EtOH Exposure Research Articles

Single essential oils and their binary mixtures protect against ethanol-induced defects in a zebrafish fetal alcohol spectrum disorder model at the same level as folic acid

This study evaluated protective effects of clove (SEO), thyme white (TEO), oregano (OEO), and caraway (CEO) essential oils (EOs), and their binary mixtures, in a zebrafish fetal alcohol spectrum disorder model. Furthermore, folic acid (FA) was used for comparison as it had previously shown protection against ethanol (EtOH)-induced defects. The co-exposure of zebrafish embryos to EtOH (150 mM) and FA (75 μM) or EOs and their binary mixtures (0.5-1 mg/L) was carried out during 6 or 22 hours postfertilization (hpf). Different developmental endpoints (epiboly measurement, survival rate at 24 hpf, embryonic developmental progression measurement at 24 hpf, larval development at 48-96 hpf, and hatching rate at 72-96 hpf) were evaluated at 8-96 hpf. EtOH exposure reduced epiboly. Only FA and the SEO + TEO binary mixture protected against these defects, and SEO and TEO single exposure showed partial protection. Therefore, these groups were chosen for subsequent experiments. At 24 hpf, EtOH showed developmental delay and hatching rate was delayed at 72 hpf. FA, SEO, TEO, and SEO + TEO partially protected against these defects. This study supports the conclusion that FA partially protects against EtOH-induced defects. SEO and TEO single exposure partially protect against EtOH-induced defects. However, the binary mixture of SEO + TEO was more effective, showing similar efficacy as FA.

Alcohol Activates Cannabinoid Receptor 1 and 2 in a Model of Pathogen Induced Pulmonary Inflammation

Alcohol use disorder (AUD) is defined as patterns of alcohol misuse and affects over 30 million people in the US. AUD is a systemic disease with the epidemiology of acute lung injury and excessive alcohol use established in the literature. However, the distinct mechanisms by which alcohol induces the risk of pulmonary inflammation are less clear. A compelling body of evidence shows that cannabinoid receptors (CB1R and CB2R) play a relevant role in AUD. For this study, we investigated the role of CBR signaling in pulmonary immune activation. Using a human macrophage cell line, we evaluated the expression of CBR1 and CBR2 after cells were exposed to EtOH, +/- cannabinoid agonists and antagonists by flow cytometry. We also evaluated the expression of cannabinoid receptors from the lungs of adolescent mice exposed to acute binge EtOH +/- cannabinoid agonists and antagonists at both resting state and after microbial challenge via western blot, rt-PCR, cytokine analysis, and histology. Our results suggest that EtOH exposure modulates the expression of CBR1 and CBR2. Second, EtOH may contribute to the release of DAMPs and other proinflammatory cytokines, Finally, microbial challenge induces pulmonary inflammation in acute binge EtOH-exposed mice, and this observed immune activation may be CBR-dependent. We have shown that adolescent binge drinking primes the lung to subsequent microbial infection in adulthood and this response can be mitigated with cannabinoid antagonists. These novel findings may provide a framework for developing potential novel therapeutics in AUD research.

Hepatic GPx1 and GIT1 expression altered by ethanol exposure during third trimester-equivalent development

Background Ethanol (EtOH) exposure throughout gestation and breastfeeding leads to multiple adverse outcomes in the hepatic system. Under oxidative stress, alterations in the liver are related to the inhibition of induced nitric oxide synthase activity in sinusoidal cells as a consequence of low expression of G-protein-coupled receptor (GPCR)-kinase interacting (GIT1). Here, we hypothesized that both glutathione peroxidase 1 (GPx1) and GIT1 could be altered by EtOH exposure during the third trimester of human equivalent development. Methods We exposed rats during the third trimester equivalent [postnatal days (PD) 2-8] to moderate levels of maternal EtOH (20%). GPx1 and GIT1 expression was detected by western blotting, and the antioxidant activity of glutathione peroxidase GPx and the concentration of hepatic carbonyl groups (CG were determined by spectrophotometry. Serum biochemistry parameters such as alanine aminotransferase (ALT), glucose (gluc), cholesterol (chol), and triglycerides (TG) were also measured. Results We found that ethanol decreased both GIT1 and GPx1 selenoprotein expression, affecting GPx antioxidant activity and increasing protein oxidation. Conclusions These results demonstrate for the first time that the GPx antioxidant system altered by EtOH exposure during the third trimester of development is related to a parallel decrease in GIT1 expression [1].

