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Reproductive toxicology testing is essential to safeguard public health of current and future generations. Traditional toxicological testing of male reproduction has focused on evaluating substances for acute toxicity to the reproductive system, with fertility assessment as a main endpoint and infertility a main adverse outcome. Newer studies in the last few decades have significantly widened our understanding of what represents an adverse event in reproductive toxicology, and thus changed our perspective of what constitutes a reproductive toxicant, such as endocrine disrupting chemicals that affect fertility and offspring health in an intergenerational manner. Besides infertility or congenital abnormalities, adverse outcomes can present as increased likelihood for various health problems in offspring, including metabolic syndrome, neurodevelopmental problems like autism and increased cancer predisposition, among others. To enable toxicologic studies to accurately represent the population, toxicologic testing designs need to model changing population characteristics and exposure circumstances. Current trends of increasing importance in human reproduction include increased paternal age, with an associated decline of nicotinamide adenine dinucleotide (NAD), and a higher prevalence of obesity, both of which are factors that toxicological testing study design should account for. In this perspective article, we highlighted some limitations of standard testing protocols, the need for expanding the assessed reproductive endpoint by including genetic and epigenetic sperm parameters, and the potential of recent developments, including mixture testing, novel animal models, in vitro systems like organoids, multigenerational testing protocols, as well as in silico modelling, machine learning and artificial intelligence.

Knowledge of the reproductive biology of fishes is essential for effective fisheries management. Information derived from an understanding of fish reproduction, such as size and age at maturity, is used in models to assess fish stocks and can affect estimates of important ecological processes such as recruitment, abundance, and trophic interactions. Common practices for determining the reproductive status of teleost fishes include macroscopic evaluation of gonads as well as histological analysis. However, macroscopic evaluation can be biased and histological analysis is time-consuming, resulting in limitations to spatial and temporal data availability. Here, we explore Raman spectroscopy of ovaries as a novel approach to rapidly determine the reproductive status of walleye pollock (Gadus chalcogrammus), a commercially and ecologically important species in the North Pacific. We used a two-stage partial least-squares (PLS) regression analysis followed by a linear discriminant analysis (LDA) to classify walleye pollock ovary samples as physiologically mature or immature and to subsequently predict their histologically-determined reproductive stage based on the Raman spectra. Biologically mature samples with visible yolk differentiated from mature and immature samples (non-yolked; 99% accuracy). Non-yolked ovaries that were physiologically mature (either mature non-developing or previously spawned) were further differentiated from physiologically immature ovaries (93% accuracy). In addition, detailed, histologically-determined reproductive stages of yolked samples also differentiated via Raman spectroscopy, but with reduced accuracy (79% - 86% accuracy). Our results indicate that accurate identification of maturity status and the reproductive staging of oocytes of walleye pollock based on spectral data from ovaries is possible. This can provide a fast and efficient way to increase the availability of a key component of reproductive data to inform fisheries research and management.

Abstract Rapid advancements in technology often yield research inquiry into novel applications and drone (i.e., unoccupied aircraft systems or UAS) applications in wildlife management are no exception. We questioned the time lag between drone‐related research and end‐user assessments. We implemented an online, cross‐sectional survey of wildlife professionals to better understand current drone use and benefits or concerns, complemented by a review of contemporary peer‐reviewed and gray literature. We found little disparity between scientific inquiry and end‐user experiences (i.e., similar trends among concerns in published literature and survey results). Exploring new (i.e., advancements in computer vision) and refining original drone applications (i.e., evaluating animal behavior responses during monitoring) were strong among pilots of relatively minimal experience (1–5 years). Advancements in drone technology and changes in drone‐related legislation will continue to offer benefits and challenges.

Abstract Background Tissue cryopreservation requires saturation of the structure with cryoprotectants (CPAs) that are also toxic to cells within a short timeframe unless frozen. The race between CPA delivery and cell death is the main barrier to realizing transplantation banks that can indefinitely preserve tissues and organs. Unrealistic cost and urgency leaves less life‐threatening ailments unable to capitalize on traditional organ transplantation systems that immediately match and transport unfrozen organs. For instance, human intervertebral discs (IVD) could be transplanted to treat back pain or used as ex vivo models for studying regenerative therapies, but both face logistical hurdles in organ acquisition and transport. Here we aimed to overcome those challenges by cryopreserving intact IVDs using compressive loading and swelling to accelerate CPA delivery. Methods CPAs were tested on bovine nucleus pulposus cells to determine the least cytotoxic solution. Capitalizing on our CPAs Computed Tomography (CT) contrast enhancement, we imaged and quantified saturation time in intact bovine IVDs under different conditions in a bioreactor. Finally, the entire protocol was tested, including 1 week of frozen storage, to confirm tissue viability in multiple IVD regions after thawing. Results Results showed cryopreserving medium containing dimethyl sulfoxide and ethylene glycol gave over 7.5 h before cytotoxicity. While non‐loaded IVDs required over 3 days to fully saturate, a dynamic loading protocol followed by CPA addition and free‐swelling decreased saturation time to

The ubiquitin-proteasome system (UPS) controls the majority of protein degradation in cells and dysregulation of the UPS has been implicated in the pathophysiology of numerous neurodegenerative disorders, including Alzheimer’s disease. Further, strong evidence supports a critical role for the UPS in synaptic plasticity and memory formation. However, while proteasome function is known to decrease broadly in the brain across the lifespan, whether it changes in the hippocampus, a region critical for memory storage and among the first impacted in Alzheimer’s disease, at rest and following learning in the aged brain remains unknown. Further, which proteins have altered targeting for protein degradation in the aged hippocampus has yet to be explored and whether learning in advanced age interacts with changes in ubiquitin-proteasome function across the lifespan remains unknown. Here, using proteasome activity assays and unbiased proteomic analyses, we report age-dependent changes in proteasome activity and degradation-specific K48 polyubiquitin protein targeting in the hippocampus and retrosplenial cortex of male and female rats across the lifespan. In the hippocampus, the targets of altered protein degradation were involved in transcription and astrocyte structure or G-protein and Interferon signaling in males and females, respectively. Importantly, we found that contextual fear conditioning led to an increase in proteasome activity and K48 polyubiquitin protein targeting in the hippocampus of aged male rats, a result in direct contrast to what was previously reported in young adult animals. Together, these data suggest that changes in protein degradation in the hippocampus across the lifespan may be contributing to age-related memory loss.

Abstract Loss-of-function mutations have provided crucial insights into the immunoregulatory actions of Foxp3+ regulatory T cells (Tregs). By contrast, we know very little about the consequences of defects that amplify aspects of Treg function or differentiation. Here we show that mice heterozygous for an Ikbkb gain-of-function mutation develop psoriasis. Doubling the gene dose (Ikbkb GoF/GoF) results in dactylitis, spondylitis, and characteristic nail changes, which are features of psoriatic arthritis. Ikbkb GoF mice exhibit a selective expansion of Foxp3 + CD25+ Tregs of which a subset express IL-17. These modified Tregs are enriched in both inflamed tissues, blood and spleen, and their transfer is sufficient to induce disease without conventional T cells. Single-cell transcriptional and phenotyping analyses of isolated Tregs reveal expansion of non-lymphoid tissue (tissue-resident) Tregs expressing Th17-related genes, Helios, tissue-resident markers including CD103 and CD69, and a prominent NF-κB transcriptome. Thus, IKK2 regulates tissue-resident Treg differentiation, and overactivity drives dose-dependent skin and systemic inflammation.

