Drosophila melanogaster Toll-9 elicits antiviral immunity against Drosophila C virus.

The Toll pathway plays a pivotal role in innate immune responses against pathogens. The evolutionary conserved pathogen recognition receptors (PRRs), including Toll like receptors (TLRs), play a crucial role in recognition of pathogen associated molecular patterns (PAMPs). The Drosophila genome encodes nine Toll receptors that are orthologous to mammalian TLRs. While mammalian TLRs directly recognize PAMPs, most Drosophila Tolls recognize the proteolytically cleaved ligand Spätzle to activate downstream signaling cascades. In this study, we demonstrated that Toll-9 is crucial for antiviral immunity against Drosophila C virus (DCV), a natural pathogen of Drosophila . A transposable element insertion in the Toll-9 gene renders the flies more susceptible to DCV. The stable expression of Toll-9 in S2 cells confers resistance against DCV infection by upregulation of the RNAi pathway. Toll-9 promotes the dephosphorylation of AKT, resulting in the induction of antiviral RNAi genes to inhibit DCV replication. Toll-9 localizes to the endosome where it binds dsRNA, suggesting its role to detect viral dsRNA. Toll-9 also induces apoptosis during DCV infection, contributing to its antiviral role. Together, this work identifies the role of Toll-9 in antiviral immunity against DCV infection through its ability to bind dsRNA and induce AKT-mediated RNAi antiviral immunity. IMPORTANCE: Insects rely on innate immunity and RNA interference (RNAi) to combat viral infections. Our study underscores the pivotal role of Drosophila Toll-9 in antiviral immunity, aligning with findings in Bombyx mori , where Toll-9 activation upregulates the RNAi component Dicer2 . We demonstrate that Drosophila Toll-9 functions as a pattern recognition receptor (PRR) for double-stranded RNA (dsRNA) during Drosophila C virus (DCV) infection, akin to mammalian TLRs. Toll-9 activation leads to the upregulation of key RNAi components, Dicer2 and Argonaute2 , and dephosphorylation of AKT triggers apoptosis via induction of proapoptotic genes Hid and Reaper . This study also reveals that Toll-9 localizes in endosomal compartments where it interacts with dsRNA. These insights enhance our understanding of Drosophila innate immune mechanisms, reflecting the evolutionary conservation of immune responses across diverse species and providing impetus for further research into the conserved roles of TLRs across the animal kingdom.

"Modeling Diffusive Search by Non-Adaptive Sperm: Empirical and Computational Insights".

During fertilization, mammalian sperm undergo a winnowing selection process that reduces the candidate pool of potential fertilizers from ∼10 6 -10 11 cells to 10 1 -10 2 cells (depending on the species). Classical sperm competition theory addresses the positive or 'stabilizing' selection that acts on sperm phenotypes within populations of organisms but does not strictly address the developmental consequences of sperm traits among individual organisms that are under purifying selection during fertilization. It is the latter that is of utmost concern for improving assisted reproductive technologies (ART) because 'low fitness' sperm may be inadvertently used for fertilization during interventions that rely heavily on artificial sperm selection, such as intracytoplasmic sperm injection (ICSI). Importantly, some form of sperm selection is used in nearly all forms of ART (e.g., differential centrifugation, swim-up, or hyaluronan binding assays, etc.). To date, there is no unifying quantitative framework (i.e., theory of sperm selection) that synthesizes causal mechanisms of selection with observed natural variation in individual sperm traits. In this report, we reframe the physiological function of sperm as a collective diffusive search process and develop multi-scale computational models to explore the causal dynamics that constrain sperm 'fitness' during fertilization. Several experimentally useful concepts are developed, including a probabilistic measure of sperm 'fitness' as well as an information theoretic measure of the magnitude of sperm selection, each of which are assessed under systematic increases in microenvironmental selective pressure acting on sperm motility patterns.

Intelligibility of Natively and Nonnatively Produced English Speech Presented in Noise to a Large Cohort of United States Service Members.

