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Using a prion amplification assay, we identified prions in tissues from wild pigs (Sus scrofa) living in areas of the United States with variable chronic wasting disease (CWD) epidemiology. Our findings indicate that scavenging swine could play a role in disseminating CWD and could therefore influence its epidemiology, geographic distribution, and interspecies spread.

Volatile organic compounds, colloquially referred to as “terpenes”, have been proposed to impact the therapeutic qualities that are traditionally ascribed to cannabis. However, the contribution of these terpenes in anxiety, at relevant levels and exposure methods common with cannabis use, is lacking empirical assessment. We tested the anxiolytic properties of two prominent cannabis terpenes, linalool and β-myrcene, in male and female mice using short duration vapor pulls to model human inhalation when combusting flower or vaping cannabis oil. We observed sex differences in the locomotor effects in the open field and anxiolytic properties in the elevated plus maze of these terpenes that depended on their exposure characteristics. Both linalool and β-myrcene had anxiolytic effects in female mice when delivered in discrete vapor pulls over the course of 30 min. In male mice, only a single vapor hit containing linalool or β-myrcene had anxiolytic effects. The combination of sub-effective levels of linalool and the phytocannabinoid, cannabidiol (CBD), had synergistic anxiolytic effects in females, but these entourage effects between CBD and terpenes were absent with β-myrcene for females and for either terpene in males. Together, our findings reveal sex differences in the anxiolytic properties of common cannabis terpenes and highlight the potential benefits of unique combinations of CBD and terpenes in expanding the therapeutic dose window.

Abstract Digital health tools are emerging as a promising solution for optimizing Deep Brain Stimulation (DBS) therapy for neurodegenerative diseases such as Parkinson’s Disease (PD), addressing challenges of therapy maintenance, care access, and discrete assessments. Wearable technology and mobile health platforms can offer remote monitoring capabilities, allowing for real-time virtual programming with optimization of patients’ therapy, use of digital biomarkers that can help identify the onset, symptoms, medication-related fluctuations and side-effects, and response to DBS treatment. Through an investigational remote monitoring application (RM app) integrating patient-reported outcomes (PROs) and objective data using wearables, we aimed to develop an accessible, data-driven digital tool to monitor patient symptoms and deliver low-burden and easy-to-access DBS therapy. The platform is designed to be compatible with several consumer-grade wearables and ensures patient data privacy and security. We conducted a feasibility study to test the tool’s effectiveness in a large-scale DBS study, where subjects received a home-monitoring kit consisting of an iPhone and Apple Watch, pre-configured with the RM app to enable remote data collection. Compliance in this paper is defined as adherence to the RM app and Apple watch usage. Analysis of 67 subjects from the study demonstrated an average compliance rate of 63.3% for daily check-ins, 85.5% for monthly surveys and 61.9% for passive data with an average of 55 days of post-implant data. The study highlights the feasibility of frequent monitoring in the early post-implant period that could lead to optimal outcomes for patients. The platform developed can further optimize study execution and be extended to other chronic conditions treated with neuromodulation implants such as depression and pain.