Thermal sensors mounted on drones (unoccupied aircraft systems) are popular and effective tools for monitoring cryptic animal species, although few studies have quantified sampling error of animal counts from thermal images. Using decoys is one effective strategy to quantify bias and count accuracy; however, plastic decoys do not mimic thermal signatures of representative species. Our objective was to produce heat signatures in animal decoys to realistically match thermal images of live animals obtained from a drone-based sensor. We tested commercially available methods to heat plastic decoys of three different size classes, including chemical foot warmers, manually heated water, electric socks, pad, or blanket, and mini and small electric space heaters. We used criteria in two categories, 1) external temperature differences from ambient temperatures (ambient difference) and 2) color bins from a palette in thermal images obtained from a drone near the ground and in the air, to determine if heated decoys adequately matched respective live animals in four body regions. Three methods achieved similar thermal signatures to live animals for three to four body regions in external temperatures and predominantly matched the corresponding yellow color bins in thermal drone images from the ground and in the air. Pigeon decoys were best and most consistently heated with three-foot warmers. Goose and deer decoys were best heated by mini and small space heaters, respectively, in their body cavities, with a heated sock in the head of the goose decoy. The materials and equipment for our best heating methods were relatively inexpensive, commercially available items that provide sustained heat and could be adapted to various shapes and sizes for a wide range of avian and mammalian species. Our heating methods could be used in future studies to quantify bias and validate methodologies for drone surveys of animals with thermal sensors. • We determined optimal heating methods for plastic animal decoys with inexpensive and commercially available equipment to mimic thermal signatures of live animals. • Methods could be used to quantify bias and improve thermal surveys of animals with drones in future studies.

Traditional approaches employing natural plant products to treat a wide array of ailments have been documented and described for thousands of years. However, there remains limited scientific study of the therapeutic potential or effectiveness of ethnobotanical applications. Increases in the incidence of cancer and emerging infectious diseases demonstrate a growing need for advances in the development of therapeutic options. In this study, we evaluate the therapeutic potential of aqueous extracts prepared from four plants, purple aster (Symphyotrichum novae-angliae (L.) Nemsom), common sage (Salvia lyrata (L.)), northern spicebush (Lindera benzoin (L.) Blume), and lamb’s ear (Stachys byzantina (K.) Koch)) traditionally used in Native American medicine in Indiana, USA. Using a combination of cytotoxicity assays, immunofluorescence microscopy, and antiviral assays, we found that sage and spicebush extracts exhibit cytotoxic and antiproliferative effects on HeLa cell proliferation and that sage, spicebush, and aster extracts were capable of significantly inhibiting human respiratory syncytial virus (hRSV), a major respiratory pathogen of infants and the elderly. Chemical analysis of the four extracts identified four major compounds which were subsequently evaluated to identify the responsible constituents in the extracts. While none of the identified compounds were shown to induce significant impacts on HeLa cell proliferation, two of the compounds, (1S)-(-)-Borneol and 5-(hydroxymethyl)-furfural, identified in sage and spicebush, respectively, were shown to have antiviral activities. Our data suggest that several of the extracts tested exhibited either anti-proliferative or antiviral activity supporting future further analysis.

Abstract Visible and thermal images acquired from drones (unoccupied aircraft systems) have substantially improved animal monitoring. Combining complementary information from both image types provides a powerful approach for automating detection and classification of multiple animal species to augment drone surveys. We compared eight image fusion methods using thermal and visible drone images combined with two supervised deep learning models, to evaluate the detection and classification of white-tailed deer (Odocoileus virginianus), domestic cow (Bos taurus), and domestic horse (Equus caballus). We classified visible and thermal images separately and compared them with the results of image fusion. Fused images provided minimal improvement for cows and horses compared to visible images alone, likely because the size, shape, and color of these species made them conspicuous against the background. For white-tailed deer, which were typically cryptic against their backgrounds and often in shadows in visible images, the added information from thermal images improved detection and classification in fusion methods from 15 to 85%. Our results suggest that image fusion is ideal for surveying animals inconspicuous from their backgrounds, and our approach uses few image pairs to train compared to typical machine-learning methods. We discuss computational and field considerations to improve drone surveys using our fusion approach.

Background Marijuana leaf vaporizers, which heat plant material and sublimate Δ‐9‐tetrahydrocannabinol without combustion, are popular alternatives to smoking cannabis that are generally perceived to be less harmful. We have shown that smoke from tobacco and marijuana, as well as aerosol from e‐cigarettes and heated tobacco products, impair vascular endothelial function in rats measured as arterial flow‐mediated dilation (FMD). Methods and Results We exposed 8 rats per group to aerosol generated by 2 vaporizer systems (Volcano and handheld Yocan) using marijuana with varying Δ‐9‐tetrahydrocannabinol levels, in a single pulsatile exposure session of 2 s/min over 5 minutes, and measured changes in FMD. To model secondhand exposure, we exposed rats for 1 minute to diluted aerosol approximating release of uninhaled Volcano aerosol into typical residential rooms. Exposure to aerosol from marijuana with and without cannabinoids impaired FMD by ≈50%. FMD was similarly impaired by aerosols from Yocan (237 °C), and from Volcano at both its standard temperature (185 °C) and the minimum sublimation temperature of Δ‐9‐tetrahydrocannabinol (157 °C), although the low‐temperature aerosol condition did not effectively deliver Δ‐9‐tetrahydrocannabinol to the circulation. Modeled secondhand exposure based on diluted Volcano aerosol also impaired FMD. FMD was not affected in rats exposed to clean air or water vapor passed through the Volcano system. Conclusions Acute direct exposure and modeled secondhand exposure to marijuana leaf vaporizer aerosol, regardless of cannabinoid concentration or aerosol generation temperature, impair endothelial function in rats comparably to marijuana smoke. Our findings indicate that use of leaf vaporizers is unlikely to reduce the vascular risk burden of smoking marijuana.

Abstract Surveying animal populations using drones (unoccupied aircraft systems [UAS]) provides numerous advantages; however, few best practices exist to survey animal communities with drones. Among myriad factors that can affect human identification and counts of animals from drone images, we focused on three factors typically controlled in the study design or by the drone pilot: flight altitude, camera angle, and time of day. Identifying interactions and patterns among these three variables represents an important first step to determining best survey practices. We used a drone to survey known numbers of eight animal decoy species, representing a range of body sizes and colors, at four ground sampling distance (GSD) values (0.35, 0.70, 1.06, and 1.41 cm/pixel) representing equivalent flight altitudes (15.2, 30.5, 45.7, and 61.0 m) at two camera angles (45° and 90°) and across a range of times of day (morning to late afternoon). Expert human observers identified and counted animals from drone images to determine how the three controllable factors affected accuracy and precision. Observer precision was high and unaffected by tested factors. However, results for observer accuracy revealed an interaction among all three controllable factors. Increasing flight altitude resulted in decreased accuracy in animal counts overall; however, accuracy was best at midday than during the morning and afternoon, when decoy and structure shadows were present or more pronounced. Surprisingly, the 45° camera enhanced accuracy more than the 90° camera, but only when animals were most difficult to identify and count, such as at higher flight altitudes or during the early morning and late afternoon. We provide recommendations for improving human accuracy in identifying and counting animals from drone images used in monitoring animal populations and communities. These results should be incorporated into drone survey design in addition to considering funding, logistical, and animal behavior constraints.

