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Abstract A deep learning model using attention-based multiple instance learning (aMIL) and self-supervised learning (SSL) was developed to perform pathologic classification of neuroblastic tumors and assess MYCN-amplification status using H&E-stained whole slide images from the largest reported cohort to date. The model showed promising performance in identifying diagnostic category, grade, mitosis-karyorrhexis index (MKI), and MYCN-amplification with validation on an external test dataset, suggesting potential for AI-assisted neuroblastoma classification.

Abstract Since the 5th Assessment Report of the Intergovernmental Panel on Climate Change (AR5) an extended concept of the energetic analysis of climate change including forcings, feedbacks and adjustment processes has become widely adopted. Adjustments are defined as processes that occur in response to the introduction of a climate forcing agent, but that are independent of global‐mean surface temperature changes. Most considered are the adjustments that impact the Earth energy budget and strengthen or weaken the instantaneous radiative forcing due to the forcing agent. Some adjustment mechanisms also impact other aspects of climate not related to the Earth radiation budget. Since AR5 and a following description by Sherwood et al. (2015, https://doi.org/10.1175/bams‐d‐13‐00167.1), much research on adjustments has been performed and is reviewed here. We classify the adjustment mechanisms into six main categories, and discuss methods of quantifying these adjustments in terms of their potentials, shortcomings and practicality. We furthermore describe aspects of adjustments that act beyond the energetic framework, and we propose new ideas to observe adjustments or to make use of observations to constrain their representation in models. Altogether, the problem of adjustments is now on a robust scientific footing, and better quantification and observational constraint is possible. This allows for improvements in understanding and quantifying climate change.

Abstract Aggregation of misfolded α-synuclein (α-syn) is a key characteristic feature of Parkinson’s disease (PD) and related synucleinopathies. The nature of these aggregates and their contribution to cellular dysfunction is still not clearly elucidated. We employed mass spectrometry-based total and phospho-proteomics to characterize the underlying molecular and biological changes due to α-syn aggregation using the M83 mouse primary neuronal model of PD. We identified gross changes in the proteome that coincided with the formation of large Lewy body-like α-syn aggregates in these neurons. We used protein-protein interaction (PPI)-based network analysis to identify key protein clusters modulating specific biological pathways that may be dysregulated and identified several mechanisms that regulate protein homeostasis (proteostasis). The observed changes in the proteome may include both homeostatic compensation and dysregulation due to α-syn aggregation and a greater understanding of both processes and their role in α-syn-related proteostasis may lead to improved therapeutic options for patients with PD and related disorders.

Abstract The mechanisms by which clouds impact the variability of the mid‐latitude atmosphere are poorly understood. We use an idealized, dry atmospheric model to investigate the relationship between Atmospheric Cloud Radiative Effects (ACRE) and annular mode persistence. We force the model with time‐varying diabatic heating that mimics the observed ACRE response to the Southern Annular Mode (SAM). Realistic ACRE forcing reduces annular mode persistence by 5 days (−16%), which we attribute to a weakening of low‐frequency eddy forcing via modified low‐level temperature gradients, though this effect is partly compensated by reduced frictional damping due to zonal wind anomalies becoming more top‐heavy. The persistence changes are nonlinear with respect to the amplitude of ACRE forcing, reflecting nonlinearities in the response of the eddy forcing. These results highlight the ACRE's impact on low‐frequency eddy forcing as the dominant cause of changes in annular mode persistence.

As technological advancements continue to redefine the landscape of adult education and training, virtual reality (VR) has emerged as a potent tool for enhancing skill acquisition. This literature review synthesizes existing research on the utilization of immersive and discrete VR in training adults in the medical domain. The primary focus was on understanding the effectiveness, challenges, and potential applications of VR-based training programs. Specifically, we reviewed studies related to practical skills critical to safety that target a non-surgical, discrete medical procedure (e.g., diabetes care procedures, how to correctly set a G-tube, CPR, correct personal protective equipment [PPE] usage) using an immersive VR technology as a training modality. Further, the studies reviewed had to include a comparison of immersive VR training to that of a business-as-usual (BAU) method. We conducted a review of the six studies that met the criteria and coded variables related to what technology was used, targeted skills being trained, social validity, effectiveness of the intervention, and whether generalization occurred. Key themes explored in the literature include the role of immersive experiences in enhancing learning outcomes when comparing VR training to BAU and the adaptability of VR platforms to different skill sets. Special attention was given to identifying factors that contribute to the success or limitation of VR-based training initiatives, including individual differences, technology acceptance, and effectiveness.

Abstract Aerosol‐cloud interactions (ACI) in warm clouds are the primary source of uncertainty in effective radiative forcing (ERF) during the historical period and, by extension, inferred climate sensitivity. The ERF due to ACI (ERFaci) is composed of the radiative forcing due to changes in cloud microphysics and cloud adjustments to microphysics. Here, we examine the processes that drive ERFaci using a perturbed parameter ensemble (PPE) hosted in CAM6. Observational constraints on the PPE result in substantial constraints in the response of cloud microphysics and macrophysics to anthropogenic aerosol, but only minimal constraint on ERFaci. Examination of cloud and radiation processes in the PPE reveal buffering of ERFaci by the interaction of precipitation efficiency and radiative susceptibility.

Adult-born granule cells (abGCs) project to the CA2 region of the hippocampus, but it remains unknown how this circuit affects behavioral function. Here, we show that abGC input to the CA2 of adult mice is involved in the retrieval of remote developmental memories of the mother. Ablation of abGCs impaired the ability to discriminate between a caregiving mother and a novel mother, and this ability returned after abGCs were regenerated. Chemogenetic inhibition of projections from abGCs to the CA2 also temporarily prevented the retrieval of remote mother memories. These findings were observed when abGCs were inhibited at 4–6 weeks old, but not when they were inhibited at 10–12 weeks old. We also found that abGCs are necessary for differentiating features of CA2 network activity, including theta-gamma coupling and sharp wave ripples, in response to novel versus familiar social stimuli. Taken together, these findings suggest that abGCs are necessary for neuronal oscillations associated with discriminating between social stimuli, thus enabling retrieval of remote developmental memories of the mother by their adult offspring.

