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Exposure to alcohol during pregnancy produces Fetal Alcohol Spectrum Disorders, which in its most severe form is characterized by physical dysmorphology and neurobehavioral alterations. Moderate prenatal alcohol exposure (mPAE) is known to produce deficits in discrimination of spatial locations in adulthood. However, the impact of mPAE on higher-order sensory representations, such as discrimination of perceptually similar stimuli, is currently unknown. In the present study, we tested the hypothesis that mPAE would disrupt performance on hippocampal-sensitive tasks that require discrimination between perceptually similar objects or discrimination between spatial locations in a radial arm maze. Here we report that male mPAE rats exhibited intact performance on three types of object discrimination tasks: one in which rats discriminated between distinct toy objects, a second in which discrimination was made between distinct and similar LEGO objects, and a mnemonic similarity task in which rats discriminated between randomly presented LEGO objects that varied in similarity with a learned object. Although adult male mPAE rats performed similarly to control rats on all three object discrimination tasks, they showed deficits when tested in a radial arm maze spatial discrimination task. Specifically, male mPAE rats expressed a significantly higher number of working memory errors (returns to previously visited arms) and were more likely to use non-spatial strategies during training. Together, the findings of the present study support the conclusion that mPAE produces specific deficits in the online processing of spatial information and executing spatial navigation strategies, but spares the ability to discriminate between perceptually similar stimuli., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Excessive alcohol consumption poses a significant threat to human health, leading to cellular dehydration, degeneration, and necrosis. Alcohol-induced cellular damage is closely linked to alterations in cellular mechanical properties. However, characterizing these changes following alcohol-related injury remains challenging. Moreover, current research on single-cell mechanics often struggles to culture and measure cells within a controlled microenvironment, leading to complex experimental procedures and imprecise results. (63)., Results: In this study, we developed a novel single cell measurement method that combines cell microculture in alcohol-containing solutions with cytomechanics assessment within microdroplets. This approach integrates key operations, including single-cell encapsulation and culture in droplets, droplet reinjection, and cell deformation analysis within droplets, enabling high-throughput and multi-parameter quantification of single-cell mechanical properties. The use of droplets provides a precisely regulated microculture environment, effectively avoiding channel clogging issues. Additionally, the integration of cytomechanics measurement simplifies the analytical process by eliminating the need for complex techniques within the droplets. Gastric mucosal epithelial cells (GES-1) and human umbilical vein endothelial cells (HUVECs) were selected as models for ethanol-induced injury to validate the proposed technique. The results demonstrate a bidirectional response in cellular deformability following ethanol treatment, with cells becoming stiffer at lower ethanol concentrations and softer at higher concentrations. (136)., Significance: The integration of droplet microfluidics and cell mechanics offers a powerful platform for investigating the underlying mechanisms of ethanol-induced cellular damage. This approach is also applicable for studying changes in cellular mechanical properties by precisely modulating the microculture environment, providing a reliable tool for drug screening and disease modeling in biochemistry and biomedical engineering. (54)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)

The present study investigated the impact of chronic intermittent ethanol (CIE) exposure and single-prolonged stress (SPS) on the acquisition of fear memories in both male and female Wistar rats. Adult rats were first subjected to CIE by vapor inhalation followed by SPS. Following a subsequent 8-day incubation period, the rats underwent a Pavlovian fear conditioning procedure (tone-shock pairings) followed by cued-tone extinction training, and then testing of extinction recall memory and fear renewal memory. In control animals that had not been exposed to either CIE or SPS, female rats exhibited significantly lower levels of freezing compared to male rats during tone-shock pairings. This lower level of freezing in female rats during conditioning was associated with an increased speed of movement compared to males. Also compared to males, female rats exhibited lower levels of fear extinction, recall, and renewal. Exposure to CIE, SPS, or CIE+SPS had no effect on freezing during the cued-conditioning, extinction, or extinction recall phases of the testing procedure in either sex. In fear renewal, CIE exposure decreased freezing in male but not female rats, while SPS increased freezing in female but not male rats. CIE exposure significantly reduced freezing during the fear renewal phase. Taken together, these results provide further evidence that male and female rats adopt different avoidance strategies for threat responding. These results also revealed that prior exposure to CIE, SPS, or CIE+SPS had minimal effects on threat responding using conditioned freezing as an indicator of fear responsivity., (Copyright © 2024. Published by Elsevier B.V.)

Episodic consumption of high doses of alcohol in a short period (binge drinking - BD) among adolescents is known to be harmful to their brain development. Chronic use of taurine increases voluntary alcohol consumption and shows an anxiolytic-like effect in rats. In this study, we evaluated the differential effects of chronic and intermittent taurine administration on alcohol consumption and behavioral changes in adolescent and young adult rats subjected to the BD model. Male Wistar rats (35 days old) were divided into 4 groups for daily intraperitoneal administration of saline (SAL); taurine, 100 mg/kg (TAU); taurine on BD days and saline on intervals (TAU/SAL); and saline on BD days and taurine on intervals (SAL/TAU). They were exposed to 4 cycles of BD, with free access to alcoholic solution (20 % w/v), for 2 h, 3 days per week. At the end of the 3rd cycle, anxiety-like behaviors were assessed using the light-dark task. After euthanasia, plasma and prefrontal cortex samples were collected to measure corticosterone and BDNF levels, respectively. Chronic taurine treatment did not alter alcohol consumption in rats, whereas intermittent administration increased alcohol intake after 4 BD exposures (TAU/SAL: +19.4 % and SAL/TAU: +21.6 %). No anxiolytic-like effects were found by taurine administration, nor were there changes in serum corticosterone or BDNF levels in the frontal cortex of young adult rats. Intermittent taurine, but not chronic treatment, increased alcohol intake among rats after the second week of exposure. The translation of these results to humans is concerning since the combination of alcohol and drinks containing taurine is common among adolescent and young adult individuals., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Antioxidant dipeptide Phe-Cys (FC) could dramatically improve yeast cells resistance to ethanol-oxidation cross-stress, but the regulatory mechanisms remain unclear. Therefore, transcriptomic and proteomic analyses were conducted to investigate the effects of FC treatment on yeast under ethanol-oxidation cross-stress. Following FC supplementation, 875 differential expressed genes (DEGs) and 1296 differential expressed proteins (DEPs) were identified. Integrated analysis revealed a substantial enrichment of DEGs and DEPs in the KEGG pathways of carbon metabolism, amino acid biosynthesis, cofactor biosynthesis, and glycolysis/gluconeogenesis. Furthermore, FC improved yeast cell membrane integrity by promoting fatty acids and steroids biosynthesis, and implemented a high-energy strategy by upregulating glycolysis and oxidative phosphorylation. Additionally, alterations in DEGs and DEPs levels associated with amino acids metabolism accelerated protein synthesis and enhanced cell viability. In conclusion, this study elucidated the response mechanisms of yeast to FC treatment under ethanol-oxidation cross-stress, providing a theoretical basis for the application of FC in high-gravity brewing., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Obstructive sleep apnea (OSA) is a breathing disorder in which airway obstruction during sleep leads to periodic bouts of inadequate (hypopneic) or absent (apneic) ventilation despite neurorespiratory effort. Repetitive apneic and hypopneic exposures can induce intermittent hypoxemia and lead to a host of maladaptive behavioral and physiological outcomes. Intermittent hypoxia treatment (IH), which consists of alternating exposure to hypoxic and normal air, can induce a long-lasting increase in breathing motor outputs called long term facilitation (LTF). IH models key aspects of the hypoxemia experienced during OSA and LTF might serve to prevent OSA or ameliorate its severity by stimulating ventilatory output during or after apnea/hypopnea. Ethanol consumption prior to sleep exacerbates existing OSA, but it is unknown how ethanol affects LTF expression. Thus, we hypothesized that ethanol treatment would attenuate LTF expression and the magnitude of the ventilatory response during acute hypoxic exposure. We administered either low-dose (0.8 g/kg) or high-dose (3 g/kg) ethanol or saline to adult female Sprague-Dawley rats through intraperitoneal injection and then measured subjects' ventilatory output by whole-body plethysmography during baseline, a 5 by 3-minute moderate IH protocol (hypoxia: F i O 2 = 0.11, Normoxia: room air), and for one hour following the end of IH. Results indicate that low-dose ethanol abolishes LTF of respiratory rate and minute ventilation and trends suggest that low-dose ethanol might attenuate respiratory rate and minute ventilation during acute hypoxic exposure. While high-dose ethanol significantly diminished subjects' respiratory rate and minute ventilation during hypoxia, LTF expression was not significantly different between high-dose ethanol and saline-treated subjects. Overall, data indicate that ethanol exposure dramatically attenuates LTF expression following IH treatment and impairs ventilatory responses to hypoxia in a dose-dependent manner. Such findings inspire further consideration of ethanol's negative effects upon endogenous compensatory mechanisms for repeated hypoxic exposure, both in the context of OSA and beyond., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Mu-opioid receptors (MORs) in the amygdala and striatum are important in addictive and rewarding behaviors. The transcription factor Foxp2 is a genetic marker of intercalated (ITC) cells in the amygdala and a subset of striatal medium spiny neurons (MSNs), both of which express MORs in wild-type mice and are neuronal subpopulations of potential relevance to alcohol-drinking behaviors. For the current series of studies, we characterized the behavior of mice with genetic deletion of the MOR gene Oprm1 in Foxp2-expressing neurons (Foxp2-Cre/Oprm1 fl/fl ). Male and female Foxp2-Cre/Oprm1 fl/fl mice were generated and heterozygous Cre+ (knockout) and homozygous Cre- (control) animals were tested for aversion-resistant alcohol consumption using an intermittent access (IA) task, operant responding for a sucrose reward, conditioned place aversion (CPA) to morphine withdrawal, and locomotor sensitization to morphine. The results demonstrate that deletion of MOR on Foxp2-expressing neurons renders mice more sensitive to quinine-adulterated alcohol. Mice with the deletion (vs. Cre- controls) also consumed less alcohol during the final sessions of the IA task, were less active at baseline and following morphine injection, and there was a trend toward less responding for sucrose under an FR3 schedule. Foxp2-MOR deletion did not impair the ability to learn to respond for reward or develop a conditioned aversion to morphine withdrawal. Together, these investigations demonstrate that Foxp2-expressing neurons may be involved in escalation of alcohol consumption and the development of compulsive-like alcohol drinking., Competing Interests: Declaration of competing interest The authors declare no conflicting interests in this work., (Copyright © 2024. Published by Elsevier Inc.)

