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Fibrillar collagen deposition, stiffness and downstream signalling support the development of leiomyomas (LMs), common benign mesenchymal tumours of the uterus, and are associated with aggressiveness in multiple carcinomas. Compared with epithelial carcinomas, however, the impact of fibrillar collagens on malignant mesenchymal tumours, including uterine leiomyosarcoma (uLMS), remains elusive. In this study, we analyse the network morphology and density of fibrillar collagens combined with the gene expression within uLMS, LM and normal myometrium (MM). We find that, in contrast to LM, uLMS tumours present low collagen density and increased expression of collagen‐remodelling genes, features associated with tumour aggressiveness. Using collagen‐based 3D matrices, we show that matrix metalloproteinase‐14 (MMP14), a central protein with collagen‐remodelling functions that is particularly overexpressed in uLMS, supports uLMS cell proliferation. In addition, we find that, unlike MM and LM cells, uLMS proliferation and migration are less sensitive to changes in collagen substrate stiffness. We demonstrate that uLMS cell growth in low‐stiffness substrates is sustained by an enhanced basal yes‐associated protein 1 (YAP) activity. Altogether, our results indicate that uLMS cells acquire increased collagen remodelling capabilities and are adapted to grow and migrate in low collagen and soft microenvironments. These results further suggest that matrix remodelling and YAP are potential therapeutic targets for this deadly disease.

Abstract Resource pulses are ecologically important phenomenon that occur in most ecosystems globally. Following optimal foraging theory, many consumers switch to pulsatile foods when available, examples of which include fruit mast and vulnerable young prey. Yet how the availability of resource pulses shapes the ecology of predators is still an emerging area of research; and how much individual variation there is in response to pulses is not well understood. We hypothesized that resource pulses would lead to dietary convergence in our population, which we tested by tracking both population‐level and individual coyote diets for 3 years in South Carolina, USA. We (1) described seasonal dietary shifts in relation to resource pulses; (2) compared male and female diets across seasons; and (3) tested this dietary convergence hypothesis by quantifying individual dietary variation both across and within periods when resource pulses were available. We found that pulses of white‐tailed deer fawns and blackberries composed over half of coyote diet in summer, and persimmon fruits were an important component in fall. Male and female coyotes generally had similar diets, but males consumed more deer in fall, perhaps driven by scavenging more. We found support for our dietary convergence hypothesis, where individuals had more similar diets during resource pulses compared to a non‐pulse period. We also found that this convergence happened before peak availability, suggesting a non‐symmetric response to pulse availability. We show that nearly all coyotes eat fawns, suggesting that targeted efforts to remove “fawn killers” would be in vain. Instead, given how quickly coyotes collectively converge on resource pulses, our findings show that resource pulses could potentially be used by managers to alter the behavior of apex predators. More broadly, we open a new line of inquiry into how variation in individual foraging decisions scales up to shape the effects of resource pulses on ecological communities.

Abstract Tropical cyclones (TCs) that undergo rapid intensification (RI) before landfall are notoriously difficult to predict and have caused tremendous damage to coastal regions in the United States. Using downscaled synthetic TCs and physics‐based models for storm tide and rain, we investigate the hazards posed by TCs that rapidly intensify before landfall under both historical and future mid‐emissions climate scenarios. In the downscaled synthetic data, the percentage of TCs experiencing RI is estimated to rise across a significant portion of the North Atlantic basin. Notably, future climate warming causes large increases in the probability of RI within 24 hr of landfall. Also, our analysis shows that RI events induce notably higher rainfall hazard levels than non‐RI events with equivalent TC intensities. As a result, RI events dominate increases in 100‐year rainfall and storm tide levels under climate change for most of the US coastline.

Abstract Cowpea, Vigna unguiculata L. Walp., is a diploid warm‐season legume of critical importance as both food and fodder in sub‐Saharan Africa. This species is also grown in Northern Africa, Europe, Latin America, North America, and East to Southeast Asia. To capture the genomic diversity of domesticates of this important legume, de novo genome assemblies were produced for representatives of six subpopulations of cultivated cowpea identified previously from genotyping of several hundred diverse accessions. In the most complete assembly (IT97K‐499‐35), 26,026 core and 4963 noncore genes were identified, with 35,436 pan genes when considering all seven accessions. GO terms associated with response to stress and defense response were highly enriched among the noncore genes, while core genes were enriched in terms related to transcription factor activity, and transport and metabolic processes. Over 5 million single nucleotide polymorphisms (SNPs) relative to each assembly and over 40 structural variants >1 Mb in size were identified by comparing genomes. Vu10 was the chromosome with the highest frequency of SNPs, and Vu04 had the most structural variants. Noncore genes harbor a larger proportion of potentially disruptive variants than core genes, including missense, stop gain, and frameshift mutations; this suggests that noncore genes substantially contribute to diversity within domesticated cowpea.

Background Annual heart transplant (HT) volumes have increased, as have post‐HT outpatient care needs. Data on HT‐related emergency department (ED) visits are limited. Methods and Results A retrospective analysis of 177 450 HT patient ED visits from the 2009 to 2018 Nationwide Emergency Department Sample was conducted. HT recipients, primary diagnoses, and comorbidities associated with ED visits were identified via International Classification of Diseases, Ninth Revision (ICD‐9) and International Classification of Diseases, Tenth Revision (ICD‐10) codes. Multivariable logistic regression was used to predict outcomes of hospital admission and death. HT volumes and HT‐related ED visits increased from 2009 to 2018. Infection was the most common primary diagnosis (24%), and cardiac primary diagnoses represented 10% of encounters. Hospital admissions occurred in 48% of visits, but overall mortality was low (1.6%). Length of stay was 3.1 days (interquartile range, 1.6–5.9 days), and comorbidity burden was high: 42% had hypertension, 38% had diabetes, and 31% had ≥2 comorbidities. Those aged ≥65 years had significantly higher odds of admission (odds ratio [OR], 2.14 [95% CI, 1.97–2.33]) and death (OR, 2.06 [95% CI, 1.61–2.62]). Comorbidities increased odds of admission (OR, 1.62 [95% CI, 1.51–1.75]) but not death. Renal primary diagnosis had the highest risk of admission (OR, 4.1 [95% CI, 3.6–4.6]), but cardiac primary diagnosis had the highest odds of death (OR, 11.6 [95% CI, 9.1–14.8]). Conclusions HT‐related ED visits increased from 2009 to 2018 with high admission rates but low in‐hospital mortality, suggesting an opportunity to improve prehospital care. Older patients and those with cardiac primary diagnoses had the highest risk of death. The observed contrast between predictors of admission and mortality signals a need for further study to improve risk stratification and outpatient care strategies.

Abstract Background Refugees and migrants face suboptimal involvement in spaces for primary healthcare decision‐making. Given the rising numbers of resettled refugees and migrants in primary care settings in the United States, there is an urgent need for patient‐centred outcome research in practice‐based research networks (PBRNs) with diverse ethnolinguistic communities. This study explored whether researchers, clinicians and patients would achieve consensus on (1) a common set of clinical problems that were applicable across a PBRN and (2) potential clinical interventions to address those problems to inform a patient‐centred outcomes research (PCOR) study in a similar research network. Methods In this qualitative participatory health research study, patients from diverse ethnolinguistic communities and clinicians from seven practices in a US PBRN discussed preferences for PCOR responsive to patients and the clinicians who serve them in language‐discordant settings. Researchers and an advisory panel that included patients and clinicians from each participating practice held regular advisory meetings to monitor progress on project milestones and solve emerging problems. Participants took part in 10 sessions using Participatory Learning in Action and the World Café methods to identify and prioritise their ideas, using questions set for them by the advisory panel. Data were analysed based on principles of qualitative thematic content analysis. Results Participants identified common barriers in language‐discordant healthcare settings, principally patient‐clinician communication barriers and suggestions to overcome these barriers. A key finding was an unanticipated consensus about the need for attention to healthcare processes rather than a clinical research priority. Negotiation with research funders enabled further analysis of potential interventions for care processes to improve communication and shared decision‐making in consultations and the practice as a whole. Conclusion PCOR studies should examine interventions for improving communication between patients from diverse ethnolinguistic communities and primary care staff if the sorts of harms experienced by patients experiencing language‐discordant healthcare are to be reduced or prevented. Flexibility and responsiveness from funders to unanticipated findings are key structural supports for participatory health research in primary care clinical settings with this population and others who experience marginalisation and exclusion. Patient or Public Contribution Patients and clinicians participated in the study both in the formulation of the study question, data collection, analysis and dissemination of these results; consented to their individual participation; and reviewed early drafts of the manuscript.