Mohan Prasath Mani,1,2 Hemanth Ponnambalath Mohanadas,3 Ahmad Athif Mohd Faudzi,4,5 Ahmad Fauzi Ismail,6 Nick Tucker,7 Shahrol Mohamaddan,8 Manikandan Ayyar,9 Tamilselvam Palanisamy,1 Rajasekar Rathanasamy,10 Saravana Kumar Jaganathan11– 13 1Department of Mechanical Engineering, SNS College of Technology, Coimbatore, TN, India; 2School of Biomedical Engineering and Health Sciences, Faculty of Engineering, Universiti Teknologi Malaysia, Skudai, JB, Malaysia; 3Device Development Group, New Product Research and Development, Abbott Diabetes Care, Alameda, CA, USA; 4School of Electrical Engineering, Faculty of Engineering, UniversitiTeknologi Malaysia, Skudai, JB, Malaysia; 5Centre for Artificial Intelligence and Robotics, Universiti Teknologi Malaysia, Kuala Lumpur, SG, Malaysia; 6Advanced Membrane Technology Research Centre (AMTEC), School of Chemical and Energy Engineering, Universiti Teknologi Malaysia, Skudai, JB, Malaysia; 7School of Engineering and Physical Sciences, College of Health and Sciences, University of Lincoln, Lincoln, LS, UK; 8Innovative Global ProgramCollege of Engineering Shibaura Institute of Technology Tokyo, Tokyo, Japan; 9Department of Chemistry, Centre for Materials Chemistry, Karapagam Acdemy of Higher Education, Coimbatore, TN, India; 10Department of Mechanical Engineering, Kongu Engineering College, Perunduari, TN, India; 11Institute of Research and Development, Duy Tan University, Da Nang, Vietnam; 12School of Engineering & Technology, Duy Tan University, Da Nang, Vietnam; 13Biomaterials and Tissue Engineering, School of Engineering and Physical Sciences, College of Health and Sciences, University of Lincoln, Lincoln, LS, UKCorrespondence: Saravana Kumar Jaganathan, Institute of Research and Development, Duy Tan University, Da Nang, Vietnam, Email jaganathansaravanakumar@dtu.edu.vn; sjaganathan@lincoln.ac.ukPurpose: Wound patches are essential for wound healing, yet developing patches with enhanced mechanical and biological properties remains challenging. This study aimed to enhance the mechanical and biological properties of polyurethane (PU) by incorporating magnesium chloride (MgCl2) into the patch.Methodology: The composite patch was fabricated using the electrospinning technique, producing nanofibers from a mixture of PU and MgCl2 solutions. The electrospun PU/MgCl2 was then evaluated for various physico-chemical characteristics and biological properties to determine its suitability for wound healing applications.Results: Tensile strength testing showed that the mechanical properties of the composite patch (10.98 ± 0.18) were significantly improved compared to pristine PU (6.66 ± 0.44). Field scanning electron microscopy (FESEM) revealed that the electrospun nanofiber patch had a smooth, randomly oriented non-woven structure (PU – 830 ± 145 nm and PU/MgCl2 – 508 ± 151 nm). Fourier infrared spectroscopy (FTIR) confirmed magnesium chloride’s presence in the polyurethane matrix via strong hydrogen bond formation. Blood compatibility studies using coagulation assays, including activated partial thromboplastin time (APTT), prothrombin time (PT), and hemolysis assays, demonstrated improved blood compatibility of the composite patch (APTT – 174 ± 0.5 s, PT – 91 ± 0.8s, and Hemolytic percentage – 1.78%) compared to pristine PU (APTT – 152 ± 1.2s, PT – 73 ± 1.7s, and Hemolytic percentage – 2.55%). Antimicrobial testing showed an enhanced zone of inhibition (Staphylococcus aureus – 21.5 ± 0.5 mm and Escherichia coli – 27.5 ± 2.5 mm) compared to the control, while cell viability assays confirmed the non-cytotoxic nature of the developed patches on fibroblast cells.Conclusion: The study concludes that adding MgCl2 to PU significantly improves the mechanical, biological, and biocompatibility properties of the patch. This composite patch shows potential for future wound healing applications, with further studies needed to validate its efficacy in-vivo.Keywords: wound dressings, polyurethane, MgCl2, biomaterial, nanofibers

ABSTRACT: The primary objective of this cross-sectional observational study was to determine whether bulk tank milk quality, udder health, udder hygiene, and milk production outcomes were associated with facility type on organic dairies. A secondary objective was to identify other management-related risk factors associated with bulk tank milk quality, udder health, udder hygiene, and milk production on organic dairy herds in Vermont. We aimed to enroll 40 farms, to compare herds using the 2 most common systems (freestalls, tiestalls) for housing organic dairy cattle in the state with those using a bedded pack during the nongrazing season (typically November–May). Two general styles of bedded packs were observed: cultivated bedded packs and untilled deep-bedded packs. Due to the limited number of herds using bedded packs to house lactating dairy cattle in Vermont, we combined untilled and cultivated bedded packs to describe udder hygiene, milk quality, and udder health on these loose-housing systems deeply bedded with organic material. The study was completed on 21 farms (5 bedded packs, 6 freestalls, 10 tiestalls) before interruption due to the COVID-19 pandemic. Data captured from DHIA records from the test closest to the date of the farm visit included average SCS, standardized 150-d milk (pounds), percentage of cows with current high SCS (SCS ≥4.0), percentage of cows with newly elevated SCS (previous SCS

Abstract Image segmentation of the liver is an important step in treatment planning for liver cancer. However, manual segmentation at a large scale is not practical, leading to increasing reliance on deep learning models to automatically segment the liver. This manuscript develops a generalizable deep learning model to segment the liver on T1-weighted MR images. In particular, three distinct deep learning architectures (nnUNet, PocketNet, Swin UNETR) were considered using data gathered from six geographically different institutions. A total of 819 T1-weighted MR images were gathered from both public and internal sources. Our experiments compared each architecture’s testing performance when trained both intra-institutionally and inter-institutionally. Models trained using nnUNet and its PocketNet variant achieved mean Dice-Sorensen similarity coefficients>0.9 on both intra- and inter-institutional test set data. The performance of these models suggests that nnUNet and PocketNet liver segmentation models trained on a large and diverse collection of T1-weighted MR images would on average achieve good intra-institutional segmentation performance.

