Adaptations in glutathione-based redox protein signaling pathways and alcohol drinking across species.

Alcohol use disorder (AUD) is the most prevalent substance use disorder but there is incomplete knowledge of the underlying molecular etiology. Here, we examined the cytosolic proteome from the nucleus accumbens core (NAcC) of ethanol drinking rhesus macaques to identify ethanol-sensitive signaling proteins. The targets were subsequently investigated using bioinformatics, genetic, and pharmacological manipulations in mouse models of ethanol drinking. Of the 1000+ cytosolic proteins identified in our screen, 50 proteins differed significantly between control and ethanol drinking macaques. Gene Ontology analysis of the differentially expressed proteins identified enrichment in pathways regulating metabolic processes and proteasome activity. Because the family of Glutathione S-transferases (GSTs) was enriched in these pathways, validation studies targeted GSTs using bioinformatics and genetically diverse mouse models. Gstp1 and Gstm2 were identified in Quantitative Trait Loci and published gene sets for ethanol-related phenotypes (e.g., ethanol preference, conditioned taste aversion, differential expression), and recombinant inbred strains that inherited the C57BL/6J allele at the Gstp2 interval consumed higher amounts of ethanol than those that inherited the DBA/2J allele. Genetic deletion of Gstp1/2 led to increased ethanol consumption without altering ethanol metabolism or sucrose preference. Administration of the pharmacologic activator of Gstp1/2, carnosic acid, decreased voluntary ethanol drinking. Proteomic analysis of the NAcC cytosolic of heavy drinking macaques that were validated in mouse models indicate a role for glutathione-mediated redox regulation in ethanol-related neurobiology and the potential of pharmacological interventions targeting this system to modify excessive ethanol drinking.

Safety of a 4-Dose 20-Valent Pneumococcal Conjugate Vaccine Series in Infants: A Randomized Trial.

BACKGROUND AND OBJECTIVES: The 20-valent pneumococcal conjugate vaccine (PCV20) was developed to expand protection for pneumococcal disease. It contains all 13-valent pneumococcal conjugate vaccine (PCV13) components plus conjugates for 7 additional serotypes. Our primary objective with this study was to evaluate PCV20 tolerability and safety. METHODS: In this phase 3, multi-country, double-blind study, healthy infants born at ≥34 weeks' gestation were randomly assigned 2:1 to receive PCV20 or PCV13 at 2, 4, 6, and 12 to 15 months of age. Safety assessments included local reactions and systemic events within 7 days after each vaccination, adverse events (AEs) from dose 1 to 1 month after dose 3 and from dose 4 to 1 month after dose 4, and serious AEs and newly diagnosed chronic medical conditions from dose 1 through 6 months after the last dose. RESULTS: Participants received PCV20 (N = 1000) or PCV13 (N = 504); 91.7% received all 4 doses. The frequencies of local reactions and systemic events were generally similar in PCV20 and PCV13 groups, with most reported as mild or moderate. The most common local reaction was injection site pain (PCV20, 24.7% to 40.5%; PCV13, 26.8% to 42.0%); irritability was the most common systemic event (PCV20, 54.8% to 68.2%; PCV13, 54.7% to 68.5%). AE frequencies were similar in both groups. No serious AEs were related to study vaccines. Few newly diagnosed chronic medical conditions were reported (2.8% in both groups). PCV20 was safe across multiple countries, in late preterm infants, and when administered with other vaccines. CONCLUSIONS: A 4-dose series of PCV20 had a tolerability and safety profile similar to that of PCV13.

Novel Technique for Biopsying Osteofibrous Dysplasia Using a Vacuum-assisted Bone Harvester: A Case Report.

Introduction: Osteofibrous dysplasia (OFD) is a rare benign bone lesion primarily affecting the tibia, characterized by fibrous tissue proliferation with varying osseous involvement. Diagnosis involves clinical evaluation, imaging, and histopathological analysis. Traditional bone biopsies for OFD can be challenging due to the lesion's nature. Case Report: We present a case report of a young patient presenting with pain concerning for OFD and describe a novel technique for biopsy and curettage using a vacuum-assisted bone harvester. Conclusion: The vacuum-assisted bone harvester allows the surgeon to effectively obtain a biopsy and curettage.