Sex Differences in Contextual Extinction Learning After Single Binge-Like EtOH Exposure in Adolescent C57BL/6J Mice.

This study investigates the impact of binge-like alcohol exposure on context extinction learning, aiming to identify previously unrecognized risks associated with this common drinking pattern and the development of trauma- and stress-related disorders, such as PTSD. Our findings reveal that binge-like alcohol exposure impairs extinction learning in male adolescent mice by disrupting molecular mechanisms within fear memory circuits, suggesting novel therapeutic and preventive targets. Dysregulated candidates include the canonical Wnt/β-catenin signaling proteins, β-catenin and GSK-3β, along with FKBP5, a key player in glucocorticoid signaling and part of a gene network linked to PTSD. These alterations, found in the dorsal hippocampus (dHPC), basolateral amygdala (BLA), striatum, and nucleus accumbens (NAc) core and shell, may serve as promising targets for future pharmacological intervention.

Decreased antioxidant-related superoxide dismutase 1 expression in peripheral immune cells indicates early ethanol exposure

Alcohol consumption increases oxidative stress and imbalances in the antioxidant system, even with ethanol (EtOH) exposure at a young age. This study assessed changes in the antioxidant system following young EtOH exposure in peripheral immunity and measured sensitive indicators of heavy alcohol consumption. We used peripheral blood mononuclear cells (PBMCs) from 197 male university students without smoking habits to examine changes in antioxidant-related gene expression in vitro and in PBMCs. In vitro, the antioxidant system was impaired by EtOH. Next, we examined the expression of 84 antioxidant-related genes in the PBMCs of 162 young adults, among which the superoxide dismutase (SOD) 1 expression was most negatively correlated with alcohol consumption degree. The plasma SOD1 level had the highest area under the curve value (0.806) for heavy alcohol consumption. Our data demonstrated that a decreased SOD1 level is a sensitive indicator of an impaired antioxidant system and heavy alcohol consumption with early EtOH exposure.

Curcumin attenuates smoking and drinking activated NF-κB/IL-6 inflammatory signaling axis in cervical cancer

BackgroundHigh-risk strains of HPV are known to cause cervical cancer. Multiple clinical studies have emphasized that smoking and drinking are critical risk factors for cervical cancer and its high-grade precursors. In this study, we investigated if smoking and/or drinking augment the molecular mechanisms of cervical carcinogenesis and defined a potential therapeutic approach for their attenuation.MethodsThe impact of benzo[a]pyrene (B[a]P) and/or ethanol (EtOH) exposure on cervical cancer cells was assessed by measuring changes in their cell migration and invasion characteristics. Expression of HPV16 E6/E7, NF-κB, cytokines, and inflammation mediators was determined using qRT-PCR, immunoblotting, ELISA, luciferase reporter assay, and confocal microscopy. Herein, we used curcumin (Cur), and PLGA nanoparticle formulation of curcumin (PLGA-Cur) and determined effectiveness of free Cur and PLGA-Cur formulation on smoking and drinking activated NF-κB/IL-6 mediated inflammatory signaling pathways using in vitro cervical cancer models.ResultsTreatments with B[a]P and/or EtOH altered the expression of HPV16 E6/E7 oncogenes and EMT markers in cervical cancer cells; it also enhanced migration and invasion. In addition, B[a]P and/or EtOH exposure promoted inflammation pathways through TNF-α and NF-κB signaling, leading to IL-6 upregulation and activation of VEGF. The molecular effects caused by B[a]P and/or EtOH exposure were effectively attenuated by curcumin (Cur)/PLGA-Cur treatment.ConclusionsThese data suggest a molecular link between smoking, drinking, and HPV infectivity in cervical carcinogenesis. In addition, attenuation of these effects by treatment with Cur/PLGA-Cur treatment, implies the role of curcumin in cervical cancer prevention and treatment.