Rationale & Objectives: A core symptom of alcohol use disorder (AUD) is a progressively increased choice of alcohol over alternative rewards despite negative consequences. Here, we investigated choice between personalized alcohol vs. natural rewards in a laboratory setting, and compared this behavior between non-treatment-seeking heavy drinkers and light social drinkers., Methods: 30 light social drinkers (15 men drinking < 15 drinks/week and 15 women drinking < 10 drinks/week) and 30 heavy, non-treatment-seeking drinkers (drinking more than these levels; 15 women). In the Concurrent Choice Alcohol Food (CCAF) task, participants chose between individually tailored images of alcohol and snack rewards and collected points towards the respective reward. To assess cost sensitivity, points associated to the images varied so that they favored alcohol or snack, or were equal, creating three relative point levels., Results: Choice preference for alcohol was strongly correlated with Alcohol Use Disorder Identification Test (AUDIT) scores, supporting the external validity of the choice procedure. Compared to light drinkers, heavy drinkers showed increased choice preference for alcohol, as indicated by a between-group difference in points of subjective equality, a metric that quantifies the relative point level at which alcohol and snacks were equally likely to be chosen. In both groups, choice preference strongly depended on the relative point level of alcohol compared to snacks, suggesting that responding for alcohol in heavy drinkers was sensitive to costs., Conclusions: Our results replicate previous findings of a relationship between self-reported alcohol use and choice preference for alcohol. We also found that choice behavior was strongly dependent on relative cost of alcohol in both groups, although price sensitivity was lower in heavy compared to light drinkers. An increased choice preference for alcohol in heavy drinkers suggests that they attribute a higher relative reinforcing value to alcohol compared to natural rewards., Competing Interests: Declarations. Conflict of interest: MH has received consulting fees, research support or other compensation from Indivior, Camurus, Molteni, BrainsWay, Aelis Farma, and Janssen Pharmaceuticals., (© 2024. The Author(s).)

Stress is a major contributing factor to binge drinking and development of alcohol use disorders (AUD), particularly in women. Both stress and chronic ethanol can enhance neuroinflammatory processes, which may dysregulate limbic circuits involved in ethanol reinforcement. Clinical and preclinical studies have identified sex differences in alcohol intake in response to neuroinflammatory triggers. Since both cyclic AMP (cAMP) signaling and microglial activation contribute to neuroinflammation, we explored their contribution to stress-induced ethanol drinking in mice. To this end, we first trained C57BL/6J male and female mice to volitionally drink ethanol through a modified version of the "Drinking-in-the-Dark" paradigm. We then assessed whether exposure to foot shock stress followed by repeated exposure to the previously stress-paired context might alter volitional ethanol drinking. We observed that stress exposure resulted in a delayed increase in ethanol intake, but only in female mice. The anti-inflammatory drug Apremilast, an inhibitor of phosphodiesterase type 4 (PDE4; the primary enzyme for cAMP degradation in the brain), reduced ethanol intake and decreased preference for ethanol regardless of stress exposure in females. In contrast, a partial pharmacological depletion of microglia via PLX3397 treatment did not significantly alter baseline ethanol drinking or stress-induced ethanol drinking in female mice. This study shows that female mice are more susceptible to stress-induced ethanol drinking than males, and that this occurs even after partial microglial depletion. In addition, modulation of cAMP signaling by Apremilast administration reduced ethanol drinking regardless of stress exposure, supporting the idea that it might be useful for treatment of AUD., Competing Interests: Declaration of competing interest The authors have no competing interests to declare., (Copyright © 2024 Elsevier Inc. All rights reserved.)

This daily diary study expands knowledge of the pharmacological alcohol-sleep relationship using a multilevel modeling approach. The interplay between alcohol and sleep on hangover susceptibility is also explored. College students (n = 337; 52 % female) provided 2976 days of self-reported alcohol use. We regressed sleep duration onto accumulated sleep debt, prior night sleep duration, and estimated blood alcohol concentration (eBAC) at bedtime; linear mixed models disaggregated day and person-level effects. Binomial models, assessing days after drinking when eBAC = 0 % versus when eBAC>0 % at waketime, regressed hangover susceptibility onto the same predictors plus sleep duration. More accumulated sleep debt predicted slightly longer same-night sleep. Greater than average bedtime intoxication predicted longer than average same-night sleep when drinking ceased early, but later drinking attenuated the relationship. People who typically stopped drinking later in the night reported typically shorter sleep durations on drinking nights. When waketime eBAC = 0 %, higher eBAC at bedtime and drinking later on a given night predicted greater next-day hangover susceptibility. Typical bedtime eBAC and typically later drinking predicted typically greater hangover susceptibility. When waketime eBAC>0 %, longer sleep duration predicted more likely hangovers. Bedtime eBAC and sleep debt interacted, such that more sleep debt attenuated the positive association between intoxication and next-day hangover susceptibility. Late-night drinking appeared to reduce sleep duration and increase hangover susceptibility. Accumulated sleep debt complicated the alcohol-sleep-hangover relationship. External factors influencing sleep behaviors were not assessed, but the results highlight the need to deconstruct sleep into acute and chronic processes. Future studies should better subdivide physiological processes related to hangovers., Competing Interests: Declaration of competing interest None., (Published by Elsevier Inc.)

RNA viruses have high genetic diversity, allowing rapid adaptation to environmental pressures, such as disinfection. This diversity increases the likelihood of mutations influencing the viral sensitivity to disinfectants. Ethanol is widely used to control viral transmission; however, insufficient disinfection facilitates the survival of less-sensitive viruses. Further, the underlying mechanisms of ethanol-induced changes in viral sensitivity remain unclear. Here, we assessed the genetic characteristics of ethanol-adapted murine norovirus (MNV) and associated changes in viral sensitivity. Experimental ethanol-facilitated MNV adaptation and subsequent genetic characteristic evaluation of the whole genome sequence was performed. MNV was exposed to 70% ethanol for 5 s to achieve ± 3-log 10 inactivation. Twelve MNV populations were identified as "less sensitive," consisting of nine treated and three control populations. Less-sensitive MNV populations exhibited significantly higher synonymous nucleotide diversity (πS) in ORF1 (p = 0.001), which encodes the non-structural protein, than sensitive populations. Ethanol sensitivity and πS were negatively correlated in ORF1 (R = - 0.49, p = 0.003), indicating that high genetic diversity in ORF1 could be linked to reduced ethanol sensitivity. This study demonstrates an association between nucleotide diversity in specific coding regions of the MNV genome and ethanol sensitivity. These findings are vital for improving disinfection methods and anticipating emerging viruses that are more resistant to disinfectants. KEY POINTS: • Several MNV populations reduced sensitivity to ethanol. • Higher synonymous diversity in ORF1 linked to reduced ethanol sensitivity. • Synonymous mutations can influence viral adaptation to ethanol., Competing Interests: Declarations. Ethics approval: This article does not contain any studies with human participants or animals performed by any of the authors. Conflict of interest: The authors declare no competing interests., (© 2025. The Author(s).)

Clinical and preclinical studies suggest that early life stress can increase the risk of developing ethanol use disorder later in life. Although the endocannabinoid (eCB) system plays a role in stress-related behaviors and ethanol consumption, it remains unclear whether the eCB system is affected in response to a combination of both factors. By using male and female adolescent C57BL/6J mice subjected to a maternal separation (MS) stress paradigm from postnatal day (PND) 1 to 14, we explored (1) the consequences of early life stress experiences on ethanol consumption in adolescent mice and (2) how these events affect the eCB system and neuronal activation in brain regions associated with the reward system. In Experiment 1, we found that MS increased involuntary ethanol consumption specifically during the first exposure to the drug (during a 24 h-long trial on PND 28) and decreased the active/inactive nose poke ratio (discrimination index) specifically when mice were subjected to 1 h-sessions (PND 82-86) in an operant ethanol self-administration paradigm. In Experiment 2, during a two-bottle free choice paradigm, we found that MS increased mice preference for high ethanol concentrations (15 % and 20 %) but not lower ethanol concentrations (5 % and 10 %). Except for Mgll gene expression in the dorsal striatum (DS) in Experiment 2, no statistically significant effects of MS were observed regarding neuronal activation on the prefrontal cortex, DS, globus pallidus, and substantia nigra following a binge operant ethanol self-administration session (Experiment 1) or the eCB system molecules (Cnr1 and Faah gene expression) in the DS (Experiment 2)., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 International Brain Research Organization (IBRO). Published by Elsevier Inc. All rights reserved.)