The electron density ( n _e ) of the interstellar medium (ISM) in star-forming galaxies is intimately linked to star formation and ionization condition. Using the high-resolution spectra obtained from the JWST Near-Infrared Spectrograph (NIRSpec) microshutter assembly (MSA) as part of the GLASS-JWST program, we have assembled the largest sample to date (34 galaxies) with individual n _e measurements derived from the [O ii ] λλ 3726, 29 and/or [S ii ] λλ 6718, 32 doublets at 0.7 ≲ z ≲ 9.3. The gravitational lensing magnification by the foreground A2744 cluster allows us to probe n _e in galaxies with stellar masses ( M _* ) down to ≃10 ^7.5 M _⊙ across the entire redshift range. Our analysis reveals that the [O ii ] flux ratios are marginally anticorrelated with a specific star formation rate (sSFR) within a 1 σ confidence interval, whereas the [S ii ] flux ratios show no significant correlation with sSFR. Despite a clear correlation between sSFR and redshift within our sample, we find no apparent redshift evolution of n _e at z ≃ 1–9. Our data set also includes 13 galaxies where n _e can be measured from both [O ii ] and [S ii ]. Contrary to findings at lower redshifts, we observe considerable scatter in n _e measurements from [O ii ] and [S ii ], indicating a complex gaseous environment with significant variations in n _e in high-redshift galaxies. This work highlights the unique capability of JWST NIRSpec/MSA high-resolution spectroscopy to characterize the detailed physical properties of the ISM in individual high-redshift galaxies.

The radial gradient of gas-phase metallicity is a powerful probe of the chemical and structural evolution of star-forming galaxies, closely tied to disk formation and gas kinematics in the early Universe. We present spatially resolved chemical and dynamical properties for a sample of 25 galaxies at 0.5 ≲ z ≲ 1.7 from the MSA-3D survey. These innovative observations provide 3D spectroscopy of galaxies at a spatial resolution approaching JWST’s diffraction limit and a high spectral resolution of R ≃ 2700. The metallicity gradients measured in our galaxy sample range from −0.03 to 0.02 dex kpc ^−1 . Most galaxies exhibit negative or flat radial gradients, indicating lower metallicity in the outskirts or uniform metallicity throughout the entire galaxy. We confirm a tight relationship between stellar mass and metallicity gradient at z ∼ 1 with small intrinsic scatter of 0.02 dex kpc ^−1 . Our results indicate that metallicity gradients become increasingly negative as stellar mass increases, likely because the more massive galaxies tend to be more “disky.” This relationship is consistent with the predictions from cosmological hydrodynamic zoom-in simulations with strong stellar feedback. This work presents the effort to harness the multiplexing capability of the JWST NIRSpec microshutter assembly in slit-stepping mode to map the chemical and kinematic profiles of high-redshift galaxies in large samples and at high spatial and spectral resolution.

Objective: Compare the differences in cost, complications, new intra-articular diagnoses, and reoperations among patients with ankle instability submitted to lateral ankle ligament repair/reconstruction with or without arthroscopic procedures. Methods: A retrospective analysis of 2,428 patients from the PearlDiver Humana dataset with ankle sprain or instability codes and compared outcomes between those submitted to lateral ankle ligament repair/reconstruction with or without arthroscopy. Results: Patients without arthroscopy had higher complication rates (9.87% vs. 5.41%; χ2[1, n = 1,236] = 5.83, p = 0.01), while the difference in reconstruction groups was insignificant (p = 0.09). Arthroscopy groups had higher rates of newly diagnosed intra-articular pathology: repair with arthroscopy (57.0%) vs. without (35.6%; χ2[1, n = 1,236] = 44.47, p < 0.001); reconstruction with arthroscopy (63.0%) vs. without (39.8%; χ2[1, n = 1,211] = 61.90, p < 0.001). Reoperation rates for intra-articular pathology were higher in the arthroscopy group (6.89% vs. 4.18%; χ2[1, n = 2,433] = 8.09, p = 0.006), with significantly shorter time to reoperation (303 vs. 474 days, p = 0.045). Conclusions: Arthroscopy does not increase complication rates and allows for earlier diagnosis and treatment of intra-articular pathology, potentially leading to earlier reoperation. Level of evidence III; Retrospective Comparative Study.