The ability of ticks to adapt to different ecological zones, coupled with the spread of infectious pathogens negatively affects livestock production and thus, there is a need for better control strategies. However, control measures within a geographical region can only be effective if there is available information on tick population dynamics and ecology. This study focused on ticks infesting livestock in the Kassena-Nankana Districts of the Upper East Region of Ghana. The ticks were morphologically identified, variables such as season, animal host, and predilection sites were recorded, and the data were analyzed using STATA version 13. Out of 448 livestock examined, tick infestation in cattle was (78.60%), followed by sheep (25%) and goats (5.88%). A total of 1,550 ticks including nymphs (303) and adults (1,247) were collected. Adult ticks were found to be significantly associated with season (p

Abstract Over the past three decades, China's government has implemented many projects under its ecological compensation policy, including paying compensation fees for habitat creation to redress natural habitat losses caused by development. However, a critical evaluation of both the policy design and its ecological outcomes, has not previously been carried out. We assemble diverse data sources to provide the first evaluation of China's eco‐compensation policy and practice, identifying several challenges. In policy, the pricing of forest restoration fees is insufficient in several provinces, and there is no requirement for use of biodiversity metrics or for ecological equivalence of compensation and losses. In practice, only 23% of a sample of 31 developments applied quantitative biodiversity metrics, and fewer than 1% of China's local governments have disclosed information regarding compensation implementation. Thus, to improve the validity of its compensation policy and practice to better secure biodiversity, China may need to embrace higher compensation standards, having first prevented ecological losses where possible. Equally important, China may also need to improve compensation governance for data tracking and conservation effectiveness monitoring.

Abstract Background Lineage‐tracing experiments have established that the central region of the mature intervertebral disc, the nucleus pulposus (NP), develops from the embryonic structure called “the notochord”. However, changes in the cells derived from the notochord which form the NP (i.e., notochordal cells [NCs]), in terms of their phenotype and functional identity from early developmental stages to skeletal maturation are less understood. These key issues require further investigation to better comprehend the role of NCs in homeostasis and degeneration as well as their potential for regeneration. Progress in utilizing NCs is currently hampered due to poor consistency and lack of consensus methodology for in vitro NC extraction, manipulation, and characterization. Methods Here, an international group has come together to provide key recommendations and methodologies for NC isolation within key species, numeration, in vitro manipulation and culture, and characterization. Results Recommeded protocols are provided for isolation and culture of NCs. Experimental testing provided recommended methodology for numeration of NCs. The issues of cryopreservation are demonstrated, and a pannel of immunohistochemical markers are provided to inform NC characterization. Conclusions Together we hope this article provides a road map for in vitro studies of NCs to support advances in research into NC physiology and their potential in regenerative therapies.

Abstract Myocardial infarction (MI) is one of the leading causes of death and disability. Recently developed cardiac patches provide mechanical support and additional conductive paths to promote electrical signal propagation in the MI area to synchronize cardiac excitation and contraction. Cardiac patches based on conductive polymers offer attractive features; however, the modest levels of elasticity and high impedance interfaces limit their mechanical and electrical performance. These structures also operate as permanent implants, even in cases where their utility is limited to the healing period of tissue damaged by the MI. The work presented here introduces a highly conductive cardiac patch that combines bioresorbable metals and polymers together in a hybrid material structure configured in a thin serpentine geometry that yields elastic mechanical properties. Finite element analysis guides optimized choices of layouts in these systems. Regular and synchronous contraction of human induced pluripotent stem cell‐derived cardiomyocytes on the cardiac patch and ex vivo studies offer insights into the essential properties and the bio‐interface. These results provide additional options in the design of cardiac patches to treat MI and other cardiac disorders.

Abstract Large carcasses often attract multiple carnivore species, so subordinate carnivores must weigh the reward of a profitable meal with the risk of being attacked by dominant carnivores. These risk–reward trade‐offs are likely influenced by a variety of factors, including scale‐dependent risk from dominant carnivores (e.g., short‐ vs. long‐term risk) and the amount of carcass remaining. In the southeastern United States, human hunters provision a large amount of white‐tailed deer carrion, which appears to be an important food source for coyotes (a novel top predator), but we know little about how coyotes influence the scavenging behavior of smaller carnivores. In this study, we evaluated the relative importance of risk from coyotes, vulture activity, forest structure, and remaining food on bobcat, gray fox, raccoon, and opossum scavenging by deploying 71 deer carcasses within a managed forest in South Carolina during January 2020 and 2021. We found that coyotes only had direct effects on bobcat behavior, suggesting that competition for carcasses was greatest between these two species. However, the relative importance of long‐ versus short‐term risk from coyotes was dependent on the stage in the scavenging process. Effects from forest structure were also stage‐dependent, where tree density and age were related to carcass discovery for bobcats and raccoons, while minimal understory cover facilitated bobcat, gray fox, and opossum scavenging, despite short‐term risk from coyotes. Vulture activity appeared to serve as a cue for gray foxes to discover carcasses. Ultimately, we found that risk from coyotes had species‐specific and context‐dependent effects on smaller carnivore scavenging. This represents some of the first direct evidence of how coyotes alter smaller carnivore behavior in a region where coyotes are a novel top predator. However, forest structure (particularly understory cover) seemed to mediate risk from coyotes, highlighting how habitat can influence predator–predator interactions. Future research should also investigate these interactions during other times of the year and try to quantify how human‐provisioned carcasses influence populations and communities.

Abstract Wild pigs (Sus scrofa), which are invasive in many regions globally, can alter ecosystems and compete with native species through interference competition and resource exploitation. Wild pig impacts on other species may increase with greater niche overlap, which could vary over time based on environmental conditions, resource availability, or biological traits like diet, especially as seasonal variation in wild pig diet has been widely documented. A limited number of studies have assessed spatial or temporal overlap between native species and invasive wild pigs, with only a handful simultaneously assessing overlap in these niche dimensions. We investigated the potential for interspecific interactions involving invasive wild pigs in the Piedmont region of South Carolina, USA, by examining seasonal spatiotemporal overlap with other wildlife using N‐mixture models and diel activity overlap analyses. Site use by white‐tailed deer (Odocoileus virginianus) and coyote (Canis latrans) was negatively associated with wild pig activity in the fall, when the species had high diel activity overlap, indicating spatial partitioning could reduce interference competition with wild pigs in this season. Conversely, white‐tailed deer site use was positively associated with wild pig activity in the winter, suggesting higher spatial overlap may be necessary if resources are limited. Site use by bobcat (Lynx rufus) and nine‐banded armadillo (Dasypus novemcinctus) in the spring, along with raccoon (Procyon lotor) and wild turkey (Meleagris gallopavo) site use in the summer, was positively associated with wild pig activity. With the exception of diurnal wild turkey, diel activity overlap between these species and wild pigs was high, although temporal partitioning could have occurred at finer spatiotemporal scales than we examined. Our results collectively emphasize the importance of accounting for seasonal spatial and temporal responses by individual species to invasive wild pigs, with special consideration given to species in seasons where high niche overlap with wild pigs is anticipated.