Objective: We used a framework to assess the value implications of thoracic surgeon operative volume within an 8-hospital health system. Methods: Surgical cases for non–small cell lung cancer were assessed from March 2015 to March 2021. High-volume (HV) surgeons performed >25 pulmonary resections annually. Metrics include length of stay, infection rates, 30-day readmission, in-hospital mortality, median 30-day charges and direct costs, and 3-year recurrence-free and overall survival. Multivariate regression-based propensity scores matched patients between groups. Metrics were graphed on radar charts to conceptualize total value. Results: All 638 lung resections were performed by 12 surgeons across 6 hospitals. Two HV surgeons performed 51% (n = 324) of operations, and 10 low-volume surgeons performed 49% (n = 314). Median follow-up was 28.8 months (14.0-42.3 months). Lobectomy was performed in 71% (n = 450) of cases. HV surgeons performed more segmentectomies (33% [n = 107] vs 3% [n = 8]; P

Abstract The type VI secretion system (T6SS) of Gram-negative bacteria inhibits competitor cells through contact-dependent translocation of toxic effector proteins. In Proteobacteria, the T6SS is anchored to the cell envelope through a megadalton-sized membrane complex (MC). However, the genomes of Bacteroidota with T6SSs appear to lack genes encoding homologs of canonical MC components. Here, we identify five genes in Bacteroides fragilis (tssNQOPR) that are essential for T6SS function and encode a Bacteroidota-specific MC. We purify this complex, reveal its dimensions using electron microscopy, and identify a protein-protein interaction network underlying the assembly of the MC including the stoichiometry of the five TssNQOPR components. Protein TssN mediates the connection between the Bacteroidota MC and the conserved baseplate. Although MC gene content and organization varies across the phylum Bacteroidota, no MC homologs are detected outside of T6SS loci, suggesting ancient co-option and functional convergence with the non-homologous MC of Pseudomonadota.

No abstract available.

Abstract Proximal spinal muscular atrophy (SMA) is a leading genetic cause for infant death in the world and results from the selective loss of motor neurons in the spinal cord. SMA is a consequence of low levels of SMN protein and small molecules that can increase SMN expression are of considerable interest as potential therapeutics. Previous studies have shown that both 4-phenylbutyrate (4PBA) and trichostatin A (TSA) increase SMN expression in dermal fibroblasts derived from SMA patients. AR42 is a 4PBA-tethered TSA derivative that is a very potent histone deacetylase inhibitor. SMA patient fibroblasts were treated with either AR42, AR19 (a related analogue), 4PBA, TSA or vehicle for 5 days and then immunostained for SMN localization. AR42 as well as 4PBA and TSA increased the number of SMN-positive nuclear gems in a dose-dependent manner while AR19 did not show marked changes in gem numbers. While gem number was increased in AR42-treated SMA fibroblasts, there were no significant changes in FL-SMN mRNA or SMN protein. The neuroprotective effect of this compound was then assessed in SMNΔ7 SMA (SMN2 +/+ ;SMNΔ7 +/+ ;mSmn −/− ) mice. Oral administration of AR42 prior to disease onset increased the average lifespan of SMNΔ7 SMA mice by ~ 27% (20.1 ± 1.6 days for AR42-treated mice vs. 15.8 ± 0.4 days for vehicle-treated mice). AR42 treatment also improved motor function in these mice. AR42 treatment inhibited histone deacetylase (HDAC) activity in treated spinal cord although it did not affect SMN protein expression in these mice. AKT and GSK3β phosphorylation were both significantly increased in SMNΔ7 SMA mouse spinal cords. In conclusion, presymptomatic administration of the HDAC inhibitor AR42 ameliorates the disease phenotype in SMNΔ7 SMA mice in a SMN-independent manner possibly by increasing AKT neuroprotective signaling.

One of the salient nuclear fuel performance parameters for new fuel types under development is changes in fuel thickness. To test the new commercially fabricated U-10Mo monolithic plate-type fuel, an irradiation experiment was designed that consisted of multiple mini-plate capsules distributed within the Advanced Test Reactor (ATR) core, the mini-plate 1 (MP-1) experiment. Each capsule contains eight mini-plates that were either fueled or “dummy” plates. Fuel thickness changes within a fuel assembly can be characterized by measuring the gaps between the plates ultrasonically. The channel gap probe (CGP) system is designed to measure the gaps between the plates and will provide information that supports qualification of U-10Mo monolithic fuel. This study will discuss the design and the results from the use of a custom-designed CGP system for characterizing the gaps between mini-plates within the MP-1 capsules. To ensure accurate and repeatable data, acceptance and calibration procedures have been developed. Unfortunately, there is no “gold” standard measurement to compare to CGP measurements. An effort was made to use plate thickness obtained from post-irradiation measurements to derive channel gap estimates for comparison with the CGP characterization.

ABSTRACT Developing software tools that leverage biological data sets to accelerate drug discovery is an important aspect of bioinformatic research. Here, we present a novel example: a web application called Rocket-miR that applies an existing bioinformatic algorithm (IntaRNA) to predict cross-species miRNA-mRNA interactions and identify human miRNAs with potential antimicrobial activity against antibiotic-resistant bacterial infections. Rocket-miR is the logical extension of our prior finding that human miRNA let-7b-5p impairs the ability of the ubiquitous opportunistic pathogen Pseudomonas aeruginosa to form biofilms and resist the bactericidal effect of β-lactam antibiotics. Rocket-miR’s point and click interface enables researchers without programming expertise to predict additional human-miRNA-pathogen interactions. Identified miRNAs can be developed into novel antimicrobials effective against the 24 clinically relevant pathogens, implicated in diseases of the lung, gut, and other organs, that are included in the application. The paper incorporates three case studies contributed by microbiologists that study human pathogens to demonstrate the usefulness and usability of the application. Rocket-miR is accessible at the following link: http://scangeo.dartmouth.edu/RocketmiR/.IMPORTANCEAntimicrobial-resistant infections contribute to millions of deaths worldwide every year. In particular, the group of bacteria collectively known as ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter sp.) pathogens are of considerable medical concern due to their virulence and exceptional ability to develop antibiotic resistance. New kinds of antimicrobial therapies are urgently needed to treat patients for whom existing antibiotics are ineffective. The Rocket-miR application predicts targets of human miRNAs in bacterial and fungal pathogens, rapidly identifying candidate miRNA-based antimicrobials. The application’s target audience are microbiologists that have the laboratory resources to test the application’s predictions. The Rocket-miR application currently supports 24 recognized human pathogens that are relevant to numerous diseases including cystic fibrosis, chronic obstructive pulmonary disease (COPD), urinary tract infections, and pneumonia. Furthermore, the application code was designed to be easily extendible to other human pathogens that commonly cause hospital-acquired infections.

Abstract The pancreatic islet β-cell’s preference for release of newly synthesized insulin requires careful coordination of insulin exocytosis with sufficient insulin granule production to ensure that insulin stores exceed peripheral demands for glucose homeostasis. Thus, the cellular mechanisms regulating insulin granule production are critical to maintaining β-cell function. In this report, we utilized the synchronous protein trafficking system, RUSH, in primary β-cells to evaluate proinsulin transit through the secretory pathway leading to insulin granule formation. We demonstrate that the trafficking, processing, and secretion of the proinsulin RUSH reporter, proCpepRUSH, are consistent with current models of insulin maturation and release. Using both a rodent dietary and genetic model of hyperglycemia and β-cell dysfunction, we show that proinsulin trafficking is impeded at the Golgi and coincides with the decreased appearance of nascent insulin granules at the plasma membrane. Ultrastructural analysis of β-cells from diabetic leptin receptor deficient mice revealed gross morphological changes in Golgi structure, including shortened and swollen cisternae, and partial Golgi vesiculation, which are consistent with defects in secretory protein export. Collectively, this work highlights the utility of the proCpepRUSH reporter in studying proinsulin trafficking dynamics and suggests that altered Golgi export function contributes to β-cell secretory defects in the pathogenesis of Type 2 diabetes.