Ethanol (EtOH) is one of the most widely consumed substance, affecting neurobehavioral functions depending on multiple environmental and biological factors. Although EtOH modulates zebrafish (Danio rerio) anxiety-like behaviors in novelty-based paradigms, the potential role of biological sex and populational variability in the exploratory dynamics in the open field test (OFT) is unknown. Here, we explored whether a repeated EtOH exposure protocol modulates the spatio-temporal exploration and homebase-related parameters in a population- and sex-dependent manner. Male and female fish from the short-fin (SF) and leopard (LEO) phenotypes were exposed to EtOH for 7 days (1 % v/v, 20 min per day). On the 8th day, the OFT was performed to assess locomotor and exploratory behaviors. We verified significant populational differences in the baseline spatio-temporal exploration patterns, supporting a pronounced anxiety in LEO with a higher homebase index compared to SF. We also found sex-dependent differences in EtOH sensitivity, where SF was more sensitive to EtOH, especially in females, which showed marked alterations in thigmotaxis and homebase occupancy. Conversely, only LEO female subjects showed increased center occupancy following EtOH. Principal component analysis (PCA) showed the main components that explained data variability, which were sex- and population-dependent. Overall, our novel findings support the utility of zebrafish-based models to assess how EtOH influences the exploratory profile in the OFT, as well as to elucidate potential differences of sex and population in the neurobehavioral responses of alcohol exposure in a translational perspective., Competing Interests: Declaration of competing interest The authors declare no conflict of interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Prenatal alcohol exposure (PAE) induces a wide range of neurodevelopmental disabilities that are grouped under the term 'fetal alcohol spectrum disorders' (FASD). The effects of PAE on brain development are dependent on complex neurochemical events, including modification of AMPA receptors (AMPARs). We have recently found that chronic ethanol (EtOH) exposure decreases AMPA-mediated neurotransmission and expression through the overexpression of the specific microRNA (miR)137 and 501-3p, which target GluA1 AMPA subunit, in the developing hippocampus in vitro. Here, we explored how PAE mice may alter AMPAergic synapses in the hippocampus, and its effects on behavior., Methods: To model PAE, we exposed C57Bl/6 pregnant mice to 10 % EtOH during during the first 10 days of gestation (GD 0-10; equivalent to the first trimester of pregnancy in humans). AMPA subunits postsynaptic expression in the hippocampus, electrical properties of CA1 neurons, memory recognition, and locomotor functions were then analyzed in adolescent PAE-exposed offspring., Results: PAE adolescent mice showed dysregulation of AMPAergic neurotransmission, and increased miR 501-3p expression, associated with a significant reduction of spontaneous AMPA currents and intrinsic somatic excitability. In addition, PAE reduced the phosphorylation of AMPAR-containing GluA1 subunit, despite an increase in its total levels. Of note, the total levels of GluA2 and GluA3 AMPA receptors were enhanced as well. Consistently, at behavioral level, PAE reduced object recognition without altering locomotor activity., Conclusions: Our study shows that PAE leads to dysfunctional formation of AMPAergic synapses that could be responsible for neurobehavioral impairments, contributing to the understanding of the pathogenesis of FASD., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Alcohol and cannabis are often taken in combination, and extensive co-use has been linked to enduring changes in cognitive and metabolic functioning. The underlying mechanisms for these effects are unclear, but we recently demonstrated that co-administration of ethanol and delta-9-tetrahydrocannbinol (THC) to adolescent rats caused lasting adaptations in GABA and glycogen synthase kinase 3ß (GSK3ß) signaling in the medial prefrontal cortex (mPFC). As a ubiquitous protein kinase, GSK3ß is downstream to the protein kinase B (also known as AKT) pathway that is activated by insulin receptor signaling in a main control center for metabolism and energy homeostasis, the mediobasal hypothalamus (MBH). Our goal here was to investigate if volitional co-use of low to moderate levels of ethanol and THC would impact the total and phosphorylated levels (p) of AKT and GSK3ß in the mPFC and MBH. Peri-adolescent Long Evans rats [postnatal day (P) 30-47] consumed 10 % ethanol, cookies laced with THC (3-10 mg/kg/day), both drugs, or vehicle controls. On P114, we modeled re-exposure to a behaviorally relevant dose of THC by challenging rats (i.p.) with 5 mg/kg THC (or vehicle) and sacrificed them 30 min later. Western blot analysis revealed that THC challenge increased pAKT and pGSK3ß compared to control similarly across all treatment groups, sexes, and brain regions; no effects on total levels of AKT or GSK3ß were found. Previously reported behavioral results from these rats showed no differences in working memory assessed in adulthood. Although future studies will be necessary to determine the role of exposure dose on drug-induced adaptations in AKT and GSK3ß signaling, the current findings suggest that moderate volitional co-use of alcohol and THC may not produce long-term deficits that persist into adulthood., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

About 296 million people worldwide are living with chronic hepatitis B viral (HBV) infection, and outcomes to end-stage liver diseases are potentiated by alcohol. HBV replicates in hepatocytes, but other liver non-parenchymal cells can sense the virus. In this study, we aimed to investigate the regulatory effects of macrophages on HBV marker and interferon-stimulated genes (ISGs) expressions in hepatocytes. This study was performed on HBV-replicating HepG2.2.15 cells and human monocyte-derived macrophages (MDMs). We found that exposure of HepG2.2.15 cells to an acetaldehyde-generating system (AGS) increased HBV RNA, HBV DNA, and cccDNA expressions and suppressed the activation of ISGs, APOBEC3G , ISG15 , and OAS1 . Supernatants collected from IFNα-activated MDMs decreased HBV marker levels and induced ISG activation in AGS-treated and untreated HepG2.215 cells. These effects were reversed by exposure of MDMs to ethanol and mimicked by treatment with exosome release inhibitor GW4869. We conclude that exosome-mediated crosstalk between IFN-activated macrophages and HBV-replicating hepatocytes plays a protective role via the up-regulation of ISGs and suppression of HBV replication. However, ethanol exposure to macrophages breaks this protection.

Rationale: Glucagon-like peptide-1 (GLP-1) receptor agonists reduce alcohol consumption in rodents and non-human primates. Semaglutide is a new long-acting GLP-1 receptor agonist, widely used in the clinic against type 2 diabetes and obesity. It is also reported to reduce alcohol intake in rodents., Objectives: This study investigates the possible inhibitory effect of semaglutide on alcohol intake in alcohol-preferring African green monkeys., Methods: We performed a vehicle-controlled study on male monkeys that had demonstrated a preference for alcohol. In the monkeys selected for voluntary alcohol drinking, alcohol consumption was measured for ten days at baseline (Monday to Friday for two weeks). During this period, the monkeys had access to alcohol 4 h per day and free access to water 24 h per day. After two weeks of baseline measurements, the monkeys were randomized to semaglutide or vehicle. Each group consisted of ten monkeys, and the two groups were balanced with respect to baseline alcohol intake. Following the baseline period, the monkeys were treated with escalating doses of semaglutide (up to 0.05 mg/kg) or vehicle subcutaneously twice weekly for two weeks during which period alcohol was not available. After uptitration, the monkeys had access to alcohol 4 h daily for 20 days (Monday to Friday for 4 weeks), and alcohol consumption was measured. During this alcohol exposure period, treatment with semaglutide (0.05 mg/kg twice weekly) or vehicle continued for three weeks followed by a one-week washout period., Results: Compared to the vehicle, semaglutide significantly reduced alcohol intake. There were no signs of emetic events or changes in water intake., Conclusions: These data demonstrate for the first time the potent effect of semaglutide in reducing voluntary alcohol intake in non-human primates and further substantiate the need for clinical trials investigating the effect of semaglutide in patients with alcohol-use disorder., Competing Interests: Declarations. Competing interests: AFJ has received two unrestricted research grants from Novo Nordisk A/S for two clinical studies investigating the effects of liraglutide (finished) and semaglutide (ongoing) in prediabetic schizophrenia patients on antipsychotic medication (Larsen et al. (2017), JAMA Psychiatry, 74(7):719–728) and (Reeberg Sass et al. (2023), BMJ Open, 13(1):e068652, respectively. BH is a board member and cofounder of Bainan Biotech. JJH has served on scientific advisory panels and/or speaker for Novo Nordisk, Eli Lilly, and Zealand Pharma. He has given lectures and received financial support for travel from Novo Nordisk, Novo Nordisk Pharma, Novo Nordisk Scandinavia AB, and Mayo Clinic. He has served as a consultant for Alphasights, Eli Lilly, Shouti/Structure Therapeutics X, and Zealand Pharma. JJH is currently consulting for GV Management L.L.C. He is cofounder and on the board of directors of Antag Therapeutics and Bainan Biotech and sits on the board of directors of Antag Therapeutics and Bainan Biotech, which is unpaid. JJH serves as an investigator for Boehringer Ingelheim and Scohia. The other authors declare no competing financial interests concerning the work described., (© 2024. The Author(s).)

Conditioned preference paradigms like conditioned colour preference tests (CCP) can be used to investigate addictive drug seeking in zebrafish (Danio rerio), but many aspects of this procedure require further study. Conditioned preference can be tested with either biased or unbiased conditioning methods, each with their own strengths and weaknesses. The present study used unbiased stimuli to test seeking behaviour in ethanol-exposed zebrafish at different durations of drug withdrawal. Zebrafish were exposed to one of two equally preferred colours (red or yellow) while dosed with 0.8 % vol/vol ethanol or with habitat water (controls) for 1 h each day for 21 days. Next, fish experienced withdrawal for either 2-, 4-, or 8-days then were tested in a two-way red and yellow task for 10 min with their movement recorded via motion-tracking software. Fish conditioned to red showed a main effect of ethanol and a significant preference for red compared to yellow at 8-days of withdrawal but not at 2-days or 4-days of withdrawal. Fish conditioned to yellow did not show any colour preference during the 2-, 4-, or 8-days of withdrawal, but did show a main effect of withdrawal duration. This work expands our understanding of CCP paradigms in zebrafish and highlights the capacity of zebrafish to develop an association to red but not yellow under our experimental conditions., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background and Purpose: GABAergic neurons in mouse ventral tegmental area (VTA) exhibit elevated activity during withdrawal following chronic ethanol exposure. While increased glutamatergic input and decreased GABA A receptor sensitivity have been implicated, the impact of inhibitory signaling in VTA GABA neurons has not been fully addressed., Experimental Approach: We used electrophysiological and ultrastructural approaches to assess the impact of chronic intermittent ethanol vapour exposure in mice on GABAergic transmission in VTA GABA neurons during withdrawal. We used CRISPR/Cas9 ablation to mimic a somatodendritic adaptation involving the GABA B receptor (GABA B R) in ethanol-naïve mice to investigate its impact on anxiety-related behaviour., Key Results: The frequency of spontaneous inhibitory postsynaptic currents was reduced in VTA GABA neurons following chronic ethanol treatment and this was reversed by GABA B R inhibition, suggesting chronic ethanol strengthens the GABA B R-dependent suppression of GABAergic input to VTA GABA neurons. Similarly, paired-pulse depression of GABA A receptor-dependent responses evoked by optogenetic stimulation of nucleus accumbens inputs from ethanol-treated mice was reversed by GABA B R inhibition. Somatodendritic currents evoked in VTA GABA neurons by GABA B R activation were reduced following ethanol exposure, attributable to the suppression of GIRK (K ir 3) channel activity. Mimicking this adaptation enhanced anxiety-related behaviour in ethanol-naïve mice., Conclusions and Implications: Chronic ethanol weakens the GABAergic regulation of VTA GABA neurons in mice via pre- and postsynaptic mechanisms, likely contributing to their elevated activity during withdrawal and expression of anxiety-related behaviour. As anxiety can promote relapse during abstinence, interventions targeting VTA GABA neuron excitability could represent new therapeutic strategies for treatment of alcohol use disorder., (© 2024 The Author(s). British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of British Pharmacological Society.)