Abstract Background In vitro studies using nucleus pulposus (NP) cells are commonly used to investigate disc cell biology and pathogenesis, or to aid in the development of new therapies. However, lab‐to‐lab variability jeopardizes the much‐needed progress in the field. Here, an international group of spine scientists collaborated to standardize extraction and expansion techniques for NP cells to reduce variability, improve comparability between labs and improve utilization of funding and resources. Methods The most commonly applied methods for NP cell extraction, expansion, and re‐differentiation were identified using a questionnaire to research groups worldwide. NP cell extraction methods from rat, rabbit, pig, dog, cow, and human NP tissue were experimentally assessed. Expansion and re‐differentiation media and techniques were also investigated. Results Recommended protocols are provided for extraction, expansion, and re‐differentiation of NP cells from common species utilized for NP cell culture. Conclusions This international, multilab and multispecies study identified cell extraction methods for greater cell yield and fewer gene expression changes by applying species‐specific pronase usage, 60–100 U/ml collagenase for shorter durations. Recommendations for NP cell expansion, passage number, and many factors driving successful cell culture in different species are also addressed to support harmonization, rigor, and cross‐lab comparisons on NP cells worldwide.

Abstract Biodiversity may play a key role in generating the well‐being benefits of visiting green‐spaces. The ability of people to accurately perceive variation in biodiversity is, however, unclear and evidence supporting links between biodiversity exposure and well‐being outcomes remains equivocal. In part, this is due to the paucity of controlled experimental studies that deal adequately with confounding factors that covary with biodiversity. Attention restoration theory (ART) proposes that natural environments contain many softly fascinating stimuli that provide visitors with a sense of separation from their normal settings and routines, switching off direct attention and allowing recovery from attention fatigue. Increased biodiversity could increase these stimuli, and ART therefore potentially provides a mediating effect linking biodiversity to well‐being. Here, we conduct a controlled experiment in which participants virtually experience urban green‐space containing high and low levels of avian biodiversity (altered by manipulating bird song). Respondents accurately identified the contrast in biodiversity and reported greater enjoyment of the high biodiversity treatment than the low diversity control. Higher biodiversity did not, however, elicit greater self‐reported stimulation or restoration, and did not increase perceived restorativeness scores or attentional capacity (quantified using the Digit Span Backwards attention test). Respondents that were more connected to nature, however, had greater attentional capacity following exposure to green‐space. Our study provides rare experimental evidence that people can accurately detect variation in biodiversity, that high avian diversity boosts visitor perceptions of urban green‐space quality, and that people with increased nature connectedness show enhanced attentional capacity following an exposure to green‐space. A free Plain Language Summary can be found within the Supporting Information of this article.

Abstract Filanesib is a first‐in‐class kinesin spindle protein inhibitor which demonstrated safety and encouraging activity in combination with bortezomib and dexamethasone in relapsed/refractory multiple myeloma in a preliminary analysis of dose‐escalation phase results. This multicenter study included first a dose‐escalation phase to determine maximum tolerated dose of two schedules of filanesib, bortezomib, and dexamethasone and a subsequent dose‐expansion phase using the maximum tolerated doses. In the dose‐expansion phase, 28 patients were evaluable for safety and efficacy. The most common grade ≥3 adverse events were neutropenia (21%) and anemia (18%), which were noncumulative and reversible, and hypertension (18%). The overall response rate was 43% with median duration of response not yet reached (range, 2.8–23.7+ months) with median follow‐up of 6.3 months. A post hoc analysis incorporated 29 dose‐escalation phase patients who received therapeutic filanesib doses, with an overall response rate of 39% and median duration of response of 18.0 months among the 57 total patients with median progression‐free survival of 8.5 months. Notably, the PFS of high risk patients was comparable at 8.5 months, driven by the patients with 1q21 gain, characterized by increased MCL‐1 expression, with a PFS of 9.1 months versus 3.5 months for the remainder of high risk patients. Patients with t(11;14) also had an encouraging PFS of 15.0 months. The combination of filanesib, bortezomib, and dexamethasone continues to show safety and encouraging activity in relapsed/refractory multiple myeloma, particularly in those patients with 1q21 gain and t(11;14).

Old world monkeys separated from the great apes, including the ancestor of humans, about 25 million years ago, but most of the genes in humans and various nonhuman primates are quite similar even though their anatomical appearances are quite different. Like other mammals, primates have four tropomyosin genes (TPM1, TPM2, TPM3, and TPM4) each of which generates a multitude of TPM isoforms via alternative splicing. Only TPM1 produces two sarcomeric isoforms (TPM1α and TPM1κ), and TPM2, TPM3, and TPM4 each generate one sarcomeric isoform. We have cloned and sequenced TPM1α, TPM1κ, TPM2α, TPM3α, and TPM4α with RNA from cynomolgus (Cyn) monkey hearts and skeletal muscle. We believe this is the first report of directly cloning and sequencing of these monkey transcripts. In the Cyn monkey heart, the rank order of TPM isoform expression is TPM1α > TPM2α > TPM1κ > TPM3α > TPM4α. In the Cyn monkey skeletal muscle, the rank order of expression is TPM1α > TPM2α > TPM3α > TPM1κ > TPM4α. The major differences in the human heart are the increased expression of TPM1κ, although TPM1α is still the dominant transcript. In the Cyn monkey heart, the only sarcomeric TPM isoform at the protein level is TPM1α. This is in contrast to human hearts where TPM1α is the major sarcomeric isoform but a lower quantity of TPM1κ, TPM2α, and TPM3α is also detected at the protein level. These differences of tropomyosin and/or other cardiac protein expression in human and Cyn monkey hearts may reflect the differences in physiological activities in daily life.

Abstract Background Intervertebral disc (IVD) degeneration continues to be a major global health challenge, with strong links to lower back pain, while the pathogenesis of this disease is poorly understood. In cartilage, much more is known about mechanotransduction pathways involving the strain‐generated potential (SGP) and function of voltage‐gated ion channels (VGICs) in health and disease. This evidence implicates a similar important role for VGICs in IVD matrix turnover. However, the field of VGICs, and to a lesser extent the SGP, remains unexplored in the IVD. Methods A two‐step process was utilized to investigate the role of VGICs in the IVD. First, immunohistochemical staining was used to identify and localize several different VGICs in bovine and human IVDs. Second, a pilot study was conducted on the function of L‐type voltage gated calcium channels (VGCCs) by inhibiting these channels with nifedipine (Nf) and measuring calcium influx in monolayer or gene expression from 3D cell‐embedded alginate constructs subject to dynamic compression. Results Several VGICs were identified at the protein level, one of which, Cav2.2, appears to be upregulated with the onset of human IVD degeneration. Inhibiting L‐type VGCCs with Nf supplementation led to an altered cell calcium influx in response to osmotic loading as well as downregulation of col 1a, aggrecan and ADAMTS‐4 during dynamic compression. Conclusions This study demonstrates the presence of several VGICs in the IVD, with evidence supporting a role for L‐type VGCCs in mechanotransduction. These findings highlight the importance of future detailed studies in this area to fully elucidate IVD mechanotransduction pathways and better inform treatment strategies for IVD degeneration.

Abstract Population models often require detailed information on sex‐, age‐, or size‐specific abundances, but population monitoring programs cannot always acquire data at the desired resolution. Thus, state uncertainty in monitoring data can potentially limit the demographic resolution of management decisions, which may be particularly problematic for stage‐ or size‐structured species subject to consumptive use. American alligators (Alligator mississippiensis; hereafter alligator) have a complex life history characterized by delayed maturity and slow somatic growth, which makes the species particularly sensitive to overharvest. Though alligator populations are subject to recreational harvest throughout their range, the most widely used monitoring method (nightlight surveys) is often unable to obtain size class‐specific counts, which limits the ability of managers to evaluate the effects of harvest policies. We constructed a Bayesian integrated population model (IPM) for alligators in Georgetown County, SC, USA, using records of mark–recapture–recovery, clutch size, harvest, and nightlight survey counts collected locally, and auxiliary information on fecundity, sex ratio, and somatic growth from other studies. We created a multistate mark–recapture–recovery model with six size classes to estimate survival probability, and we linked it to a state‐space count model to derive estimates of size class‐specific detection probability and abundance. Because we worked from a count dataset in which 60% of the original observations were of unknown size, we treated size class as a latent property of detections and developed a novel observation model to make use of information where size could be partly observed. Detection probability was positively associated with alligator size and water temperature, and negatively influenced by water level. Survival probability was lowest in the smallest size class but was relatively similar among the other five size classes (>0.90 for each). While the two nightlight survey count sites exhibited relatively stable population trends, we detected substantially different patterns in size class‐specific abundance and trends between each site, including 30%–50% declines in the largest size classes at the site with greater harvest pressure. Here, we illustrate the use of IPMs to produce high‐resolution output of latent population structure that is partially observed during the monitoring process.