Sirtuin 6 (SIRT6) is a member of the mammalian sirtuin family of NAD+-dependent protein deacylases, homologues of the yeast silent information regulator 2 (Sir2). SIRT6 has remarkably diverse functions and plays a key role in a variety of biological processes for maintaining cellular and organismal homeostasis. In this review, our primary aim is to summarize recent progress in understanding SIRT6’s functions in the retina and retinal pigment epithelium (RPE), with the hope of further drawing interests in SIRT6 to increase efforts in exploring the therapeutic potential of this unique protein in the vision field. Before describing SIRT6’s role in the eye, we first discuss SIRT6’s general functions in a wide range of biological contexts. SIRT6 plays an important role in gene silencing, metabolism, DNA repair, antioxidant defense, inflammation, aging and longevity, early development, and stress response. In addition, recent studies have revealed SIRT6’s role in macrophage polarization and mitochondrial homeostasis. Despite being initially understudied in the context of the eye, recent efforts have begun to elucidate the critical functions of SIRT6 in the retina and RPE. In the retina, SIRT6 is essential for adult retinal function, regulates energy metabolism by suppressing glycolysis that affects photoreceptor cell survival, protects retinal ganglion cells from oxidative stress, and plays a role in Müller cells during early neurodegenerative events in diabetic retinopathy. In the RPE, SIRT6 activates autophagy in culture and protects against oxidative stress in mice. Taken together, this review demonstrates that better understanding of SIRT6’s functions and their mechanisms, both in and out of the context of the eye, holds great promise for the development of SIRT6-targeted strategies for prevention and treatment of blinding eye diseases.

Background: Stuttering support groups (SSGs) have been a long-standing invaluable resource for people who stutter (PWS) but research into SSGs is only emerging. Speech-language therapists (SLTs) need further insight to successfully facilitate SSGs. Objectives: To determine PWS’ perspectives regarding why they attend SSGs in South Africa. Method: Thirteen PWS who attend SSGs, between 20 and 58 years old, were a part of this qualitative study. Purposive sampling was utilised. Semi-structured telephonic interviews were used and data was analysed thematically. Results: Four themes, namely, ‘altered perceptions’, ‘increased sense of community’, ‘support group reciprocity’, and ‘support group environment, participants and topics’, were identified. The results yielded clinical implications which included SLTs encouraging: (1) improved perceptions of being a PWS through education and self-empowerment, (2) PWS’ connections between meetings to increase the sense of community, (3) reciprocity in meetings, (4) sharing personal stories to promote learning and general self-management and (5) support, praise and education to empower and encourage PWS. This study’s findings show that SSGs helped PWS accept their stutter and gain confidence. This study showcased how SSGs can help PWS manage their fluency and gain confidence. Additionally, this study supports current research which suggests that dysfluency and social-emotional well-being should be equally addressed. Conclusion: Recommendations were generated from PWS’ perspectives and included focusing discussions on fluency, emotions and sharing personal stories. Insights from PWS helped better inform SLTs of their role within SSGs including guiding and facilitating conversations. Contribution: People who stutters’ perspectives can be used in clinical practice to help SLTs meet the needs of PWS and guide best practice when facilitating SSGs.

Aggressive diagnostic testing remains an indispensable strategy for health and aged care facilities to prevent the transmission of SARS-CoV-2 in vulnerable populations. The preferred diagnostic platform has shifted towards COVID-19 rapid antigen tests (RATs) to identify the most infectious individuals. As such, RATs are being manufactured faster than at any other time in our history yet lack the relevant quantitative analytics required to inform on absolute analytical sensitivity enabling manufacturers to maintain high batch-to-batch reproducibility, and end-users to accurately compare brands for decision making. Here, we describe a novel reference standard to measure and compare the analytical sensitivity of RATs using a recombinant GFP-tagged nucleocapsid protein (NP-GFP). Importantly, we show that the GFP tag does not interfere with NP detection and provides several advantages affording streamlined protein expression and purification in high yields as well as faster, cheaper and more sensitive quality control measures for post-production assessment of protein solubility and stability. Ten commercial COVID-19 RATs were evaluated and ranked using NP-GFP as a reference standard. Analytical sensitivity data of the selected devices as determined with NP-GFP did not correlate with those reported by the manufacturers using the median tissue culture infectious dose (TCID50) assay. Of note, TCID50 discordance has been previously reported. Taken together, our results highlight an urgent need for a reliable reference standard for evaluation and benchmarking of the analytical sensitivity of RAT devices. NP-GFP is a promising candidate as a reference standard that will ensure that RAT performance is accurately communicated to healthcare providers and the public.

We report on the detection of radio bursts from the Galactic bulge using the real-time transient detection and localization system, realfast . The pulses were detected commensally on the Karl G. Jansky Very Large Array during a survey of unidentified Fermi γ -ray sources. The bursts were localized to subarcsecond precision using realfast fast-sampled imaging. Follow-up observations with the Green Bank Telescope detected additional bursts from the same source. The bursts do not exhibit periodicity in a search up to periods of 480 s, assuming a duty cycle of

Fast radio bursts (FRBs) are a powerful and mysterious new class of transients that are luminous enough to be detected at cosmological distances. By associating FRBs to host galaxies, we can measure intrinsic and environmental properties that test FRB origin models, in addition to using them as precise probes of distant cosmic gas. The Deep Synoptic Array (DSA-110) is a radio interferometer built to maximize the rate at which it can simultaneously detect and localize FRBs. Here, we present the first sample of FRBs and host galaxies discovered by the DSA-110. This sample of 11 FRBs is the largest, most uniform sample of localized FRBs to date, as it is selected based on association with host galaxies identified in optical imaging by Pan-STARRS1. These FRBs have not been observed to repeat, and their radio properties (dispersion, temporal scattering, energy) are similar to that of the known nonrepeating FRB population. Most host galaxies have ongoing star formation, as has been identified before for FRB hosts. Two hosts of the new sample are massive, quiescent galaxies. The distribution of star formation history across this host-galaxy sample shows that the delay time distribution is wide, with a power-law model that spans from ∼100 Myr to ≳2 Gyr. This requires the existence of one or more progenitor formation channels associated with old stellar populations, such as the binary evolution of compact objects.

Detecting fast radio bursts (FRBs) with frequency-dependent intensity remains a challenge, as existing search algorithms do not account for the spectral shape, potentially leading to nondetections. We propose a novel detection statistic, which we call the Kalman detector, that improves the sensitivity of FRB signal detection by incorporating spectral shape information. The detection statistic is based on an optimal matched filter, marginalizing over all possible intensity functions, weighted by a random walk probability distribution, considering some decorrelation bandwidth. Our analysis of previously detected FRBs demonstrates that the Kalman score provides a comparable yet independent source of information for bursts with significant spectral structure, and the sensitivity improvement is of the order 0%–200% with a median improvement of 20%. We also applied the Kalman detector to existing data from FRB 20201124A and detected two new repeat bursts that were previously missed. Furthermore, we suggest a practical implementation for real-time surveys by employing a low significance soft-trigger from initial flux integration-based detection algorithms. The Kalman detector has the potential to significantly enhance FRB detection capabilities and enable new insights into the spectral properties of these enigmatic astrophysical phenomena.