Methods from Machine Learning (ML) and Computer Vision (CV) have proven powerful tools for quickly and accurately analyzing behavioral recordings. The computational complexity of these techniques, however, often precludes applications that require real-time analysis: for example, experiments where a stimulus must be applied in response to a particular behavior or samples must be collected soon after the behavior occurs. Here, we describe SARTAB (Scalable Automated Real-Time Analysis of Behavior), a system that achieves automated real-time behavior detection by continuously monitoring animal positions relative to behaviorally relevant Regions Of Interest (ROIs). We then show how we used this system to detect infrequent courtship behaviors in Pseudotropheus demasoni (a species of Lake Malawi African cichlid fish) to collect neural tissue samples from actively behaving individuals for multiomic profiling at single nucleus resolution. Within this experimental context, we achieve high ROI and animal detection accuracies (mAP@[.5 : .95] of 0.969 and 0.718, respectively) and 100% classification accuracy on a set of 32 manually selected behavioral clips. SARTAB is unique in that all analysis runs on low-cost, edge-deployed hardware, making it a highly scalable and energy-efficient solution for real-time experimental feedback. Although our solution was developed specifically to study cichlid courtship behavior, the intrinsic flexibility of neural network analysis ensures that our approach can be adapted to novel species, behaviors, and environments.

Abstract Immunohistochemistry is a crucial method for detecting specific proteins within tissue samples, yet constrained to one biomarker per tissue section. Multiplexed immunofluorescence, while allowing simultaneous visualization of multiple proteins, faces limitations in the number of simultaneous fluorescent labels due to spectral overlap. Although cyclic immunofluorescence techniques have successfully broadened antibody staining capacities in a single tissue sample, they are plagued by time-consuming and labor-intensive procedures, sample degradation risks, and inability to scale beyond thin sections. In this study, we introduce the use of 3D confocal Fluorescence Lifetime Imaging Microscopy as a high-throughput, multiplexed immunofluorescence platform that can differentiate 11 or more biomarkers in 3D tissue volumes. Leveraging both spectral and lifetime information, this approach allows for practical spatial biology in thin sections that can readily scale to larger volumes of tissue. We believe that this highly multiplexed and versatile biomarker imaging platform will significantly expedite cancer research and enable new translational approaches in the future.

Introduction: Medical and nonmedical ketamine use is increasing in the United States. This will likely lead to an increase in emergency department (ED) visits in individuals experiencing associated side effects. Physicians will need to be able to effectively recognize and manage ketamine-related complications. Case Report: A 31-year-old male with a three-year history of inhalational, intramuscular, and intravenous nonmedical ketamine use presented to the ED twice within a week with symptoms of severe atraumatic back pain, abdominal pain, and dyspepsia. A comprehensive workup, including advanced imaging, was unrevealing for identifiable causes, and the patient was discharged with instructions for primary care follow-up for further evaluation. The patient used information shared on Reddit, an online forum and social network, to identify that the cause of his pain was related to chronic ketamine use. Subsequently, upon discontinuation of ketamine, the pain improved in 24 hours. The patient self-navigated to addiction treatment. Conclusion: Emergency physicians should consider sequelae of chronic ketamine use as a possible cause for gastrointestinal and urologic symptoms in the ED. In addition to thorough examination and assessment for other acute medical problems, patients should be offered education, symptomatic treatment, and linkage to harm reduction and substance use disorder treatment services.

Abstract While quiescent galaxies have comparable amounts of cool gas in their outer circumgalactic medium (CGM) compared to star-forming galaxies, they have significantly less interstellar gas. However, open questions remain on the processes causing galaxies to stop forming stars and stay quiescent. Theories suggest dynamical interactions with the hot corona prevent cool gas from reaching the galaxy, therefore predicting the inner regions of quiescent galaxy CGMs are devoid of cool gas. However, there is a lack of understanding of the inner regions of CGMs due to the lack of spatial information in quasar-sightline methods. We present integral-field spectroscopy probing 10–20 kpc (2.4–4.8 Re) around a massive quiescent galaxy using a gravitationally lensed star-forming galaxy. We detect absorption from Magnesium (MgII) implying large amounts of cool atomic gas (108.4–109.3 M⊙ with T~104 Kelvin), in comparable amounts to star-forming galaxies. Lens modeling of Hubble imaging also reveals a diffuse asymmetric component of significant mass consistent with the spatial extent of the MgII absorption, and offset from the galaxy light profile. This study demonstrates the power of galaxy-scale gravitational lenses to not only probe the gas around galaxies, but to also independently probe the mass of the CGM due to it’s gravitational effect.