Rationale: A recent study by our group found that women displayed greater attentional bias to alcohol-related cues during the late versus early follicular phase in both sober and intoxicated states, suggesting a greater risk of excessive drinking among women during this phase. Changes in attentional bias as a function of menstrual cycle phase raise questions about potential sex differences in the relative consistency by which women and men display attentional bias to alcohol over time., Objectives: The present study tested sex differences in attentional bias to alcohol by comparing the change in women's attentional bias from early to late follicular phase to that observed in men over the same period., Methods: Twenty-five men and 25 women aged 21-32 participated in a placebo-controlled study examining sex differences in the rewarding properties of alcohol. Participants completed measures of attentional bias to alcohol-related cues during two sessions following both 0.6 g/kg alcohol and placebo. Test sessions occurred one week apart, and for female participants coincided with the early and late follicular phases., Results: Men consistently displayed attentional bias to alcohol-related cues across sessions under both doses. By contrast, women showed attentional bias only during the late follicular phase, at a magnitude greater than that observed in men, and persistent under both doses., Conclusions: These findings highlight the potential role of sex and menstrual cycle phase in sensitizing drinkers to rewarding properties of alcohol-related cues. Men's motivation to drink may remain relatively consistent, whereas women may be most motivated during the late follicular phase., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The ability of environmental cues to trigger alcohol-seeking behaviours is thought to facilitate problematic alcohol use. Individuals' tendency to attribute incentive salience to cues may increase the risk of addiction. We sought to study the relationship between incentive salience and alcohol addiction using non-preferring rats to model the heterogeneity of human alcohol consumption, investigating both males and females. Adult rats were subjected to the alcohol deprivation effect (ADE) paradigm, where they were given voluntary access to different alcohol solutions with repeated interruptions by deprivation and reintroduction phases over a protracted period (five Alcohol Deprivation Cycles). Before each Alcohol Deprivation Cycle, rats were tested in the Pavlovian Conditioned Approach (PCA) paradigm, which quantifies the individual salience toward a conditional cue and the reward, thus allowing us to trace the process of attributing incentive salience to reward cues. During the final Alcohol Deprivation Cycle (ADE5), animals were tested for compulsive-like behaviour using quinine taste adulteration. We investigated sex differences in drinking behaviour and PCA performance. We observed thatb females drank significantly more alcohol than males and displayed more sign-tracking (ST) behaviour in the PCA, whereas males showed goal-tracking (GT) behaviour. Furthermore, we found that high drinkers exhibited more ST behaviour. The initial PCA phenotype was correlated with later alcohol consumption. Our findings indicate a complex relationship between incentive salience and alcohol addiction and emphasize the importance of considering both sexes in preclinical research., (© 2025 The Author(s). Addiction Biology published by John Wiley & Sons Ltd on behalf of Society for the Study of Addiction.)

The consumption of ethanol has frequently been seen as largely restricted to humans. Here, we take a broad eco-evolutionary approach to understanding ethanol's potential impact on the natural world. There is growing evidence that ethanol is present in many wild fruits, saps, and nectars and that ethanol ingestion offers benefits that favour adaptations for its use in multiple taxa. Explanations for ethanol consumption span both the nutritional and non-nutritional, with potential medicinal value or cognitive effects (with social-behavioural benefits) explored. We conclude that ethanol is ecologically relevant and that it has shaped the evolution of many species and structured symbiotic relationships among organisms, including plants, yeast, bacteria, insects, and mammals., Competing Interests: Declaration of interests No interests are declared., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Alcohol is the most abused substance among adolescents and has a profound impact on health, society, and the economy. Alcohol intoxication is linked to neuroinflammation and neuronal damage, which result in behavioral alterations such as motor dysfunction, neuronal injury, cognitive deficits, and inflammation. Alcohol-induced neuroinflammation is associated with the activation of central nervous system cells, including astrocytes, and the release of proinflammatory cytokines. In this study, we investigated the role of the NLRP6 inflammasome signaling pathway in inducing cellular activation and neuroinflammation in human primary astrocytes exposed to ethanol. Our results demonstrated that ethanol upregulates the expression of NLRP6 inflammasome signaling mediators, including NLRP6, caspase 1, and proinflammatory cytokines IL-1β and IL-18, in human primary astrocytes. Gene silencing studies using NLRP6 siRNA further validate ethanol-mediated activation of NLRP6, cleavage of caspase 1, IL-1β, and IL-18 in human primary astrocytes. miR array analysis of ethanol-exposed human primary astrocytes reveals decreased levels of miR-339, accompanied by an upregulation of NLRP6 inflammasome signaling and astrocyte activation. Through bioinformatics analyses, Argonaute immunoprecipitation assays, and miR-339 overexpression experiments, we identify NLRP6 as a novel 3'-UTR target of miR-339. Overall, our findings confirmed the involvement of miR-339 in NLRP6 inflammasome signaling and its association with cellular activation and neuroinflammation in human primary astrocytes exposed to ethanol and provide novel insights highlighting a previously unrecognized mechanism in alcohol-induced neuroinflammation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Alcohol is widely consumed worldwide and its abuse can cause cognitive dysfunction, affecting memory and learning due to several neurophysiological changes. An imbalance in several neurotransmitters, including the cholinergic and glutamatergic systems, have been implicated in these effects. Zebrafish are sensitive to alcohol, respond to reward stimuli, and tolerate and exhibit withdrawal behaviors. Therefore, we investigated the effects of repetitive exposure to ethanol (REE) and the NMDA receptor antagonist dizocilpine (MK-801) on memory acquisition and glutamatergic and cholinergic neurotransmission. Memory was assessed using the inhibitory avoidance and object recognition tasks. Brain glutamate levels and the activity of Na + -dependent transporters were evaluated as indexes of glutamatergic activity, while acetylcholinesterase (AChE) and choline acetyltransferase (ChAT), enzyme activity were evaluated as indexes of cholinergic activity. Behavioral assessments showed that REE impaired aversive and spatial memory, an effect that MK-801 mimicked. Glutamate levels, but not transporter activity, were significantly lower in the REE group; similarly, REE increased the activity of AChE, but not ChAT, activity. These findings suggest that intermittent exposure to ethanol leads to impairments in zebrafish memory consolidation, and that these effects could be associated with alterations in parameters related to neurotransmission systems mediated by glutamate and acetylcholine. These results provide a better understanding of the neurophysiological and behavioral changes caused by repetitive alcohol use., Competing Interests: Declaration of competing interest The authors have no relevant conflict of interest., (Copyright © 2024. Published by Elsevier Inc.)

Alcohol abuse damages the brain and triggers cognitive impairment. Intestinal dysbiosis has recently been shown to be involved in psychiatric disorders, which suggests the possibility of intestine-to-brain interactions in the development of alcohol abuse. In this study, chronic intermittent alcohol exposure (CIAE) model was established in C57BL/6 male mice and the spatial memory were detected by Barnes maze (n = 16/group). The fecal microbiota and its metabolites were detected by 16S rDNA sequencing and non-target liquid chromatograph mass spectrometer (LC-MS) (n = 8/group). Effects of alcohol on intestinal barrier and blood-brain barrier (BBB) permeability were detected by Evens blue leakage assay (n = 4/group), and the activation state of microglia and TLR4 expression were conducted by immunofluorescence co-localization (n = 4/group). The morphological changes of microglia were analyzed with Image J Analyze Skeleton software, and the protein levels of TLR4 and inflammatory factors were detected by Western Blot (n = 8/group). Results indicated that alcohol alters the components of fecal microbiota and metabolites, and damages the intestinal barrier and BBB, leading to spatial memory impairment in mice. By giving mice specific prebiotics (n = 16/group), we pointed out that increased endotoxin coming from Gram negative bacteria such as lipopolysaccharides (LPS) cross the BBB to activate microglia and inflammatory pathways in the prefrontal cortical (PFC) and hippocampus (HIP), releasing inflammatory factors and resulting in neuroinflammation. Thus, the fecal microbiota seems to be a potential target in the management of alcoholic brain disease., Competing Interests: Declaration of competing interest The authors declare no conflict of interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The pathogenesis of alcohol-related liver disease (ALD) is closely linked to mitochondrial dysfunction and impaired cellular energy metabolism. In this study, we explored how ethanol triggers inflammation, oxidative stress, and mitochondrial dysfunction in Kupffer cells, i.e.hepatic resident macrophages, primarily focusing on the mitochondrial unfolded protein response (UPR mt ) using immortalized mouse Kupffer cells (ImKCs) and mouse primary KCs. The UPR mt is a cellular defense mechanism activated in response to the perturbation of mitochondrial proteostasis to maintain mitochondrial integrity and function by upregulating the expression of mitochondrial chaperones and proteases. We also determined whether nicotinamide riboside (NR), a NAD + precursor, could mitigate ethanol-triggered cellular damage. When ImKCs were exposed to 80 mm ethanol for 72 h, they displayed inflammation, oxidative stress, and impaired mitochondrial function with decreased mitochondrial content and deformed mitochondrial crista structure. NR, however, counteracted the effects of ethanol. Furthermore, ethanol increased mRNA and protein levels of UPR mt genes, such as mitochondrial chaperones and proteases, which were attenuated by NR. Notably, the ethanol-induced shift in the entry of activating transcription factor 5 (ATF5), a putative transcriptional regulator of UPR mt , to the nucleus from the mitochondria was abolished by NR. The induction of UPR mt genes by ethanol was significantly repressed when Atf5 was knocked down, indicating the role of ATF5 in the induction of UPR mt genes in ImKCs exposed to ethanol. We also confirmed the induction of UPR mt gene expression in mouse and human livers exposed to alcohol. Our findings demonstrate the ability of NR to alleviate ethanol-induced oxidative stress, inflammation, and mitochondrial dysfunction, partly by modulating the ATF5-dependent UPR mt pathway in ImKCs, suggesting its potential for ALD therapy. © 2024 The Pathological Society of Great Britain and Ireland., (© 2024 The Pathological Society of Great Britain and Ireland.)

