. Furthermore, the biological processes controlled by DHA in the eye remain poorly defined as well. Here, we combined genetic manipulations with lipidomic analysis in mice to demonstrate that acyl-CoA synthetase 6 (Acsl6) serves as a regulator of the unique composition of retinal membranes. Inactivation of Acsl6 reduced the levels of DHA-containing phospholipids, led to progressive loss of light-sensitive rod photoreceptor neurons, attenuated the light responses of these cells, and evoked distinct transcriptional response in the retina involving the Srebf1/2 (sterol regulatory element binding transcription factors 1/2) pathway. This study identifies one of the major enzymes responsible for DHA enrichment in the retinal membranes and introduces a model allowing an evaluation of rod functioning and pathology caused by impaired DHA incorporation/retention in the retina.

Abstract The 2022 eruption of the Hunga volcano was a major event that propelled aerosols and water vapor up to an altitude of 53–57 km. It caused an unprecedented stratospheric hydration that is expected to affect composition, thermal structure, circulation and dynamics for years. Using vertically high resolved satellite observations from radio occultation, we focus on the temperature impact in the stratosphere from the eruption in January 2022 until December 2023. Separating the signals of the Hunga eruption from the broader stratospheric variability reveals a strong persistent radiative cooling of up to –4 K in the tropical and subtropical middle stratosphere from early after the eruption until mid-2023, clearly corresponding to the water vapor distribution. Our results provide new insights from observations into both the localized temperature changes and the persistent stratospheric cooling caused by the Hunga eruption and document this exceptional climatic effect not seen for previous volcanic eruptions.

Abstract Imaging and characterizing the dynamics of cellular adhesion in blood samples is of fundamental importance in understanding biological function. In vitro microscopy methods are widely used for this task but typically require diluting the blood with a buffer to allow for transmission of light. However, whole blood provides crucial signaling cues that influence adhesion dynamics, which means that conventional approaches lack the full physiological complexity of living microvasculature. We can reliably image cell interactions in microfluidic channels during whole blood flow by motion blur microscopy (MBM) in vitro and automate image analysis using machine learning. MBM provides a low cost, easy to implement alternative to intravital microscopy, for rapid data generation where understanding cell interactions, adhesion, and motility is crucial. MBM is generalizable to studies of various diseases, including cancer, blood disorders, thrombosis, inflammatory and autoimmune diseases, as well as providing rich datasets for theoretical modeling of adhesion dynamics.

Abstract Sacituzumab Govitecan (SG) is an antibody-drug conjugate that has demonstrated efficacy in patients with TROP-2 expressing epithelial cancers. In a xenograft model of intracranial breast cancer, SG inhibited tumor growth and increased mouse survival. We conducted a prospective window-of-opportunity trial (NCT03995706) at the University of Texas Health Science Center at San Antonio to examine the intra-tumoral concentrations and intracranial activity of SG in patients undergoing craniotomy for breast cancer with brain metastases (BCBM) or recurrent glioblastoma (rGBM). We enrolled 25 patients aged ≥18 years diagnosed with BCBM and rGBM to receive a single intravenous dose of SG at 10 mg/kg given one day before resection and continued on days 1 and 8 of 21-day cycles following recovery. The PFS was 8 months and 2 months for BCBM and rGBM cohorts, respectively. The OS was 35.2 months and 9.5 months, respectively. Grade≥3 AE included neutropenia (28%), hypokalemia (8%), seizure (8%), thromboembolic event (8%), urinary tract infection (8%) and muscle weakness of the lower limb (8%). In post-surgical tissue, the median total SN-38 was 249.8 ng/g for BCBM and 104.5 ng/g for rGBM, thus fulfilling the primary endpoint. Biomarker analysis suggests delivery of payload by direct release at target site and that hypoxic changes do not drive indirect release. Secondary endpoint of OS was 35.2 months for the BCBM cohort and 9.5 months for rGBM. Non-planned exploratory endpoint of ORR was 38% for BCBM and 29%, respectively. Exploratory endpoint of Trop-2 expression was observed in 100% of BCBM and 78% of rGBM tumors. In conclusion, SG was found to be well tolerated with adequate penetration into intracranial tumors and promising preliminary activity within the CNS. Trial Registration: Trial (NCT03995706) enrolled at Clinical Trials.gov as Neuro/Sacituzumab Govitecan/Breast Brain Metastasis/Glioblastoma/Ph 0: https://clinicaltrials.gov/study/NCT03995706?cond=NCT03995706 .