Chromatin is dynamically reorganized spatially and temporally, and the post-translational modification of histones is a key component of this regulation. The basic subunit of chromatin is the nucleosome core particle, consisting of two copies each of the histones H2A, H2B, H3, and H4 around which ∼147 base pairs of DNA wrap. The intrinsically disordered histone termini, or tails, protrude from the core and are heavily post-translationally modified. Previous studies have shown that the histone tails exist in dynamic ensembles of DNA-bound states within the nucleosome. Histone tail interactions with DNA are involved in nucleosome conformation and chromatin organization. Charge-modulating histone post-translational modifications (PTMs) are poised to perturb the dynamic interactions between histone tails and DNA. Arginine side chains form favorable interactions with DNA and are sites of charge-modulating PTMs such as citrullination. Our current focus is on the H3 tail, the longest histone tail. Four arginine residues are relatively evenly spaced along the H3 tail sequence, suggesting multivalent interactions with DNA poised for regulation by PTMs. In this study, we use NMR nuclear spin relaxation experiments to investigate the contribution of arginine residues to H3 tail dynamics within the nucleosome core particle. By neutralizing arginine via mutation to glutamine, we begin to work towards a comprehensive understanding of the contribution of individual residues to H3 tail dynamics. We find that neutralization of arginine residues results in increased regional mobility of the H3 tails, with implications for understanding the direct effects of arginine citrullination. Altogether, these studies support a role for dynamics within the histone language and emphasize the importance of charge-modulating histone PTMs in regulating chromatin dynamics, starting at the level of the basic subunit of chromatin.

While robotics have been widely used in many agricultural practices such as harvesting, seeding, cattle monitoring, etc., aquaculture farming is an important, fast-growing sector of agriculture that has not seen significant adoption of advanced technologies such as robotics and the Internet of Things (IoT). In particular, dissolved oxygen (DO) monitoring, a practice in pond aquaculture essential to the health of the fish crops, remains labor-intensive and time-consuming. The Hybrid Aerial Underwater robotiCs System (HAUCS) is an IoT framework that aims to bring transformative changes to pond aquaculture. This paper focuses on the latest development in the HAUCS mobile sensing platform and field deployment. To address some shortcomings with the current implementation, the development of a novel rigid Kirigami-based robotic extension subsystem that can expand the functionality of the HAUCS platform is also being discussed.

Abstract Background Autistic individuals frequently experience social communication challenges. Girls are diagnosed with autism less often than boys even when their symptoms are equally severe, which may be due to insufficient understanding of the way autism manifests in girls. Differences in the behavioral presentation of autism, including how people talk about social topics, could contribute to these persistent problems with identification. Despite a growing body of research suggesting that autistic girls and boys present distinct symptom profiles in a variety of domains, including social attention, friendships, social motivation, and language, differences in the way that autistic boys and girls communicate verbally are not yet well understood. Closely analyzing boys’ and girls’ socially-focused language during semi-structured clinical assessments could shed light on potential sex differences in the behavioral presentation of autistic individuals that may prove useful for identifying and effectively supporting autistic girls. Here, we compare social word use in verbally fluent autistic girls and boys during the interview sections of the ADOS-2 Module 3 and measure associations with clinical phenotype. Methods School-aged girls and boys with autism (N = 101, 25 females; aged 6–15) were matched on age, IQ, and parent/clinician ratings of autism symptom severity. Our primary analysis compared the number of social words produced by autistic boys and girls (normalized to account for differences in total word production). Social words are words that make reference to other people, including friends and family. Results There was a significant main effect of sex on social word production, such that autistic girls used more social words than autistic boys. To identify the specific types of words driving this effect, additional subcategories of friend and family words were analyzed. There was a significant effect of sex on friend words, with girls using significantly more friend words than boys. However, there was no significant main effect of sex on family words, suggesting that sex differences in social word production may be driven by girls talking more about friends compared to boys, not family. To assess relationships between word use and clinical phenotype, we modeled ADOS-2 Social Affect (SA) scores as a function of social word production. In the overall sample, social word use correlated significantly with ADOS-2 SA scores, indicating that participants who used more social words were rated as less socially impaired by clinicians. However, when examined in each sex separately, this result only held for boys. Limitations This study cannot speak to the ways in which social word use may differ for younger children, adults, or individuals who are not verbally fluent; in addition, there were more autistic boys than girls in our sample, making it difficult to detect small effects. Conclusions Autistic girls used significantly more social words than boys during a diagnostic assessment—despite being matched on age, IQ, and both parent- and clinician-rated autism symptom severity. Sex differences in linguistic markers of social phenotype in autism are especially important in light of the late or missed diagnoses that disproportionately affect autistic girls. Specifically, heightened talk about social topics could complicate autism referral and diagnosis when non-clinician observers expect a male-typical pattern of reduced social focus, which autistic girls may not always exhibit.

Abstract Background Prior studies have assessed provider knowledge and factors associated with opioid misuse; similar studies evaluating patient knowledge are lacking. The purpose of this study was to assess the degree of understanding regarding opioid use in orthopaedic trauma patients. We also sought to determine the demographic factors and clinical and personal experiences associated with level of understanding. Methods One hundred and sixty-six adult orthopaedic trauma surgery patients across two clinical sites of an academic institution participated in an internet-based survey (2352 invited, 7.1% response rate). Demographic, clinical, and personal experience variables, as well as perceptions surrounding opioid use were collected. Relationships between patient characteristics and opioid perceptions were identified using univariate and multivariable logistic regressions. Alpha = 0.05. Results Excellent recognition (> 85% correct) of common opioids, side effects, withdrawal symptoms, and disposal methods was demonstrated by 29%, 10%, 30%, and 2.4% of patients; poor recognition (

Abstract A modified Chromium 10x droplet-based protocol that subsamples cells for both short-read and long-read (nanopore) sequencing together with a new computational pipeline (FLAMES) is developed to enable isoform discovery, splicing analysis, and mutation detection in single cells. We identify thousands of unannotated isoforms and find conserved functional modules that are enriched for alternative transcript usage in different cell types and species, including ribosome biogenesis and mRNA splicing. Analysis at the transcript level allows data integration with scATAC-seq on individual promoters, improved correlation with protein expression data, and linked mutations known to confer drug resistance to transcriptome heterogeneity.

miR-126 is highly expressed in inv(16) Acute myeloid leukemia (AML) but its role is unclear. Here, the authors show that the aberrant expression of miR-126 in inv(16) AML is directly due to the CBFB-MYH11 fusion gene and that it can promote AML development and leukemia stem cell maintenance, highlighting miR-126 as a therapeutic target for inv(16) AML patients