Abstract Coyotes (Canis latrans) colonized the southeastern United States over the last century as large predators, including the red wolf (Canis rufus) and eastern cougar (Puma concolor), were extirpated from the region. As a generalist carnivore, the coyote preys on white‐tailed deer (Odocoileus virginianus) and various smaller mammals, birds, and vegetation. While resource selection by coyotes has been well documented at the home‐range scale, little is known about their foraging behavior, which is an important factor in thoroughly understanding influences of coyotes on prey and sympatric carnivores. We assessed third‐order resource selection of coyotes at sites across Alabama, Georgia, and South Carolina during 2015–2016. Using GPS collars, we tracked 41 resident coyotes across four calendar seasons and identified suspected foraging areas using recursive analysis where individuals repeatedly returned to known locations. We found that resident coyotes selected for open landcover types throughout the year, while avoiding primary and secondary roads. Additionally, resident coyotes avoided forested landcover types while selecting for forest edges except from April to June when they foraged within interior forest away from edges. Previous studies have documented substantive predation rates on white‐tailed deer neonates by coyotes, and that fawn mortality may increase in forested landscapes away from forest edge. Our findings indicate that foraging coyotes may select forest cover types during spring where fawns are more vulnerable to predation. Additionally, there has been debate in the literature as to how coyotes obtain consistent levels of deer in their diets outside of fawning and fall hunting seasons. Our study indicates that use of road‐kill carcasses by coyotes was an unlikely explanation for the presence of deer in coyote diets throughout the year, as coyotes in our study were not observed using roads during foraging excursions.

Abstract Difelikefalin, a selective kappa opioid receptor agonist designed to limit central nervous system (CNS) penetration, is under development for the treatment of pruritus. Its hydrophilic, small‐peptidic structure limits CNS entry, minimizing potential CNS‐mediated adverse events (AEs). This study assessed the effect of difelikefalin on key relevant measures of respiratory depression in healthy volunteers. This single‐center, randomized, double‐blind, placebo‐controlled, three‐way crossover study enrolled healthy, nonsmoking volunteers. Subjects were randomized to 1 of 3 treatment sequences of difelikefalin (1.0 or 5.0 mcg/kg i.v.) or placebo on sequential days with an intervening 24 (±2) h washout period. The primary end points included incidence of increased end‐tidal carbon dioxide (ETCO2) greater than or equal to 10 mm Hg versus baseline or a level greater than 50 mm Hg sustained greater than or equal to 30 seconds, and incidence of reduction in saturation of peripheral oxygen (SpO2) to less than 92% sustained greater than or equal to 30 seconds. Secondary end points included incidence of reduced respiratory rate and other safety assessments. Fifteen subjects were randomized and completed the study. No subject on placebo or difelikefalin met the increased ETCO2 or reduced SpO2 primary end point criteria for respiratory depression. All respiratory measures in each group remained near baseline values during 4‐h postdose observations. No subject met the reduced respiratory rate criterion or experienced clinically significant changes in ETCO2, SpO2, or respiratory rate. The most commonly reported treatment‐emergent AEs (TEAEs; ≥20% of subjects) were paresthesia, hypoesthesia, and somnolence in the difelikefalin arms. All TEAEs were mild and resolved without intervention. Difelikefalin 1.0 and 5.0 mcg/kg i.v. did not produce respiratory depression.

Abstract Fibrillar collagens promote cell proliferation, migration, and survival in various epithelial cancers and are generally associated with tumor aggressiveness. However, the impact of fibrillar collagens on soft tissue sarcoma behavior remains poorly understood. Unexpectedly, this study finds that fibrillar collagen‐related gene expression is associated with favorable patient prognosis in rhabdomyosarcoma. By developing and using collagen matrices with distinct stiffness and in vivo‐like microarchitectures, this study uncovers that the activation of DDR1 has pro‐apoptotic and of integrin β1 pro‐survival function, specifically in 3D rhabdomyosarcoma cell cultures. It demonstrates that rhabdomyosarcoma cell‐intrinsic or extrinsic matrix remodeling promotes cell survival. Mechanistically, the 3D‐specific collagen‐induced apoptosis results from a dual DDR1‐independent and a synergistic DDR1‐dependent TRPV4‐mediated response to mechanical confinement. Altogether, these results indicate that dense microfibrillar collagen‐rich microenvironments are detrimental to rhabdomyosarcoma cells through an apoptotic response orchestrated by the induction of DDR1 signaling and mechanical confinement. This mechanism helps to explain the preference of rhabdomyosarcoma cells to grow in and metastasize to low fibrillar collagen microenvironments such as the lung.

Abstract Desmoplasia is characteristic of pancreatic ductal adenocarcinoma (PDAC), which exhibits 5‐year survival rates of 3%. Desmoplasia presents physical and biochemical barriers that contribute to treatment resistance, yet depleting the stroma alone is unsuccessful and even detrimental to patient outcomes. This study is the first demonstration of targeted photoactivable multi‐inhibitor liposomes (TPMILs) that induce both photodynamic and chemotherapeutic tumor insult, while simultaneously remediating desmoplasia in orthotopic PDAC. TPMILs targeted with cetuximab (anti‐EGFR mAb) contain lipidated benzoporphyrin derivative (BPD‐PC) photosensitizer and irinotecan. The desmoplastic tumors comprise human PDAC cells and patient‐derived cancer‐associated fibroblasts. Upon photoactivation, the TPMILs induce 90% tumor growth inhibition at only 8.1% of the patient equivalent dose of nanoliposomal irinotecan (nal‐IRI). Without EGFR targeting, PMIL photoactivation is ineffective. TPMIL photoactivation is also sixfold more effective at inhibiting tumor growth than a cocktail of Visudyne‐photodynamic therapy (PDT) and nal‐IRI, and also doubles survival and extends progression‐free survival by greater than fivefold. Second harmonic generation imaging reveals that TPMIL photoactivation reduces collagen density by >90% and increases collagen nonalignment by >103‐fold. Collagen nonalignment correlates with a reduction in tumor burden and survival. This single‐construct phototoxic, chemotherapeutic, and desmoplasia‐remediating regimen offers unprecedented opportunities to substantially extend survival in patients with otherwise dismal prognoses.

Abstract To mitigate coyote (Canis latrans) introgression in the wild red wolf (Canis rufus) genome, the United States Fish and Wildlife Service (USFWS) Red Wolf Recovery Program used a combination of reproductive sterilization and lethal removal of coyotes to minimize hybridization and increase the endangered red wolf population. Although sterilization assisted in limiting coyote introgression to ≤4% in the wild red wolf genome, its potential negative effect on coyote and hybrid abundance and density is unknown. Using long‐term capture–mark–recapture and radio‐telemetry data collected on red wolves, coyotes, and hybrids under the USFWS Red Wolf Adaptive Management Plan implemented during 2000–2013, we explored three areas of research: (1) spatial modeling to correlate land cover characteristics with the relative probabilities of capture for red wolves, coyotes, and hybrids; (2) survival analysis of radio‐marked canids; and (3) annual population estimates for the three Canis taxa. We detected no differences in the relative probability of capture among Canis taxa. Red wolves, coyotes, and hybrids were most frequently captured in areas proximate to road networks with low canopy cover (i.e., cropland) and away from coastal bottomland forests. Annual apparent survival for red wolves and hybrids was greater than survival for coyotes; however, wolves and hybrids exhibited similar annual survival. Mortality of coyotes and hybrids was predominately attributed to deliberate take through lethal control by the USFWS biologists and harvest by hunters and trappers. We observed annual densities of coyotes ranging between 2.5 and 21.5 coyotes/1000 km2, with densities annually increasing during 2005–2013 when the red wolf population plateaued before declining after 2013. Despite the increase in coyote density, our density estimates are less than most estimates reported throughout the coyote's geographic range, and similar to those reported in areas where coyote populations are limited by extreme environments such as their northern range limits in Alaska, United States, and Canada. Our findings indicate that red wolf presence and federal management of coyotes using fertility control may have limited coyote densities in northeastern North Carolina.