Abstract Background Small unoccupied aircraft systems (UAS) are replacing or supplementing occupied aircraft and ground-based surveys in animal monitoring due to improved sensors, efficiency, costs, and logistical benefits. Numerous UAS and sensors are available and have been used in various methods. However, justification for selection or methods used are not typically offered in published literature. Furthermore, existing reviews do not adequately cover past and current UAS applications for animal monitoring, nor their associated UAS/sensor characteristics and environmental considerations. We present a systematic map that collects and consolidates evidence pertaining to UAS monitoring of animals. Methods We investigated the current state of knowledge on UAS applications in terrestrial animal monitoring by using an accurate, comprehensive, and repeatable systematic map approach. We searched relevant peer-reviewed and grey literature, as well as dissertations and theses, using online publication databases, Google Scholar, and by request through a professional network of collaborators and publicly available websites. We used a tiered approach to article exclusion with eligible studies being those that monitor (i.e., identify, count, estimate, etc.) terrestrial vertebrate animals. Extracted metadata concerning UAS, sensors, animals, methodology, and results were recorded in Microsoft Access. We queried and catalogued evidence in the final database to produce tables, figures, and geographic maps to accompany this full narrative review, answering our primary and secondary questions. Review findings We found 5539 articles from our literature searches of which 216 were included with extracted metadata categories in our database and narrative review. Studies exhibited exponential growth over time but have levelled off between 2019 and 2021 and were primarily conducted in North America, Australia, and Antarctica. Each metadata category had major clusters and gaps, which are described in the narrative review. Conclusions Our systematic map provides a useful synthesis of current applications of UAS-animal related studies and identifies major knowledge clusters (well-represented subtopics that are amenable to full synthesis by a systematic review) and gaps (unreported or underrepresented topics that warrant additional primary research) that guide future research directions and UAS applications. The literature for the use of UAS to conduct animal surveys has expanded intensely since its inception in 2006 but is still in its infancy. Since 2015, technological improvements and subsequent cost reductions facilitated widespread research, often to validate UAS technology to survey single species with application of descriptive statistics over limited spatial and temporal scales. Studies since the 2015 expansion have still generally focused on large birds or mammals in open landscapes of 4 countries, but regulations, such as maximum altitude and line-of-sight limitations, remain barriers to improved animal surveys with UAS. Critical knowledge gaps include the lack of (1) best practices for using UAS to conduct standardized surveys in general, (2) best practices to survey whole wildlife communities in delineated areas, and (3) data on factors affecting bias in counting animals from UAS images. Promising advances include the use of thermal sensors in forested environments or nocturnal surveys and the development of automated or semi-automated machine-learning algorithms to accurately detect, identify, and count animals from UAS images.

Abstract Ring‐billed (Larus delawarensis) and herring (L. argentatus) gulls are numerous and widespread in North America. These gulls rank among the top 9 species for risk of bird‐aircraft collisions (hereafter strikes). The ubiquitous presence of gulls in urban coastal environments, including rooftop nesting behavior, are factors impacting strike risk. Our purpose was to assess gull response to a small uncrewed aircraft system (UAS) in hazing flights at night during the nest‐building phase. We hypothesized that nocturnal UAS operation, like nocturnal predator disturbance, might reduce gull numbers and, thus, strike risk to aircraft. In spring 2021, we conducted UAS treatments over target roofs at least once every hour from 2000 until 0200, weather permitting, for 15 min and over a 14‐day period for each site. The UAS flew directly above (~4 m) and then descended (~4 m/s) within 1 m of loafing gulls. No gulls interacted with the UAS and most flushed within 6 minutes. Generally, the first treatment of a night dispersed all gulls (min–max = 1–130 individuals) from the target roof for an extended period. Our operations were often grounded because of weather and our gull response data were limited because of few individuals present. We discuss our observations with particular attention to feasibility and possible implications such as shifting birds to other sites which, potentially, could be counterproductive for management.

Encapsulation of bioactive peptides in slow-release particles is complex and relies on organic solvents. Here, the authors absorb peptides in a polymer phase from water, creating a simple low-cost encapsulation process in a class of polymer depot.

A 25-year-old male was admitted to the neurological intensive care unit for neurologic deterioration, likely caused by a paraneoplastic syndrome secondary to testicular malignancy. He experienced spontaneous rupture and hemorrhage of his testicular mass arising from an undescended testis while admitted. The tumor was excised, revealing a mixed germ cell tumor. Serum tumor markers began to rise after 4 cycles of chemotherapy. Surveillance scans 32 weeks after mass rupture revealed numerous tumor deposits throughout his peritoneum concerning for teratoma. We review a case of intraperitoneal metastasis of a testicular mixed germ cell tumor following intra-abdominal mass rupture.

IntroductionThe blue orchard bee, Osmia lignaria, is a widespread North American native pollinator that can be employed for commercial fruit and nut crop production. The largest supplies of these bees are collected in the wildlands ssssof Utah and Washington, United States. How O. lignaria from different geographic regions respond to current recommended management practices or translocation to novel environments is not fully understood.MethodsUtah- and Washington-originated O. lignaria were reared in laboratory incubators under two thermal regimens: (1) constant temperatures used to manage bees through immature development, adult winter dormancy, and for spring emergence, and (2) hourly fluctuating temperatures programmed to mimic the natural daily (24 h) thermal cycles of the nearest cherry orchard growing region through their life cycles.ResultsIn comparison to rearing bees at orchard temperatures, we found that rearing bees at a constant temperature increased survival and shortened egg–adult development periods. Washington bees were more adversely affected by the consistent warm rearing temperatures than were Utah bees, possibly due to their adaptations to Washington’s relatively moderate climate. At orchard temperatures, Utah bees suffered high prepupal and pupal mortality, while Washington bees suffered high pupal and adult mortality. These late life stages coincided with the hottest maximum temperatures in their respective thermal regimens. Adult females from both states naturally emerged in synchrony with local bloom time, but their emergence period overall was prolonged compared to bees in the constant thermal regimen that were induced to emerge at orchard bloom times.DiscussionOur data support that bees originating from cool montane habitats of different U.S. states suffer from the warmer climatic conditions at lower altitudes of their respective crop-growing regions. A better understanding of optimal management temperatures for O. lignaria from different geographic regions and the effect of bee origin and temperature on survival and development timing is needed for best managing these pollinators when they are translocated or when climate change results in increased temperatures during bee development periods.