Abstract The role of calcium in blood pressure has been widely studied among hypertensive patients; however, no study has explored the role of calcium in hypertensive crises. The primary objective of this study is to evaluate the differences in serum calcium levels between hypertensive crises patients and a 1:1 random matched controls (age‐, sex‐, race‐, diabetes, and body mass index matched). This study is a single‐center, retrospective, chart review, case‐control study of patients with hypertensive crises (case group) and patients without hypertensive crises (control group). Patients were included in the case group if they were 18 years of age or older with hypertensive crises and have a documented calcium level. The control group patients were required to be 18 years of age or older, have a documented calcium level, and have no diagnosis of hypertensive crises. The primary outcome of the study was to compare the mean serum calcium in patients with hypertensive crises vs patients without hypertensive crises. Five hundred and sixty‐six patients were included in the study: 283 patients in both the case group and control group. The primary outcome results showed that serum calcium concentration was not significantly different between the case group (8.99 ± 0.78 mg/dL) and control group (8.96 ± 0.75 mg/dL) (P = .606). This study found no significant difference in serum calcium levels in patients with hypertensive crises compared to a random matched control group. Larger observational or experimental studies may be useful to evaluate the effect of calcium on blood pressure in hypertensive crises.

Category: Ankle Arthritis; Ankle Introduction/Purpose: Choosing a treatment course can be challenging for patients with ankle osteoarthritis given the multiple operative and nonoperative options. We developed and validated the Ankle Arthritis Patient Decision Aid as a tool to improve shared decision-making for such patients. Methods: Eighty patients of 2 surgeons at a US academic foot and ankle practice were recruited from October 2020 to June 2021 using an electronic medical record messaging system. Participants reviewed the tool and answered survey questions evaluating its quality and factors affecting treatment choice in a hypothetical patient scenario. We evaluated the tool using decisional conflict scores (maximum, 40 points, with higher scores indicating greater uncertainty), pre- and post-knowledge test scores (maximum, 8 points), and helpfulness scores (maximum, 7 points). We compared pre- and post-test knowledge scores using paired Student t- tests. Alpha=.05. Results: Mean (+- standard deviation) knowledge scores improved from 4.8 +- 1.2 (pre-test) to 6.7 +- 1.3 (post-test), for a mean improvement of 1.9 +- 1.4 (p

Category: Sports; Basic Sciences/Biologics Introduction/Purpose: Achilles tendon ruptures have increased 10-fold in the past three decades, leading to long-term functional deficits in nearly two-thirds of patients. To counter this, rehabilitation protocols have been developed to strike a balance between protecting the healing tendon from re-rupture while allowing patients to return to activities of daily living as early as possible. However, Achilles tendon loading is neither prescribed nor quantified during immobilization and there is no standardized method of immobilization. Therefore, the purpose of this study was to quantify the amount of tendon loading experienced while walking in 3 different kinds immobilizing boot with respect to prescribed ankle angle and walking speed. Methods: Two healthy-young adults (2M; Age: 29.6 +- 2.3 y/o; BMI: 30 +- 7) walked over flat ground while wearing 3 different immobilizing boots and athletic shoes after providing written informed consent in this IRB approved study. We calculated Achilles tendon loading while subjects walked in 3 boots that positioned the ankle in 30 degrees of plantar flexion (Figure A, Air Cam Walker, United Ortho; AirSelect Walker, Aircast, VACOped, OPED). In all three boots, an instrumented insole (Loadsol, Novel) was placed under the foot to quantify load experienced by the tendon. We calculated the Achilles tendon loading profiles during gait for each immobilizing boot and compared them against the normal shod condition. We also provided the surgical repair threshold of 0.5 bodyweights to visualize the potential efficacy of each immobilizing boot for early rehabilitation when the tendon is early in the healing phase. Results: As expected, tendon loading was highest in the shod condition with no ankle bracing and lowest in the fully plantarflexed adjustable boot (Figure B). On average, the immobilizing boots reduced tendon loading decreased by 68% compared to unsupported walking in shoes but there were differences among the boots. The 3 different immobilizing boots provided differing levels of support. The 2 walking boots that use heel wedges to support the ankle in plantar flexion provided the least support, reducing loading by an average of 60-68%. The more rigid boot that constrained ankle angle using a posterior strut reduced tendon loading by 77% compared to gait in a normal walking shoe. Based on biomechanical studies of Achilles tendon repair strength in cadaveric experiments, only the rigid boot with the posterior strut reduced tendon loading biomechanics to levels that could be resisted by the surgical repair (Figure B). Conclusion: In this study, we used an instrumented insole to quantify Achilles tendon loading in a variety of immobilizing boots used by rupture patients. Our initial findings show that tendon loading is varies greatly between boot types and patients, highlighting the importance of understanding how these loads change in patients. Our future work centers around identifying the tendon loading profiles that promote tendon healing and optimize patient outcomes. These loading profiles will then provide the quantitative data needed to personalize loading for patients and guide rehabilitation to improve outcomes.

Background: Severe acute malnutrition (SAM) is associated with cognitive and motor deficits. Little is known about the swallowing, feeding and communication characteristics of hospitalised toddlers with SAM, limiting the abilities of speech-language therapists to provide effective early intervention. Objective: To explore the background, swallowing, feeding and communication characteristics of toddlers with SAM during in-patient nutritional rehabilitation. Method: An exploratory, prospective, collective case-study was conducted with three hospitalised toddlers who were 12–18 months old and independently diagnosed with SAM, at least 1 week after transitioning to oral feeding. Detailed case histories were compiled through medical file perusal and parent interviews. Cross-sectional clinical bedside assessments were completed with the Rossetti Infant-Toddler Language Scale and Schedule for Oral-Motor Assessment. Results: All three participants had a history of feeding difficulties before admission. Despite intact pharyngeal swallows, heterogeneous oral-sensorimotor dysfunction and disruptive feeding behaviours were identified. Risk for oropharyngeal dysphagia indicates the need to modify dietary consistencies to prevent prolonging recovery or SAM relapse. Participants had mild-to-moderate language delays, particularly in interaction-attachment, play and language comprehension, with an atypical moderate receptive and mild expressive language delay profile. None of the participants were referred for speech-language therapy. Conclusion: This exploratory research showed the oral-sensorimotor skills, swallowing and communication characteristics of children with SAM. Speech-language therapists could address oral-sensorimotor functioning, feeding difficulties and communication interaction delays before discharge to community-based management for SAM. Further investigation with a larger sample size is recommended. Contribution: Novel description of the oral-sensorimotor skills for feeding and the communication development of three severely malnourished toddlers with HIV and tuberculosis co-infection was presented. The complexity of the three cases is highlighted.