Abstract In canids, resident breeders hold territories but require different resources than transient individuals (i.e., dispersers), which may result in differential use of space, land cover, and food by residents and transients. In the southeastern United States, coyote (Canis latrans) reproduction occurs during spring and is energetically demanding for residents, but transients do not reproduce and therefore can exhibit feeding behaviors with lower energetic rewards. Hence, how coyotes behave in their environment likely differs between resident and transient coyotes. We captured and monitored 36 coyotes in Georgia during 2018–2019 and used data from 11 resident breeders, 12 predispersing residents (i.e., offspring of resident breeders), and 11 transients to determine space use, movements, and relationships between these behaviors and landcover characteristics. Average home range size for resident breeders and predispersing offspring was 20.7 ± 2.5 km² and 50.7 ± 10.0 km², respectively. Average size of transient ranges was 241.4 ± 114.5 km². Daily distance moved was 6.3 ± 3.0 km for resident males, 5.5 ± 2.7 km for resident females, and 6.9 ± 4.2 km for transients. We estimated first‐passage time values to assess the scale at which coyotes respond to their environment, and used behavioral change‐point analysis to determine that coyotes exhibited three behavioral states. We found notable differences between resident and transient coyotes in regard to how landcover characteristics influenced their behavioral states. Resident coyotes tended to select for areas with denser vegetation while resting and foraging, but for areas with less dense vegetation and canopy cover when walking. Transient coyotes selected areas closer to roads and with lower canopy cover while resting, but for areas farther from roads when foraging and walking. Our findings suggest that behaviors of both resident and transient coyotes are influenced by varying landcover characteristics, which could have implications for prey.

Abstract Context Migration is one of the most politically pressing issues of the 21st century but migrant health remains an under‐researched area. The International Collaboration for Participatory Health Research (ICPHR) working group on migration developed this position statement to address opportunities and challenges in relation to migrant health. It aims to contribute to a shift from a deficit model that sees migrants as passively affected by policies to their reconceptualization as citizens who are engaged in the co‐creation of solutions. Methods This paper examines the opportunities and challenges posed by the use of PHR with migrants. It draws on a broad literature to provide examples of successful PHR with migrants and highlights critical issues for consideration. Findings Successful initiatives illustrate the value of engaging migrants in the definition of the research agenda, the design and implementation of health interventions, the identification of health‐protective factors and the operationalization and validation of indicators to monitor progress. Within increasingly super diverse contexts, fragmented community landscapes that are not necessarily constructed along ethnicity traits, inadequate structures of representation, local tensions and operational barriers can hamper meaningful PHR with migrants. Conclusion For each research context, it is essential to gauge the ‘optimal’ level and type of participation that is more likely to leverage migrants’ empowerment. The development of Monitoring and Evaluation tools and methodological strategies to manage inter‐stakeholder discrepancies and knowledge translation gaps are steps in this direction. Patient or public contribution This paper draws from contributions of migrant populations and other stakeholders to policymaking.

Abstract Future coastal flood hazard at many locations will be impacted by both tropical cyclone (TC) change and relative sea‐level rise (SLR). Despite sea level and TC activity being influenced by common thermodynamic and dynamic climate variables, their future changes are generally considered independently. Here, we investigate correlations between SLR and TC change derived from simulations of 26 Coupled Model Intercomparison Project Phase 6 models. We first explore correlations between SLR and TC activity by inference from two large‐scale factors known to modulate TC activity: potential intensity (PI) and vertical wind shear. Under the high emissions SSP5‐8.5, SLR is strongly correlated with PI change (positively) and vertical wind shear change (negatively) over much of the western North Atlantic and North West Pacific, with global mean surface air temperature (GSAT) modulating the co‐variability. To explore the impact of the joint changes on flood hazard, we conduct climatological–hydrodynamic modeling at five sites along the US East and Gulf Coasts. Positive correlations between SLR and TC change alter flood hazard projections, particularly at Wilmington, Charleston and New Orleans. For example, if positive correlations between SLR and TC changes are ignored in estimating flood hazard at Wilmington, the average projected change to the historical 100 years storm tide event is under‐estimated by 12%. Our results suggest that flood hazard assessments that neglect the joint influence of these factors and that do not reflect the full distribution of GSAT change may not accurately represent future flood hazard.

Abstract Objectives The ECOG‐ACRIN Cancer Research Group trial E2511 recently demonstrated a potential benefit for the addition of veliparib to cisplatin‐etoposide (CE) in patients with extensive stage small cell lung cancer (ES‐SCLC) in a phase II randomized controlled trial. Secondary trial endpoints included comparison of the incidence and severity of neurotoxicity, hypothesized to be lower in the veliparib arm, and tolerability of the addition of veliparib to CE. Physician‐rated and patient‐reported neurotoxicity was also compared. Materials and Methods Patients randomized to veliparib plus CE (n = 64) or placebo plus CE (n = 64) completed the 11‐item Functional Assessment of Cancer Therapy Gynecologic Oncology Group Neurotoxicity (questionnaire pre‐treatment, end of cycle 4 [ie 3 months after randomization] and 3 months post‐treatment [ie 6‐months]). Adherence analysis was based on treatment forms. Results and Conclusion No significant differences in mean or magnitude of change in neurotoxicity scores were observed between treatment arms at any time point. However, patients in the placebo arm reported worsening neurotoxicity from baseline to 3‐months (M difference = −1.5, P = .045), compared to stable neurotoxicity in the veliparib arm (M difference = −0.2, P = .778). Weakness was the most common treatment‐emergent (>50%) and moderate to severe (>16%) symptom reported, but did not differ between treatment arms. The proportion of adherence to oral therapy in the overall sample was 75%. Three percent of patients reported clinically significant neurotoxicity that was not captured by physician assessment. Neurotoxicity scores were not different between treatment arms. The addition of veliparib to CE appeared tolerable, though weakness should be monitored. ClinicalTrials.gov Identifier NCT01642251.

Abstract Objective This study aimed to explore whether positive impacts were sustained and unanticipated ripple effects had occurred four years after the implementation of interventions to improve cross‐cultural communication in primary care. Background Sustaining the implementation of change using complex interventions is challenging. The EU‐funded “RESTORE” study implemented guidelines and training on cross‐cultural communication in five Primary Care sites in Europe, combining implementation theory (Normalisation Process Theory) with participatory methodology (participatory learning and action—PLA). There were positive impacts on knowledge, skills and clinical routines. Design, setting and participants Four of the five original sites (England, Ireland, Greece, The Netherlands) were available for this qualitative follow‐up study. The study population (N = 44) was primary healthcare staff and migrants, most of whom had participated in RESTORE. Intervention; main outcome measures PLA‐style focus groups and interviews explored routine practice during consultations with migrants. Etic cards based on the effects of RESTORE stimulated the discussion. Deductive framework analysis was performed in each country followed by comparative data analysis and synthesis. Results Changes in knowledge, attitudes and behaviour with regard to consultations with migrants were sustained and migrants felt empowered by their participation in RESTORE. There were ongoing concerns about macro level factors, like the political climate and financial policies, negatively affecting migrant healthcare. Conclusion There were sustained effects in clinical settings, and additional unanticipated positive ripple effects, due in part, from the participatory approach employed.