Objectives: We evaluated the effects of four treatments for pathogen inactivation in bone tissue (ethanol storage, formalin fixation, gamma irradiation, and heat treatment via autoclave) on stiffness and anisotropy values in bone samples., Materials and Methods: Cortical bone samples from the humerus of 14 bovine specimens were subjected to Knoop microindentation analysis in longitudinal and transverse planes of section and four indenter orientations within each section. From each specimen, individual samples were assigned to one of five treatment conditions: 50% ethanol-saline solution, formalin immersion, gamma irradiation, autoclave, and buffered saline (controls). Each sample was microindented 100 times and the resultant stiffness data were analyzed by a resampled factorial analysis of variance., Results: First- and second-order interactions as well as main effects of indenter orientation, treatment, and specimen were significant for both transverse and longitudinal sections, with the sole exception that indenter orientation-treatment interaction was nonsignificant for longitudinal sections. Interaction plots reveal that thermal (autoclave) and irradiation treatments depress stiffness values the most, while patterns of indentation stiffness at different orientations are relatively unaffected., Discussion: Patterns of anisotropy are relatively unaffected by preservative treatments, but elastic modulus changes are consistent and unambiguous. Formalin and ethanol treatments are most comparable to controls and represent the best preservative media for mechanical testing. These options, however, are likely to be the least effective for ensuring the inactivation or sterilization of potentially contaminated samples., (© 2024 Wiley Periodicals LLC.)

Anxiety is an emotion that represents a crucial anticipatory reaction of aversive stimuli, with clinical relevance in cases of disproportional and severe occurrences. Although distinct animal models have contributed to elucidate anxiety-related mechanisms, the influence of anxiogenic and anxiolytic modulations on both locomotion and exploration-related parameters in the open field test (OFT) is not fully elucidated. Here, we aimed to assess the influence of anxiogenic and anxiolytic manipulations on the exploratory dynamics of adult zebrafish (Danio rerio) focusing on homebase-related behaviors. As anxiogenic manipulations, we used the morphine (1.5 mg/L) withdrawal protocol (MOR); 3.5 mL/L conspecific alarm substance (CAS) for 5 min; and 100 mg/L caffeine (CAF) for 15 min. To evoke anxiolytic-like responses, animals were acutely exposed to 0.5 % (v/v) ethanol (ETOH) for 1 h; 100 μg/L fluoxetine (FLU) for 15 min; and 0.006 mg/L clonazepam (CZP) for 10 min. Then, fish were individually exposed to the 30-min OFT trial, with posterior analysis of behavioral activity. While MOR induced hyperlocomotion and increased periphery occupancy, CAS and CAF groups showed higher immobility and increased latency to homebase formation, respectively. Conversely, ETOH and FLU reduced homebase occupancy, supporting anxiolytic-like behaviors, while CZP did not change zebrafish behavior in the OFT. Cluster analysis was used to reconfirm the remarkable similarities and discrepancies between treatments, thus contributing to characterize the distinct responses measured. Overall, our novel data show the relevance of homebase-related analysis as a sensitive tool to reflect affective-like states in zebrafish, providing innovative approaches to unravel the spatio-temporal dynamics of anxiety-like behaviors in vertebrates., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Adolescent intermittent ethanol (AIE) exposure in rats leads to social deficits. Parvalbumin (PV) expressing fast-spiking interneurons in the prelimbic cortex (PrL) contribute to social behavior, and perineuronal nets (PNNs) within the PrL preferentially encompass and regulate PV interneurons. AIE exposure increases PNNs, but it is unknown if this upregulation contributes to AIE-induced social impairments. The current study was designed to determine the effect of AIE exposure on PNN expression in the PrL and to assess whether PNN dysregulation contributes to social deficits elicited by AIE. cFos-LacZ male and female rats were exposed every other day to tap water or ethanol (4 g/kg, 25% w/v) via intragastric gavage between postnatal day (P) 25-45. We evaluated neuronal activation by β-galactosidase expression and PNN levels either at the end of the exposure regimen on P45 and/or in adulthood on P70. In addition, we used Chondroitinase ABC (ChABC) to deplete PNNs following adolescent exposure (P48) and allowed for PNN restoration before social testing in adulthhod. AIE exposure increased PNN expression in the PrL of adult males, but decreased PNNs immediately following AIE. Vesicular glutamate transporter 2 (vGlut2) and vesicular GABA transporter (vGat) near PNNs were downregulated only in AIE-exposed females. Gene expression of PNN components was largely unaffected by AIE exposure. Removal and reestablishment of PrL PNNs by ChABC led to upregulation of PNNs and social impairments in males, regardless of adolescent exposure. These data suggest that AIE exposure in males upregulates PrL PNNs that likely contribute to social impairments induced by AIE., Competing Interests: Declaration of competing interest All co-authors have read and approved the manuscript for submission and have made substantial contributions to the conception, design, gathering, analysis and/or interpretation of the data as well as contributed to the writing and/or intellectual content of the article. All authors have exercised due care in ensuring the integrity of the work, and all animal procedures were conducted according to NIH guidelines and approved by our institutional animal care and use committee. None of the original material contained in the manuscript, outside of conference abstracts, has been submitted for consideration elsewhere. None of the authors have any financial declarations or conflicts of interest related to the submitted study., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Ethanol infusion into the vein of Marshall (EIVOM) has been performed as an adjunctive atrial fibrillation therapy. However, the time course change, quantitative lesion investigation, and effects on epicardial fat pads and fractionated atrial electrograms created by EIVOM have never been investigated., Objective: This study aimed to perform a quantitative analysis of lesions created by EIVOM., Methods: We created voltage maps using a 3-dimensional mapping system immediately before and 30 minutes and 60 minutes after performing EIVOM to study the time course change in the lesions. We compared differences in the average contact force value required for successful conduction block in the Marshall vein area of patients with and without EIVOM. We also investigated effects of EIVOM on the area of complex fractionated atrial electrograms before and after EIVOM. We measured the total epicardial fat pad volume before and after EIVOM by computed tomography., Results: Voltage was significantly reduced after EIVOM, and there were significant differences in voltage reduction between the control status and 30 minutes and 60 minutes after EIVOM (P < .05). The average contact force value was significantly lower with vs without EIVOM (P < .05). The total epicardial fat volume and complex fractionated atrial electrogram area also significantly decreased after EIVOM (P < .05)., Conclusion: EIVOM provided significant therapeutic effects on the left atrial tissue perpetuating atrial fibrillation, which was demonstrated by a quantitative analysis., Competing Interests: Disclosures The authors have no conflicts of interest to disclose., (Copyright © 2024 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.)

Alcohol misuse increases infections and cancer fatalities, but mechanisms underlying its toxicity are ill-defined. We show that alcohol treatment of human tracheobronchial epithelial cells leads to inactivation of giantin-mediated Golgi targeting of glycosylation enzymes. Loss of core 2 N-acetylglucosaminyltransferase 1, which uses only giantin for Golgi targeting, coupled with shifted targeting of other glycosylation enzymes to Golgi matrix protein 130-Golgi reassembly stacking protein 65, the site normally used by core 1 enzyme, results in loss of sialyl Lewis x and increase of sialyl Lewis a and α2-6sialo mucin O-glycans. The α2-6sialo mucin O-glycans induced by alcohol cause death of U937 macrophages mediated by sialic acid-binding immunoglobulin-like lectin 7. These results provide a mechanistic insight into the cause of the toxic effects of alcohol and might contribute to the development of therapies to alleviate its toxicity., (© 2024 The Author(s). FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

The co-use of alcohol and cannabis is rising in prevalence, yet the mechanisms driving individuals to co-use are not well understood. Subjective response to alcohol or cannabis may predict the desire to use either substance. However, which specific facets of subjective response predict cross-substance craving remains unclear. The present study investigated whether acute administration of alcohol or cannabis facilitates cue-induced craving for the other substance, with an emphasis on the underlying subjective response mechanisms contributing to co-use. This is a secondary analysis of a behavioral pharmacology study that combined alcohol/cannabis administration with a cross-substance cue-reactivity paradigm in individuals who were heavy alcohol and heavy cannabis co-users. Over two sessions, twenty-nine individuals (17M/12F) self-administered alcohol or cannabis (in a crossover design), and then completed a cue-reactivity exercise with the other substance. Analyses tested how changes in subjective response variables following substance administration predicted cross-substance cue-induced craving. Following alcohol administration, greater subjective ratings of positive mood predicted significantly greater cue-induced cannabis craving (β = 1.14, SE = 0.41, t = 2.80, p = 0.010). Following cannabis administration, lower subjective effects ratings of positive mood/arousal predicted significantly greater cue-induced alcohol craving (β = -1.08, SE = 0.38, t = -2.85, p = 0.009; β = -2.38, SE = 1.13, t = -2.10, p = 0.047). This study identified subject response mechanisms contributing to cross-substance cue induced craving. These mechanisms include increases in positive mood following alcohol use and decreases in positive mood and arousal, akin to increases in relaxation, following cannabis use., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