Dual Coaching of Medical Clerkship Students' History Taking Skills by Volunteer Inpatients at the Bedside and Faculty Physicians on Zoom During the COVID-19 Pandemic.

Purpose: Coaching is a well-described means of providing real-time, actionable feedback to learners. We aimed to determine whether dual coaching from faculty physicians and real inpatients led to an improvement in history-taking skills of clerkship medical students. Patients and Methods: Expert faculty physicians (on Zoom) directly observed 13 clerkship medical students as they obtained a history from 26 real, hospitalized inpatients (in person), after which students received immediate feedback from both the physician and the patient. De-identified audio-video recordings of all interviews were scored by independent judges using a previously validated clinical rating tool to assess for improvement in history-taking skills between the two interviews. Finally, all participants completed a survey with Likert scale questions and free-text prompts. Results: Students' history-taking skills - specifically in the domains of communication, medical knowledge and professional conduct - on the validated rating tool, as evaluated by the independent judges, did not significantly improve between their first and second patient interviews. However, students rated the dual coaching as overwhelmingly positive (average score of 1.43, with 1 being Excellent and 5 being Poor), with many appreciating the specificity and timeliness of the feedback. Patients also rated the experience very highly (average score of 1.23, with 1 being Excellent and 5 being Poor), noting that they gained new insights into medical training. Conclusion: Students value receiving immediate and specific feedback and real patients enjoy participating in the feedback process. Dual physician-patient coaching is a unique way to incorporate more direct observation into undergraduate medical education curricula.

Scalable, compressed phenotypic screening using pooled perturbations.

High-throughput phenotypic screens using biochemical perturbations and high-content readouts are constrained by limitations of scale. To address this, we establish a method of pooling exogenous perturbations followed by computational deconvolution to reduce required sample size, labor and cost. We demonstrate the increased efficiency of compressed experimental designs compared to conventional approaches through benchmarking with a bioactive small-molecule library and a high-content imaging readout. We then apply compressed screening in two biological discovery campaigns. In the first, we use early-passage pancreatic cancer organoids to map transcriptional responses to a library of recombinant tumor microenvironment protein ligands, uncovering reproducible phenotypic shifts induced by specific ligands distinct from canonical reference signatures and correlated with clinical outcome. In the second, we identify the pleotropic modulatory effects of a chemical compound library with known mechanisms of action on primary human peripheral blood mononuclear cell immune responses. In sum, our approach empowers phenotypic screens with information-rich readouts to advance drug discovery efforts and basic biological inquiry.

Firearm screening and intervention beliefs and practices among U.S. Air Force (USAF) mental health providers and behavioral health technicians.

We examined beliefs and practices regarding firearm assessment and lethal means safety counseling (LMSC) among U.S. Air Force (USAF) mental health providers (MHPs) and behavioral health technicians (BHTs). Data were collected from 204 USAF MHPs (74.0%; n = 151) and BHTs (26.0%; n = 53) via an anonymous, voluntary survey. A modest proportion indicated they strongly/extremely believe that firearm ownership (42.2%) and storage practices (58.3%) are related to suicide risk. A minority indicated they "strongly"/"extremely" believe that LMSC will yield changes in storage practices (30.9%) and decreases in suicide risk (29.9%). Across patient scenarios, most indicated that "most of the time"/"always" they assess for firearm access (74.5%-99.5%) and provide LMSC (57.8%-95.6%). About half (52.5%) reported having distributed cable locks. Most (59.3%) indicated they are somewhat interested/very interested in receiving additional training on LMSC. MHPs, compared with BHTs, were significantly more likely to report believing a link between firearm ownership and storage practices and suicide risk, believing LMSC is effective at reducing suicide risk, providing LMSC to female patients and patients with current suicidal ideation, and having distributed cable locks. Findings suggest that there is not widespread agreement among USAF MHPs and BHTs that personal firearm ownership and nonsecure storage practices are associated with elevated suicide risk, and there were low levels of confidence in the effectiveness of LMSC. Yet, most USAF MHPs and BHTs reported they integrate firearm access assessment and LMSC as part of their routine clinical care, particularly for patients with identified suicide risk. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

LSD1 and CoREST2 Potentiate STAT3 Activity to Promote Enteroendocrine Cell Differentiation in Mucinous Colorectal Cancer.