Background Access to language assistance is a patient's right under federal law. Despite this, underuse of language services persists. Objective The aim of this study was to explore the interest in obtaining bilingual certification and to describe perspectives on language services by resident physicians. Methods Between May and August 2021, we conducted a cross-sectional survey of residents at a public, urban hospital serving mostly patients with limited English proficiency (LEP). We assessed resident perspectives on language services, exposure to language-related trainings, non-English language (NEL) skills, and interest in bilingual certification. Results A total of 214 residents of 289 completed the survey (a 74% response rate). Of the 95 residents who used their NEL for patient care, 65 (68%) would be interested in bilingual certification. Sixty-nine (33%), 65 (31%), and 95 (45%) residents disagreed or strongly disagreed with being satisfied with the language services available, convenience, and sufficient equipment, respectively. Furthermore, 28 (13%) disagreed or strongly disagreed that they could achieve bi-directional communication with LEP patients. Conclusions Over a quarter of the residents expressed interest in bilingual certification and were likely to pass the certification exam. Many reported using their own NEL skills without certification and held negative views on services and trainings.

PURPOSEIncreased automation has been identified as one approach to improving global cancer care. The Radiation Planning Assistant (RPA) is a web-based tool offering automated radiotherapy (RT) contouring and planning to low-resource clinics. In this study, the RPA workflow and clinical acceptability were assessed by physicians around the world.METHODSThe RPA output for 75 cases was reviewed by at least three physicians; 31 radiation oncologists at 16 institutions in six countries on five continents reviewed RPA contours and plans for clinical acceptability using a 5-point Likert scale.RESULTSFor cervical cancer, RPA plans using bony landmarks were scored as usable as-is in 81% (with minor edits 93%); using soft tissue contours, plans were scored as usable as-is in 79% (with minor edits 96%). For postmastectomy breast cancer, RPA plans were scored as usable as-is in 44% (with minor edits 91%). For whole-brain treatment, RPA plans were scored as usable as-is in 67% (with minor edits 99%). For head/neck cancer, the normal tissue autocontours were acceptable as-is in 89% (with minor edits 97%). The clinical target volumes (CTVs) were acceptable as-is in 40% (with minor edits 93%). The volumetric-modulated arc therapy (VMAT) plans were acceptable as-is in 87% (with minor edits 96%). For cervical cancer, the normal tissue autocontours were acceptable as-is in 92% (with minor edits 99%). The CTVs for cervical cancer were scored as acceptable as-is in 83% (with minor edits 92%). The VMAT plans for cervical cancer were acceptable as-is in 99% (with minor edits 100%).CONCLUSIONThe RPA, a web-based tool designed to improve access to high-quality RT in low-resource settings, has high rates of clinical acceptability by practicing clinicians around the world. It has significant potential for successful implementation in low-resource clinics.

Summary: Patients with cancer are at increased risk of death from COVID-19 and have reduced immune responses to SARS-CoV2 vaccines, necessitating regular boosters. We performed comprehensive chart reviews, surveys of patients attitudes, serology for SARS-CoV-2 antibodies and T cell receptor (TCR) β sequencing for cellular responses on a cohort of 982 cancer patients receiving active cancer therapy accrued between November-3-2020 and Mar-31-2023. We found that 92 · 3% of patients received the primer vaccine, 70 · 8% received one monovalent booster, but only 30 · 1% received a bivalent booster. Booster uptake was lower under age 50, and among African American or Hispanic patients. Nearly all patients seroconverted after 2+ booster vaccinations (>99%) and improved cellular responses, demonstrating that repeated boosters could overcome poor response to vaccination. Receipt of booster vaccinations was associated with a lower risk of all-cause mortality (HR = 0 · 61, p = 0 · 024). Booster uptake in high-risk cancer patients remains low and strategies to encourage booster uptake are needed.