Accurately determining the macronutrient profile of mare milk is a precursor to studying how milk composition affects foals’ growth and development. This study optimized and validated an extraction and quantification method for mare milk oligosaccharides, which make up a portion of the carbohydrate fraction of mare milk. Mare milk was extracted with chloroform and methanol, and oligosaccharides were selectively isolated from the carbohydrate fraction using porous-graphitized carbon solid-phase-extraction (SPE). Good recovery rates for milk oligosaccharides (between 70 and 100%) were achieved with the optimized method. This study also compared the use of Fourier-Transform infrared (FTIR) spectroscopy versus wet chemistry quantification methods for protein, fat, and lactose. The FTIR method produced statistically equivalent protein contents to the wet chemistry method, along with substantial savings in both analyst time and consumable consumption. FTIR analysis slightly underestimated the fat content of mare milk relative to the official wet chemistry method, with the difference between the methods increasing at higher fat contents. FTIR also overestimated the lactose content of mare milk and appeared to generate “lactose” values that included the milk oligosaccharides and thus represented the total carbohydrate (lactose and milk oligosaccharides) content of mare milk.

Human papillomavirus (HPV) is the most common sexually transmitted infection in the US and the leading cause of oropharyngeal cancer (OPC), an oral cancer most often identified by dental providers. Given the rise in HPV-associated OPC and recent Food and Drug Administration (FDA) approval of the HPV vaccine to prevent OPC, dental providers have a unique role in HPV prevention. This study assessed US adults’ comfort levels discussing HPV and OPC with dental providers. An online survey platform was used to recruit a nationally representative sample of US adults (n = 300). The questionnaire assessed participants’ knowledge, acceptability, and comfort discussing HPV-related topics with dental providers. SPSS 24 was utilized for data analyses. In general, participants reported feeling comfortable discussing HPV and OPC with dental providers. Participants reported feeling more comfortable with dentists than dental hygienists when discussing (t = 2.85, p

Abstract A challenge that conservation practitioners face is manipulating behavior of nuisance species. The turkey vulture (Cathartes aura) can cause substantial damage to aircraft if struck. The goal of this study was to assess vulture responses to unmanned aircraft systems (UAS) for use as a possible dispersal tool. Our treatments included three platforms (fixed-wing, multirotor, and a predator-like ornithopter [powered by flapping flight]) and two approach types (30 m overhead or targeted towards a vulture) in an operational context. We evaluated perceived risk as probability of reaction, reaction time, flight-initiation distance (FID), vulture remaining index, and latency to return. Vultures escaped sooner in response to the fixed-wing; however, fewer remained after multirotor treatments. Targeted approaches were perceived as riskier than overhead. Vulture perceived risk was enhanced by flying the multirotor in a targeted approach. We found no effect of our treatments on FID or latency to return. Latency was negatively correlated with UAS speed, perhaps because slower UAS spent more time over the area. Greatest visual saliency followed as: ornithopter, fixed-wing, and multirotor. Despite its appearance, the ornithopter was not effective at dispersing vultures. Because effectiveness varied, multirotor/fixed-wing UAS use should be informed by management goals (immediate dispersal versus latency).

Abstract Background Small unmanned aircraft systems (sUAS) are replacing or supplementing manned aircraft and ground-based surveys in many animal monitoring situations due to better coverage at finer spatial and temporal resolutions, access, cost, bias, impacts, safety, efficiency, and logistical benefits. Various sUAS models and sensors are available with varying features and usefulness depending on survey goals. However, justification for selection of sUAS and sensors are not typically offered in published literature and existing reviews do not adequately cover past and current sUAS applications for animal monitoring nor their associated sUAS model and sensor technologies, taxonomic and geographic scope, flight conditions and considerations, spatial distributions of sUAS applications, and reported technical difficulties. We outline a systematic map protocol to collect and consolidate evidence pertaining to sUAS monitoring of animals. Our systematic map will provide a useful synthesis of current applications of sUAS-animal related studies and identify major knowledge clusters (well-represented subtopics that are amenable to full synthesis by a systematic review) and gaps (unreported or underrepresented topics that warrant additional primary research) that may influence future research directions and sUAS applications. Methods Our systematic map will investigate the current state of knowledge using an accurate, comprehensive, and repeatable search. We will find relevant peer-reviewed and grey literature as well as dissertations and theses using online publication databases, Google Scholar, and by request through a professional network of collaborators and publicly available websites. We will use a tiered approach to article exclusion with eligible studies being those that monitor (i.e., identify, count, estimate, etc.) terrestrial vertebrate animals. Extracted data concerning sUAS, sensors, animals, methodology, and results will be recorded in Microsoft Access. We will query and catalogue evidence in the final database to produce tables, figures, and geographic maps to accompany a full narrative review that answers our primary and secondary questions.

Dynamic loading is a shared feature of tendon tissue homeostasis and pathology. Tendon cells have the inherent ability to sense mechanical loads that initiate molecular-level mechanotransduction pathways. While mature tendons require physiological mechanical loading in order to maintain and fine tune their extracellular matrix architecture, pathological loading initiates an inflammatory-mediated tissue repair pathway that may ultimately result in extracellular matrix dysregulation and tendon degeneration. The exact loading and inflammatory mechanisms involved in tendon healing and pathology is unclear although a precise understanding is imperative to improving therapeutic outcomes of tendon pathologies. Thus, various model systems have been designed to help elucidate the underlying mechanisms of tendon mechanobiology via mimicry of the in vivo tendon architecture and biomechanics. Recent development of model systems has focused on identifying mechanoresponses to various mechanical loading platforms. Less effort has been placed on identifying inflammatory pathways involved in tendon pathology etiology, though inflammation has been implicated in the onset of such chronic injuries. The focus of this work is to highlight the latest discoveries in tendon mechanobiology platforms and specifically identify the gaps for future work. An interdisciplinary approach is necessary to reveal the complex molecular interplay that leads to tendon pathologies and will ultimately identify potential regenerative therapeutic targets.

Background: Sketching exists in many disciplines and varies in how it is assessed, making it challenging to define fundamental sketching skills and the characteristics of a high-quality sketch. For instructors to apply effective strategies for teaching and assessing engineering sketching, a clear summary of the constructs, metrics, and objectives for sketching assessment across engineering education and related disciplines is needed. Purpose: This systematic literature review explores sketching assessment definitions and approaches across engineering education research. Methodology/Approach: We collected 671 papers from five major engineering and education databases at all skill levels for reported sketching constructs and metrics, cognition, and learning contexts. Based on the selection criteria, we eliminated all but 41 papers, on which we performed content analysis. Findings/Conclusions: Engineering, design, and art emerged as three major disciplines in the papers reviewed. We found that sketching assessment most often employs metrics on accuracy, perspective, line quality, annotations, and aesthetics. Most collected studies examined beginners in undergraduate engineering design sketching or drawing ability tests. Cognitive skills included perceiving the sketch subject, creatively sketching ideas, using metacognition to monitor the sketching process, and using sketching for communication. Implications: Sketching assessment varies by engineering discipline and relies on many types of feedback and scoring metrics. Cognitive theory can inform instructional activities as a foundation for sketching skills. There is a need for robust evidence of high-quality assessment practices in sketching instruction. Assessment experts can apply their knowledge toward improving sketching assessment development, implementation, and validation.