PURPOSE: A corpus of English matrix sentences produced by 60 native and nonnative speakers of English was developed as part of a multinational coalition task group. This corpus was tested on a large cohort of U.S. Service members in order to examine the effects of talker nativeness, listener nativeness, masker type, and hearing sensitivity on speech recognition performance in this population. METHOD: A total of 1,939 U.S. Service members (ages 18-68 years) completed this closed-set listening task, including 430 women and 110 nonnative English speakers. Stimuli were produced by native and nonnative speakers of English and were presented in speech-shaped noise and multitalker babble. Keyword recognition accuracy and response times were analyzed. RESULTS: General(ized) linear mixed-effects regression models found that, on the whole, speech recognition performance was lower for listeners who identified as nonnative speakers of English and when listening to speech produced by nonnative speakers of English. Talker and listener effects were more pronounced when listening in a babble masker than in a speech-shaped noise masker. Response times varied as a function of recognition score, with longest response times found for intermediate levels of performance. CONCLUSIONS: This study found additive effects of talker and listener nonnativeness when listening to speech in background noise. These effects were present in both accuracy and response time measures. No multiplicative effects of talker and listener language background were found. There was little evidence of a negative interaction between talker nonnativeness and hearing impairment, suggesting that these factors may have redundant effects on speech recognition. SUPPLEMENTAL MATERIAL: https://doi.org/10.23641/asha.26060191.

DNA mismatch and damage patterns revealed by single-molecule sequencing.

Mutations accumulate in the genome of every cell of the body throughout life, causing cancer and other diseases1,2. Most mutations begin as nucleotide mismatches or damage in one of the two strands of the DNA before becoming double-strand mutations if unrepaired or misrepaired3,4. However, current DNA-sequencing technologies cannot accurately resolve these initial single-strand events. Here we develop a single-molecule, long-read sequencing method (Hairpin Duplex Enhanced Fidelity sequencing (HiDEF-seq)) that achieves single-molecule fidelity for base substitutions when present in either one or both DNA strands. HiDEF-seq also detects cytosine deamination-a common type of DNA damage-with single-molecule fidelity. We profiled 134 samples from diverse tissues, including from individuals with cancer predisposition syndromes, and derive from them single-strand mismatch and damage signatures. We find correspondences between these single-strand signatures and known double-strand mutational signatures, which resolves the identity of the initiating lesions. Tumours deficient in both mismatch repair and replicative polymerase proofreading show distinct single-strand mismatch patterns compared to samples that are deficient in only polymerase proofreading. We also define a single-strand damage signature for APOBEC3A. In the mitochondrial genome, our findings support a mutagenic mechanism occurring primarily during replication. As double-strand DNA mutations are only the end point of the mutation process, our approach to detect the initiating single-strand events at single-molecule resolution will enable studies of how mutations arise in a variety of contexts, especially in cancer and ageing.

MicroRNAs Associated with Metformin Treatment in the Diabetes Prevention Program.

The Diabetes Prevention Program (DPP) randomized controlled trial demonstrated that metformin treatment reduced progression to type 2 diabetes (T2D) by 31% compared to placebo in adults with prediabetes. Circulating micro-ribonucleic acids (miRs) are promising biomarkers of T2D risk, but little is known about their associations with metformin regimens for T2D risk reduction. We compared the change in 24 circulating miRs from baseline to 2 years in a subset from DPP metformin intervention (n = 50) and placebo (n = 50) groups using Wilcoxon signed rank tests. Spearman correlations were used to evaluate associations between miR change and baseline clinical characteristics. Multiple linear regression was used to adjust for covariates. The sample was 73% female, 17% Black, 13% Hispanic, and 50 ± 11 years. Participants were obese, normotensive, prediabetic, and dyslipidemic. Change in 12 miR levels from baseline to 2 years was significantly different in the metformin group compared with placebo after adjusting for multiple comparisons: six (let-7c-5p, miR-151a-3p, miR-17-5p, miR-20b-5p, miR-29b-3p, and miR-93-5p) were significantly upregulated and six (miR-130b-3p, miR-22-3p, miR-222-3p, miR-320a-3p, miR-320c, miR-92a-3p) were significantly downregulated in the metformin group. These miRs help to explain how metformin is linked to T2D risk reduction, which may lead to novel biomarkers, therapeutics, and precision health strategies.