Neuroendocrine cells have been implicated in therapeutic resistance and worse overall survival in many cancer types. Mucinous colorectal cancer (mCRC) is uniquely enriched for enteroendocrine cells (EECs), the neuroendocrine cell of the normal colon epithelium, as compared to non-mCRC. Therefore, targeting EEC differentiation may have clinical value in mCRC. Here, single cell multi-omics uncovered epigenetic alterations that accompany EEC differentiation, identified STAT3 as a regulator of EEC specification, and discovered a rare cancer-specific cell type with enteric neuron-like characteristics. Furthermore, LSD1 and CoREST2 mediated STAT3 demethylation and enhanced STAT3 chromatin binding. Knockdown of CoREST2 in an orthotopic xenograft mouse model resulted in decreased primary tumor growth and lung metastases. Collectively, these results provide rationale for developing LSD1 inhibitors that target the interaction between LSD1 and STAT3 or CoREST2, which may improve clinical outcomes for patients with mCRC.

SARS-CoV-2 ferritin nanoparticle vaccines produce hyperimmune equine sera with broad sarbecovirus activity.

The rapid emergence of SARS-CoV-2 variants of concern (VoC) and the threat of future zoonotic sarbecovirus spillover emphasizes the need for broadly protective next-generation vaccines and therapeutics. We utilized SARS-CoV-2 spike ferritin nanoparticle (SpFN), and SARS-CoV-2 receptor binding domain ferritin nanoparticle (RFN) immunogens, in an equine model to elicit hyperimmune sera and evaluated its sarbecovirus neutralization and protection capacity. Immunized animals rapidly elicited sera with the potent neutralization of SARS-CoV-2 VoC, and SARS-CoV-1 pseudoviruses, and potent binding against receptor binding domains from sarbecovirus clades 1b, 1a, 2, 3, and 4. Purified equine polyclonal IgG provided protection against Omicron XBB.1.5 virus in the K18-hACE2 transgenic mouse model. These results suggest that SARS-CoV-2-based nanoparticle vaccines can rapidly produce a broad and protective sarbecovirus response in the equine model and that equine serum has therapeutic potential against emerging SARS-CoV-2 VoC and diverse sarbecoviruses, presenting a possible alternative or supplement to monoclonal antibody immunotherapies.

Downregulation of PIEZO2 in the Detrusor of Men With Bladder Outlet Obstruction and Its Association With Urinary Retention and Decreased Bladder Compliance.

PURPOSE: Recent research has highlighted the mechanotransducer PIEZO2 as a crucial factor in lower urinary tract function, demonstrating associations with bladder compliance (BC), bladder wall thickening, and elevated bladder pressure. We explored the hypothesis that PIEZO2 expression may be associated with lower urinary tract dysfunction in men with bladder outlet obstruction (BOO) due to benign prostatic hyperplasia (BPH). METHODS: The study included a consecutive series of patients undergoing open prostatectomy for BPH at our hospital between September 2014 and January 2016. All participants underwent comprehensive preoperative evaluations, including urodynamic assessments. During prostatectomy, a full-thickness fragment of the bladder wall was obtained for subsequent PIEZO2 gene expression analysis. Cadaveric organ donors served as the control group. RESULTS: PIEZO2 expression was downregulated in the detrusor muscle of men with BPH compared to the control group. Among patients with BPH, those experiencing urinary retention and requiring an indwelling catheter exhibited significantly lower PIEZO2 messenger RNA (mRNA) expression than patients capable of spontaneous voiding. PIEZO2 mRNA expression was similar in men with and without detrusor overactivity. Additionally, a positive correlation was found between PIEZO2 mRNA expression levels and BC. CONCLUSION: Our findings indicate that PIEZO2 is downregulated in the detrusor muscle of men with BPH, particularly in those experiencing urinary retention and those with reduced BC. These results suggest a potential role for PIEZO2 in BOOinduced bladder dysfunction. Further research is required to clarify the role of PIEZO mechanotransducers in the bladder and to explore their therapeutic implications.