Background The past decade has seen tremendous growth in patients with ambulatory ventricular assist devices. We sought to identify patients that present to the emergency department (ED) at the highest risk of death. Methods and Results This retrospective analysis of ED encounters from the Nationwide Emergency Department Sample includes 2010 to 2017. Using a random sampling of patient encounters, 80% were assigned to development and 20% to validation cohorts. A risk model was derived from independent predictors of mortality. Each patient encounter was assigned to 1 of 3 groups based on risk score. A total of 44 042 ED ventricular assist device patient encounters were included. The majority of patients were male (73.6%),

Abstract Background Asthma is a disease of the lung and a systemic disease. Functional disorders are associated with multiple systemic abnormalities that have been explained by complexity models. The aim was to test the similarity in type and aetiology between the extra‐pulmonary symptoms of severe asthma and the symptoms of fibromyalgia. Methods One Hundred patients recruited from a specialist severe asthma clinic and 1751 people reporting different functional disorder diagnoses recruited via the internet completed the same 60‐item questionnaire. Symptom patterns were compared between groups using a new measure, the symptom pattern similarity index where 0 = no relationship, 1 = identical patterns between groups. Results Severe asthma patients report numerous extra‐pulmonary symptoms. The similarity index between the symptom pattern of the asthma patients with other groups was irritable bowel syndrome = 0.54, chronic fatigue syndrome = 0.69, and fibromyalgia = 0.75. The index between fibromyalgia and asthma patients with the most and least frequent extra‐pulmonary symptoms was 0.81 and 0.55 respectively. Conclusions Patients with severe asthma have numerous extra‐pulmonary symptoms similar in type and pattern to the symptoms of fibromyalgia. The similarity of the symptom pattern between asthma and fibromyalgia increases as the number of extra‐pulmonary symptoms increases as predicted by network theory and previously shown to be the case with other functional disorders. These findings support the hypothesis that functional disorders and extra‐pulmonary asthma symptoms have a common complexity or network aetiology. Evidence based behavioural interventions for fibromyalgia may be helpful for patients with severe asthma reporting extra‐pulmonary symptoms.

Abstract Net outcome‐type biodiversity policies are proliferating globally as perceived mechanisms to reconcile economic development and conservation objectives. The UK government's Environment Bill will mandate that most new developments in England demonstrate that they deliver a biodiversity net gain (BNG) to receive planning permission, representing the most wide‐ranging net outcome type policy globally. However, as with many nascent net‐outcome policies, the likely outcomes of mandatory BNG have not been explored empirically. We assemble all BNG assessments (accounting for ∼6% of England's annual housebuilding and other infrastructure) submitted from January 2020 to February 2021 in six early‐adopter councils who are implementing mandatory no net loss or BNG requirements in advance of the national adoption of mandatory BNG, and analyze the aggregate habitat changes proposed. Our sample is associated with a 34% reduction in the area of nonurban habitats, generally compensated by commitments to deliver smaller areas of higher quality habitat years later in the development project cycle. Ninety‐five percent of biodiversity units delivered in our sample come from habitats within or directly‐adjacent to the development footprint managed by the developers. However, we find that these gains fall within a governance gap whereby they risk being unenforceable, a challenge that is shared with other net outcome type policies implemented internationally.

Abstract Regular contact with nature provides multiple health benefits for people, but biodiversity is declining fast in an urbanizing world. Biodiversity offsets are implemented to compensate for the negative residual impacts of economic development projects on biodiversity, but the impacts on people who stand to lose biodiversity from their local environment are rarely considered. Offsetting typically involves creating, restoring or protecting biodiversity values at a specified site that can be located some distance away from the development site. In this article, we explore whether any relocation of nature is occurring due to development and offsets in Western Australia (WA); a jurisdiction with one of the world's few spatially referenced and comprehensive public offset registers. We analyzed data from 158 projects within the WA Environmental Offsets Register. We compared the location of development sites within 50 km (the urban and peri urban zone) and 500 km (~one day's drive) of the central business district (CBD) of Perth with the associated offset sites. The development and offset process together can be considered to contribute to a loss of urban nature as the offset sites tended to be further away from urban areas than the associated development sites. The offset sites were also located in significantly lower population density areas. However, offsets increased the publicly accessible land area by changing land ownership and creating amenity benefit by improving nature values on public land. Nevertheless, it is unclear to what extent relocation of nature further from people is balanced by increased public access to nature. In order to maintain nature connectedness, ecosystem service delivery and environmental justice in cities, we argue offset policies should require spatial proximity between impact and offset sites.

Abstract Fire exclusion and mismanaged grazing are globally important drivers of environmental change in mesic C4 grasslands and savannas. Although interest is growing in prescribed fire for grassland restoration, we have little long‐term experimental evidence of the influence of burn season on the recovery of herbaceous plant communities, encroachment by trees and shrubs, and invasion by exotic grasses. We conducted a prescribed fire experiment (seven burns between 2001 and 2019) in historically fire‐excluded and overgrazed grasslands of central Texas. Sites were assigned to one of four experimental treatments: summer burns (warm season, lightning season), fall burns (early cool season), winter burns (late cool season), or unburned (fire exclusion). To assess restoration outcomes of the experiment, in 2019, we identified old‐growth grasslands to serve as reference sites. Herbaceous‐layer plant communities in all experimental sites were compositionally and functionally distinct from old‐growth grasslands, with little recovery of perennial C4 grasses and long‐lived forbs. Unburned sites were characterized by several species of tree, shrub, and vine; summer sites were characterized by certain C3 grasses and forbs; and fall and winter sites were intermediate in composition to the unburned and summer sites. Despite compositional differences, all treatments had comparable plot‐level plant species richness (range 89–95 species/1000 m2). At the local‐scale, summer sites (23 species/m2) and old‐growth grasslands (20 species/m2) supported greater richness than unburned sites (15 species/m2), but did not differ significantly from fall or winter sites. Among fire treatments, summer and winter burns most consistently produced the vegetation structure of old‐growth grasslands (e.g., mean woody canopy cover of 9%). But whereas winter burns promoted the invasive grass Bothriochloa ischaemum by maintaining areas with low canopy cover, summer burns simultaneously limited woody encroachment and controlled B. ischaemum invasion. Our results support a growing body of literature that shows that prescribed fire alone, without the introduction of plant propagules, cannot necessarily restore old‐growth grassland community composition. Nonetheless, this long‐term experiment demonstrates that prescribed burns implemented in the summer can benefit restoration by preventing woody encroachment while also controlling an invasive grass. We suggest that fire season deserves greater attention in grassland restoration planning and ecological research.

Abstract The new global biodiversity framework (GBF) being developed under the Convention on Biological Diversity must drive action to reverse the ongoing decline of the Earth's biodiversity. Explicit, measurable goals that specify the outcomes we want to achieve are needed to set the course for this action. However, the current draft goals and targets fail to set out these clear outcomes. We argue that distinct outcome goals for species, ecosystems, and genetic diversity are essential and should specify net outcomes required for each. Net outcome goals such as “no net loss” do, however, have a controversial history, and loose specification can lead to perverse outcomes. We outline seven general principles to underpin net outcome goal setting that minimize risk of such perverse outcomes. Finally, we recommend inclusion of statements of impact in action targets that support biodiversity goals, and we illustrate the importance of this with an example from the draft GBF action targets. These modifications would help reveal the specific contribution each action would make to achieving the outcome goals and provide clarity on whether the successful achievement of action targets would be adequate to achieve the outcome goals and, in turn, the 2050 vision: living in harmony with nature.