Chronic Ethanol Exposure Produces Sex-Dependent Impairments in Value Computations in the Striatum.

Value-based decision-making relies on the striatum, where neural plasticity can be altered by chronic ethanol (EtOH) exposure, but the effects of such plasticity on striatal neural dynamics during decision-making remain unclear. This study investigated the long-term impacts of EtOH on reward-driven decision-making and striatal neurocomputations in male and female rats using a dynamic probabilistic reversal learning task. Following a prolonged withdrawal period, EtOH-exposed male rats exhibited deficits in adaptability and exploratory behavior, with a preference for value updating based on rewards rather than omissions. These behavioral changes were linked to altered neural encoding in the dorsomedial striatum (DMS), where EtOH increased outcome-related signals and decreased choice-related signals. In contrast, female rats showed minimal behavioral changes with distinct EtOH-evoked alterations of neural signals, revealing significant sex differences in the impact of chronic EtOH. Our findings underscore the profound impact of chronic EtOH exposure on adaptive decision-making, revealing enduring changes in neurocomputational processes in the striatum underlying cognitive deficits that differ by sex.

Sex-dependent effects of short-term ethanol, energy drinks and acute noise exposure on hippocampal oxidative balance and glutamate transporter EAAT-1 during rat adolescence

It is known that human adolescents often consume ethanol (EtOH) alone or mixed with energy drinks (ED), especially in noisy environments. Although these agents impact the developing brain, their effects after brief exposure or when presented together remain unclear. Given that few animal studies in this subject are available, this research aimed to study the effects of a brief exposure to these stimuli on the oxidative state and EAAT-1 glutamate transporter levels in the developing rat hippocampus (HC). Adolescent Wistar rats were subjected to a two-bottle choice, limited access to drinking in the dark paradigm, for EtOH and EtOH+ED intake, for 4 days, and subsequent acute noise exposure. Next, hippocampal catalase activity, reactive oxygen species (ROS), glutaredoxin-1 (Grx-1) and glutamate transporter EAAT-1 levels were assessed. Results showed sex-dependent alterations after exposure to these stimuli: Females consuming EtOH had higher hippocampal ROS levels, which decreased when combined with noise; males showed reduced ROS levels only after noise exposure. No significant changes occurred in catalase activity, Grx-1, or EAAT-1 levels with EtOH and noise exposure in neither sex. Additionally, ED raised EtOH consumption in both sexes, normalizing ROS levels only in females when combined with EtOH. Finally, ED consumption altered Grx-1 and EAAT-1 levels in both sexes. In summary, brief exposure to these stimuli induced sex-dependent alterations, suggesting differentiated coping strategies between sexes. Whereas ED consumption may have antioxidant effects in some cases, it could also increase excitotoxicity risk. These novel findings raise questions for future research on the underlying corresponding mechanisms.

Understanding sex and populational differences in spatio-temporal exploration patterns and homebase dynamics of zebrafish following repeated ethanol exposure

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.

Effects of isolation stress and voluntary ethanol exposure during adolescence on ethanol and nicotine co-use in adulthood using male rats

BackgroundAlcohol use in adolescence may increase susceptibility to substance use disorders (SUDs) in adulthood. This study determined if voluntary ethanol (EtOH) consumption during adolescence, combined with social isolation, alters the trajectory of EtOH and nicotine intake during adulthood, as well as activating brain neuroinflammation. MethodsAdolescent male isolate- and group-housed rats were given 0.2 % saccharin/20 % EtOH (Sacc/EtOH) or water using intermittent 2-bottle choice; controls were given water in both bottles (n=17–20 per group). Some rats from each group (n=5–6) were euthanized one week later to measure autoradiographic [3H]PK-11195 binding, an indicator of microglial reactivity, and the remainder (n=11–14 per group) were tested in adulthood in 2-bottle choice, followed by nicotine self-administration using an incremental fixed ratio (FR) schedule with Sacc/EtOH and water concurrently available. ResultsIsolation housing increased adolescent intake of Sacc/EtOH, but the increase did not produce an observable neuroimmunological response in brain. Adolescent EtOH exposure decreased adult intake of both Sacc/EtOH and unsweetened EtOH, with isolate-housed rats showing a greater effect than group-housed rats. In the co-use model, a cross-price economic demand analysis revealed a substitutional relationship between Sacc/EtOH and nicotine, but no effect of adolescent Sacc/EtOH exposure. Compared to group-housed rats, isolate-housed rats were more sensitive to the changing price of nicotine and showed greater substitutability of Sacc/EtOH for nicotine. ConclusionThe current results suggest that adolescent EtOH exposure per se, with or without isolation stress, does not likely explain the enhanced risk for either alcohol or nicotine use later in life.