As the demand for alternatives to antibiotic growth promoters (AGP) increases in food animal production, phytobiotic compounds gain popularity because of their ability to mimic the desirable bioactive properties of AGP. Chestnut tannins (ChT) are one of many phytobiotic compounds used as feed additives, particularly in South America, for broilers because of its favorable antimicrobial and growth promotion capabilities. Although studies have observed the microbiological and immunologic effects of ChT, there is a lack of studies evaluating the metabolic function of ChT. Therefore, the objective of this study was to characterize the cecal metabolic changes induced by ChT inclusion and how they relate to growth promotion. A total of 200 day-of-hatch broiler chicks were separated into 2 feed treatment groups: control and 1% ChT. The ceca from all the chicks in the treatment groups were collected on day 2, 4, 6, 8, and 10 after hatch. The cytokine mRNA quantitative RT-PCR was determined using TaqMan gene expression assays for IL-1B, IL-6, IL-8, IL-10, and interferon gamma quantification. The cytokine expression showed highly significant increased expressions of IL-6 and IL-10 on day 2 and 6, whereas the other proinflammatory cytokines did not have significantly increased expression. The results from the kinome array demonstrated that the ceca from birds fed with 1% ChT had significant (P

ABSTRACT: Intestinal organoids (IO), known as “mini-guts”, derived from intestinal crypts, are self-organizing three-dimensional (3D) multicellular ex vivo models that recapitulate intestine epithelial structure and function and have been widely used for studying intestinal physiology, pathophysiology, molecular mechanisms of host-pathogen interactions, and intestinal disease in mammals. However, studies on avian IO are limited and the development of long-term cultures of IO model for poultry research is lacking. Therefore, the objectives of this study were to generate crypt-derived organoids from chicken intestines and to optimize conditions for cell growth and enrichments, passages, and cryopreservation. Crypts were collected from the small intestines of birds at embryonic d-19 and ceca from layer and broiler chickens with ages ranging from d 1 to 20 wk, embedded in a basement membrane matrix, and cultured with organoid growth media (OGM) prepared in house. The crypt-derived organoids were successfully grown and propagated to form 3D spheres like structures that were cultured for up to 3 wk. Organoids were formed on d one, budding appeared on d 3, and robust budding was observed on d 7 and beyond. For cryopreservation, dissociated organoids were resuspended in a freezing medium. The characteristics of IO upon extended passages and freeze-thaw cycles were analyzed using reverse transcription (RT)-PCR, immunoblotting, and live cell imaging. Immunoblotting and RT-PCR using E-cadherin (the marker for epithelial cells), leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5, the marker for stem cells), chromogranin A (the marker for enteroendocrine cells), lysozyme (the marker for Paneth cells), and mucin (the biomarker for goblet cells) confirmed that IO were composed of heterogeneous cell populations, including epithelial cells, stem cells, enteroendocrine cells, Paneth cells, and goblet cells. Furthermore, OGM supplemented with both valproic acid and CHIR99021, a glycogen synthase kinase 3β inhibitor and a histone deacetylase inhibitor, increased the size of the avian IO (P < 0.001). To the best of our knowledge, this is the first comprehensive report for establishing long-term, organoid culture models from small intestines and ceca of layer and broiler chickens. This model will facilitate elucidation of the mechanisms impacting host-pathogen interactions, eventually leading to the discovery of pathogen intervention strategies in poultry.

BackgroundNon-invasive finger-cuff monitors measuring cardiac index and vascular tone (SVRI) classify emergency department (ED) patients with acute heart failure (AHF) into three otherwise-indistinguishable subgroups. Our goals were to validate these "hemodynamic profiles" in an external cohort and assess their association with clinical outcomes.MethodsAHF patients (n = 257) from five EDs were prospectively enrolled in the validation cohort (VC). Cardiac index and SVRI were measured with a ClearSight finger-cuff monitor (formerly NexFin, Edwards Lifesciences) as in a previous study (derivation cohort, DC, n = 127). A control cohort (CC, n = 127) of ED patients with sepsis was drawn from the same study as the DC. K-means cluster analysis previously derived two-dimensional (cardiac index and SVRI) hemodynamic profiles in the DC and CC (k = 3 profiles each). The VC was subgrouped de novo into three analogous profiles by unsupervised K-means consensus clustering. PERMANOVA tested whether VC profiles 1-3 differed from profiles 1-3 in the DC and CC, by multivariate group composition of cardiac index and vascular tone. Profiles in the VC were compared by a primary outcome of 90-day mortality and a 30-day ranked composite secondary outcome (death, mechanical cardiac support, intubation, new/emergent dialysis, coronary intervention/surgery) as time-to-event (survival analysis) and binary events (odds ratio, OR). Descriptive statistics were used to compare profiles by two validated risk scores for the primary outcome, and one validated score for the secondary outcome.ResultsThe VC had median age 60 years (interquartile range {49-67}), and was 45% (n = 116) female. Multivariate profile composition by cardiac index and vascular tone differed significantly between VC profiles 1-3 and CC profiles 1-3 (p = 0.001, R2 = 0.159). A difference was not detected between profiles in the VC vs. the DC (p = 0.59, R2 = 0.016). VC profile 3 had worse 90-day survival than profiles 1 or 2 (HR = 4.8, 95%CI 1.4-17.1). The ranked secondary outcome was more likely in profile 1 (OR = 10.0, 1.2-81.2) and profile 3 (12.8, 1.7-97.9) compared to profile 2. Diabetes prevalence and blood urea nitrogen were lower in the high-risk profile 3 (pConclusionsHemodynamic profiles in ED patients with AHF, by non-invasive finger-cuff monitoring of cardiac index and vascular tone, were replicated de novo in an external cohort. Profiles showed significantly different risks of clinically-important adverse patient outcomes.

Research into the functions of nicotinamide adenine dinucleotide (NAD) has intensified in recent years due to the insight that abnormally low levels of NAD are involved in many human pathologies including metabolic disorders, neurodegeneration, reproductive dysfunction, cancer, and aging. Consequently, the development and validation of novel NAD-boosting strategies has been of central interest, along with the development of models that accurately represent the complexity of human NAD dynamics and deficiency levels. In this review, we discuss pioneering research and show how modern researchers have long since moved past believing that pellagra is the overt and most dramatic clinical presentation of NAD deficiency. The current research is centered on common human health conditions associated with moderate, but clinically relevant, NAD deficiency. In vitro and in vivo research models that have been developed specifically to study NAD deficiency are reviewed here, along with emerging strategies to increase the intracellular NAD concentrations.

Abstract Introduction In this study, magnetic resonance imaging data was used to (1) model IVD‐specific gradients of glucose, oxygen, lactate, and pH; and (2) investigate possible effects of covariate factors (i.e., disc geometry, and mean apparent diffusion coefficient values) on the IVD’s microenvironment. Mathematical modeling of the patient’s specific IVD microenvironment could be important when selecting patients for stem cell therapy due to the increased nutrient demand created by that treatment. Materials and Methods Disc geometry and water diffusion coefficients were extracted from MRIs of 37 patients using sagittal T1‐weighted images, T2‐weighted images, and ADC Maps. A 2‐D steady state finite element mathematical model was developed in COMSOL Multiphysics® 5.4 to compute concentration maps of glucose, oxygen, lactate and pH. Results Concentration of nutrients (i.e., glucose, and oxygen) dropped with increasing distance from the cartilaginous endplates (CEP), whereas acidity levels increased. Most discs experienced poor nutrient levels along with high acidity values in the inner annulus fibrosus (AF). The disc’s physiological microenvironment became more deficient as degeneration progressed. For example, minimum glucose concentration in grade 4 dropped by 31.1% compared to grade 3 (p < 0.0001). The model further suggested a strong effect of the following parameters: disc size, AF and CEP diffusivities, metabolic reactions, and cell density on solute concentrations in the disc (p < 0.05). Conclusion The significance of this work implies that the individual morphology and physiological conditions of each disc, even among discs of the same Pfirrmann grade, should be evaluated when modeling IVD solute concentrations.