Abstract Polyethylene terephthalate (PET), the most abundantly produced polyester plastic, can be depolymerized by the Ideonella sakaiensis PETase enzyme. Based on multiple PETase crystal structures, the reaction has been proposed to proceed via a two-step serine hydrolase mechanism mediated by a serine-histidine-aspartate catalytic triad. To elucidate the multi-step PETase catalytic mechanism, we use transition path sampling and likelihood maximization to identify optimal reaction coordinates for the PETase enzyme. We predict that deacylation is likely rate-limiting, and the reaction coordinates for both steps include elements describing nucleophilic attack, ester bond cleavage, and the “moving-histidine” mechanism. We find that the flexibility of Trp185 promotes the reaction, providing an explanation for decreased activity observed in mutations that restrict Trp185 motion. Overall, this study uses unbiased computational approaches to reveal the detailed reaction mechanism necessary for further engineering of an important class of enzymes for plastics bioconversion.

This is the first in a series of interviews with those closely tied to the Hachette v. Internet Archive lawsuit. In March 2023, the court ruled against the Internet Archive and its use of the Emergency Lending Library causing a ripple throughout the library and education fields. Below, find the answers to some of the questions that the case elicited by JCEL contributors and copyright scholars Dave Hansen, Michelle Wu, and Kyle Courtney.

The practical application of new single molecule protein sequencing (SMPS) technologies requires accurate estimates of their associated sequencing error rates. Here, we describe the development and application of two distinct parameter estimation methods for analyzing SMPS reads produced by fluorosequencing. A Hidden Markov Model (HMM) based approach, extends whatprot, where we previously used HMMs for SMPS peptide-read matching. This extension offers a principled approach for estimating key parameters for fluorosequencing experiments, including missed amino acid cleavages, dye loss, and peptide detachment. Specifically, we adapted the Baum-Welch algorithm, a standard technique to estimate transition probabilities for an HMM using expectation maximization, but modified here to estimate a small number of parameter values directly rather than estimating every transition probability independently. We demonstrate a high degree of accuracy on simulated data, but on experimental datasets, we observed that the model needed to be augmented with an additional error type, N-terminal blocking. This, in combination with data pre-processing, results in reasonable parameterizations of experimental datasets that agree with controlled experimental perturbations. A second independent implementation using a hybrid of DIRECT and Powell's method to reduce the root mean squared error (RMSE) between simulations and the real dataset was also developed. We compare these methods on both simulated and real data, finding that our Baum-Welch based approach outperforms DIRECT and Powell's method by most, but not all, criteria. Although some discrepancies between the results exist, we also find that both approaches provide similar error rate estimates from experimental single molecule fluorosequencing datasets.

Background and purpose: In many clinics, positron-emission tomography is unavailable and clinician time extremely limited. Here we describe a deep-learning model for autocontouring gross disease for patients undergoing palliative radiotherapy for primary lung lesions and/or hilar/mediastinal nodal disease, based only on computed tomography (CT) images. Materials and methods: An autocontouring model (nnU-Net) was trained to contour gross disease in 379 cases (352 training, 27 test); 11 further test cases from an external centre were also included. Anchor-point-based post-processing was applied to remove extraneous autocontoured regions. The autocontours were evaluated quantitatively in terms of volume similarity (Dice similarity coefficient [DSC], surface Dice coefficient, 95th percentile Hausdorff distance [HD95], and mean surface distance), and scored for usability by two consultant oncologists. The magnitude of treatment margin needed to account for geometric discrepancies was also assessed. Results: The anchor point process successfully removed all erroneous regions from the autocontoured disease, and identified two cases to be excluded from further analysis due to ‘missed’ disease. The average DSC and HD95 were 0.8 ± 0.1 and 10.5 ± 7.3 mm, respectively. A 10-mm uniform margin-distance applied to the autocontoured region was found to yield “full coverage” (sensitivity > 0.99) of the clinical contour for 64 % of cases. Ninety-seven percent of evaluated autocontours were scored by both clinicians as requiring no or minor edits. Conclusions: Our autocontouring model was shown to produce clinically usable disease outlines, based on CT alone, for approximately two-thirds of patients undergoing lung radiotherapy. Further work is necessary to improve this before clinical implementation.

