Alcohol dependence or alcohol use disorder is a chronic mental disorder that imposes a heavy global burden. Small extracellular vesicles (EVs) are emerging as a promising source of real-time information about brain status, yet existing studies are predominantly cross-sectional. We aimed to characterize longitudinal changes in EV properties and immunophenotype during alcohol withdrawal treatment. In our study plasma-derived EVs were analyzed and immunophenotyping via flow cytometry was performed to measure the concentrations of vesicle fractions carrying erythrocyte-, platelet-, endothelial-, and neuronal-associated markers in patients undergoing alcohol withdrawal and in healthy volunteers. Patient-derived EVs exhibited greater size heterogeneity, and microscopy images revealed vesicles with altered morphology. Concentrations of EVs bearing NLGN3 were lower in patients. CD62P+ and CD146+ EVs were also less represented in patients. Notably, CD31+ EVs separated into two distinct clusters; their concentrations were higher in patients with a binge drinking pattern but markedly lower in those with a steady pattern. The findings reveal dynamic phenotypic shifts in circulating EVs during alcohol withdrawal. The divergent patterns according to alcohol use history highlight the potential of EV profiling for personalized monitoring and underscore the necessity of longitudinal designs in vesicle-based biomarker research.

Alcohol use contributes substantially to global morbidity and mortality, necessitating reliable biomarkers for detection and monitoring. Existing biomarkers, including indirect markers and direct markers, have important limitations related to specificity, sensitivity and temporal detection windows. As a biomarker, creatine kinase (CK) demonstrates strong biological plausibility, as alcohol exerts direct and indirect myotoxic effects through oxidative stress, impaired protein synthesis and metabolic disruption, leading to muscle injury and enzyme leakage. However, its diagnostic specificity is also limited, as CK elevation can result from multiple non-alcohol-related conditions, including trauma, seizures and metabolic disturbances. Despite these limitations, CK shows clinical utility in identifying complications such as rhabdomyolysis, severe withdrawal states and risk of acute kidney injury. It correlates with withdrawal severity and may aid in risk stratification. Overall, CK is best positioned as a supplementary biomarker reflecting alcohol-related complications rather than a primary indicator of alcohol consumption.

Background Dolutegravir (DTG)-based combination antiretroviral therapy (cART), specifically the tenofovir disoproxil fumarate, lamivudine, and dolutegravir (TLD) regimen, is being widely prescribed for the management of people living with HIV (PLHIV) in India and other low- and middle-income countries. Although DTG is well-tolerated, concerns persist regarding its hepatic and metabolic safety. Prospective real-world data characterizing the longitudinal hepatic and metabolic trajectories following DTG initiation in antiretroviral therapy (ART)-naïve South Asian PLHIV remain limited. Therefore, we conducted a prospective study to determine the incidence and grading of liver enzyme elevation (LEE) and characterize the longitudinal trajectories of alanine aminotransferase (ALT), anthropometric, and glycemic parameters over six months in ART-naïve PLHIV initiated on the TLD regimen. Methodology We conducted a six-month prospective cohort study and followed 101 ART-naïve adult PLHIV initiated on the TLD regimen at a tertiary care hospital in Bengaluru, India. Serum ALT, weight, body mass index (BMI), waist circumference, and random blood sugar (RBS) were measured at baseline and at one, two, three, and six months. Repeated-measures analysis of variance (rmANOVA) was used to assess longitudinal trends; paired-samples t-test compared baseline and six-month values for waist circumference; chi-square tests examined categorical associations. Results All 101 enrolled participants completed the six-month follow-up. The mean age was 36.5 ± 11.2 years, and 67 (66.3%) participants were male. Any-grade LEE occurred in 22 (21.8%) participants. Mean ALT rose transiently from 26 ± 14 IU/L at baseline to a peak of 32 IU/L at one month, then declined progressively to 27 ± 17 IU/L at six months; the overall trajectory was not statistically significant (F(1.94, 192.29) = 1.114, P = 0.329). All participants with LEE remained asymptomatic; no jaundice, coagulopathy, or hepatic encephalopathy was observed in this cohort. Statistically significant increases were noted in weight +1.79 kg (F(2.02, 201.87) = 12.549, P < 0.001), and mean BMI +0.71 kg/m[2] (F(2.00, 199.90) = 13.117, P < 0.001). Mean RBS rose by only +1.06 mg/dL (F(3.71, 370.96) = 2.274, P = 0.066) and waist circumference by +0.17 cm (t(100) = 1.42, P = 0.159) - neither change reached statistical significance. LEE was not significantly associated with age (t(99) = 0.298, P = 0.766), sex (χ[2](1) = 3.02, P = 0.082, φ = 0.17), concurrent anti-tuberculosis therapy (Fisher's exact P = 0.116), or alcohol use (χ[2](1) = 0.001, P = 0.973, φ = 0.003). Conclusions In ART-naïve South Indian PLHIV initiating DTG-based cART, LEEs were relatively common but were predominantly mild, asymptomatic, and self-limited. LEE was characterized by an early rise in ALT followed by a spontaneous decline in most cases by six months despite uninterrupted therapy, a pattern that may reflect hepatic adaptation. In contrast, body weight and BMI increased progressively throughout follow-up, suggesting that hepatic and metabolic responses to DTG may follow distinct trajectories. Overall, ALT elevations were predominantly mild and self-limited, indicating favorable hepatic tolerability of DTG-based cART in this cohort while highlighting the need for continued metabolic monitoring.