Abstract Metabolic reprogramming, a hallmark of tumorigenesis, involves alterations in glucose and fatty acid metabolism. Here, we investigate the role of Carnitine palmitoyl transferase 1a (Cpt1a), a key enzyme in long-chain fatty acid (LCFA) oxidation, in ErbB2-driven breast cancers. In ErbB2+ breast cancer models, ablation of Cpt1a delays tumor onset, growth, and metastasis. However, Cpt1a-deficient cells exhibit increased glucose dependency that enables survival and eventual tumor progression. Consequently, these cells exhibit heightened oxidative stress and upregulated nuclear factor erythroid 2-related factor 2 (Nrf2) activity. Inhibiting Nrf2 or silencing its expression reduces proliferation and glucose consumption in Cpt1a-deficient cells. Combining the ketogenic diet, composed of LCFAs, or an anti-ErbB2 monoclonal antibody (mAb) with Cpt1a deficiency significantly perturbs tumor growth, enhances apoptosis, and reduces lung metastasis. Using an immunocompetent model, we show that Cpt1a inhibition promotes an antitumor immune microenvironment, thereby enhancing the efficacy of anti-ErbB2 mAbs. Our findings underscore the importance of targeting fatty acid oxidation alongside HER2-targeted therapies to combat resistance in HER2+ breast cancer patients.

Objective: Thrombocytopenia is a common complication of hematopoietic stem-cell transplantation (HSCT), though many patients will become immune refractory to platelet transfusions over time. We built and evaluated an electronic health record (EHR)-integrated, standards-based application that enables blood-bank clinicians to match platelet inventory with patients using data previously not available at the point-of-care, like human leukocyte antigen (HLA) data for donors and recipients. Materials and methods: The web-based application launches as an EHR-embedded application or as a standalone application. The application coalesces disparate data streams into a unified view, including platelet count, HLA data, demographics, and real-time inventory. We looked at application usage over time and developed a multivariable logistic regression model to compute odds ratios that a patient undergoing HSCT would have a complicated thrombocytopenia course, with several model covariates including pre-/post-application deployment. Results: Usage of the application has been consistent since launch, with a slight dip during the first COVID wave. Our model, which included 376 patients in the final analysis, did not demonstrate a significantly decreased odds that a patient would have a complicated thrombocytopenia course after application deployment as compared to before application deployment. Discussion: We built an EHR-integrated application to improve platelet transfusion processes. Whereas our model did not demonstrate decreased odds of a patient having a complicated thrombocytopenia course, there are other workflow and clinical benefits that will benefit from future evaluation. Conclusion: A web-based, EHR-integrated application was built and integrated into our EHR system and is now part of the standard operating procedures of our blood bank.

In “Measuring Physical Functioning Using Wearable Sensors in Parkinson Disease and Chronic Obstructive Pulmonary Disease (the Accuracy of Digital Assessment of Performance Trial Study): Protocol for a Prospective Observational Study” (JMIR Res Protoc 2024 May 7:13:e55452) the authors noticed that one of the authors was not named correctly.

This paper considers a method for estimating bounce-averaged quasi-linear diffusion coefficients due to whistler-mode waves for a specified ratio of plasma frequency to gyrofrequency, ωp/Ωe, using values precomputed for a different value of that ratio. This approach was recently introduced to facilitate calculations associated with the “POES technique,” generalized to infer both wave intensity and cold plasma density from measurements of particle fluxes near the loss cone. The original derivation was justified on the basis of parallel-propagating waves but applied to calculations with much more general models of the waves. Here, we justify the estimates, which are based on equating resonant frequencies for differing values of ωp/Ωe and energy, for wide ranges of wave normal angle, resonance number, energy, and equatorial pitch angle. Refinements of the original estimates are obtained and tested numerically against full calculations of the diffusion coefficients for representative wave models. The estimated diffusion coefficients can be calculated rapidly and generally give useful estimates for energies in the 30-keV–300-keV range, especially when both relevant values of the ratio ωp/Ωe are large.