Metrics to assess relative adult bee body size have included both mass and morphometrics, but these metrics may not equally or reliably estimate body size for all bee species and in all situations, due to bee age, diet, and/or environment. Understanding the relationships between different metrics and possible redundancies in the information they afford is important but not always known. Body size measurements provide valuable data for interpreting research outcomes for managed solitary bees, including Osmia lignaria Say and Megachile rotundata F. (Hymenoptera: Megachilidae). Applied studies of these important and readily available U.S. crop pollinators focus on refining commercial management practices, and basic empirical studies in various scientific disciplines (from genomics to ecology) employ them as model systems to study solitary bees. To examine common metrics of body size, we measured head capsule width (HCW), intertegular distance (ITD), and fresh and dry weights of newly emerged adults of both species. Using linear and exponential models, we determined relationships between these body size metrics. For M. rotundata, linear models best described relationships between ITD and all other metrics, and between HCW and fresh and dry weights. For O. lignaria, linear models best fit relationships between all metrics except for fresh weight with both ITD and HCW, which were fitted better with exponential models. For both species, model fits were strongest when males and females were pooled. Depending on the study question, knowing that only one metric may reliably measure body size can simplify evaluations of O. lignaria and M. rotundata responses to artificial or environmental variables.

Protein phosphorylation helps to control many important cellular activities. Here the authors describe a genetic selection strategy to isolate designed ankyrin repeat proteins that bind specifically to phosphomodified targets.

Summary: Heterozygous gain-of-function (GOF) mutations of hypoxia-inducible factor 2α (HIF2A), a key hypoxia-sensing regulator, are associated with erythrocytosis, thrombosis, and vascular complications that account for morbidity and mortality of patients. We demonstrated that the vascular pathology of HIF2A GOF mutations is independent of erythrocytosis. We generated HIF2A GOF-induced pluripotent stem cells (iPSCs) and differentiated them into endothelial cells (ECs) and smooth muscle cells (SMCs). Unexpectedly, HIF2A-SMCs, but not HIF2A-ECs, were phenotypically aberrant, more contractile, stiffer, and overexpressed endothelin 1 (EDN1), myosin heavy chain, elastin, and fibrillin. EDN1 inhibition and knockdown of EDN1-receptors both reduced HIF2-SMC stiffness. Hif2A GOF heterozygous mice displayed pulmonary hypertension, had SMCs with more disorganized stress fibers and higher stiffness in their pulmonary arterial smooth muscle cells, and had more deformable pulmonary arteries compared with wild-type mice. Our findings suggest that targeting these vascular aberrations could benefit patients with HIF2A GOF and conditions of augmented hypoxia signaling.

Introduction: Few emergency department (ED)-specific fall-risk screening tools exist. The goals of this study were to externally validate Tiedemann et al’s two-item, ED-specific fall screening tool and test handgrip strength to determine their ability to predict future falls. We hypothesized that both the two-item fall screening and handgrip strength would identify older adults at increased risk of falling. Methods: A convenience sample of patients ages 65 and older presenting to a single-center academic ED were enrolled. Patients were asked screening questions and had their handgrip strength measured during their ED visit. Patients were given one point if they answered “yes” to “Are you taking six or more medications?” and two points for answering “yes” to “Have you had two or more falls in the past year?” to give a cumulative score from 0 to 3. Participants had monthly follow- ups, via postcard questionnaires, for six months after their ED visit. We performed sensitivity and specificity analyses, and used likelihood ratios and frequencies to assess the relationship between risk factors and falls, fall-related injury, and death. Results: In this study, 247 participants were enrolled with 143 participants completing follow-up (58%). During the six-month follow-up period, 34% of participants had at least one fall and 30 patients died (12.1%). Fall rates for individual Tiedemann scores were 14.3%, 33.3%, 60.0% and 72.2% for scores of 0,1, 2 and 3, respectively. Low handgrip strength was associated with a higher proportion of falls (46.3%), but had poor sensitivity (52.1%). Conclusion: Handgrip strength was not sensitive in screening older adults for future falls. The Tiedemann rule differentiated older adults who were at high risk for future falls from low risk individuals, and can be considered by EDs wanting to screen older adults for future fall risk.

Ninety-three percent of all reported bird strikes occur below 1,067 m, which based on the typical approach and departure angles of aircraft is within 8–13 km of an airport. Concomitantly, the Federal Aviation Administration and the International Civil Aviation Organization recommend that any feature that would attract hazardous wildlife to the approach and departure airspace be restricted. Thus, preventing the establishment of wildlife attractants, such as municipal solid waste landfills (MSWLFs) within 8 km or 13 km extents (U.S. and international recommendations, respectively) of airports, has been recommended to mitigate the risk of bird–aircraft collisions (strikes). However, robust evidence linking wildlife attractants at these spatial scales to an increase in strikes is lacking. We investigated the effect of densities of MSWLFs and construction and demolition (C&D) landfills, landscape diversity, and human population density on the adverse effect (AE; strikes that caused damage or had a negative effect on flight) bird strike rate involving species broadly associated with MSWLFs. We predicted that airports surrounded by a high density of MSWLFs, high human population densities, and high landscape diversity would increase the AE strike rate. We evaluated our predictions via generalized linear mixed models with bird strike data from 2009 through 2017 at 111 Part 139 airports. Only U.S. airports were used because of high wildlife strike reporting rates. Part 139 certificated airports are those that facilitate air carriers with >30 seats for passengers and crew. Our average model included density of MSWLFs and C&D landfills for both the 8- and 13-km extent from the airports. We found no significant contribution by any variable to the AE strike rate variance. Our results indicated that the effects of landfills on AE strike rates are inconclusive. Possible explanations for our findings include the influence of unmeasured landscape features and lack of fine-scale data on bird habitat use at landfill facilities. Future research should investigate bird 3-dimensional space use in addition to bird and habitat survey techniques.