Ethanol disrupts hepatocellular lipophagy by altering Rab5-centric LD-lysosome trafficking.

Previous reports suggest that lipid droplets (LDs) in the hepatocyte can be catabolized by a direct engulfment from nearby endolysosomes (microlipophagy). Further, it is likely that this process is compromised by chronic ethanol (EtOH) exposure leading to hepatic steatosis. This study investigates the hepatocellular machinery supporting microlipophagy and EtOH-induced alterations in this process with a focus on the small, endosome-associated, GTPase Rab5. Here we report that this small Ras-related GTPase is a resident component of LDs, and its activity is important for hepatocellular LD-lysosome proximity and physical interactions. We find that Rab5 siRNA knockdown causes an accumulation of LDs in hepatocytes by inhibiting lysosome dependent LD catabolism. Importantly, Rab5 appears to support this process by mediating the recruitment of early endosomal and or multivesicular body compartments to the LD surface before lysosome fusion. Interestingly, while wild-type or a constituently active GTPase form (Q79L) of Rab5 supports LD-lysosome transport, this process is markedly reduced in cells expressing a GTPase dead (S34N) Rab5 protein or in hepatocytes exposed to chronic EtOH. These findings support the novel premise of an early endosomal/multivesicular body intermediate compartment on the LD surface that provides a "docking" site for lysosomal trafficking, not unlike the process that occurs during the hepatocellular degradation of endocytosed ligands that is also known to be compromised by EtOH exposure.

Fetal Brain-Derived Exosomal miRNAs from Maternal Blood: Potential Diagnostic Biomarkers for Fetal Alcohol Spectrum Disorders (FASDs).

Fetal alcohol spectrum disorders (FASDs) are leading causes of neurodevelopmental disability but cannot be diagnosed early in utero. Because several microRNAs (miRNAs) are implicated in other neurological and neurodevelopmental disorders, the effects of EtOH exposure on the expression of these miRNAs and their target genes and pathways were assessed. In women who drank alcohol (EtOH) during pregnancy and non-drinking controls, matched individually for fetal sex and gestational age, the levels of miRNAs in fetal brain-derived exosomes (FB-Es) isolated from the mothers' serum correlated well with the contents of the corresponding fetal brain tissues obtained after voluntary pregnancy termination. In six EtOH-exposed cases and six matched controls, the levels of fetal brain and maternal serum miRNAs were quantified on the array by qRT-PCR. In FB-Es from 10 EtOH-exposed cases and 10 controls, selected miRNAs were quantified by ddPCR. Protein levels were quantified by ELISA. There were significant EtOH-associated reductions in the expression of several miRNAs, including miR-9 and its downstream neuronal targets BDNF, REST, Synapsin, and Sonic hedgehog. In 20 paired cases, reductions in FB-E miR-9 levels correlated strongly with reductions in fetal eye diameter, a prominent feature of FASDs. Thus, FB-E miR-9 levels might serve as a biomarker to predict FASDs in at-risk fetuses.