Rumen methanogens predominantly fall into two physiological groups: hydrogenotrophs which use hydrogen (H2) to reduce carbon dioxide (CO2) to methane (CH4), and methylotrophs which use H2 to reduce methanol and methylamines as substrates for methanogenesis. We used a dilution to extinction approach to isolate two hydrogenotrophic Methanocatella spp. and four cultures of methylotrophic methanogens from sheep rumen contents. Three of the methylotrophs were stable mixed cultures containing methanogens belonging to different lineages of the order Methanomassiliicoccales and one was a pure Methanosphaera culture. Methanosphaera sp. ISO3-F5 has a comparatively large genome (2.68 Mb) comprised of two replicons, a chromosome and a megaplasmid. The genome has an average G + C content of 30.5 % and encodes 2360 putative protein-coding genes. Cells of ISO3-F5 have a spherical shape, 0.6–1.2 µm in diameter, usually occurring in pairs or loose clumps, and have no flagellum. Cells stain Gram positive, have a single thick cell wall and divide by the formation of a cross wall. The optimum temperature for growth was 39°C to 42°C and the optimum pH was 6.7–6.8. Acetate was required for growth, but CH4 was not produced from acetate, formate, ethanol, methylamine, or isopropanol with or without H2/CO2. Volatile fatty acids and rumen fluid were also found to enhance the growth of ISO3-F5, while coenzyme M did not. ISO3-F5 produced CH4 from methanol in the presence of H2 and the genes encoding the necessary methanogenesis pathway have been identified. Based on morphological, physiological, and genomic characteristics, ISO3-F5 is a new species of the genus Methanosphaera. Our study shows that simple isolation methods allowed us to culture diverse and significant members of the rumen methanogen community.

New residents are often unprepared to respond to medical emergencies. To address this gap, we implemented a simulated cross-cover paging curriculum. All senior medical students enrolled in a required specialty-specific (internal medicine, procedures, emergency medicine [EM], obstetrics and gynecology [OBGYN], family medicine and pediatrics) residency preparation course (RPC) in 2020–2021 participated. Students received 3–6 specialty-specific pages that represented an urgent change in clinical status about a simulated patient. For each page, students first called a standardized registered nurse (SRN) to ask additional questions, then recommended next steps in evaluation and management. The SRNs delivered immediate verbal feedback, delayed written feedback, and graded clinical performance using a weighted rubric. Some items were categorized as ‘must do,’ which represented the most clinically important actions. Trends in clinical performance over time were analyzed using the Jonckheere–Terpstra test. Of the 315 eligible students, 265 (84.1%) consented for their data to be included in the analysis. Clinical performance improved from a median (interquartile range) of 59.4% (46.9%, 75.0%) on case 1 to 80.0% (68.0%, 86.7%) on case 6 (p

Food and agricultural waste contribute to resource depletion, waste accumulation and climate change. Chicken egg and almond hull waste streams have favourable nutritional profiles but are underexplored for environmentally friendly livestock feed applications. We report extrusion processing and nutritional analysis of blends containing liquid egg and almond hulls. Compositions showed favourable processing, protein content comparable to grain silages and relatively low neutral detergent fibre. No differences in crude protein or fat were observed between Hard-variety and Nonpareil almond hull compositions. Hard-variety compositions contained nearly twice the crude fibre as Nonpareil mixtures. Crude protein increased from 9.3 to 13.8% with 30–50% egg content. Bacterial proliferation in extrudates was minimal due to the low water activity (aw = 0.80–0.82 in 30% egg formulations). No Salmonella or mycotoxins (aflatoxin, vomitoxin, zearalenone) were detected. This work provides a method for repurposing high moisture, high protein food waste into animal feed using abundant, low-cost almond crop residues and demonstrates the feasibility of creating an animal feed from liquid egg waste and almond hulls.

Introduction Over the past decade, the growth of accelerated three-year MD (3YMD) programs has flourished. In 2015, with support from the Josiah Macy Jr. Foundation, the Consortium of Medical Pathway Programs (CAMPP) started with eight North American medical schools. The objective of this paper is to evaluate the current state of the 3YMD programs.Material and Methods Since 2015, the CAMPP has tracked new and prospective 3YMD programs. An electronic survey collecting curricular and programmatic information about the programs was disseminated to all members of the CAMPP in August 2023. The survey included elements related to year of initiation, number of graduates, and curricular elements.Results Of the schools with known established three-year MD programs, 29 of 32 programs responded (response rate 90%). There is growth of Accelerated Medical Pathway Programs over time with almost 20% of United States Allopathic Medical Schools having or developing an accelerated program. There have been 817 graduates from these programs from 2013–2023. Most schools include an opportunity for a ‘directed pathway’ experience for students. A directed pathway is where a student completes the MD degree in three-years and then has a direct placement into an affiliated residency program, provided they meet the goals and objectives of the curriculum. Most of the schools report a mission to reduce medical student debt and build a workforce for a specialty, for a population of patients, or geographical distribution.Conclusions Accelerated three-year medical pathway programs have grown significantly over the last decade, consistent with an overall effort to redesign medical curricula, reduce debt and contribute to the workforce.