AAV gene therapy for hereditary spastic paraplegia type 50: a phase 1 trial in a single patient.

There are more than 10,000 individual rare diseases and most are without therapy. Personalized genetic therapy represents one promising approach for their treatment. We present a road map for individualized treatment of an ultra-rare disease by establishing a gene replacement therapy developed for a single patient with hereditary spastic paraplegia type 50 (SPG50). Through a multicenter collaboration, an adeno-associated virus-based gene therapy product carrying the AP4M1 gene was created and successfully administered intrathecally to a 4-year-old patient within 3 years of diagnosis as part of a single-patient phase 1 trial. Primary endpoints were safety and tolerability, and secondary endpoints evaluated efficacy. At 12 months after dosing, the therapy was well tolerated. No serious adverse events were observed, with minor events, including transient neutropenia and Clostridioides difficile gastroenteritis, experienced but resolved. Preliminary efficacy measures suggest a stabilization of the disease course. Longer follow-up is needed to confirm the safety and provide additional insights on the efficacy of the therapy. Overall, this report supports the safety of gene therapy for SPG50 and provides insights into precision therapy development for rare diseases. Clinical trial registration: NCT06069687 .

Exome-wide evidence of compound heterozygous effects across common phenotypes in the UK Biobank.

The phenotypic impact of compound heterozygous (CH) variation has not been investigated at the population scale. We phased rare variants (MAF ∼0.001%) in the UK Biobank (UKBB) exome-sequencing data to characterize recessive effects in 175,587 individuals across 311 common diseases. A total of 6.5% of individuals carry putatively damaging CH variants, 90% of which are only identifiable upon phasing rare variants (MAF < 0.38%). We identify six recessive gene-trait associations (p < 1.68 × 10-7) after accounting for relatedness, polygenicity, nearby common variants, and rare variant burden. Of these, just one is discovered when considering homozygosity alone. Using longitudinal health records, we additionally identify and replicate a novel association between bi-allelic variation in ATP2C2 and an earlier age at onset of chronic obstructive pulmonary disease (COPD) (p < 3.58 × 10-8). Genetic phase contributes to disease risk for gene-trait pairs: ATP2C2-COPD (p = 0.000238), FLG-asthma (p = 0.00205), and USH2A-visual impairment (p = 0.0084). We demonstrate the power of phasing large-scale genetic cohorts to discover phenome-wide consequences of compound heterozygosity.

Dysanapsis Genetic Risk Predicts Lung Function Across the Lifespan.

Rationale Dysanapsis refers to a mismatch between airway tree caliber and lung size arising early in life. Dysanapsis assessed by computed tomography (CT) is evident by early adulthood and associated with chronic obstructive pulmonary disease (COPD) risk later in life. Objective By examining the genetic factors associated with CT-assessed dysanapsis, we aimed to elucidate its molecular underpinnings and physiological significance across the lifespan. Methods We performed a genome-wide association study (GWAS) of CT-assessed dysanapsis in 11,951 adults, including individuals from two population-based and two COPD-enriched studies. We applied colocalization analysis to integrate GWAS and gene expression data from whole blood and lung. Genetic variants associated with dysanapsis were combined into a genetic risk score that was applied to examine association with lung function in children from a population-based birth cohort (n=1,278) and adults from the UK Biobank (n=369,157). Measurements and Main Results CT-assessed dysanapsis was associated with genetic variants from 21 independent signals in 19 gene regions, implicating HHIP, DSP, and NPNT as potential molecular targets based on colocalization of their expression. Higher dysanapsis genetic risk score was associated with obstructive spirometry among 5 year old children and among adults in the 5th, 6th and 7th decades of life. Conclusions CT-assessed dysanapsis is associated with variation in genes previously implicated in lung development and dysanapsis genetic risk is associated with obstructive lung function from early life through older adulthood. Dysanapsis may represent an endo-phenotype link between the genetic variations associated with lung function and COPD.

A Neuroradiologist's Guide to Operationalizing the Response Assessment in Neuro-Oncology (RANO) Criteria Version 2.0 for Gliomas in Adults.