ABSTRACT The high-consequence pathogen Bacillus anthracis causes human anthrax and often results in lethal infections without the rapid administration of effective antimicrobial treatment. Antimicrobial resistance profiling is therefore critical to inform post-exposure prophylaxis and treatment decisions, especially during emergencies such as outbreaks or where intentional release is suspected. Whole-genome sequencing using a rapid long-read sequencer can uncover antimicrobial resistance patterns if genetic markers of resistance are known. To identify genomic markers associated with antimicrobial resistance, we isolated B. anthracis derived from the avirulent Sterne strain with elevated minimal inhibitory concentrations to clarithromycin. Mutants were characterized both phenotypically through broth microdilution susceptibility testing and observations during culturing, as well as genotypically with whole-genome sequencing. We identified two different in-frame insertions in the L22 ribosomal protein-encoding gene rplV, which were subsequently confirmed to be involved in clarithromycin resistance through the reversion of the mutant gene to the parent (drug-susceptible) sequence. Detection of the rplV insertions was possible with rapid long-read sequencing, with a time-to-answer within 3 h. The mutations associated with clarithromycin resistance described here will be used in conjunction with known genetic markers of resistance for other antimicrobials to strengthen the prediction of antimicrobial resistance in B. anthracis.IMPORTANCEThe disease anthrax, caused by the pathogen Bacillus anthracis, is extremely deadly if not treated quickly and appropriately. Clarithromycin is an antibiotic recommended for the treatment and post-exposure prophylaxis of anthrax by the Centers for Disease Control and Prevention; however, little is known about the ability of B. anthracis to develop resistance to clarithromycin or the mechanism of that resistance. The characterization of clarithromycin-resistant isolates presented here provides valuable information for researchers and clinicians in the event of a release of the resistant strain. Additionally, knowledge of the genetic basis of resistance provides a foundation for susceptibility prediction through rapid genome sequencing to inform timely treatment decisions.

Abstract Introduction This study aimed to develop a modified frailty index (MFI) to predict the risks of revision total hip arthroplasty (THA). Methods Data from the American College of Surgeons - National Surgical Quality Improvement Program were analyzed for patients who underwent revision THA from 2015 to 2020. An MFI was composed of the risk factors, including severe obesity (body mass index > 35), osteoporosis, non-independent function status prior to surgery, congestive heart failure within 30 days of surgery, hypoalbuminemia (serum albumin < 3.5), hypertension requiring medication, type 1 or type 2 diabetes, and a history of chronic obstructive pulmonary disease or pneumonia. The patients were assigned based on the MFI scores (MFI0, no risk factor; MFI1, 1–2 risk factors; MFI2, 3–4 risk factors; and MFI3, 5+ risk factors). Confidence intervals were set at 95% with a P value less than or equal to 0.05 considered statistically significant. Results A total of 17,868 patients (45% male, 55% female) were included and had an average age of 68.5 ± 11.5 years. Odds of any complication, when compared to MFI0, were 1.4 (95% CI [1.3, 1.6]) times greater for MFI1, 3.2 (95% CI [2.8, 3.6]) times greater for MFI2, and 10.8 (95% CI [5.8, 20.0]) times greater for MFI3 (P < 0.001). Odds of readmission, when compared to MFI0, were 1.4 (95% CI [1.3, 1.7]) times greater for MFI1, 2.5 (95% CI [2.1, 3.0]) times greater for MFI2, and 4.1 (95% CI [2.2, 7.8]) times greater for MFI3 (P < 0.001). Conclusion Increasing MFI scores correlate with increased odds of complication and readmission in patients who have undergone revision THA. This MFI may be used to predict the risks after revision THA.

Abstract Parkinson’s disease (PD) is the second most prevalent neurodegenerative disease and arises from dopamine (DA) neuron death selectively in the substantia nigra pars compacta (SNc). Rit2 is a reported PD risk allele, and recent single cell transcriptomic studies identified a major RIT2 cluster in PD DA neurons, potentially linking Rit2 expression loss to a PD patient cohort. However, it is still unknown whether Rit2 loss itself impacts DA neuron function and/or viability. Here we report that conditional Rit2 silencing in mouse DA neurons drove motor dysfunction that occurred earlier in males than females and was rescued at early stages by either inhibiting the DA transporter (DAT) or with L-DOPA treatment. Motor dysfunction was accompanied by decreased DA release, striatal DA content, phenotypic DAergic markers, DA neurons, and DAergic terminals, with increased pSer129-alpha synuclein and pSer935-LRRK2 expression. These results provide clear evidence that Rit2 loss is causal for SNc cell death and motor dysfunction, and reveal key sex-specific differences in the response to Rit2 loss.