The mechanism(s) underlying gut microbial metabolite (GMM) contribution towards alcohol-mediated cardiovascular disease (CVD) is unknown. Herein we observe elevation in circulating phenylacetylglutamine (PAGln), a known CVD-associated GMM, in individuals living with alcohol use disorder. In a male murine binge-on-chronic alcohol model, we confirm gut microbial reorganization, elevation in PAGln levels, and the presence of cardiovascular pathophysiology. Fecal microbiota transplantation from pair-/alcohol-fed mice into naïve male mice demonstrates the transmissibility of PAGln production and the CVD phenotype. Independent of alcohol exposure, pharmacological-mediated increases in PAGln elicits direct cardiac and vascular dysfunction. PAGln induced hypercontractility and altered calcium cycling in isolated cardiomyocytes providing evidence of improper relaxation which corresponds to elevated filling pressures observed in vivo. Furthermore, PAGln directly induces vascular endothelial cell activation through induction of oxidative stress leading to endothelial cell dysfunction. We thus reveal that the alcohol-induced microbial reorganization and resultant GMM elevation, specifically PAGln, directly contributes to CVD., Competing Interests: Competing interests: Z.W. reports being named as co-inventor on pending and issued patents held by the Cleveland Clinic relating to cardiovascular diagnostics and therapeutics. Z.W. also reports having received royalty payments for inventions or discoveries related to cardiovascular diagnostics or therapeutics from Cleveland Heart Lab, a fully owned subsidiary of Quest Diagnostics and Procter & Gamble. C.M.T. consults for Abbott Laboratories. S.L.H. reports being named as co-inventor on pending and issued patents held by the Cleveland Clinic relating to cardiovascular diagnostics and therapeutics, being a paid consultant for Procter & Gamble and Zehna Therapeutics, having received research funds from Procter & Gamble, Zehna Therapeutics, and Roche Diagnostics, and being eligible to receive royalty payments for inventions or discoveries related to cardiovascular diagnostics or therapeutics from Cleveland HeartLab and Procter & Gamble. All other authors: Z.L., M.G., A.Z., X.F., K.M., M.M., X.S.L., Z.Y., J.M., T.T., K.C., M.H., L.H., K.G., T.T.V., D.W., A.M.G., P.Q., C.A.M., J.M.B., D.J.L., D.A.W., and T.E.S. have no competing interests., (© 2024. The Author(s).)

Psychoactive drugs such as alcohol and stimulants are typically used in social settings such as bars, parties or small groups. Yet, relatively little is known about how social contexts affect responses to drugs, or how the drugs alter social interactions. It is possible that positive social contexts enhance the rewarding properties of drugs, perhaps increasing their potential for repeated use and abuse. In addition, drugs may enhance the rewarding effects of social interactions by increasing feelings of social closeness and connectedness. To examine these relations, we investigated the effects of several drugs (MDMA, methamphetamine, alcohol) on feelings of connection between two strangers engaged in a conversation. We also investigated feelings of connection between two participants who discussed either 'shallow' or deeper topics in two conversations, without any drugs. All four conditions: deeper conversations, MDMA, methamphetamine and alcohol significantly increased feelings of connection and closeness compared with control conditions (small talk or placebo). We postulate that these feelings of connection could contribute to the drugs' rewarding effects when the drugs are used in social contexts., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Disclosures: HdW is on the Board of Directors of PharmAla Biotech, and on scientific advisory committees of Gilgamesh Pharmaceuticals and MIND Foundation. These activities are unrelated to the present report. Other authors declare no conflicts., (© 2024. The Author(s).)

Background: Propolis is a natural substance produced by honeybees that has various biological properties including, anti-inflammatory, antioxidant and antimicrobial properties. Although previous studies have evaluated the antimicrobial effects of propolis in dentistry, its effects on dental pulp stem cell (DPSC) viability, migration, and differentiation are yet not well understood. The objective of this study was to investigate the effects of Chinese propolis on viability/proliferation, migration, differentiation and cytokine expression in DPSCs., Methods: Commercially available DPSCs (Lonza) were treated with aqueous extract of propolis (AEP) or ethanolic extract of propolis (EEP), and viability/proliferation was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays and quantification of nuclear staining. DPSC differentiation into mineralizing cells was evaluated with Alizarin red staining and cell migration was assessed using Boyden Chamber Transwell inserts. Cytokine expression was measured by RT-qPCR. AEP and EEP at 0.03 and 0.1 mg/mL did not affect DPSC viability/proliferation for up to 7-days treatment., Results: Higher doses (0.33-33 mg/mL) induced a dose dependent decrease in DPSC viability/proliferation with a more prominent effect with EEP at 7 days. Neither AEP nor EEP induced DPSC differentiation into mineralizing cells, but both AEP and EEP (0.03-0.1 mg/ml) induced a dose dependent increase in DPSC migration. In addition, EEP prevents the upregulation of IL1b and IL6 but not IL8 and CCL2 in response to lipopolysaccharide stimulation. AEP has less potent anti-inflammatory effects and prevents only IL1b upregulation., Conclusion: This study provides new information about the biologic properties of ethanolic and aqueous extracts of propolis and shows that propolis, at doses that do not affect cell viability, induces DPSC migration and has anti-inflammatory properties. These data highlight the potential use of propolis as an alternative intra-canal medicament for regenerative endodontic procedures., Competing Interests: The authors declare that they have no competing interests., (© 2024 Park et al.)

Background: Concomitant use of alcohol and benzodiazepines are described among elderly, raising concerns about their combined impact on memory. We aimed to evaluate the long-term impact of chronic diazepam use associated with ethanol intoxication on memory in aging mice., Methods: Twelve-month-old male C57BL6 mice were assigned into 4 groups: ethanol (OH), diazepam (DIA), diazepam + ethanol (DOH) and control (CTL). For 16 weeks, ethanol was available ad libitum and diazepam was mixed with food. Behavioral testing, performed during and after treatment cessation included working memory and visual recognition memory assessment. The second session was implemented with spatial reference learning and memory assessment in the Barnes maze test. In vivo magnetic resonance spectroscopy (MRS) acquisitions were performed to quantify hippocampal metabolites during and after cessation treatment., Results: During treatment, visual recognition memory was significantly different between groups with the DIA group exhibiting the worst performance. MRS acquisition highlighted higher glutamate and choline levels in OH and DOH groups in comparison to CTL and DIA groups. After treatment wash-out, there was no difference between in the different memories evaluated. Only the learning phase of the spatial reference memory test differed significantly with worst performance in OH groups. Three months after treatment cessation, there was no remanent effect of diazepam + ethanol on hippocampal metabolites changes., Conclusions: We did not evidence additive effect of ethanol and diazepam on memory and hippocampal metabolite levels. The disturbances observed during treatment were no remanent, highlighting the benefits of discontinuing these substances., Competing Interests: Declaration of competing interest The authors have no conflict of interest to disclose., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

There has been a recent renewed interest in the potential use of psychedelic drugs as therapeutics for certain neuropsychiatric disorders, including substance use disorders. The psychedelic drug 2,5-dimethoxy-4-iodoamphetamine (DOI) has demonstrated therapeutic efficacy in preclinical models of opioid use disorder (OUD). Alcohol is commonly co-used in individuals with OUD, but preclinical models that recapitulate this comorbidity are lacking. We developed a polydrug model wherein male and female rats were allowed to self-administer intravenous heroin and oral alcohol (or saccharin control solution) over weeks of behavioral training, and then we conducted a series of progressive ratio tests to assess the animals' motivational state for heroin and alcohol. In this model, motivation for heroin is higher than alcohol, and DOI (0.4 mg/kg) administered prior to testing significantly reduced heroin motivation measured as the animals' break point, or maximum effort the animal is willing to expend to obtain a single infusion of heroin. The 5-HT2A receptor antagonist MDL 100,907 (0.3 mg/kg), but not the 5-HT2C receptor antagonist SB-242084 (0.5 mg/kg), blocked the therapeutic effect of DOI on heroin motivation. No significant effects on alcohol break points were observed, nor did MDL 100,907 or SB-242084 have any effect on break points on their own. These data support the view that psychedelic drugs like DOI may have therapeutic effects on opioid use in individuals with OUD and comorbid alcohol use, by acting as a 5-HT2A receptor agonist., Competing Interests: Declaration of competing interest J.P. is a consultant for Delix Therapeutics, Inc. D.E.O. is a co-founder of Delix Therapeutics, Inc., serves as the Chief Innovation Officer and Head of the Scientific Advisory Board, and has sponsored research agreements with Delix Therapeutics., (Copyright © 2024. Published by Elsevier Ltd.)

Although most adults in the United States will drink alcohol in their life, only ∼6% will go on to develop an alcohol use disorder (AUD). While a great deal of work has furthered our understanding of the cycle of addiction, it remains unclear why certain people transition to disordered drinking. Altered activity in regions implicated in AUDs, like the basolateral amygdala (BLA), has been suggested to play a role in the pathophysiology of AUDs, but how these networks contribute to alcohol misuse remains unclear. Here we investigated how the impact of alcohol on the BLA network relates to alcohol exposure. We first examined the effect of acute ethanol administration on the BLA and frontal cortical networks and the relationship with subsequent voluntary ethanol consumption using the Intermittent Access paradigm. In addition, we recorded network activity from the BLA and frontal cortex throughout the Drinking-in-the-Dark-Multiple Scheduled Access paradigm to assess the impact of voluntary alcohol consumption on network states during binge and abstinence periods. Finally, we investigated the impact of acute withdrawal from chronic alcohol exposure on BLA and frontal cortex network states using the Chronic Intermittent Ethanol (vapor) paradigm. We demonstrate that across paradigms, ethanol alters low gamma band (40-70 Hz) oscillations and remarkably correlates with the extent of future voluntary ethanol consumption in the IA paradigm. These data suggest that BLA network states play a role in the mechanisms influencing voluntary alcohol intake., (Copyright © 2024 DiLeo et al.)