Abstract In the USA, Species Conservation Banking is a prominent example of compensatory biodiversity impact mitigation, with an annual market value estimated at US$354.2 million. Species Conservation Banking represents a useful case study of a well‐established program that can provide empirical insights into the practicalities of implementing quantitative compensatory biodiversity mitigation on‐the‐ground. Using semi‐structured key‐informant interviews structured around well‐established technical challenges to compensatory mitigation, this study aimed to understand (i) how and why these challenges are or are not addressed in practice; and (ii) how these challenges relate to practical challenges faced by conservation banking stakeholders on‐the‐ground. Challenges identified included: (i) defining trading currencies and equivalence, (ii) regulatory and political uncertainty, (iii) regulatory agency capacity, will and knowledge, (iv) lack of policies, standards, and competition with other mitigation mechanisms, (v) long‐term uncertainty/longevity, and (vi) lack of species knowledge and data transparency. These challenges are numerous, diverse, interlinked and transdisciplinary, and collectively inhibit the ability of practitioners to resolve underlying technical challenges—a finding likely applicable to related biodiversity offset programs. To help address challenges and navigate this complexity, we formulate several recommendations for conservation banking stakeholders to improve the chances of beneficial biodiversity outcomes being achieved.

Abstract Anthropogenic carbon dioxide (CO2) emissions dominate uncertainties in the global carbon budget. Global inventories, such as the National Greenhouse Gas Inventories, have latencies of 12–24 months and may not keep pace with rapidly changing infrastructure, particularly in the developing world. Our work reveals that airborne and satellite imaging spectrometers provide 3–30 m spatial resolution and accurate quantification of CO2 emissions at the facility scale. Examples from 17 coal and gas fired power plants across the United States demonstrate robust correlation and 21% agreement on average between our remotely sensed estimates and simultaneous in situ measured emissions. We highlight four examples of coal‐fired power plants in India, Poland, and South Korea, where we quantify significant carbon dioxide emissions from power plants where limited public emissions data exist. Leveraging previous work on methane (CH4) plume detection, we present a strategy to exploit joint CO2 and CH4 plume imaging to quantify carbon emissions across widely distributed industrial infrastructure, including facilities that co‐emit CO2 and CH4. We show an example of a coal operation, where we attribute 25% of greenhouse gas emissions to coal extraction (CH4) and the remaining 75% to energy generation (CO2). Satellite spectrometers could track high emitting coal‐fired power plants that collectively contribute to 60% or more of global coal CO2 emissions. Multiple revisits and coordinated targeting of these high emitting facilities by multiple spaceborne instruments will be key to reducing uncertainties in global anthropogenic CO2 emissions and supporting emissions mitigation strategies.

Background Pediatric cardiomyopathy is a genetically heterogeneous disease with substantial morbidity and mortality. Current guidelines recommend genetic testing in children with hypertrophic, dilated, or restrictive cardiomyopathy, but practice variations exist. Robust data on clinical testing practices and diagnostic yield in children are lacking. This study aimed to identify the genetic causes of cardiomyopathy in children and to investigate clinical genetic testing practices. Methods and Results Children with familial or idiopathic cardiomyopathy were enrolled from 14 institutions in North America. Probands underwent exome sequencing. Rare sequence variants in 37 known cardiomyopathy genes were assessed for pathogenicity using consensus clinical interpretation guidelines. Of the 152 enrolled probands, 41% had a family history of cardiomyopathy. Of 81 (53%) who had undergone clinical genetic testing for cardiomyopathy before enrollment, 39 (48%) had a positive result. Genetic testing rates varied from 0% to 97% between sites. A positive family history and hypertrophic cardiomyopathy subtype were associated with increased likelihood of genetic testing (P=0.005 and P=0.03, respectively). A molecular cause was identified in an additional 21% of the 63 children who did not undergo clinical testing, with positive results identified in both familial and idiopathic cases and across all phenotypic subtypes. Conclusions A definitive molecular genetic diagnosis can be made in a substantial proportion of children for whom the cause and heritable nature of their cardiomyopathy was previously unknown. Practice variations in genetic testing are great and should be reduced. Improvements can be made in comprehensive cardiac screening and predictive genetic testing in first‐degree relatives. Overall, our results support use of routine genetic testing in cases of both familial and idiopathic cardiomyopathy. Registration URL: https://www.clinicaltrials.gov; Unique identifier: NCT01873963.

Abstract Purpose Notch signaling dysregulation is implicated in the development of pancreatic adenocarcinoma (PDAC). Tarextumab is a fully human IgG2 antibody that inhibits Notch2/3 receptors. Patients and Methods Aphase 2, randomized, placebo‐controlled, multicenter trial evaluated the activity of tarextumab in combination with nab‐paclitaxel and gemcitabine in patients with metastatic PDAC. Patients were stratified based on ECOG performance score and Ca 19‐9 level and randomized 1:1 to nab‐paclitaxel, gemcitabine with either tarextumab or placebo. Based on preclinical and phase Ib results suggesting a positive correlation between Notch3 gene expression and tarextumab anti‐tumor activity, patients were also divided into subgroups of low, intermediate, and high Notch3 gene expression. Primary endpoint was overall survival (OS) in all and in patients with the three Notch3 gene expression subgroups (≥25th, ≥50% and ≥75% percentiles); secondary end points included progression‐free survival (PFS), 12‐month OS, overall response rate (ORR), and safety and biomarker investigation. Results Median OS was 6.4 months in the tarextumab group vs 7.9 months in the placebo group (HR = 1.34 [95% CI = 0.95, 1.89], P = .0985). No difference observed in OS in the Notch3 gene expression subgroups. PFS in the tarextumab‐treated group (3.7 months) was significantly shorter compared with the placebo group (5.5 months) (hazard ratio was 1.43 [95% CI = 1.01, 2.01]; P = .04). Grade 3 diarrhea and thrombocytopenia were more common in the tarextumab group. Conclusions The addition of tarextumab to nab‐paclitaxel and gemcitabine did not improve OS, PFS, or ORR in first‐line metastatic PDAC, and PFS was specifically statistically worse in the tarextumab‐treated patients. Clinical trial registry no NCT01647828.

Abstract Rationale Tick paralysis has not been reported in horses in North America. Clinical Findings Two American Miniature horses were examined for progressive weakness and recumbency. Numerous ticks (Dermacentor variabilis) were found on both horses. Horse 1 was recumbent (grade 5/5 gait deficit) on presentation, whereas Horse 2 was standing but ataxic (grade 4/5 gait deficit) and tetraparetic. Both horses had decreased tongue and tail muscle tone, and had normal spinal reflexes. Cerebrospinal fluid cytology was normal. Equine herpesvirus‐1 testing was negative. Pertinent Interventions Ticks were removed within 24 hours of presentation. Both horses were treated topically with permethrin. Supportive care included fluid therapy, treatment for corneal ulceration, and frequent repositioning during recumbency. Outcome Within 48 hours of tick removal, both horses were neurologically normal. Clinical Relevance Ours is the first reported case of presumptive tick paralysis in horses in North America. Although rare, tick paralysis should be considered in horses presented with acute‐onset weakness progressing to recumbency.

Objectives: To describe the extent to which irritable bowel syndrome (IBS), fibromyalgia syndrome (FMS), and chronic fatigue syndrome (CFS) exhibit symptom overlap, and to validate a patient-derived, generic symptom questionnaire. Methods: A patient-derived 61-item symptom-frequency questionnaire was completed by participants recruited through IBS, FMS and CFS self-help websites. Principal axis factor analysis with oblimin rotation was performed separately for those reporting an IBS, FMS or CFS diagnosis. Results: Questionnaires were completed by 1751 participants of whom 851 reported more than one of the three diagnoses. Stomach pain on at least a weekly basis was reported by 79% of IBS, 52% of FMS, and 43% of CFS single diagnosis participants. Pain increasing the day after activity was reported by 32% of IBS, 94% of FMS, and 85% of CFS single diagnosis participants. Waking still tired at least once weekly was reported by 75% of IBS, 97% of FMS, and 95% of CFS single diagnosis participants. Exploratory factor analysis produced consistent results across all three diagnostic groups, the 61 items loading on 12 correlated factors with a single higher order factor on which all items loaded. Frequency analysis led to the rejection of one item (cold sores on or near lips), and freeform reporting by participants of additional symptoms identified an additional five, namely, restless legs, hair loss/brittle hair/thinning, dizziness/balance problems, blurred vision and urination problems. Conclusions: IBS, FMS and CFS are polysymptomatic spectrum disorders with a wide range of overlapping symptoms, many of which are unrelated to diagnostic criteria. Frequency analysis and factor analysis confirm the validity of using the same questionnaire across different diagnostic categories. The 65-item general symptom questionnaire (GSQ-65) is a valid generic symptom scale suitable for assessing the many different symptoms of people with IBS, FMS and CFS. Keywords: Functional disorder, Fibromyalgia, Irritable bowel syndrome, Chronic fatigue syndrome, Symptom measurement