Intermittent ethanol vapor exposure enhances ferroptosis following repeated mild traumatic brain injury in the frontal lobe of adolescent rats; potential role of mitochondrial dysregulation

Adolescents involved in sports have increased risk of traumatic brain injuries (TBI), and frequently participate in binge alcohol consumption in comparison to non-athletic peers. Recovery from a TBI requires substantial energy, and effcient metabolic activity, which depend on intact mitochondria structure and function. Alcohol impairs mitochondrial function, including mitochondrial dynamics and metabolism); that may ultimately lead to cell death. Among cell death pathways, the iron-dependent programmed cell death known as ferroptosis; which has been shown to be predominant in TBI is especially sensitive to changes in mitochondrial energy production. Whether alcohol affects TBI-associated alterations in mitochondrial function and promotes ferroptosis is not known. The aim of this study was to determine how alcohol contributes to repeated-mild TBI (rmTBI)-induced alterations in mitochondrial function (dynamics & metabolism) and ferroptosis signaling in the prefrontal cortex of adolescent male rats. Adolescent male Wistar rats were randomly assigned to i) Sham + Air (control), ii) Sham + ethanol (EtOH), iii) rmTBI + Air, or iv) rmTBI + EtOH. Starting on postnatal day 45, the animals received 3 days of intermittent EtOH vapor exposure (14 hrs on/10 hrs off). Twenty-four hours after the final EtOH exposure, animals were given an mTBI or sham procedure. This sequence was repeated for four cycles. Seven days after the final rmTBI, animals were euthanized, and brains excised for analysis, specifically the frontal lobe. Our preliminary results demonstrate that mitochondrial fusion protein, OPA1, expression was not altered in the PFC by either rmTBI or by EtOH exposure alone. However, rmTBI+EtOH resulted in a significant increase in OPA1 expression. EtOH exposure produced a significant increase in COX-IV (a subunit of ETC complex IV) expression. However, COX-IV expression was not significantly different between rmTBI or rmTBI + EtOH and controls. Mitochondrial complex I activity was not altered by rmTBI, EtOH, or rmTBI + EtOH. Expression of transferrin receptor (TfR1), a specific marker of ferroptosis was significantly increased by rmTBI and EtOH this effect was further exacerbated in the rmTBI + EtOH group. Overall, our study shows that the combination of rmTBI with EtOH exposure affects OPA1-mediated mitochondrial fusion. Additionally, exposure to EtOH increases complex IV expression. Moreover, both rmTBI and EtOH exposure independently triggered signaling pathways associated with ferroptosis, and this effect was synergistically increased when with rmTBI+ EtOH. These findings underscore the intricate interplay between EtOH exposure and rmTBI, influencing crucial mitochondrial proteins and markers of ferroptosis in the frontal lobe of the adolescent rat brain. Further research is needed to fully comprehend the underlying mechanisms and implications of these observations. This research was supported by NIH/NIAAA R01 AA025792 (PEM, NWG) & T32 AA007577 (PEM). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Abstract 1151: Concurrent Obesity And Chronic Alcohol Consumption Exacerbates Skeletal Muscle Mitochondrial Dysfunction In A Murine Model Of Peripheral Artery Disease (PAD).