Blood loss during three column osteotomies: influence on outcomes and mitigation strategies.

Background: Three column osteotomies (3CO) are a powerful tool for the correction of rigid sagittal spinal deformities. However, 3CO surgeries are technically demanding procedures that are associated with prolonged operative times, large amounts of intraoperative blood loss, and high rates of post-operative neurological and medical complications. The purpose of this retrospective cross-sectional study is to delineate the effects of blood loss, blood transfusion, and tranexamic acid use on perioperative outcomes following 3CO. Methods: Patients undergoing 3CO between 2014 and 2021 were identified using current procedural terminology (CPT) codes, and charts were reviewed for demographics, surgical characteristics, and post-operative length of stay (LOS). Statistical analysis included T-tests for continuous characteristics and χ2 testing for categorical characteristics. Regression models were used to further investigate relationships between outcomes and variables of interest. Results: Forty-two patients met inclusion criteria. Increased intensive care unit (ICU) LOS was positively correlated with increased operative length (P=0.02), percent estimated blood volume (EBV) lost (P=0.02), and number of units of red blood cell transfusion (P<0.001). Increased hospital LOS was associated with increased number of intraoperative blood transfusions (P=0.01). Linear regression analyses controlling for age, gender, and American Society of Anesthesiologists (ASA) class showed that each 21% increase in EBV loss was associated with a one day increase in ICU LOS (P=0.01). Similarly, each additional unit of packed red blood cells transfused was associated with 0.728 day increase in hospital LOS. Conclusions: Increased intraoperative blood loss was associated with longer ICU stays. Red blood cell transfusions were also associated with increased hospital and ICU LOS. No variables of interest related to intraoperative blood loss or blood transfusion were associated with readmission or reoperation at any time point. These findings highlight the need for continued focus on surgical techniques and adjuncts that can minimize blood loss and transfusion requirements.

Diabetes and heart disease do not affect short-term lumbar fusion outcomes accounting for other risk factors in a matched cohort analysis.

Objectives: Comprehensive preoperative management involves the identification and optimization of medical comorbidities while avoiding excessive healthcare utilization. While diabetes and heart disease are major causes of morbidity that can worsen surgical outcomes, further study is needed to evaluate how well current perioperative strategies mitigate their risks. This study employs an exact matching protocol to isolate the effects of both diabetes and cardiovascular disease on spine surgery outcomes. Methods: 4680 consecutive patients undergoing single-level, posterior-only lumbar fusion were retrospectively enrolled. Univariate logistic regression was performed on comorbidity subgroups, then coarsened exact matching (CEM) was employed for patients with diabetes or cardiovascular disease. Patients were matched 1:1 on ten patient and procedural characteristics known to affect neurosurgical outcomes. Separate pairs of exact-matched cohorts were generated to isolate both cardiovascular disease (matched n = 192), and diabetes (matched n = 380). Primary outcomes were surgical complications; length of stay; discharge disposition (home vs. non-home); and 30- and 90-day Emergency Department (ED) visits, readmissions, reoperations, and mortality. Results: Cardiovascular disease and diabetes subgroups were not associated with short term outcomes after matching to control for confounders. Compared to univariate statistics, this method demonstrates that confounding control variables may drive outcomes more than these comorbidities themselves. Conclusion: Between otherwise exactly matched patients undergoing lumbar fusion, diabetes and cardiovascular disease posed no greater risk of short-term adverse outcomes. This suggests proper selection criteria for surgical candidates and effective current perioperative strategies to mitigate these common comorbidities. Further studies are warranted to assess and optimize the cost-effectiveness of preoperative management for patients with common comorbidities.

Short-Course TNT Improves Rectal Tumor Downstaging in a Retrospective Study of the US Rectal Cancer Consortium.