Radiographic assessment plays a crucial role in the management of patients with central nervous system (CNS) tumors, aiding in treatment planning and evaluation of therapeutic efficacy by quantifying response. Recently, an updated version of the Response Assessment in Neuro-Oncology (RANO) criteria (RANO 2.0) was developed to improve upon prior criteria and provide an updated, standardized framework for assessing treatment response in clinical trials for gliomas in adults. This article provides an overview of significant updates to the criteria including (1) the use of a unified set of criteria for high and low grade gliomas in adults; (2) the use of the post-radiotherapy MRI scan as the baseline for evaluation in newly diagnosed high-grade gliomas; (3) the option for the trial to mandate a confirmation scan to more reliably distinguish pseudoprogression from tumor progression; (4) the option of using volumetric tumor measurements; and (5) the removal of subjective non-enhancing tumor evaluations in predominantly enhancing gliomas (except for specific therapeutic modalities). Step-by-step pragmatic guidance is hereby provided for the neuroradiologist and imaging core lab involved in operationalization and technical execution of RANO 2.0 in clinical trials, including the display of representative cases and in-depth discussion of challenging scenarios.ABBREVIATIONS: BTIP = Brain Tumor Imaging Protocol; CE = Contrast-Enhancing; CNS = Central Nervous System; CR = Complete Response; ECOG = Eastern Cooperative Oncology Group; HGG = High-Grade Glioma; IDH = Isocitrate Dehydrogenase; IRF = Independent Radiologic Facility; LGG = Low-Grade Glioma; KPS = Karnofsky Performance Status; MR = Minor Response; mRANO = Modified RANO; NANO = Neurological Assessment in Neuro-Oncology; ORR = Objective Response Rate; OS = Overall Survival; PD = Progressive Disease; PFS = Progression-Free Survival; PR = Partial Response; PsP = Pseudoprogression; RANO = Response Assessment in Neuro-Oncology; RECIST = Response Evaluation Criteria In Solid Tumors; RT = Radiation Therapy; SD = Stable Disease; Tx = Treatment.

Evaluating the immunologically "cold" tumor microenvironment after treatment with immune checkpoint inhibitors utilizing PET imaging of CD4 + and CD8 + T cells in breast cancer mouse models.

BACKGROUND: Immune-positron emission tomography (PET) imaging with tracers that target CD8 and granzyme B has shown promise in predicting the therapeutic response following immune checkpoint blockade (ICB) in immunologically "hot" tumors. However, immune dynamics in the low T-cell infiltrating "cold" tumor immune microenvironment during ICB remain poorly understood. This study uses molecular imaging to evaluate changes in CD4 + T cells and CD8 + T cells during ICB in breast cancer models and examines biomarkers of response. METHODS: [89Zr]Zr-DFO-CD4 and [89Zr]Zr-DFO-CD8 radiotracers were used to quantify changes in intratumoral and splenic CD4 T cells and CD8 T cells in response to ICB treatment in 4T1 and MMTV-HER2 mouse models, which represent immunologically "cold" tumors. A correlation between PET quantification metrics and long-term anti-tumor response was observed. Further biological validation was obtained by autoradiography and immunofluorescence. RESULTS: Following ICB treatment, an increase in the CD8-specific PET signal was observed within 6 days, and an increase in the CD4-specific PET signal was observed within 2 days in tumors that eventually responded to immunotherapy, while no significant differences in CD4 or CD8 were found at the baseline of treatment that differentiated responders from nonresponders. Furthermore, mice whose tumors responded to ICB had a lower CD8 PET signal in the spleen and a higher CD4 PET signal in the spleen compared to non-responders. Intratumoral spatial heterogeneity of the CD8 and CD4-specific PET signals was lower in responders compared to non-responders. Finally, PET imaging, autoradiography, and immunofluorescence signals were correlated when comparing in vivo imaging to ex vivo validations. CONCLUSIONS: CD4- and CD8-specific immuno-PET imaging can be used to characterize the in vivo distribution of CD4 + and CD8 + T cells in response to immune checkpoint blockade. Imaging metrics that describe the overall levels and distribution of CD8 + T cells and CD4 + T cells can provide insight into immunological alterations, predict biomarkers of response to immunotherapy, and guide clinical decision-making in those tumors where the kinetics of the response differ.