Light alcohol consumption (LAC) may reduce the incidence and improve the prognosis of ischemic stroke. Recently, we found that LAC promotes cerebral angiogenesis and inhibits early inflammation following ischemic stroke. In addition, LAC upregulates lipocalin-type prostaglandin D2 synthase (L-PGDS) in the brain. Thus, we determined the role of endothelial L-PGDS in the protective effect of LAC. In in vitro studies, chronic exposure to low-concentration ethanol upregulated L-PGDS and significantly increased the proliferation in cultured C57BL/6J mouse brain microvascular endothelial cells (MBMVECs). AT-56, a selective L-PGDS inhibitor, abolished low-concentration ethanol exposure-induced proliferation. In in vivo studies, 8-week gavage feeding with 0.7 g/kg/day ethanol, defined as LAC, promoted cerebral angiogenesis under physiological conditions and following ischemic stroke in male C57BL/6J mice. In addition, LAC inhibited the post-ischemic expression of adhesion molecules, neutrophil infiltration, and microglial activation. AT-56 and endothelial cell (EC)-specific L-PGDS conditional knockout did not significantly alter cerebral angiogenesis and post-ischemic inflammation in the control mice but eliminated the pro-angiogenic and anti-inflammatory effects of LAC. Furthermore, EC-specific L-PGDS conditional knockout alleviated the neuroprotective effect of LAC against cerebral ischemia/reperfusion injury. These findings suggest that endothelial L-PGDS may be crucial in the pro-angiogenic and anti-inflammatory effects of LAC against ischemic stroke.

Background: It is mandatory to demonstrate the removal or inactivation of potential viral contaminants in the manufacturing processes of pharmaceuticals derived from biomaterials. Porcine-derived gelatin is used in various medical fields, including regenerative medicine, tissue engineering, and medical devices. However, the steps of virus inactivation in the gelatin manufacturing process are poorly defined. In this study we evaluated virus inactivation in two steps of the gelatin manufacturing process., Methods: Pig skin (4.5 g), including solid pieces as intermediate products, was spiked with model viruses, including CPV (canine parvovirus), BAV (bovine adenovirus), BPIV3 (bovine parainfluenza type 3), PRV (pseudorabies virus), BReoV3 (bovine reovirus type 3), and PPV (porcine parvovirus), and underwent chemical treatment with alkaline ethanol or heat treatment at 62 °C followed by inoculation in relevant cell cultures. Viral titers in the samples were calculated based on the Behrens-Kärber method., Results: Model viruses were inactivated at different rates; however, effective inactivation of all model viruses was demonstrated by an LRV (log reduction value) over 4 by both chemical and heat treatment, and chemical treatment demonstrated rapid inactivation compared to heat treatment., Conclusion: The chemical and heat treatment steps exhibited meaningful viral inactivation capacity. They are integrated parts in the extraction and manufacturing process of porcine-derived gelatin, ensuring virus safety for use in medical applications., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: All authors are employees of Nitta Gelatin Inc., (© 2024. The Author(s).)

Background: In approximately 80% of colorectal cancer cases, mutations in the adenomatous polyposis coli ( APC ) gene disrupt the Wingless-related integration site (Wnt)/β-catenin signaling pathway, a crucial factor in carcinogenesis. This disruption may result in consequences such as aberrant spindle segregation and mitotic catastrophe. This study aimed to analyze the effectiveness of the ethanolic extract of red okra ( Abelmoschus esculentus ) pods (EEROP) in inducing apoptosis in colorectal cancer cells (SW480) by inhibiting the Wnt/β-catenin signaling pathway., Methods: The IC 50 of EEROP in SW480 cells was determined by treating the cells with varying doses of EEROP, ranging from 0 to 1000 µg/mL. Apoptosis assay and signaling pathway analysis were performed through immunofluorescence staining and Western Blotting on SW480 cells treated with 250 µg/mL of EEROP for 72 hours., Results: EEROP treatment induced apoptosis in SW480 cells, marked by elevated levels of active caspase-3 (P<0.001) and cleaved poly-(ADP-ribose) polymerase (PARP)-1. Moreover, it notably decreased β-catenin protein levels, resulting in an augmented occurrence of cells displaying abnormal spindle segregation during mitosis (P=0.04)., Conclusion: EEROP treatment reduces β-catenin protein levels, promotes abnormal spindle apparatus segregation, and finally leads to apoptotic cell death in CRC cells., Competing Interests: None declared., (Copyright: © Iranian Journal of Medical Sciences.)

Dengue hemorrhagic fever (DHF) is a major health concern in tropical and subtropical countries. Indonesia has DHF cases perennially every year. On the other hand, Indonesia is abundant with seaweed (Sargassum duplicatum ), which can be found across its seashore. The macroalgae contains secondary metabolites with ovicidal activity; hence, it has the potential to be utilized in suppressing the Aedes aegypti . The aim of this study was to determine the ovicidal activity of S. duplicatum against the Ae. aegypti eggs. The algae were macerated with ethanol 70% before being subjected to qualitative phytochemical screenings. The ovicidal tests were conducted with an extract concentration of 50 ppm, 100 ppm, 500 ppm, 1000 ppm, and 1500 ppm, while distilled water was used as the control. The hatchability of Ae. aegypti was observed 24 hours a day for 4 days and the larval development was investigated under a microscope. Phytochemical screenings revealed that the extract was positive containing alkaloids, flavonoids, steroids, saponins, and phenols. The hatchability of Ae. aegypti eggs were significantly reduced following the S. duplicatum extract exposure for four days ( p =0.000). The extract had LC 50 of 828.653 ppm and LC 50 of 1786.09 ppm for the ovicidal activity against Ae. aegypti eggs. The concentration of ethanol extract of S. duplicatum did not affect the mosquito development from larvae to adult stage ( p =0.263). Further research is needed to explore the effect of specific compounds contained in the S. duplicatum and investigate their ovicidal potential., Competing Interests: The authors declare that they have no conflict of interest in the writing or publication of this article., (© 2024 The Author(s).)

Alcohol use is widespread across many societies. While most people can control their alcohol use, a vulnerable sub-population develops alcohol use disorder, characterized by continued alcohol use despite negative consequences. We used a rat model of alcohol self-administration despite negative consequences to identify brain activity associated with this addiction-like behaviour. We and others have previously shown that response-contingent punishment of alcohol self-administration with mild footshock reliably identifies two sub-populations. One group substantially decreases alcohol self-administration in the face of punishment (punishment-sensitive, controlled) and another group continues alcohol self-administration despite negative consequences (punishment-resistant, addiction-like behaviour). In this study, we aimed to validate this model in females and identify associated brain regions. We trained Long-Evans outbred rats (n = 96) to self-administer 20% ethanol, and then introduced response-contingent footshock. We found that female rats consumed more alcohol in unpunished and punished sessions compared to male rats. In one group of rats (n = 24, m/f), we identified neuronal activity associated with punishment-resistant alcohol self-administration using the neurobiological marker of activity cFos. We found lower cFos expression in NAcSh associated with punishment-resistant alcohol self-administration. In another group of rats (n = 72, m/f), we used chemogenetic inhibition of NAcSh during punished alcohol self-administration. We found that chemogenetic NAcSh inhibition had no effect on unpunished alcohol self-administration but selectively increased punished alcohol self-administration in punishment-resistant female rats. These results indicate that more female rats develop punishment-resistant alcohol consumption, and that NAcSh hypofunction may underlie this phenotype., (© 2024. The Author(s), under exclusive licence to American College of Neuropsychopharmacology.)

Background: Zuranolone is an oral, once-daily, 14-day treatment course approved for adults with postpartum depression in the United States., Aims: To assess cognitive effects, pharmacokinetics, and safety of zuranolone, alone or with alprazolam/ethanol., Methods: This was a phase 1, two-part, two-period, randomized, double-blind, placebo-controlled crossover trial. Participants received zuranolone 50 mg or placebo once daily for 9 days, and additionally received alprazolam (1 mg, Part A), ethanol (males: 0.7 g/kg; females: 0.6 g/kg, Part B), or corresponding placebo on days 1, 5, and 9. Within each part, participants received all treatment combinations. Cognition was assessed using a computerized test battery; pharmacokinetics and safety were also evaluated., Results: All participants (Part A, N  = 24; Part B, N  = 25) received ⩾1 dose of zuranolone/placebo. Compared to placebo, zuranolone produced small-to-moderate cognitive decline (Cohen's | d | = 0.126-0.76); effects were larger with alprazolam (Cohen's | d | = 0.523-0.93) and ethanol (Cohen's | d | = 0.345-0.88). Zuranolone coadministration with alprazolam (Cohen's | d | = 0.6-1.227) or ethanol (Cohen's | d | = 0.054-0.5) generally worsened cognitive decline when compared with zuranolone alone. Maximal pharmacodynamic effects occurred at approximately 5 h and were resolved by 12 h postbaseline. No pharmacokinetic interactions were observed. Incidence of adverse events was similar between groups; most events were mild or moderate in severity., Conclusion: A general small-to-moderate magnitude decline in cognition occurred with zuranolone alone. Coadministration with alprazolam/ethanol increased the magnitude, but not the duration, of effects compared with single-agent administration. Zuranolone prescribers and patients should be aware of the potential for increased central nervous system-depressant effects if coadministered with GABAergic active compounds such as alprazolam and ethanol., Competing Interests: Declaration of conflicting interestsThe author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: David P Walling has received grants or contracts from Acadia, Alkermes, Allergan, Avanir, Biogen, Boehringer Ingelheim, Cerevel, Indivior, IntraCellular, Janssen, J&J PRD, Karuna, Lundbeck, Lupin, Lyndra, Merck, Neurocrine, Novartis, Noven, Otsuka, Pfizer, Roche, Sage, Sunovion, Takeda; and consulting fees from Biogen, Boehringer Ingelheim, Janssen, Lyndra, Merck, and Otsuka.Howard A Hassman is a current employee of CenExel Hassman Research Institute, an independent research site that conducts investigator-initiated and industry-sponsored pharmaceutical trials. He has no conflicts of interest or bias in the conclusions of the current investigation or promotion of the current study results.Paul Maruff is an employee of Cogstate Ltd. and may hold stock and/or stock options.Rakesh Jain reports research funding from AbbVie, Lilly, Lundbeck, Otsuka, Pfizer, Shire, and Takeda; participation on advisory boards for Adamas, Alkermes, Corium, Eisai, Janssen, Lilly, Lundbeck, Merck, Neos Therapeutics, Neurocrine Biosciences, Otsuka, Pamlab, Pfizer, Sage, Shire, Sunovion, Supernus, Takeda, Teva, and Usona; honoraria for speakers’ bureaus from AbbVie, Alkermes, Almatica, Axsome, Corium, Eisai, Intra-Cellular Therapies, Ironshore Pharmaceuticals, Janssen, Lilly, Lundbeck, Merck, Neos Therapeutics, Otsuka, Pamlab, Pfizer, Shire, Sumitomo, Sunovion, Takeda, Tris Pharmaceuticals, and Viatris; and consulting fees from AbbVie, Acadia, Adamas, Alfasigma, Alkermes, Almatica, Axsome, Biogen, Boehringer Ingleheim, Cingulate Therapeutics, Corium, Eisai, Evidera, Impel, Janssen, Lilly, Lundbeck, Merck, Neos Therapeutics, Neurocrine Biosciences, Osmotica, Otsuka, Pamlab, Pfizer, Sage, Shire, Sumitomo, Sunovion, Supernus, Takeda, Teva, Transcend Therapeutics, and Viatris.Andy Czysz was an employee of Sage Therapeutics, Inc., at the time that the study was completed, and may hold stock and/or stock options.Indrani Nandy, Victor Ona, and Joi Dunbar are employees of Sage Therapeutics, Inc., and may hold stock and/or stock options.Margaret K Moseley and Seth Levin are employees of Biogen Inc. and may hold stock.