Tessa Fischer,1,2 Jenna Tucker,1,2 Charlie Li,3 Kyungmi Min,3 Dadong Xu,3 Laurence R Upjohn,1 Madhur Kumar1 1Food and Drug Branch, California Department of Public Health, Sacramento, CA, USA; 2California Epidemiologic Investigation Service (Cal EIS) Fellowship Program, California Department of Public Health, Sacramento, CA, USA; 3Food and Drug Laboratory Branch, California Department of Public Health, Richmond, CA, USACorrespondence: Madhur Kumar, Food and Drug Branch, California Department of Public Health, 1500 Capitol Avenue, MS 7602, Sacramento, CA, 95814, USA, Email madhur.kumar@cdph.ca.govPurpose: The Food and Drug Administration (FDA) has warned against tainted products for sexual enhancement containing prescription phosphodiesterase-5 (PDE5) inhibitors marketed as dietary supplements. The California Department of Public Health (CDPH)’s Food and Drug Branch (FDB) initiated a study to assess the presence of such adulterated products, specifically to identify the presence of PDE5 inhibitors in dietary supplements marketed for sexual enhancement, sampled at local retail locations.Methods: A convenience sample of products marketed as sexual enhancement dietary supplements was purchased from retail stores, within a 15-mile radius of downtown Sacramento, California. Samples were submitted to the Food and Drug Laboratory Branch (FDLB) and screened for 19 different PDE5 inhibitors using liquid chromatography-mass spectrometry. Samples were collected and analyzed from 2016 to 2018.Results: One hundred and two different products were purchased from 28 different retail locations. Sixty-seven percent were found to be adulterated with at least one PDE5 inhibitor. Of the positive samples, 40% were found to contain one PDE5 inhibitor and 60% were found to contain two or more. Sildenafil was the most common PDE5 inhibitor identified (74%), followed by tadalafil (59%). Eighteen percent of the tested samples had been associated with previous FDA warnings listed in their Tainted Products Database. Of these warning-associated samples, 72% were found to contain PDE5 inhibitors and 46% of these contained at least one of the same adulterants reported by the FDA.Conclusion: FDA reports and warnings have focused on dietary supplements found online or through screenings of international mail shipments. FDB results add to FDA findings by demonstrating that such adulterated products marketed for sexual enhancement are also sold at retail locations in California. These products have the potential to cause severe adverse health effects and therefore, it is essential to raise awareness of this significant public health concern.Keywords: dietary supplements, sexual enhancement products, adulteration

Abstract Planning for palliative radiotherapy is performed without the advantage of MR or PET imaging in many clinics. Here, we investigated CT-only GTV delineation for palliative treatment of head and neck cancer. Two multi-institutional datasets of palliative-intent treatment plans were retrospectively acquired: a set of 102 non-contrast-enhanced CTs and a set of 96 contrast-enhanced CTs. The nnU-Net auto-segmentation network was chosen for its strength in medical image segmentation, and five approaches separately trained: (1) heuristic-cropped, non-contrast images with a single GTV channel, (2) cropping around a manually-placed point in the tumor center for non-contrast images with a single GTV channel, (3) contrast-enhanced images with a single GTV channel, (4) contrast-enhanced images with separate primary and nodal GTV channels, and (5) contrast-enhanced images along with synthetic MR images with separate primary and nodal GTV channels. Median Dice similarity coefficient ranged from 0.6 to 0.7, surface Dice from 0.30 to 0.56, and 95th Hausdorff distance from 14.7 to 19.7 mm across the five approaches. Only surface Dice exhibited statistically-significant difference across these five approaches using a two-tailed Wilcoxon Rank-Sum test (p ≤ 0.05). Our CT-only results met or exceeded published values for head and neck GTV autocontouring using multi-modality images. However, significant edits would be necessary before clinical use in palliative radiotherapy.

Abstract Many polymer-based medical devices are sterilized by gamma irradiation. To reduce the use of cobalt-60 gamma-ray sources, transition from gamma ray to alternative irradiation technologies was proposed, namely electron beam (e-beam) and X-ray. A major impediment for such a transition is the knowledge gap in material compatibility with the different radiation sources. In this study, multi-layer films consisting of ethylene vinyl acetate (EVA) and ethylene vinyl alcohol (EVOH) components were irradiated to target doses of 30, 45, and 60 kGy by gamma-ray, e-beam, and X-ray sources. Effects of irradiation were evaluated on 12 material properties, and statistical comparisons between gamma irradiation and alternative technologies were conducted using the two one-sided t-test (or “equivalence test”) and classic t-test. Melting temperature and UV absorbance below 300 nm showed dose dependencies, while other investigated properties such as discoloration and mechanical durability did not change with dose up to 60 kGy. Based on these results, there is no material compatibility issue associated with the transition from gamma to e-beam or to X-ray as source of sterilization radiation of the studied multi-layer film.