BACKGROUND AND OBJECTIVES: The RAPIDO trial showed promising rates of pathologic complete response (pCR) after neoadjuvant short-course radiation with consolidation chemotherapy (total neoadjuvant therapy [SC TNT]) for rectal cancer. Only single-center reviews comparing tumor downstaging between SC TNT and long-course chemoradiation (LCRT) have been published in the United States. We reviewed our multi-institutional experience with both. METHODS: The US Rectal Cancer Consortium database (2007-2018) including data from six high-volume rectal cancer care centers was reviewed. Patients with nonmetastatic, rectal adenocarcinoma who had neoadjuvant LCRT alone or SC TNT before excision or definitive nonoperative management were included. The primary outcome was the rate of complete response (CR), including pCR or durable (12 month) clinical complete response. RESULTS: Of 857 included patients, 175 (20%) received SC TNT and 682 (80%) received LCRT. The LCRT group had more low tumors (51.8% vs. 37.1%, p < 0.0001) and more clinically node-negative disease (31.8% vs. 22.3%, p < 0.0001). The CR rate was higher after SC TNT (34.1% vs. 20.3%, p = 0.0001). SC TNT was a predictor of CR (OR: 2.52, CI: 1.68-3.78). SC TNT patients completing 5-6 months of consolidation chemotherapy had a CR rate of 42.9%. There was no difference in 3-year PFS. CONCLUSIONS: SC TNT increases CR rate when compared to LCRT. For patients seeking nonoperative options or fewer radiation treatments, SC TRT should be preferred over LCRT alone.

Information About Sexual and Gender Minority Services and Policies on US Hospital Websites.

This cross-sectional study assesses the extent to which hospitals provide information related to sexual and gender minority policies and health services on their websites.

Trends in Initial Antihypertensive Medication Prescribing Among >2.8 Million Veterans Newly Diagnosed With Hypertension, 2000 to 2019.

BACKGROUND: Among patients diagnosed with high blood pressure (BP), initial dual therapy has been recommended for patients with high pretreatment systolic BP (≥160 mm Hg) since 2003, and first-line ß-blocker use without a compelling condition has fallen out of favor in US guidelines. METHODS AND RESULTS: This serial cross-sectional study of national Veterans Health Administration data included adult Veterans with incident hypertension initiating antihypertensive medication between January 1, 2000, and December 31, 2019. We assessed annual trends in initial regimens dispensed (index date: first antihypertensive dispense date) by number of classes and unique class combinations used overall and by pretreatment systolic BP (<140, 140 to <160, and ≥160 mm Hg), as well as trends in subgroups (age, sex, race and ethnicity, and comorbidities warranting ß-blocker use). Among 2 832 684 eligible Veterans (average age 61 years, 95% men, 65% non-Hispanic White, and 8% with cardiovascular disease), from 2000-2004 to 2015-2019, initial monotherapy increased across all pretreatment systolic BP levels (<140 mm Hg: 62.1% to 66.4%; 140 to <160 mm Hg: 70.7% to 76.8%; ≥160 mm Hg: 64.2% to 69.7%). Initiation of dual therapy decreased across all pretreatment systolic BP levels (<140 mm Hg: 25.0% to 24.2%; 140 to <160 mm Hg: 20.4% to 17.6%; ≥160 mm Hg: 22.7% to 22.0%). Among 2 521 696 Veterans (89% of overall) without a ß-blocker-indicated condition in 2015 to 2019, 20% initiated a ß-blocker, most commonly as monotherapy. CONCLUSIONS: More than half of US Veterans diagnosed with hypertension with a pretreatment systolic BP ≥160 mm Hg were started on antihypertensive monotherapy. There are disparities between guideline-recommended first-line treatments and the actual regimens initiated for newly diagnosed Veterans with hypertension.

Medium to Long-Term Ventricular Assist Device Support in Adults with Congenital Heart Disease.

Heart failure is the leading cause of morbidity and mortality in adults with congenital heart disease. Though for many in this population, heart transplantation is not possible or requires longer wait times necessitating prolonged circulatory support. Medium to long-term durable ventricular assist device therapy provides a possible solution. We analyzed outcomes of nine patients with congenital heart disease and at least 3 years of durable mechanical support, all age 18 or older at time of ventricular assist device implantation at our affiliated pediatric and adult hospitals. Palliated congenital anatomies varied and included biventricular physiologies as well as single ventricle with Fontan circulation. Median duration of support was 4.2 years including three patients successfully bridged to transplant averaging 2.1 years on the waitlist. Device-related complications were infrequent with HeartMate 3, a feasible and sustainable option for either bridge to transplant or destination strategies in adults with congenital heart disease.