Throughout history, the influenza A virus has caused numerous devastating global pandemics. Macrophages, as pivotal innate immune cells, exhibit a wide range of immune functions characterized by distinct polarization states, reflecting their intricate heterogeneity. In this study, we employed the time-resolved single-cell sequencing technique coupled with metabolic RNA labelling to elucidate the dynamic transcriptional changes in distinct polarized states of bone marrow-derived macrophages (BMDMs) upon infection with the influenza A virus. Our approach not only captures the temporal dimension of transcriptional activity, which is lacking in conventional scRNA-seq methods, but also reveals that M2-polarized Arg1_macrophage cluster is the sole state supporting successful replication of influenza A virus. Furthermore, we identified distinct antigen presentation capabilities to CD4+ T and CD8+ T cells across diverse polarized states of macrophages. Notably, the M1 phenotype, exhibited by (BMDMs) and murine alveolar macrophages (AMs), demonstrated superior conventional and cross-presentation abilities for exogenous antigens, with a particular emphasis on cross-presentation capacity. Additionally, as CD8+ T cell differentiation progressed, M1 polarization exhibited an enhanced capacity for cross-presentation. All three phenotypes of BMDMs, including M1, demonstrated robust presentation to CD4+ regulatory T cells, while displaying limited ability to present to naive CD4+ T cells. These findings offer novel insights into the immunological regulatory mechanisms governing distinct polarized states of macrophages, particularly their roles in restricting the replication of influenza A virus and modulating antigen-specific T cell responses through innate immunity.

Novel T-cell immunotherapies such as bispecific T-cell engagers (BiTEs) are emerging as promising therapeutic strategies for prostate cancer. BiTEs are engineered bispecific antibodies containing two distinct binding domains that allow for concurrent binding to tumor-associated antigens (TAAs) as well as immune effector cells, thus promoting an immune response against cancer cells. Prostate cancer is rich in tumor associated antigens such as, but not limited to, PSMA, PSCA, hK2, and STEAP1 and there is strong biologic rationale for employment of T-cell redirecting BiTEs within the prostate cancer disease space. Early generation BiTE constructs employed in clinical study have demonstrated meaningful antitumor activity, but challenges related to drug delivery, immunogenicity, and treatment-associated adverse effects limited their success. The ongoing development of novel BiTE constructs continues to address these barriers and to yield promising results in terms of efficacy and safety. This review will highlight some of most recent developments of BiTE therapies for patients with advanced prostate cancer and the evolving data surrounding BiTE constructs undergoing clinical evaluation.

Epithelial cell adhesion molecule (EpCAM) is a transmembrane glycoprotein that plays several roles in cancer biology. EpCAM is an attractive therapeutic target because of its expression in most solid tumors. However, targeting EpCAM has been challenging because it is also highly expressed in normal epithelial tissues. Initial attempts to develop EpCAM-specific T-cell engagers were unsuccessful due to severe cytokine release effects, as well as serious on-target, off-tumor drug-related toxicities. We developed novel, conditionally active biological (CAB) bispecific antibodies that bind to both EpCAM and CD3 in an acidic tumor microenvironment. In healthy tissues, binding to EpCAM and CD3 is greatly reduced by a novel, dual CAB selection, where each binding domain is independently blocked by the presence of physiological chemicals known as Protein-associated Chemical Switches (PaCS). The CAB anti-EpCAM T-cell engagers displayed the anticipated bispecific binding properties and mediated the potent lysis of EpCAM-positive cancer cell lines through the recruitment of T cells in the tumor microenvironment. Xenograft studies showed that the efficacy of CAB bispecific antibodies is similar to that of a non-CAB anti-EpCAM bispecific antibody, but they have markedly reduced toxicity in non-human primates, indicating an unprecedentedly widened therapeutic index of over 100-fold. These preclinical results indicate that the dual CAB bispecific antibody is potentially both a powerful and safe therapeutic platform and a promising T cell-engaging treatment for patients with EpCAM-expressing tumors.

Low molecular weight proteins and protein assemblies can now be investigated using cryo-electron microscopy (EM) as a complement to traditional structural biology techniques. It is important, however, to not lose sight of the dynamic information inherent in macromolecules that give rise to their exquisite functionality. As computational methods continue to advance the field of biomedical imaging, so must strategies to resolve the minute details of disease-related entities. Here, we employed combinatorial modeling approaches to assess flexible properties among low molecular weight proteins (~100 kDa or less). Through a blend of rigid body refinement and simulated annealing, we determined new hidden conformations for wild type p53 monomer and dimer forms. Structures for both states converged to yield new conformers, each revealing good stereochemistry and dynamic information about the protein. Based on these insights, we identified fluid parts of p53 that complement the stable central core of the protein responsible for engaging DNA. Molecular dynamics simulations corroborated the modeling results and helped pinpoint the more flexible residues in wild type p53. Overall, the new computational methods may be used to shed light on other small protein features in a vast ensemble of structural data that cannot be easily delineated by other algorithms.