Summary: Background: Though case fatality rate (CFR) is widely used to reflect COVID-19 fatality risk, its use is limited by large temporal and spatial variation. Hospital mortality rate (HMR) is also used to assess the severity of COVID-19, but HMR data is not directly available globally. Alternative metrics are needed for COVID-19 severity and fatality assessment. Methods: We introduce new metrics for COVID-19 fatality risk measurements/monitoring and a new mathematical model to estimate average hospital length of stay for deaths (Ldead) and discharges (Ldis). Multiple data sources were used for our analyses. Findings: We propose three, new metrics: hospital occupancy mortality rate (HOMR), ratio of total deaths to hospital occupancy (TDHOR), and ratio of hospital occupancy to cases (HOCR), for dynamic assessment of COVID-19 fatality risk. Estimated Ldead and Ldis for 501,079 COVID-19 hospitalizations in 34 US states between 7 August 2020 and 1 March 2021 were 18·2(95%CI:17·9-18·5) and 14·0(95%CI:13·9-14·0) days, respectively. We found the dramatic changes in COVID-19 CFR observed in 27 countries during early stages of the pandemic were mostly caused by undiagnosed cases. Compared to the first week of November 2021, the week mean HOCRs (mimics hospitalization-to-case ratio) for Omicron variant (58·6% of US new cases as of 25 December 2021) decreased 65·16% in the US as of 16 January 2022. Interpretation: The new and reliable measurements described here could be useful for COVID-19 fatality risk and variant-associated risk monitoring. Funding: No specific funding was associated with the present study.

Abstract Although the role of magnesium in blood pressure has been well studied among hypertensive patients, no study has explored the role of magnesium in hypertensive crises. The primary objective of this study is to evaluate the differences in serum magnesium levels between hypertensive crises patients and matched controls (age‐, sex‐, race‐, and diabetes‐matched) in a 1:1 random match. This study is a single‐center, retrospective, chart review, case‐control study of patients with hypertensive crises (case group) and patients without hypertensive crises (control group). Patients were included in the case group if they were 18 years of age or older with hypertensive crises and have a documented magnesium level. The control group patients were required to be 18 years of age or older, have no diagnosis of hypertensive crises, and have a documented magnesium level. The primary outcome of the study was to compare the mean serum magnesium in patients with hypertensive crises versus patients without hypertensive crises. Three hundred and fifty‐eight patients were included in the study: 179 patients in both the case group and control group. The primary outcome results showed that serum magnesium concentration was not significantly different between the case group (1.89 ± 0.29 mg/dl) and control group (1.90 ± 0.31 mg/dl) (p = .787). This study found no significant difference in serum magnesium levels in patients with hypertensive crises compared to a random matched control group. Larger observational or experimental studies may be useful to evaluate the effect of magnesium on blood pressure in hypertensive crises.

The ex-copula penile dorsiflexion reflex (PDFR) is an established measure of sexual dysfunction in male rat models of spinal cord injury. Although the PDFR after complete spinal transection is well described, information regarding the more clinically relevant incomplete spinal contusion injury model is limited. This study examined, using two-dimensional (2D) kinematic analysis, the relationship between the PDFR and degree of white matter sparing (WMS). Male Wistar rats received a T9 contusion with varying degrees of impactor forces. Weekly kinematic recordings of the PDFR were made 3–8 weeks postinjury. Sexual reflex components examined included maximum angle of penile dorsiflexion, total penile event duration, and penile ascent speed. Post hoc comparison between animals grouped based upon injury severity (moderate–severe: 13.33%–17.15% WMS vs moderate: 20.85%–33.50% WMS) indicated PDFR effects. Specifically, the numbers of animals with more moderate contusions having data points above the median in both maximum angle of penile dorsiflexion and penile ascent speed were significantly lower than animals with more severe injuries. Total penile event duration was also affected but only at more chronic time points (6–8 weeks). Thus, 2D kinematic analysis of the PDFR allows for more consistent and quantifiable analysis of the subtle differences that can occur between injury severity groups in the rat contusion model.

Background. Parathyroid allotransplantation is a burgeoning treatment for severe hypoparathyroidism. Deceased donor parathyroid gland (PTG) procurement can be technically challenging due to lack of normal intraoperative landmarks and exposure constraints in the neck of organ donors. In this study, we assessed standard 4-gland exposure in situ and en bloc surgical techniques for PTG procurement and ex vivo near-infrared autofluorescence (NIRAF) imaging for identification of PTGs during organ recovery. Methods. Research tissue consent was obtained from organ donors or donor families for PTG procurement. All donors were normocalcemic, brain-dead, solid organ donors between 18 and 65 y of age. PTGs were procured initially using a standard 4-gland exposure technique in situ and subsequently using a novel en bloc resection technique after systemic organ preservation flushing. Parathyroid tissue was stored at 4 °C in the University of Wisconsin solution up to 48 h post-procurement. Fluoptics Fluobeam NIRAF camera and Image J software were utilized for quantification of NIRAF signal. Results. Thirty-one brain-dead deceased donor PTG procurements were performed by abdominal transplant surgeons. In the initial 8 deceased donors, a mean of 1.75 glands (±1.48 glands SD) per donor were recovered using the 4-gland in situ technique. Implementation of combined en bloc resection with ex vivo NIRAF imaging in 23 consecutive donors yielded a mean of 3.60 glands (±0.4 SD) recovered per donor (P < 0.0001). Quantification of NIRAF integrated density signal demonstrated >1-fold log difference in PTG (2.13 × 105 pixels) versus surrounding anterior neck structures (1.9 × 104 pixels; P < 0.0001). PTGs maintain distinct NIRAF signal from the time of recovery (1.88 × 105 pixels) up to 48 h post-procurement (1.55 × 105 pixels) in organ preservation cold storage (P = 0.34). Conclusions. The use of an en bloc surgical technique with ex vivo NIRAF imaging significantly enhances the identification and recovery of PTG from deceased donors.

Abstract Background Individuals with autism spectrum disorder (ASD) are characterized by social communication challenges and repetitive behaviors that may be quickly detected by experts (Autism Res 10:653–62, 2017; American Psychiatric Association, Diagnostic and statistical manual of mental disorders, 2013). Recent research suggests that even naïve non-experts judge a variety of human dimensions using narrow windows of experience called “first impressions.” Growing recognition of sex differences in a variety of observable behaviors in ASD, combined with research showing that some autistic girls and women may “camouflage” outward symptoms, suggests it may be more difficult for naïve conversation partners to detect ASD symptoms in girls. Here, we explore the first impressions made by boys and girls with ASD and typically developing (TD) peers. Methods Ninety-three school-aged children with ASD or TD were matched on IQ; autistic girls and boys were additionally matched on autism symptom severity using the ADOS-2. Participants completed a 5-minute “get-to-know-you” conversation with a new young adult acquaintance. Immediately after the conversation, confederates rated participants on a variety of dimensions. Our primary analysis compared conversation ratings between groups (ASD boys, ASD girls, TD boys, TD girls). Results Autistic girls were rated more positively than autistic boys by novel conversation partners (better perceived social communication ability), despite comparable autism symptom severity as rated by expert clinicians (equivalent true social communication ability). Boys with ASD were rated more negatively than typical boys and typical girls by novel conversation partners as well as expert clinicians. There was no significant difference in the first impressions made by autistic girls compared to typical girls during conversations with a novel conversation partner, but autistic girls were rated lower than typical girls by expert clinicians. Limitations This study cannot speak to the ways in which first impressions may differ for younger children, adults, or individuals who are not verbally fluent; in addition, there were more autistic boys than girls in our sample, making it difficult to detect small effects. Conclusions First impressions made during naturalistic conversations with non-expert conversation partners could—in combination with clinical ratings and parent report—shed light on the nature and effects of behavioral differences between girls and boys on the autism spectrum.