Multimodel Approaches Are Not the Best Way to Understand Multifactorial Systems.

Information-theoretic (IT) and multi-model averaging (MMA) statistical approaches are widely used but suboptimal tools for pursuing a multifactorial approach (also known as the method of multiple working hypotheses) in ecology. (1) Conceptually, IT encourages ecologists to perform tests on sets of artificially simplified models. (2) MMA improves on IT model selection by implementing a simple form of shrinkage estimation (a way to make accurate predictions from a model with many parameters relative to the amount of data, by "shrinking" parameter estimates toward zero). However, other shrinkage estimators such as penalized regression or Bayesian hierarchical models with regularizing priors are more computationally efficient and better supported theoretically. (3) In general, the procedures for extracting confidence intervals from MMA are overconfident, providing overly narrow intervals. If researchers want to use limited data sets to accurately estimate the strength of multiple competing ecological processes along with reliable confidence intervals, the current best approach is to use full (maximal) statistical models (possibly with Bayesian priors) after making principled, a priori decisions about model complexity.

Integration of Electrospun Scaffolds and Biological Polymers for Enhancing the Delivery and Efficacy of Mesenchymal Stem/Stromal Cell Therapies.

Mesenchymal stem/stromal cells (MSCs) have emerged as a promising therapeutic approach for a variety of diseases due to their immunomodulatory and tissue regeneration capabilities. Despite their potential, the clinical application of MSC therapies is hindered by limited cell retention and engraftment at the target sites. Electrospun scaffolds, with their high surface area-to-volume ratio and tunable physicochemical properties, can be used as platforms for MSC delivery. However, synthetic polymers often lack the bioactive cues necessary for optimal cell-scaffold interactions. Integrating electrospun scaffolds and biological polymers, such as polysaccharides, proteins, and composites, combines the mechanical integrity of synthetic materials with the bioactivity of natural polymers and represents a strategic approach to enhance cell-scaffold interactions. The molecular interactions between MSCs and blended or functionalized scaffolds have been examined in recent studies, and it has been shown that integration can enhance MSC adhesion, proliferation, and paracrine secretion through the activation of multiple signaling pathways, such as FAK/Src, MAPK, PI3K/Akt, Wnt/ß-catenin, and YAP/TAZ. Preclinical studies on small animals also reveal that the integration of electrospun scaffolds and natural polymers represents a promising approach to enhancing the delivery and efficacy of MSCs in the context of regenerating bone, cartilage, muscle, cardiac, vascular, and nervous tissues. Future research should concentrate on identifying the distinct characteristics of the MSC niche, investigating the processes involved in MSC-scaffold interactions, and applying new technologies in stem cell treatment and biofabrication to enhance scaffold design. Research on large animal models and collaboration among materials scientists, engineers, and physicians are crucial to translating these advancements into clinical use.

Semaglutide and Cardiovascular Outcomes by Baseline HbA1c and Change in HbA1c in People With Overweight or Obesity but Without Diabetes in SELECT.

OBJECTIVE: To evaluate the cardiovascular effects of semaglutide by baseline glycated hemoglobin (HbA1c) and change in HbA1c in a prespecified analysis of Semaglutide Effects on Cardiovascular Outcomes in People With Overweight or Obesity (SELECT). RESEARCH DESIGN AND METHODS: In SELECT, people with overweight or obesity and atherosclerotic cardiovascular disease without diabetes were randomized to weekly semaglutide 2.4 mg or placebo. The primary end point of first major adverse cardiovascular event (MACE) (cardiovascular mortality, nonfatal myocardial infarction, or stroke) was reduced by 20% with semaglutide versus placebo. Analysis of outcomes included first MACE, its individual components, expanded MACE (cardiovascular mortality, nonfatal myocardial infarction, or stroke; coronary revascularization; or hospitalization for unstable angina), a heart failure composite (heart failure hospitalization or urgent medical visit or cardiovascular mortality), coronary revascularization, and all-cause mortality by baseline HbA1c subgroup and categories of HbA1c change (<-0.3, -0.3 to 0.3, and >0.3 percentage points) from baseline to 20 weeks using the intention-to-treat principle with Cox proportional hazards. RESULTS: Among 17,604 participants (mean age 61.6 years, 72.3% male), baseline HbA1c was <5.7% for 33.5%, 5.7% to <6.0% for 34.6%, and 6.0% to <6.5% for 31.9%. Cardiovascular risk reduction with semaglutide versus placebo was not shown to be different across baseline HbA1c groups and was consistent with that of the overall study for all end points, except all-cause mortality. Cardiovascular outcomes were also consistent across subgroups of HbA1c change. CONCLUSIONS: In people with overweight or obesity and established atherosclerotic cardiovascular disease but not diabetes, semaglutide reduced cardiovascular events irrespective of baseline HbA1c or change in HbA1c. Thus, semaglutide is expected to confer cardiovascular benefits in people with established atherosclerotic cardiovascular disease who are normoglycemic at baseline and/or in those without HbA1c improvements.