PAD causes ischemic mitochondrial and consequent muscle damage in the legs, amplifying patient morbidity and mortality. While obesity and alcoholism are common comorbidities, their impact on PAD myopathy is unclear. We sought to determine the effect of concurrent hindlimb ischemia (HLI), obesity and chronic alcohol consumption on muscle oxidative stress, and mitochondrial and limb function. Methods: Twenty-four male, 24 female, 8 month old C57BL/6 mice received high-fat-sucrose (HFS) or low-fat-sucrose (LFS) diets for 16 weeks, followed by either 20% ethanol (EtOH) added to the drinking water / continued water access for another 12 weeks ( n =12 mice/4 groups). The left femoral artery was then ligated to induce HLI, and mice continued their assigned diets for another 4 weeks. The quadriceps from ischemic and non-ischemic limbs were excised at euthanasia. High-resolution respirometry measured muscle mitochondrial respiration and reactive oxygen species (H 2 O 2 ) output. Muscle oxidative stress and mitochondrial quality control (mitophagy) were assessed enzymatically and by western blot. Results: Compared to LFS animals with/without EtOH access, two-way ANOVA showed post-HLI limb function was lower in HFS+water and HFS+EtOH animals (p<0.05). Mitochondrial respiration was lower in LFS+EtOH, HFS+water, and HFS+EtOH ischemic muscle during electron transport chain Complex I, state 2 (I.2), Complex I, state 3 (I.3) and Complex III versus LFS+water controls (p<0.05), and was further reduced during Complex II and IV in LFS and HFS-ischemic muscle exposed to EtOH (p<0.01). Mitochondrial H 2 O 2 yield was increased in HFS+water and HFS+EtOH ischemic muscle at complexes I.2, I.3, II, III and IV (p<0.05), but not in LFS+EtOH muscle. Oxidative damage (carbonyl accumulation) (p<0.0001) and reduced antioxidant levels (superoxide dismutase (p<0.05) and PGC-1alpha (p<0.001)) was only evident in HFS ischemic muscle with/without EtOH exposure. However, the mitochondrial detoxifying enzyme, aldehyde dehydrogenase-2 (p=0.03), and mitophagy markers LC3II:I ratio (p=0.002) and mitofusins 1 (p=0.02) and 2 (p=0.002), were only reduced in HFS+EtOH muscle. Conclusion: Obesity and chronic alcohol consumption exacerbates muscle mitochondriopathy in mice with HLI.

Sex Differences in Alcohol-related Cardiomyopathy

Alcohol-related Cardiomyopathy (ACM) manifests in humans with a significant history of alcohol use and is characterized by ventricular dilation and cardiac function impairment. Although the most common age group for ACM is males between the ages of 30-55, women require a lesser lifetime alcohol exposure to develop ACM than men. The objective of this study is to assess sexual differences in a mouse model of chronic plus binge alcohol consumption. We hypothesize that female mice exposed to chronic plus binge alcohol will not develop as severe cardiac dysfunction as their male counterparts. In this study, male (n=19) and female (n=14) C57BL/6J mice were provided chronic plus binge alcohol feeding. After 5 days of acclimation to the liquid diet, mice are fed ad libitum 5% ethanol (EtOH) liquid or isocaloric control liquid diet for 30 days. At 10 and 30 days, mice received an oral binge dose of EtOH (5 g/kg body weight), or isocaloric maltose dextrin solution (9 g/kg body weight). Noninvasive cardiac structural and functional data were obtained via echocardiography at baseline and following binges. After 30 days, cardiac hemodynamics was measured by open chest left ventricle catheterization. Two-way ANOVA with Tukey’s post hoc analysis was performed for each cardiac parameter, and significance was defined as p<0.05. Male EtOH mice exhibited a significant decrease in stroke work (1119 ± 297 mmHg/mL when compared to male controls, 1622 ± 365 mmHg/mL; p= 0.03). Female EtOH mice had a nonsignificant decrease in stroke work (1242 ± 374 mmHg/mL when compared to female controls, 1423 ± 469 mmHg/mL; p= 0.80). Maximum left ventricular pressure was decreased 20% in male EtOH mice (p=0.003) but increased 10% in female EtOH mice (p=0.46). The maximum rate of pressure change within the ventricle was 24% lower in the male EtOH mice (p=0.02) and 9% higher in female EtOH mice (p=0.87). The minimum rate of pressure change within the ventricle was 24% higher in male EtOH mice (p=0.02) and 17% lower in female ETOH mice (p=0.44). After 30 days of EtOH exposure, male mice showed significant systolic and diastolic dysfunction. Female mice on the other hand, did not show substantial cardiac dysfunction after chronic plus binge ethanol exposure. Further experiments involving qPCR of proinflammatory and profibrotic markers will elucidate the molecular mechanism of sex differences in ACM development. R21AA029747 (JG & CB) & T32AA007577. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Ligand engineering for improving the stability and optical properties of CsPbI3 perovskite nanocrystals