Coronary artery disease (CAD) is an immune-mediated disease in which CCR2 attracts classical, intermediate, and non-classical monocytes to the arterial intima where they differentiate to macrophages. Balance between pro-inflammatory M1 and anti-inflammatory M2 macrophages contributes to CAD prevention. Moderate to vigorous intensity physical activity (MVPA) elicits an immune response and reduces the incidence of CAD, however, the impact of prior MVPA on monocyte subset CCR2 expression and macrophage polarization following acute exercise is unknown. Purpose: To determine the impact of physical activity status on monocyte subset CCR2 surface expression and macrophage polarization in response to an acute bout of moderate intensity cycle ergometry. Methods: 24 healthy women and men (12 high physically active [HIACT]: ≥1500 METmin/wk MVPA & 12 low physically active [LOACT]:

Background Animal–vehicle collisions represent substantial sources of mortality for a variety of taxa and can pose hazards to property and human health. But there is comparatively little information available on escape responses by free-ranging animals to vehicle approach versus predators/humans. Methods We examined responses (alert distance and flight-initiation distance) of focal Canada geese (Branta canadensis maxima) to vehicle approach (15.6 m·s−1) in a semi-natural setting and given full opportunity to escape. We manipulated the direction of the vehicle approach (direct versus tangential) and availability of social information about the vehicle approach (companion group visually exposed or not to the vehicle). Results We found that both categorical factors interacted to affect alert and escape behaviors. Focal geese used mostly personal information to become alert to the vehicle under high risk scenarios (direct approach), but they combined personal and social information to become alert in low risk scenarios (tangential approach). Additionally, when social information was not available from the companion group, focal birds escaped at greater distances under direct compared to tangential approaches. However, when the companion group could see the vehicle approaching, focal birds escaped at similar distances irrespective of vehicle direction. Finally, geese showed a greater tendency to take flight when the vehicle approached directly, as opposed to a side step or walking away from the vehicle. Conclusions We suggest that the perception of risk to vehicle approach (likely versus unlikely collision) is weighted by the availability of social information in the group; a phenomenon not described before in the context of animal–vehicle interactions. Notably, when social information is available, the effects of heightened risk associated with a direct approach might be reduced, leading to the animal delaying the escape, which could ultimately increase the chances of a collision. Also, information on a priori escape distances required for surviving a vehicle approach (based on species behavior and vehicle approach speeds) can inform planning, such as location of designated cover or safe areas. Future studies should assess how information from vehicle approach flows within a flock, including aspects of vehicle speed and size, metrics that affect escape decision-making.

Responsive neurostimulation for epilepsy involves an implanted device that delivers direct electrical brain stimulation in response to detection of incipient seizures. Responsive neurostimulation is a safe and effective treatment for adults with drug-resistant epilepsy, but although novel treatments are critically needed for younger patients, responsive neurostimulation is currently not approved for children with drug-resistant epilepsy. Here, we report a 16-year-old patient with seizures arising from eloquent cortex, who was successfully treated with responsive neurostimulation. This case highlights the potential utility of this therapy for pediatric patients and underscores the need for larger studies. Keywords: Neurostimulation, RNS system, Devices, Pediatric, Drug-resistant epilepsy

The U.S. swine industry is currently inadequately prepared to counteract the increasing threat of high-consequence diseases. Although approved and preferred depopulation guidelines exist, ventilation shutdown (VSD+) is currently the only method being deployed during a state of emergency to depopulate large swine populations. However, the permitted use of VSD+ during constrained circumstances has been criticized due to raised swine welfare concerns. The objective of this study was to investigate the effectiveness of carbon dioxide gas (CO2), nitrogen gas (N2), compressed air foam (CAF), compressed nitrogen foam (CAF-N2) and aspirated foam (AF) during a 15-min dwell time on adult swine in an emergency depopulation situation. A small-scale trial using 12 sows per depopulation method showed the highest efficiency to induce cessation of movement for AF and CO2 (186.0 ± 48 vs. 202.0 ± 41, s ± SD). The ease of implementation and safety favored AF for further investigation. A large-scale field study using AF to depopulate 134 sows in modified rendering trailers showed a mean fill time of 103.8 s (SD: 5.0 s) and cessation of movement of 128.0 s (SD: 18.6 s) post filling. All sows were confirmed dead post-treatment for both trials. The implementation of AF in modified rendering trailers may allow for a safe and reliable method that allows for the expedient and mobile depopulation of both small and large numbers of sows during an emergency.

Recently, a P-deficient diet caused rickets in commercial chicks within three days. This study aimed to investigate the duration of onset of rickets in chicks. Data were collected from 3–11 day old chicks raised on 88 commercial farms. Male day-old Arbor Acres Plus broilers (n = 450) were studied in three trials, with three to four treatments each. Each treatment used one of the following crumbled feeds: control feed (calcium (Ca): phosphorus (P)-1.41), slightly high Ca:P feed (SHCa:P, Ca:P-2.69), high Ca:P ratio, P deficient feed (HCa:P, Ca:P-3.08), and HCa:P feed plus 1.5% dicalcium phosphate (HCa:P + DP). Each treatment had three replicates with 15 birds each. Rickets was induced by HCa:P, and cured by HCa:P + DP, confirmed by gross anatomy, gait score, serum P concentration and growth performance. Lameness was not found in control groups, whereas, observed in the HCa:P groups as early as day 2.7 on commercial farms and day 3 in experimental farm. Serum P was reduced in HCa:P (p < 0.01). Bodyweight and feed intake started decreasing at day 3 on commercial farms and in all trials (p < 0.01). The duration of onset of hypophosphatemic rickets in broiler chicks fed HCa:P crumbled feed is approximately three days.

In recent years, small unmanned aircraft systems (sUAS) have been used widely to monitor animals because of their customizability, ease of operating, ability to access difficult to navigate places, and potential to minimize disturbance to animals. Automatic identification and classification of animals through images acquired using a sUAS may solve critical problems such as monitoring large areas with high vehicle traffic for animals to prevent collisions, such as animal-aircraft collisions on airports. In this research we demonstrate automated identification of four animal species using deep learning animal classification models trained on sUAS collected images. We used a sUAS mounted with visible spectrum cameras to capture 1288 images of four different animal species: cattle (Bos taurus), horses (Equus caballus), Canada Geese (Branta canadensis), and white-tailed deer (Odocoileus virginianus). We chose these animals because they were readily accessible and white-tailed deer and Canada Geese are considered aviation hazards, as well as being easily identifiable within aerial imagery. A four-class classification problem involving these species was developed from the acquired data using deep learning neural networks. We studied the performance of two deep neural network models, convolutional neural networks (CNN) and deep residual networks (ResNet). Results indicate that the ResNet model with 18 layers, ResNet 18, may be an effective algorithm at classifying between animals while using a relatively small number of training samples. The best ResNet architecture produced a 99.18% overall accuracy (OA) in animal identification and a Kappa statistic of 0.98. The highest OA and Kappa produced by CNN were 84.55% and 0.79 respectively. These findings suggest that ResNet is effective at distinguishing among the four species tested and shows promise for classifying larger datasets of more diverse animals.

Newly hatched male layer chicks are considered as “by-products” in the egg industry and must be humanely euthanized at the hatchery. Instantaneous mechanical destruction (maceration) is the predominant euthanasia method applied in poultry hatcheries and is approved by the American Veterinary Medical Association (AVMA). However, maceration is not perceived by the public to be a humane means of euthanasia. The effects of alternative euthanasia methods, including carbon dioxide (CO2) or nitrogen (N2) inhalation, and a commercial negative pressure stunning system on behavioral and physiological responses of day-of-hatch male layer chicks, were evaluated in a field trial. Chick behaviors, including ataxia, loss of posture, convulsions, cessation of vocalization, and cessation of movement, were monitored. Serum hormones were assessed at the end of each of the alternative euthanasia treatments, including a control group allowed to breathe normal atmospheric air. The N2 method induced unconsciousness and death later than the CO2 and negative pressure methods, and increased serum corticosterone concentrations of neonatal chicks. Carbon dioxide inhalation increased serotonin concentrations as compared to controls, as well as the N2 and the negative pressure methods. The behavioral and physical responses observed in this study suggest that both CO2 inhalation and negative pressure stunning can be employed to humanely euthanize neonatal male layer chicks.