IntroductionThis article describes the development and initial validation of a measure of implicit internalized stigma among queer people, the Implicit Internalized Sexual Orientation Stigma Affect Misattribution Procedure (Internal-SOS-AMP), a computer-administered sequential priming procedure.MethodsThe creation of the Internal-SOS-AMP involved a mixed-methods approach, including a literature review, expert interviews, stimuli selection and pilot testing, data collection from a large sample, reliability testing, correlational analyses, and confirmatory factor analysis. Psychometric testing was conducted with a national sample of 500 queer adults who completed two waves of data collection. Confirmatory factor analysis was used to evaluate two models: a one-factor model with internalized stigma specified as one overall construct and a two-factor model with internalized stigma specified as two constructs based on binary conceptions of gender (stigma regarding queer women and stigma regarding queer men).ResultsResults showed that the two-factor model best fit the data. This indicates that although implicit attitudes toward queer men and women are highly correlated, implicit internalized stigma differentiated by two gender stimuli groups (men and women) more accurately reflects the data. There was evidence of convergent validity as Internal-SOS-AMP scores showed small positive associations with explicit internalized stigma. Regarding divergent validity, Internal-SOS-AMP scores were inversely related to affirmation of a queer identity. Reliability results for the Internal-SOS-AMP showed good internal consistency and acceptable test–retest reliability.DiscussionThe creation of the Internal-SOS-AMP used best practices for measurement development. Psychometric findings show strong evidence of content validity, convergent validity, divergent validity, and reliability of the Internal-SOS-AMP.

Abstract Intranasal insulin is a putative neuroprotective therapy after cardiac arrest, but safety in humans at doses extrapolated from animal models is unknown. This phase I, open‐label adaptive dose‐escalation study explores the maximum tolerated dose of intranasal insulin in healthy human participants. Placebo or insulin at doses from 0 to 1000 units was given to healthy participants intranasally on repeated weekly visits. Serum glucose, insulin, and C‐peptide levels were measured serially at 0, 15, 30, 60, 120, 180, and 240 min after administration. Twenty‐four participants (12 female, median age 53, IQR 35–61) were enrolled. There was minimal change in average serum glucose after administration of intranasal insulin. Average serum insulin increased slightly in a dose‐dependent manner, reaching maximum concentrations at 15 min. C‐peptide decreased over time from administration in all groups. One participant had severe hypoglycemia (24 mg/dL at 45 min) and a different participant had mild hypoglycemia (51 mg/dL at 30 min), both after receiving 600 U intranasal insulin. Hypoglycemic episodes were associated with increases in serum insulin. Both participants continued in the study without hypoglycemia after additional doses. High‐dose intranasal insulin up to 1000 U was generally well tolerated, with minimal measurable systemic absorption and without significant aggregate changes in mean glucose. Idiosyncratic episodic systemic absorption and hypoglycemia require further study and additional caution in potential clinical application. Further study of its target engagement and efficacy as a neuroprotective therapy after cardiac arrest at these doses is warranted.

Abstract A practical approach for modeling field‐scale moisture flow in a highly heterogeneous unsaturated medium is described in this study. The validity of this approach is demonstrated through comparison of the numerical simulations with field observations at a semiarid site located in southcentral Washington State. The methodology is based on upscaling the core scale hydraulic properties and combining power‐law and tensorial connectivity‐tortuosity (PA‐TCT) approaches to derive macroscopic anisotropy parameters for each hydrostratigraphic unit (HSU) identified in the field. Each heterogeneous HSU is approximated by an equivalent homogeneous medium (EHM) model for which PA‐TCT parameters are used in the flow simulations. The available field data on moisture content and matric potential are compared with steady‐state flow simulations based on the mean form of Richards' equation. While the homogenization or averaging of heterogeneities, embedded in the EHM modeling approximation, cannot capture all of the field‐scale variability, the simulated steady‐state moisture and matric potential profiles capture well the central tendency of the field data. This approach is deemed practical for assessing the fate and transport of contaminants in highly heterogeneous unsaturated media at the transport scale of hundreds of meters.