Abstract The emergence of SARS-CoV-2/2019 novel coronavirus (COVID-19) has created a global pandemic with no approved treatments or vaccines. Many treatments have already been administered to COVID-19 patients but have not been systematically evaluated. We performed a systematic literature review to identify all treatments reported to be administered to COVID-19 patients and to assess time to clinically meaningful response for treatments with sufficient data. We searched PubMed, BioRxiv, MedRxiv, and ChinaXiv for articles reporting treatments for COVID-19 patients published between 1 December 2019 and 27 March 2020. Data were analyzed descriptively. Of the 2706 articles identified, 155 studies met the inclusion criteria, comprising 9152 patients. The cohort was 45.4% female and 98.3% hospitalized, and mean (SD) age was 44.4 years (SD 21.0). The most frequently administered drug classes were antivirals, antibiotics, and corticosteroids, and of the 115 reported drugs, the most frequently administered was combination lopinavir/ritonavir, which was associated with a time to clinically meaningful response (complete symptom resolution or hospital discharge) of 11.7 (1.09) days. There were insufficient data to compare across treatments. Many treatments have been administered to the first 9152 reported cases of COVID-19. These data serve as the basis for an open-source registry of all reported treatments given to COVID-19 patients at www.CDCN.org/CORONA . Further work is needed to prioritize drugs for investigation in well-controlled clinical trials and treatment protocols.

Abstract Background Staging laparoscopy and peritoneal cytology can detect occult metastatic disease prior to treatment of gastric cancer. The yield of peritoneal staging in patients with early stage disease is lacking. We assess the yield of peritoneal staging in early stage gastric cancer and its impact on survival. Methods Data were obtained from a prospective database of patients who underwent staging laparoscopy and peritoneal cytology for gastric cancer at our institution between July 1995 and July 2018. Clinical stage was determined by endoscopic ultrasound, and early stage was defined as cT1-2 and cN0. Rates of positive cytology and carcinomatosis at time of laparoscopy were obtained. Univariate analyses were used to compare groups, and Kaplan-Meier survival analyses were used to assess survival outcomes. Results Eight hundred sixty-seven patients underwent staging laparoscopy and peritoneal cytology; 56 were defined as early stage. Age was 61 ± 12 years, 66.4% were male, and 62.3% were white. Of the patients with early stage disease, 17.9% had either gross carcinomatosis (10.7%) and/or positive peritoneal cytology (10.9%). All cases of peritoneal disease were in patients with cT2 disease. There were no differences in age, gender, or race based on peritoneal disease (all p > 0.05). The presence of carcinomatosis or positive cytology significantly affected overall survival (p < 0.001), regardless of clinical T or N stage. Conclusions Peritoneal staging identifies metastatic disease in a significant number of patients with early stage disease. Given its poor prognosis and alternate therapy options, independent staging laparoscopy and peritoneal cytology should be considered in patients with early stage gastric adenocarcinoma.

Abstract The whale shark Rhincodon typus is found throughout the world's tropical and warm‐temperate ocean basins. Despite their broad physical distribution, research on the species has been concentrated at a few aggregation sites. Comparing DNA sequences from sharks at different sites can provide a demographically neutral understanding of the whale shark's global ecology. Here, we created genetic profiles for 84 whale sharks from the Saudi Arabian Red Sea and 72 individuals from the coast of Tanzania using a combination of microsatellite and mitochondrial sequences. These two sites, separated by approximately 4500 km (shortest over‐water distance), exhibit markedly different population demographics and behavioral ecologies. Eleven microsatellite DNA markers revealed that the two aggregation sites have similar levels of allelic richness and appear to be derived from the same source population. We sequenced the mitochondrial control region to produce multiple global haplotype networks (based on different alignment methodologies) that were broadly similar to each other in terms of population structure but suggested different demographic histories. Data from both microsatellite and mitochondrial markers demonstrated the stability of genetic diversity within the Saudi Arabian aggregation site throughout the sampling period. These results contrast previously measured declines in diversity at Ningaloo Reef, Western Australia. Mapping the geographic distribution of whale shark lineages provides insight into the species’ connectivity and can be used to direct management efforts at both local and global scales. Similarly, understanding historical fluctuations in whale shark abundance provides a baseline by which to assess current trends. Continued development of new sequencing methods and the incorporation of genomic data could lead to considerable advances in the scientific understanding of whale shark population ecology and corresponding improvements to conservation policy.

Bovine leukemia virus (BLV) causes enzootic bovine leukosis, the most common neoplastic disease in cattle. Previous work estimates that 78% of US beef operations and 38% of US beef cattle are seropositive for BLV. Infection by BLV in a herd is an economic concern for producers as evidence suggests that it causes an increase in cost and a subsequent decrease in profit to producers. Studies investigating BLV in dairy cattle have noted disease resistance or susceptibility, measured by a proviral load (PVL) associated with specific alleles of the bovine leukocyte antigen (BoLA) DRB3 gene. This study aims to investigate the associations between BoLA DRB3 alleles and BLV PVL in beef cattle. Samples were collected from 157 Midwest beef cows. BoLA DRB3 alleles were identified and compared with BLV PVL. One BoLA DRB3 allele, *026:01, was found to be associated with high PVL in relation to the average of the sampled population. In contrast, two alleles, *033:01 and *002:01, were found to be associated with low PVL. This study provides evidence of a relationship between BoLA DRB3 alleles and BLV PVL in US beef cows.

Understanding and anticipating the effects of climate change on fisheries social-ecological systems (FSESs) is central to proactive fisheries management in a changing global climate. With fisheries management increasingly striving to consider interactions and feedbacks among people, targeted species, and the broader ecological and human communities, fisheries managers and participants need tools to help them assess these complex systems. We developed a new climate vulnerability assessment framework for analyzing the impacts of a climate-induced trend or event on a FSES. The framework divides the FSES into four interrelated and interacting domains: Ecological Community, Fished Species, Fishery, and Human Community. The framework provides a systematic approach to account for indirect as well as direct effects, links among subsystems, and multiple climate change-induced stressors. We demonstrate the framework’s utility by applying it to three case studies: the effects of a marine heatwave on the Dungeness crab FSES, the effects of a marine heatwave on the red sea urchin FSES, and the effects of long-term climate trends on North Pacific albacore. We found that the effects of a climatic trend or event on a FSES are often indirect and can trigger diverse and important feedbacks. These examples also showed that the climatic trend or event may cause changes in the temporal and spatial distribution of fishing effort and fished species that have a more significant impact on the FSES than changes to species abundance per se. Unlike other climate vulnerability assessment frameworks and applications, ours is designed to enable consideration of the range of feedbacks within and among both the ecological and human communities. As such, it is a valuable tool to guide the holistic examination and assessment of potential impacts to FSESs.