Spiral ganglion neuron degeneration in aminoglycoside-deafened rats involves innate and adaptive immune responses not requiring complement.

Spiral ganglion neurons (SGNs) transmit auditory information from cochlear hair cells to the brain. SGNs are thus not only important for normal hearing, but also for effective functioning of cochlear implants, which stimulate SGNs when hair cells are missing. SGNs slowly degenerate following aminoglycoside-induced hair cell loss, a process thought to involve an immune response. However, the specific immune response pathways involved remain unknown. We used RNAseq to gain a deeper understanding immune-related and other transcriptomic changes that occur in the rat spiral ganglion after kanamycin-induced deafening. Among the immune and inflammatory genes that were selectively upregulated in deafened spiral ganglia, the complement cascade genes were prominent. We then assessed SGN survival, as well as immune cell numbers and activation, in the spiral ganglia of rats with a CRISPR-Cas9-mediated knockout of complement component 3 (C3). Similar to previous findings in our lab and other deafened rodent models, we observed an increase in macrophage number and increased expression of CD68, a marker of phagocytic activity and cell activation, in macrophages in the deafened ganglia. Moreover, we found an increase in MHCII expression on spiral ganglion macrophages and an increase in lymphocyte number in the deafened ganglia, suggestive of an adaptive immune response. However, C3 knockout did not affect SGN survival or increase in macrophage number/activation, implying that complement activation does not play a role in SGN death after deafening. Together, these data suggest that both innate and adaptive immune responses are activated in the deafened spiral ganglion, with the adaptive response directly contributing to cochlear neurodegeneration.

Effect of Plate Inclination and Osteotomy Positioning on Rock-back following Tibial Plateau Levelling Osteotomy in Dogs.

OBJECTIVE: Rock-back is a reported complication following tibial plateau levelling osteotomy (TPLO), whereby tibial plateau angle (TPA) increases postoperatively. The mechanism of rock-back is not fully understood, although a recent ex vivo investigation demonstrated that osteotomy exit cut angle (ECA) and inclination of the plate in the sagittal plane might be risk factors. The purpose of this study was to explore these relationships in a clinical dog population. We hypothesized that dogs with rock-back would have a higher degree of plate inclination and downward ECA compared with those without rock-back. MATERIALS AND METHODS: Medical records and radiographs of dogs that underwent TPLO between January 2021 and January 2022 were retrospectively reviewed. TPA was recorded preoperatively, postoperatively, and at follow-up. Plate inclination, ECA, and descriptive data were collected. Observers measuring TPA were blinded to other variables. Rock-back was defined as a change in TPA ≥ 2. RESULTS: Ninety-five TPLO procedures met the inclusion criteria (n = 87 dogs). Rock-back was identified in 21% of TPLOs (n = 20/95). The mean increase in TPA in the rock-back group was 3.2 ± 2.6. Plate inclination and ECA were not correlated with the presence of rock-back. Descriptive variables did not have a significant correlation with rock-back. CONCLUSION: Plate inclination and ECA did not have a relationship with rock-back when defined as a postoperative TPA change ≥2 degrees, in this clinical retrospective study.