Clinic transfers and engagement in HIV care during the perinatal period across a network of healthcare centers in Lilongwe, Malawi.

INTRODUCTION: Disengagement from HIV care during the perinatal period remains a challenge. Improving engagement in HIV care requires monitoring engagement across multiple indicators, including retention in HIV care, visit adherence, clinic transfers, and viral suppression to support improved clinical and programmatic outcomes. METHODS: We enrolled a prospective cohort of pregnant WHIV across a network of five urban clinics in Lilongwe, Malawi from February 2020-February 2021. WHIV were followed from their first antenatal care visit through 9 months postpartum across all study sites using biometric fingerprint scanning. Study visits occurred at enrollment into antenatal care, 6 weeks', 6 months, and 9 months postpartum. In addition, all usual care HIV visits were captured via medical records. Participants who missed a study visit or usual care visit were traced. We evaluated determinants of multiple indicators of engagement in care, including retention in HIV care (attending a scheduled visit or self-reported recent visit when traced), HIV visit adherence (missed scheduled HIV visits and HIV visit coverage), clinic transfers, and viral load suppression (< 1000 copies/mL) using modified Poisson regression and sub-distributional hazard ratios to account for competing events of death and loss-to-follow-up. Associations between clinic transfer and subsequent indicators of engagement in HIV care were evaluated using generalized estimating equations. RESULTS: Among 399 participants, 81% were on ART at baseline. Retention in HIV care was 87% at 6 weeks postpartum, 77% at 6 months postpartum and 89% at 9 months postpartum. At 9 months postpartum, 91% of participants were virally suppressed, 81% had missed a scheduled HIV visit, and 19% had transferred clinics. WHIV who transferred clinics were most likely to miss their subsequent scheduled HIV visit by ≥ 30 days. Transferring clinics was not associated with unsuppressed viral load or non-retention at 9 months postpartum. CONCLUSIONS: In a cohort of WHIV, retention and viral load suppression were high in the perinatal period, but missed HIV visits and clinic transfers were common. Transferring clinics was associated with an increased likelihood of missing a subsequent HIV visit. Clinic transfers may be important indicators of disruptions in clinical care for WHIV in the perinatal period.

Robotic kidney transplant has superior outcomes compared to open kidney transplant: results of a propensity match analysis.

BACKGROUND: Several studies have demonstrated the feasibility of robotic kidney transplant (RKT) as a safe alternative to open kidney transplant (OKT). However, significant selection bias in RKT patient selection limits meaningful comparison between the two techniques. METHODS: This is a single-center retrospective review of a prospectively maintained kidney transplant database (2021-2024). Outcomes after the first 50 "non-selected" RKTs are compared with a contemporary cohort of 100 OKTs after propensity score matching for age, gender, BMI and type of donation (living vs deceased). Data pertinent to recipient demographics, intraoperative parameters, and short-term post-operative outcomes were collected and compared. RESULTS: Both groups were well-matched for recipient age, gender, BMI, and donation type. RKT group had significantly longer total operative time (RKT 258 min vs. OKT 183 min; p < 0.0001) and warm ischemia time (RKT 37 min vs. OKT 31 min; p < 0.0001) but significantly less blood loss (OKT 155 ml vs. RKT 93 ml). Average length of hospital stay for both groups was 5 days, with OKT group demonstrating significantly higher rates of post-operative complications (OKT 31% vs. RKT 14%; p = 0.028), return to OR (OKT 15% vs. RKT 2%; p = 0.021), hematoma (OKT 13% vs. RKT 2%; p = 0.0355), and lymphocele (OKT 25% vs. RKT 6%; p = 0.0039). OKT group also had higher 30-day readmission rate (OKT 31% vs. RKT 14%) and post-operative opioid requirement (OKT 93 MME vs. RKT 65; p = 0.0254). There were no differences in rates of wound infection, urine leaks, delayed graft function, acute rejection, graft loss, and patient death between the two groups. CONCLUSION: RKT is a safe and viable alternative to OKT as a first-choice procedure for all patients with ESRD. RKT offers many advantages over OKT which can lead to its wider adoption in the coming years as the new standard of care for ESRD patients.