Neuronal central chromatolysis (CC) is the histopathological hallmark of pellagra encephalopathy, a neurological deficit resulting from vitamin deficiencies. Pellagrous CC neurons are morphologically similar to ballooned achromatic neurons in other conditions but the distinct pathomechanisms remain unclear. We performed a clinico-neuropathological analysis of 10 autopsy cases of pellagra encephalopathy. The pellagra encephalopathy cases were immunohistochemically compared with disease controls, including cases of axonal injury and neurodegenerative diseases. Electron microscopic evaluation and immunohistochemical examinations targeting mitochondrial fragmentation were performed for a representative case. Four of 10 pellagra encephalopathy patients exhibited prolonged impairment of consciousness distinguishable from alcohol withdrawal delirium. Pellagrous CC neurons were negative for cytoskeletal markers whereas ballooned achromatic neurons in the disease control cases were positive. Immunohistochemical analysis of mitochondrial markers revealed that CC neurons exhibited more intense immunoreactivity for COX-IV and mitochondrial fissure factor compared to the disease controls. Transmission electron microscopy of these CC neurons revealed a marked increase in the mitochondria with amorphous densities. These findings indicate that the pathomechanism of pellagrous CC is distinct from that of the ballooned achromatic neurons of other etiologies. Mitochondrial alterations in pellagrous CC neurons suggest that neuronal energy deficits resulting from nicotinamide adenine dinucleotide deficiency induce mitochondrial fragmentation.

The opioid epidemic remains a persistent public health challenge in the United States, leaving many people affected by or vulnerable to opioid use disorder (OUD). Although US Food and Drug Administration-approved medications for OUD improve the odds of recovery and reduce mortality, relapse rates remain high and access, adherence, and individual response variabilities limit their overall impact. The ongoing burden of OUD therefore underscores the need to develop additional therapeutics that target complementary biological mechanisms and support long-term recovery. One strategy of interest is to delineate the biological overlap between OUD and other motivated behaviors, particularly the neurobiology underlying appetitive and reward-driven processes. The orexigenic peptide hormone ghrelin, often referred to as the "hunger hormone," regulates energy homeostasis, hedonic feeding, and reward sensitivity by binding to the growth hormone secretagogue receptor 1α (GHS1αR), a receptor expressed in mesolimbic circuits implicated in addiction. In the present study, we used the inverse agonist/antagonist PF5190457 to assess the impact of full GHS1αR blockade on fentanyl-mediated behaviors in preclinical models. We found that PF5190457 significantly reduced fentanyl self-administration and attenuated drug-seeking during cue tests while maintaining a favorable pharmacological and safety profile in risk assessment assays, including measures of withdrawal and respiratory depression. Collectively, these findings demonstrate a functional role for ghrelin-GHS1αR signaling in the reinforcing effects of fentanyl and positions GHS1αR inverse agonism as a promising, mechanistically novel therapeutic strategy for OUD treatment. SIGNIFICANCE STATEMENT: The present work demonstrates that the growth hormone secretagogue receptor 1α (GHS1αR) inverse agonist/antagonist PF5190457 reduces fentanyl self-administration and drug-seeking while maintaining a favorable safety profile in male rats, supporting ghrelin-GHS1αR modulation as a promising therapeutic strategy for opioid use disorder.