Abstract: Aerosol‐cloud interactions (ACI) in warm clouds are the primary source of uncertainty in effective radiative forcing (ERF) during the historical period and, by extension, inferred climate sensitivity. The ERF due to ACI (ERFaci) is composed of the radiative forcing due to changes in cloud microphysics and cloud adjustments to microphysics. Here, we examine the processes that drive ERFaci using a perturbed parameter ensemble (PPE) hosted in CAM6. Observational constraints on the PPE result in substantial constraints in the response of cloud microphysics and macrophysics to anthropogenic aerosol, but only minimal constraint on ERFaci. Examination of cloud and radiation processes in the PPE reveal buffering of ERFaci by the interaction of precipitation efficiency and radiative susceptibility.

Abstract Management‐intensive grazing (MiG) on irrigated, perennial pastures has steadily increased in the western United States due to pressure for reducing public lands grazing, overall declining land available for pasture, and decreasing commodity prices. However, there are still many unknowns regarding MiG and its environmental impact, especially with regards to soil health. Over a 2‐yr period, a study evaluating the change in soil health under a full‐scale, 82‐ha pivot‐irrigated perennial pasture system grazed with ∼230 animal units (AUs) using MiG. Soil analysis included 11 soil characteristics aggregated into the Soil Management Assessment Framework (SMAF), which outputs results for soil biological, physical, nutrient, chemical, and overall soil health indices (SHIs). Over time, positive impacts were observed in the chemical and biological SHI due to decreases in salt content and increases in microbial and enzymatic activities. Soil organic C (SOC) remained unchanged, yet positive biological SHI changes are potential precursors to future SOC increases. The chemical and nutrient SHI increased in the soil surface due to reductions in salt content in conjunction with increased plant‐available soil P, as a result of salt leaching via irrigation and pre‐study inorganic P fertilizer application in conjunction with manure deposition due to MiG, respectively. Finally, a negative impact was also observed in the physical SHI, driven primarily by increasing bulk density due to hoof pressure from cattle grazing. If managed correctly, compaction issues can be avoided, with MiG systems having potential success in supporting grazing while promoting soil health for environmental and economic sustainability.

Two component systems (TCSs) are a primary mechanism of signal sensing and response in bacteria. Systematic characterization of an entire TCS could provide a mechanistic understanding of these important signal transduction systems. Here, genetic selections were employed to dissect the molecular basis of signal transduction by the HitRS system that detects cell envelope stress in the pathogen Bacillus anthracis. Numerous point mutations were isolated within HitRS, 17 of which were in a 50-residue HAMP domain. Mutational analysis revealed the importance of hydrophobic interactions within the HAMP domain and highlighted its essentiality in TCS signaling. In addition, these data defined residues critical for activities intrinsic to HitRS, uncovered specific interactions among individual domains and between the two signaling proteins, and revealed that phosphotransfer is the rate-limiting step for signal transduction. Furthermore, this study establishes the use of unbiased genetic selections to study TCS signaling and provides a comprehensive mechanistic understanding of an entire TCS.

Peroxisome proliferator-activated receptors (PPARs) are a family of three nuclear hormone receptors (PPARα, PPARδ, and PPARγ) that are known to regulate expression of lipid metabolism and oxidative stress genes. Given their role in reducing oxidative stress in a variety of tissues, these genes are likely important for retinal homeostasis. This hypothesis has been further supported by recent studies suggesting that PPAR-activating drugs are protective against retinal degenerations. The objective of the present study was to determine the role of PPARδ in the neuroretina. RNA-seq data show that Pparα and Pparδ are both expressed in the retina, but that Pparδ is expressed at 4-fold higher levels. Single-cell RNAseq data show that Pparδ is broadly expressed in all retinal cell types. To determine the importance of Pparδ to the retina, we generated retina-specific Pparδ knockout mice. We found that deletion of Pparδ had a minimal effect on retinal function or morphology out to 12 months of age and did not increase retinal sensitivity to oxidative stress induced by exposure to bright light. While data show that PPARδ levels were increased by the drug metformin, PPARδ was not necessary for metformin-induced protection from light damage. These data suggest that Pparδ either has a redundant function with Pparα or is not essential for normal neuroretina function or resistance to oxidative stress.

Abstract Shrub encroachment is transforming arid and semiarid grasslands worldwide. Such transitions should influence predator–prey interactions because vegetation cover often affects risk perception by prey and contributes to their landscape of fear. We examined how the landscape of fear of two desert lagomorphs (black‐tailed jackrabbit, Lepus californicus; desert cottontail, Sylvilagus audubonii) changes across grassland‐to‐shrubland gradients at Jornada Basin Long Term Ecological Research site in the Chihuahuan Desert of southern New Mexico. We test whether shrub encroachment shapes risk differently for these two lagomorphs because of differences in body size and predator escape tactics. We also examine whether an ecosystem engineer of grasslands (banner‐tailed kangaroo rat, Dipodomys spectabilis) mediates risk perception through the creation of escape refuge and whether trade‐offs exist between shrub encroachment and the local reduction of banner‐tailed kangaroo rats caused by shrub expansion. We measured perceived predation risk with flight initiation distances (FIDs) and then used structural equation modeling to tease apart the hypothesized direct and indirect pathways for how shrub encroachment could affect perceived risk. A total negative effect of shrub cover on FID was supported for jackrabbits and cottontails, suggesting both species perceive shrubbier habitat as safer. Increases in fine‐scale concealment also reduced risk for cottontails, but not jackrabbits, likely because cottontails rely on crypsis to avoid predator detection whereas jackrabbits rely on speed and agility to outrun predators. Perceived risk was reduced when individuals were near kangaroo rat mounds only for cottontails because the smaller species can use banner‐tailed kangaroo rat mounds as refuge. Shrub encroachment greatly reduced the availability of mounds. Thus, a trade‐off exists for cottontails in which shrub encroachment directly reduced perceived risk, but indirectly increased perceived risk through the local extirpation of an ecosystem engineer. Our work illustrates how the expansion of shrub encroachment can create a dynamic landscape of fear for populations of prey species involving direct and indirect pathways contingent on prey body size, escape tactics, and activities of an ecosystem engineer.

Summary: Ghrelin regulates both energy intake and glucose homeostasis. In the endocrine pancreas, ghrelin inhibits insulin release to prevent hypoglycemia during fasting. The mechanism through which this is accomplished is unclear, but recent studies suggest that ghrelin acts on δ cells to stimulate somatostatin release, which in turn inhibits insulin release from β cells. Recently, the Melanocortin Receptor Accessory Protein 2 (MRAP2) was identified as an essential partner of the ghrelin receptor (GHSR1a) in mediating the central orexigenic action of ghrelin. In this study we show that MRAP2 is expressed in islet δ cells and is required for ghrelin to elicit a calcium response in those cells. Additionally, we show that both global and δ cell targeted deletion of MRAP2 abrogates the insulinostatic effect of ghrelin. Together, these findings establish that ghrelin signaling within δ cells is essential for the inhibition of insulin release and identify MRAP2 as a regulator of insulin secretion.