Predictive value of joint fluid volume on advanced pre-procedure imaging related to success of arthrocentesis and presence of septic arthritis.

PURPOSE: Septic arthritis is a dangerous medical condition requiring prompt diagnosis, often via arthrocentesis. A "dry tap" occurs when no fluid is aspirated. We hypothesized that the absence of a joint effusion on pre-procedure advanced imaging would reliably predict a dry tap and exclude septic arthritis. METHODS: A cohort of 217 arthrocentesis cases of large joints (hips, shoulders, knees) from our institution, with pre-procedure advanced imaging (CT, MR, US) of the same joint performed within the previous 48 h, was analyzed. Exclusion criteria included non-native joints or inadequate imaging of the affected joint. These cases underwent blinded review by 4 radiologists who measured the deepest pocket of joint fluid on the pre-procedure imaging. Wilcoxon rank-sum test was performed comparing joint fluid pocket size to outcomes of successful aspiration and final diagnosis. RESULTS: A smaller average joint pocket fluid size was present on advanced imaging in both dry taps compared with successful arthrocenteses (p < .0001), and in uninfected joints compared with septic joints (p = .0001). However, the overlap of values was too great to allow for a perfectly predictive cutoff. 29% (5/17) of patients with no visible joint fluid on pre-aspiration imaging underwent successful arthrocentesis, one case representing septic arthritis. CONCLUSION: Volume of joint fluid on advanced pre-arthrocentesis imaging cannot reliably predict subsequent dry tap nor exclude septic arthritis.

Bluetooth Noise-Canceling Headphones Improve the Quality of Ophthalmic Exams in Patients With Hearing Loss: A Randomized Controlled Trial.

INTRODUCTION: This study tests the utilization of Bluetooth noise-canceling headphones in improving the quality of eye exams in patients with hearing loss. This prospective study was approved on ethical standards by the University of Texas Medical Branch (UTMB) Institutional Review Board (Approval No. 22-0079) and registered with the National Institutes of Health (NCT05420038). METHODS: UTMB patients above 55 years of age were screened for hearing loss using soundcheck audiometry. Twenty-nine subjects answered pre-recorded ophthalmic exam questions that solicited precise responses. As controls, subjects were randomly administered half of the questions via headphones and half via a smartphone at normal speech volume (60 decibels). Points were awarded for responses demonstrating comprehension, and a post-exam survey was collected. RESULTS: Collectively, the mean score was 1.79 with headphones versus 0.96 with control on the Amsler grid segment and 1.90 with headphones versus 0.97 with control on education questions (p=0.001). Between red zone and yellow zone hearing loss patients, the more severe red zone group answered significantly better in both Amsler (1.78 versus 0.50; p=0.0003) and education questions (1.88 versus 0.44; p<0.0001) with headphones. The yellow zone group answered better with headphones overall but failed to reach significance. Post-exam survey indicated that 28 of 29 patients (97%) preferred the headphones during ophthalmic exams. CONCLUSION: Patients with hearing loss demonstrated better comprehension with Bluetooth headphones. These low-cost devices show great promise at improving effective, compassionate communication between providers and hearing loss patients.

Evaluation of activation characteristics of a canine platelet concentrate produced by a commercial double centrifugation system.

Introduction: In veterinary medicine there are few readily available products for platelet transfusion to patients with thrombocytopenia. Commercial tabletop platelet concentrating systems have recently become available to veterinarians, primarily directed towards uses associated with regenerative medicine. These systems could potentially be used to produce fresh concentrated platelets for use in transfusion medicine. This study evaluated the concentration, activation, and sterility of a double centrifugation platelet concentrate (PC) produced by a commercial benchtop system. Methods: Ten healthy dogs were studied. Whole blood was collected and mixed with ACD-A in a 1:7.6 ratio of ACD-A to whole blood. 12 mL of this mixture was processed into PC via single centrifugation, while 60 mL of the anticoagulated whole blood was processed via a commercial double centrifugation system. Both types of PC were evaluated for platelet concentration, CD62P expression with and without thrombin stimulation, and for sterility. Results: Mean platelet count in the double centrifuged PC was 863 ± 352 × 103/µL, with very low white blood cell contamination (median of 0.47 × 103 leukocyte/µL (range 0.15-2.18 × 103/µL)). The double-centrifuged PC had similar baseline activation characteristics (as determined by P-selectin expression) as the single centrifuge PC (0.76% vs. 0.72% unstimulated, 30.5% vs. 34.9% stimulated, p = 0.432). Discussion: The benchtop PC system studied here did not cause activation of platelets during production and produced a sterile product that can be further investigated as a source of fresh platelet concentrates for transfusion purposes.