Abstract Prior to 1900, coyotes (Canis latrans) were restricted to the western and central regions of North America, but by the early 2000s, coyotes became ubiquitous throughout the eastern United States. Information regarding morphological and genetic structure of coyote populations in the southeastern United States is limited, and where data exist, they are rarely compared to those from other regions of North America. We assessed geographic patterns in morphology and genetics of coyotes with special consideration of coyotes in the southeastern United States. Mean body mass of coyote populations increased along a west‐to‐east gradient, with southeastern coyotes being intermediate to western and northeastern coyotes. Similarly, principal component analysis of body mass and linear body measurements suggested that southeastern coyotes were intermediate to western and northeastern coyotes in body size but exhibited shorter tails and ears from other populations. Genetic analyses indicated that southeastern coyotes represented a distinct genetic cluster that differentiated strongly from western and northeastern coyotes. We postulate that southeastern coyotes experienced lower immigration from western populations than did northeastern coyotes, and over time, genetically diverged from both western and northeastern populations. Coyotes colonizing eastern North America experienced different selective pressures than did stable populations in the core range, and we offer that the larger body size of eastern coyotes reflects an adaptation that improved dispersal capabilities of individuals in the expanding range.

Background With a growing population of patients supported by ventricular assist devices (VADs) and the improvement in survival of this patient population, understanding the healthcare system burden is critical to improving outcomes. Thus, we sought to examine national estimates of VAD‐related emergency department (ED) visits and characterize their demographic, clinical, and outcomes profile. Additionally, we tested the hypotheses that resource use increased and mortality improved over time. Methods and Results This retrospective database analysis uses encounter‐level data from the 2010 to 2017 Nationwide Emergency Department Sample. The primary outcome was mortality. From 2010 to 2017, >880 million ED visits were evaluated, with 44 042 VAD‐related ED visits identified. The annual mean visits were 5505 (SD 4258), but increased 16‐fold from 2010 to 2017 (824 versus 13 155). VAD‐related ED visits frequently resulted in admission (72%) and/or death (3.0%). Median inflation‐adjusted charges were $25 679 (interquartile range, $7450, $63 119) per encounter. The most common primary diagnoses were cardiac (22%), and almost 30% of encounters were because of bleeding, stroke, or device complications. From 2010 to 2017, admission and mortality decreased from 82% to 71% and 3.4% to 2.4%, respectively (P for trends

Background Children with congenital heart disease (CHD) are known to consume a disproportionate share of resources, yet there are limited data concerning trends in resource use and mortality among admitted children with CHD. We hypothesize that charges in CHD‐related admissions increased but that mortality improved over time. Methods and Results This study, including patients

Abstract 1. To be effective, the next generation of conservation practitioners and managers need to be critical thinkers with a deep understanding of how to make evidence‐based decisions and of the value of evidence synthesis. 2. If, as educators, we do not make these priorities a core part of what we teach, we are failing to prepare our students to make an effective contribution to conservation practice. 3. To help overcome this problem we have created open access online teaching materials in multiple languages that are stored in Applied Ecology Resources. So far, 117 educators from 23 countries have acknowledged the importance of this and are already teaching or about to teach skills in appraising or using evidence in conservation decision‐making. This includes 145 undergraduate, postgraduate or professional development courses. 4. We call for wider teaching of the tools and skills that facilitate evidence‐based conservation and also suggest that providing online teaching materials in multiple languages could be beneficial for improving global understanding of other subject areas.

Abstract Insulin resistance is a metabolic disorder affecting multiple tissues and is a precursor event to type 2 diabetes (T2D). As T2D affects over 425 million people globally, there is an imperative need for research into insulin resistance to better understand the underlying mechanisms. The proposed mechanisms involved in insulin resistance include both whole body aspects, such as inflammation and metabolic inflexibility; as well as cellular phenomena, such as lipotoxicity, ER stress, and mitochondrial dysfunction. Despite numerous studies emphasizing the role of lipotoxicity in the pathogenesis of insulin resistance, an understanding of the interplay between tissues and these proposed mechanisms is still emerging. Furthermore, the tissue‐specific and unique responses each of the three major insulin target tissues and how each interconnect to regulate the whole body insulin response has become a new priority in metabolic research. With an emphasis on skeletal muscle, this mini‐review highlights key similarities and differences in insulin signaling and resistance between different target‐tissues, and presents the latest findings related to how these tissues communicate to control whole body metabolism.

Background Microvesicles are cell membrane–derived vesicles that have been shown to augment inflammation. Specifically, monocyte‐derived microvesicles (MDMVs), which can express the coagulation protein tissue factor, contribute to thrombus formation and cardiovascular disease. People living with HIV experience higher prevalence of cardiovascular disease and also exhibit increased levels of plasma microvesicles. The process of microvesicle release has striking similarity to budding of enveloped viruses. The surface protein tetherin inhibits viral budding by physically tethering budding virus particles to cells. Hence, we investigated the role of tetherin in regulating the release of MDMVs during HIV infection. Methods and Results The plasma of aviremic HIV‐infected individuals had increased levels of tissue factor + MDMVs, as measured by flow cytometry, and correlated to reduced tetherin expression on monocytes. Superresolution confocal and electron microscopy showed that tetherin localized at the site of budding MDMVs. Mechanistic studies revealed that the exposure of monocytes to HIV‐encoded Tat triggered tetherin loss and subsequent rise in MDMV production. Overexpression of tetherin in monocytes led to morphologic changes in the pseudopodia directly underneath the MDMVs. Further, tetherin knockout mice demonstrated a higher number of circulating MDMVs and less time to bleeding cessation. Conclusions Our studies define a novel regulatory mechanism of MDMV release through tetherin and explore its contribution to the procoagulatory state that is frequently observed in people with HIV. Such insights could lead to improved therapies for individuals infected with HIV and also for those with cardiovascular disease.

Background Despite excellent operative survival, correction of tetralogy of Fallot frequently is accompanied by residual lesions that may affect health beyond the incident hospitalization. Measuring resource utilization, specifically cost and length of stay, provides an integrated measure of morbidity not appreciable in traditional outcomes. Methods and Results We conducted a retrospective cohort study, using de‐identified commercial insurance claims data, of 269 children who underwent operative correction of tetralogy of Fallot from January 2004 to September 2015 with ≥2 years of continuous follow‐up (1) to describe resource utilization for the incident hospitalization and subsequent 2 years, (2) to determine whether prolonged length of stay (>7 days) in the incident hospitalization was associated with increased subsequent resource utilization, and (3) to explore whether there was regional variation in resource utilization with both direct comparisons and multivariable models adjusting for known covariates. Subjects with prolonged incident hospitalization length of stay demonstrated greater resource utilization (total cost as well as counts of outpatient visits, hospitalizations, and catheterizations) after hospital discharge (P

Background In adults with heart failure, elevated heart rate is associated with lower survival. We determined whether an elevated heart rate was associated with an increased risk of death or heart transplant in children with dilated cardiomyopathy. Methods and Results The study is an analysis of the Pediatric Cardiomyopathy Registry and includes baseline data, annual follow‐up, and censoring events (transplant or death) in 557 children (51% male, median age 1.8 years) with dilated cardiomyopathy diagnosed between 1994 and 2011. An elevated heart rate was defined as 2 or more SDs above the mean heart rate of children, adjusted for age. The primary outcomes were heart transplant and death. Heart rate was elevated in 192 children (34%), who were older (median age, 2.3 versus 0.9 years; P