Monitoring antimicrobial resistance of foodborne pathogens in poultry is critical for food safety. We aimed to compare antimicrobial resistance phenotypes in Salmonella isolated from poultry samples as influenced by isolation and antimicrobial susceptibility testing methods. Salmonella isolates were cultured from a convenience sample of commercial broiler ceca with and without selective broth enrichment, and resistance phenotypes were determined for 14 antimicrobials using the Sensititre® platform and a qualitative broth breakpoint assay. The broth breakpoint method reported higher resistance to chloramphenicol, sulfisoxazole, and the combination of trimethoprim and sulfamethoxazole, and lower resistance to streptomycin as compared to the Sensititre® assay in trial one. Selective enrichment of samples containing Salmonella in Rappaport-Vassiliadis broth reported lowered detectable resistance to amoxicillin/clavulanic acid, ampicillin, azithromycin, cefoxitin, ceftriaxone, nalidixic acid, and meropenem, and increased resistance to streptomycin and tetracycline than direct-plating samples in trial one. Using matched isolates in trial two, the Sensititre® assay reported higher resistance to chloramphenicol and gentamicin, and lower resistance to nalidixic acid as compared to the broth breakpoint method. These results suggest methodology is a critical consideration in the detection and surveillance of antimicrobial resistance phenotypes in Salmonella isolates from poultry samples and could affect the accuracy of population or industry surveillance insights and intervention strategies.

Introduction: Distal forearm fractures (DFF) account for 1.5% of emergency department (ED) visits in the United States. Clinicians frequently obtain imaging above/below the location of injury to rule out additional injuries. We sought to determine the incidence of associated proximal fractures (APF) in the setting of DFF and to evaluate the imaging practices in a nationally representative sample of EDs. Methods: We queried the 2013 National Emergency Department Sample using International Classification of Diseases, 9th edition, diagnostic codes for DFF and APF. Current Procedural Technology codes identified associated imaging studies. We calculated national estimates using a weighted analysis of patient and hospital-level characteristics associated with APF and imaging practices. An analysis of costs estimated the financial impact of additional imaging in patients with DFF using Medicare reimbursement to approximate costs according to the 2018 Medicare Physician Fee Schedule. Results: In 2013, an estimated 297,755 ED visits (weighted) were associated with a DFF, of which 1.6% (4836 cases) had an APF. The incidence of APF was lower among females (odds ratio [OR] (0.76); 95% confidence interval [CI], 0.64–0.91) but higher in metropolitan teaching hospitals compared to metropolitan non-teaching hospitals (OR [2.39]; 95% CI, 1.43–3.99) and Level 1 trauma centers (OR [3.9]; 95%, 1.91–7.96) compared to non-trauma centers. Approximately 40% (n = 117,948) of those with only DFF received non-wrist radiographs and 19% (n = 55,236) underwent non-wrist/non-forearm imaging. Factors independently associated with additional imaging included gender, payer, patient and hospital rurality, hospital region, teaching status, ownership, and trauma center level. Nearly $3.6 million (2018 U.S. dollars) was spent on the aforementioned additional imaging. Conclusion: Despite the frequency of proximal imaging in patients with DFF, the incidence of APF was low. Further study to identify risk factors for APF based on mechanism and physical examination factors may result in reduced imaging and decreased avoidable healthcare spending.

Analysis of magnetic resonance image (MRI) quality after open (Op)-transforaminal interbody fusion (TLIF) and minimally invasive (MI)-TLIF with the implantation of structurally different systems has not previously been performed. The objective of this study was to conduct a comparative analysis of the postoperative MRI following MI and Op one-segment TLIF. Material and Methods: The nonrandomized retrospective single-center study included 80 patients (46 men and 24 women) aged 48 + 14.2 years. In group I (n = 20) Op-TLIF with open transpedicular screw fixation (TSF) was performed, in II group (n = 60), the MI-TLIF technique was used: IIa (n = 20)—rigid interspinous stabilizer; IIb (n = 20)—unilateral TSF and contralateral facet fixation; IIc (n = 20)—bilateral TSF. Results: Comparison of the quality of postoperative imaging in IIa and IIb subgroups showed fewer MRI artifacts and a significantly greater MR deterioration after Op and MI TSF. Comparison of the multifidus muscle area showed less atrophy after MI-TLIF and significantly greater atrophy after Op-TLIF. Conclusion: MI-TLIF and Op-TLIF with TSF have comparable postoperative MR artifacts at the operative level, with a greater degree of muscle atrophy using the Op-TLIF. Rigid interspinous implant and unilateral TSF with contralateral facet fixation have less artifacts and changes in the multifidus muscle area.

Many enzymes are known to change conformations during their catalytic cycle, but the role of these protein motions is not well understood. Escherichia coli dihydrofolate reductase (DHFR) is a small, flexible enzyme that is often used as a model system for understanding enzyme dynamics. Recently, native tryptophan fluorescence was used as a probe to study micro- to millisecond dynamics of DHFR. Yet, because DHFR has five native tryptophans, the origin of the observed conformational changes could not be assigned to a specific region within the enzyme. Here, we use DHFR mutants, each with a single tryptophan as a probe for temperature jump fluorescence spectroscopy, to further inform our understanding of DHFR dynamics. The equilibrium tryptophan fluorescence of the mutants shows that each tryptophan is in a different environment and that wild-type DHFR fluorescence is not a simple summation of all the individual tryptophan fluorescence signatures due to tryptophan–tryptophan interactions. Additionally, each mutant exhibits a two-phase relaxation profile corresponding to ligand association/dissociation convolved with associated conformational changes and a slow conformational change that is independent of ligand association and dissociation, similar to the wild-type enzyme. However, the relaxation rate of the slow phase depends on the location of the tryptophan within the enzyme, supporting the conclusion that the individual tryptophan fluorescence dynamics do not originate from a single collective motion, but instead report on local motions throughout the enzyme.

Limited research has been performed to evaluate the effects of high-frequency electrical stunning (ES) methods on the lipid oxidative stability of the meat goose livers. This study was conducted to evaluate the effects of high-frequency-ES current intensities on lipid oxidative stability and antioxidant capacity in the liver of Yangzhou goose (Anser cygnoides domesticus). Forty 92-day-old male Yangzhou geese were randomly divided into five treatments (n = 8). Geese were not stunned (control) or exposed to ES for 10 s with alternating current (AC) at 500 Hz in a water bath. Current intensities were set at 30 V/20 mA (E30V), 60 V/40 mA (E60V), 90 V/70 mA (E90V), or 120 V/100 mA (E120V), respectively. The malondialdehyde level at day 0 was the highest in 120 V (p < 0.05). Antioxidant enzymes’ activity on day 2 was the highest in E60V. The 1, 1-diphenyl-2-picrylhydrazyl free radical (DPPH·) elimination ability was lower in the E120V than that in the E60V at two days and four days postmortem (p < 0.05). A combination of 60 V/40 mA/ 500 Hz/ 10 s per bird could be applied in the ES of Yangzhou geese to improve the lipid oxidative stability and antioxidant capacity in the livers.