Clinical and Radiographic Outcomes and Graft Incorporation Rate Assessed by CT Scan After Reverse Shoulder Arthroplasty With Glenoid Structural Bone Graft Reconstruction.

INTRODUCTION: Reverse total shoulder arthroplasty (RTSA) with structural bone graft has been described as a technique in addressing glenoid bony defects. Studies have demonstrated acceptable outcomes with structural autograft or allograft. However, most of these studies are relatively small and rarely evaluate bone graft incorporation with CT scan. The aim of this study was to assess clinical and radiographic outcomes and report graft incorporation assessed on CT scan after RTSA where structural bone autograft or allograft was used to reconstruct the glenoid. METHODS: From May 2011 through June 2016, 38 patients underwent RTSA with structural bone graft. Of these, 35 were available for a minimum 2-year follow-up and retrospectively enrolled. From July 2016 through February 2019, 32 patients undergoing RTSA with structural bone graft were prospectively enrolled. Preoperative and postoperative American Shoulder and Elbow Surgeons and visual analog scale (for pain) scores and radiographs were obtained. CT scan was obtained at least 1 year postoperatively. RESULTS: Thirty-five patients were enrolled retrospectively (52.2%) and 32 prospectively (47.8%). Autograft was used in 46 cases (68.7%) and allograft in 21 cases. The mean American Shoulder and Elbow Surgeons score improved from 33.1 (SD 18.5) to 78.2 (SD 22.4), with P < 0.0001. On postoperative radiographs, 63 cases (94.0%) showed stable RTSA constructs while four cases (6.0%) developed glenoid baseplate subsidence. Postoperative CT scan demonstrated complete graft incorporation in 45 cases (90.0%) while partial incorporation was noted in 4 cases (8.0%), and in 1 case (2.0%), there was no graft incorporation. No correlation was observed between baseplate subsidence and graft type (autograft versus allograft) or primary versus revision surgery. DISCUSSION: Reverse shoulder arthroplasty with structural bone autograft and allograft is reliable for glenoid augmentation in patients undergoing RTSA in both primary and revision settings. Bony incorporation of autograft and allograft as evaluated on CT scan is predictably high.

Investigation of U.S. landfill GHG reporting program methane emission models.

As part of its commitment to the United Nations Framework Convention on Climate Change, the U.S. annually develops a national estimate of methane emissions from municipal solid waste (MSW) landfills by aggregating activity data from each facility. Since 2010, the U.S. has reported a 20 % decrease in MSW landfill emissions despite a 21 % increase in tons disposed. Operator-submitted data were investigated to understand the causes of this decline. In the U.S., operators of landfills with a gas collection and control system (GCCS) calculate their facility's emissions via two separate approaches - (1) first-order decay (FOD) and (2) collection efficiency assumption (CEA) - and select either result to feed into the annual inventory. The FOD model predicts methane generation proportional to waste disposal and that approach calculated a 19 % increase in total methane generated from 2010 to 2022, whereas generation via the CEA approach decreased by 8.9 %. The amount of measured methane collected has increased 7.5 % for the same years. Discrepancies between the two models' generated methane, assumed gas collection efficiencies, and oxidized methane compound into substantive differences in national estimates. Operators more frequently select the CEA method, which results in decreased national estimates. If only the FOD method was used, U.S. MSW landfill emissions would be 1.3-1.7 times greater than current estimates which is similar to recent extrapolations from remote sensing campaigns in the U.S. Both models contain parameters with large inherent uncertainty. Without measurement methods that continuously quantify both point-source and diffuse emissions, an assessment of either equation's accuracy cannot be made.