The Edinger-Westphal nucleus (EW) is a midbrain nucleus composed of a preganglionic, cholinergic subpopulation and a densely clustered peptidergic subpopulation (EWcp). The EWcp is one of the few brain regions that show consistent induction of FOS following voluntary alcohol intake. Previous results in rodents point to urocortin 1 (UCN1) as one of the peptides most involved in the control of ethanol intake and preference. Notably, the functions described for UCN1, such as reward processing, stress coping or the regulation of feeding behavior are similar to those described for the neuropeptide neuromedin U (NMU). Interestingly, NMU has been recently associated with the modulation of alcohol-related behaviors. However, little is known about the expression and functionality of NMU neurons in alcohol-responsive areas. In this study, we used the recently developed Nmu-Cre knock-in mouse model to examine the expression of NMU in the subaqueductal paramedian zone comprising the EWcp. We delved into the characterization and co-expression of NMU with other markers already described in the EWcp. Moreover, using FOS as a marker of neuronal activity, we tested whether NMU neurons were sensitive to acute alcohol administration. Overall, we provided novel insights on NMU expression and functionality in the EW region. We showed the presence of NMU within a subpopulation of UCN1 neurons in the EWcp and demonstrated that this partial co-expression does not interfere with the responsivity of UCN1-containing cells to alcohol. Moreover, we proposed that the UCN1 content in these neurons may be influenced by sex., (© 2024. The Author(s).)

Alcohol misuse in people with human immunodeficiency virus (HIV) (PWH) and chronic binge alcohol (CBA) administration in simian immunodeficiency virus (SIV)-infected macaques are associated with increased physical frailty and impaired functional skeletal muscle mass, respectively. Previous studies by our group demonstrate that muscle-enriched microRNAs (myomiRs) are differentially expressed in skeletal muscle (SKM) from CBA-administered SIV-infected male macaques and their altered expression contributes to impaired differentiation of SKM stem cells or myoblasts. MicroRNAs can be transported in extracellular vesicles (EVs) to mediate numerous cellular responses through intercellular communication. The present study tested the hypothesis that EV-mediated delivery of miR-206 can ameliorate CBA-mediated decreases in myoblast differentiation. Myoblasts were isolated from SKM of female SIV-infected, antiretroviral therapy-treated macaques that received either CBA (2.5 g/kg/day, CBA/SIV) or water (VEH/SIV) for 14.5 mo. Myotube and myotube-derived EV myomiR expression, including miR-206, was lower in the CBA/SIV group. Overexpression of miR-206 decreased histone deacetylase 4 ( HDAC4 ) and paired box 7 ( PAX7 ) expression in myotubes and increased fusion index, a differentiation index, in CBA/SIV-derived myotubes. Similarly, EV-mediated delivery of miR-206 increased both fusion index and myotube density of CBA/SIV-derived myoblasts. These results support the potential therapeutic utility of EVs in delivering myomiRs to improve SKM stem cell differentiation. NEW & NOTEWORTHY Alcohol decreases skeletal muscle myoblast differentiation into myotubes, which is associated with decreased expression of microRNA-206. We show that delivering exogenous miR-206 in plasma-derived extracellular vesicles (EVs) to myoblasts derived from alcohol-administered animals increases myotube differentiation. These results support the potential therapeutic utility of EVs in delivering muscle-enriched microRNAs to improve skeletal muscle stem cell differentiation.

Rationale: Alcohol consumption despite negative consequences is a core symptom of Alcohol Use Disorder. In animal models, this is studied by pairing aversive stimuli with alcohol access, and continuation of drinking under these conditions is known as aversion resistance. Previously, we found that female mice are more aversion resistant than males. Corticotropin releasing hormone (Crh) and the Crh receptor 1 (Crhr1) regulate stress-induced reinstatement, alcohol dependence, and binge-like drinking. However, the role of the Crh system in aversion resistance has not been assessed., Objectives: We aimed to identify sex differences in the Crh system during quinine-adulterated alcohol intake., Methods: We used two-bottle choice and adulterated the alcohol solution with quinine. Next, we measured Crh and Crhr1 levels in brain tissue using real-time polymerase chain reaction (RT-qPCR) and RNAscope in situ hybridization. We then infused a Crhr1 antagonist into the medial prefrontal cortex (mPFC) prior to quinine-alcohol intake., Results: After quinine-alcohol consumption, females exhibited increased mPFC Crhr1 mRNA levels as measured by RT-qPCR. This was confirmed with greater anatomical specificity using RNAscope, with females exhibiting an increased number of Crhr1 + cells in the dorsomedial PFC and the ventromedial PFC. mPFC Crhr1 antagonist treatment reduced quinine-alcohol consumption in females but did not impact consumption in males. Quinine-free alcohol intake was unaffected by Crhr1 antagonist treatment., Conclusions: Our findings suggest that Crhr1 in mPFC plays a role in aversion resistant alcohol intake in females. Future experiments will examine the sources of Crh innervation to the mPFC and their distinct roles in alcohol seeking., Competing Interests: Declarations Conflict of interest On behalf of all authors, the corresponding author states that there is no conflict of interest., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Moderate prenatal alcohol exposure (mPAE) results in structural alterations to the hippocampus. Previous studies have reported impairments in hippocampal-sensitive tasks, but have not compared performance between male and female animals. In the present study, performance in hippocampal-sensitive spatial memory and anxiety behavior tests were compared across adult male and female saccharin (SACC) control mPAE Long-Evans rat offspring. Two tests of spatial memory were conducted that were aimed at assessing memory for recently acquired spatial information: A delayed spatial alternation task using an M-shaped maze and a delayed match-to-place task in the Morris water task. In both tasks, rats in SACC and mPAE groups showed similar learning and retention of a spatial location even after a 2-h interval between encoding and retention. A separate group of adult male and female SACC and mPAE rat offspring were tested for anxiety-like behaviors in the elevated plus-maze paradigm. In this test, both male and female mPAE rats exhibited a significantly greater amount of time and a greater number of head dips in the open arms, while locomotion and open arm entries did not differ between groups. The results suggest that mPAE produces a reduction in anxiety-like behaviors in both male and female rats in the elevated plus-maze., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Prenatal alcohol exposure (PAE) can lead to a wide range of adverse effects in humans, including impaired self-control and increased impulsive behavior. Deficits in self-control can interfere with academic performance and have lasting impacts. In the present study, a rodent model of PAE was used to assess impulsivity through operant conditioning. Pregnant rats were assigned to one of three groups: ad-lib control (CON), pair-fed (PF), and alcohol-exposed (ALC). ALC rats were given a liquid diet containing 6% alcohol, PF rats were yoked to an ALC rat and given a CON liquid diet, and CON rats received ad libitum food. Operant conditioning was used to evaluate extinction in adolescents (Experiment 1) and differential reinforcement of low rates (DRL) in adolescents and adults (Experiment 2). PAE resulted in an increase in responses and resets during DRL testing, indicative of impaired self-control, an effect that was only observed in adolescent males. Females, regardless of age, did not show increased impulsivity following PAE. This indicates that children with PAE may exhibit attentional deficits similar to those with attention deficit hyperactivity disorder, with males at a higher risk., (© 2024 The Author(s). Developmental Psychobiology published by Wiley Periodicals LLC.)

Our laboratory has previously shown that chronic ethanol exposure elicits enhanced working memory performance in female, but not male, adult Sprague-Dawley rats, indicative of a fundamental sex difference in cortical plasticity. Recent studies have furthermore revealed that females display markedly reduced HCN-mediated channel activity in inhibitory Martinotti interneurons after chronic ethanol exposure that is similarly not observed in males. From these observations we hypothesized that alcohol induces facilitated working memory performance via down-regulation of these channels' activity specifically within interneurons. To test this hypothesis, we employed a Pol-II compatible shRNA expression system to elicit targeted knockdown of HCN channel activity in these cells, and measured performance on a delayed Non-Match-to-Sample (NMS) T-maze test to gauge effects on working memory performance. A significant baseline enhancement of working memory performance with HCN channel knockdown was observed, indicative of a critical role for interneuron-expressed HCNs in maintaining optimal cortical network activity during cognitively-demanding tasks. Consistent with previous observations, ethanol exposure resulted in enhanced NMS T-maze performance, however elevated working memory performance was observed in both scram- and hcn-shRNA infected groups after alcohol administration. We therefore conclude that interneuron-expressed HCN channels, despite representing a minor population of total cortical HCN expression, contribute substantially to maintaining working memory processes. Downregulated HCN channel activity, though, does not alone appear sufficient to manifest alcohol-induced enhancement of working memory performance observed in female rats during acute withdrawal., (Copyright © 2024 Elsevier Inc. All rights reserved.)