Inorganic lead halide perovskite nanocrystals (NCs) have recently become one of the research topics for optoelectronic applications due to their excellent photophysical properties. Despite their notable thermal stability over organic-inorganic halide perovskites, CsPbI3 NCs suffer from the phase instability of α-CsPbI3 phase at room temperature and under ambient conditions. Here, the effects of 4-Hydroxybenzoic acid (4-HBA) as an additive to standard oleic acid – oleylamine pair on the stability and optical properties of CsPbI3 perovskite NCs are discussed. 4-HBA addition into perovskite NC systems causes a compressive strain on perovskite lattice, which leads to the formation of a mixed phase α- and γ-CsPbI3 phases while pristine perovskite has α-CsPbI3 phase. Time-dependent stability of the perovskite NCs was tested under an ethanol (EtOH) medium. After EtOH exposure of the perovskite NCs, CsPbI3 NCs transformed to non-perovskite phase in 1 h while 4-HBA added CsPbI3 NCs still have perovskite phase after 48 h. In addition to the improved optical properties of the perovskite NCs, 4-HBA addition remarkably improves CsPbI3 perovskite stability.

Fecal microbiota transplantation from female donors restores gut permeability and reduces liver injury and inflammation in middle-aged male mice exposed to alcohol.

Alcohol misuse, binge drinking pattern, and gender-specific effects in the middle-aged population has been clearly underestimated. In the present study, we focused on understanding gender-specific effects of alcohol exposure on the gut-liver axis and the role of gut microbiota in modulating gender-specific responses to alcohol consumption. Fifty-two-week-old female and male C57BL/6 mice were fasted for 12 h, and then administered a single oral dose of ethanol (EtOH) (6 g/kg). Controls were given a single dose of PBS. Animals were sacrificed 8 h later. Alternatively, fecal microbiota transplantation (FMT) was performed in 52-week-old male mice from female donors of the same age. Permeability of the large intestine (colon), gut microbiota, liver injury, and inflammation was thoroughly evaluated in all groups. Middle-aged male mice exposed to EtOH showed a significant increase in gut permeability in the large intestine, evaluated by FITC-dextran assay and ZO-1, OCCLUDIN and MUCIN-2 immuno-staining, compared to PBS-treated animals, whilst female mice of the same age also increased their gut permeability, but displayed a partially maintained intestinal barrier integrity. Moreover, there was a significant up-regulation of TLRs and markers of hepatocellular injury, cell death (AST, TUNEL-positive cells) and lipid accumulation (ORO) in male mice after EtOH exposure. Interestingly, FMT from female donors to male mice reduced gut leakiness, modified gut microbiota composition, ameliorated liver injury and inflammation, TLR activation and the senescence phenotype of middle-aged mice. Our findings highlighted the relevance of gender in middle-aged individuals who are exposed to alcohol in the gut-liver axis. Moreover, our study revealed that gender-specific microbiota transplantation might be a plausible therapy in the management of alcohol-related disorders during aging.

Liposomal Glutathione Augments Immune Defenses against Respiratory Syncytial Virus in Neonatal Mice Exposed in Utero to Ethanol.

We previously reported that maternal alcohol use increased the risk of sepsis in premature and term newborns. In the neonatal mouse, fetal ethanol (ETOH) exposure depleted the antioxidant glutathione (GSH), which promoted alveolar macrophage (AM) immunosuppression and respiratory syncytial virus (RSV) infections. In this study, we explored if oral liposomal GSH (LGSH) would attenuate oxidant stress and RSV infections in the ETOH-exposed mouse pups. C57BL/6 female mice were pair-fed a liquid diet with 25% of calories from ethanol or maltose-dextrin. Postnatal day 10 pups were randomized to intranasal saline, LGSH, and RSV. After 48 h, we assessed oxidant stress, AM immunosuppression, pulmonary RSV burden, and acute lung injury. Fetal ETOH exposure increased oxidant stress threefold, lung RSV burden twofold and acute lung injury threefold. AMs were immunosuppressed with decreased RSV clearance. However, LGSH treatments of the ETOH group normalized oxidant stress, AM immune phenotype, the RSV burden, and acute lung injury. These studies suggest that the oxidant stress caused by fetal ETOH exposure impaired AM clearance of infectious agents, thereby increasing the viral infection and acute lung injury. LGSH treatments reversed the oxidative stress and restored AM immune functions, which decreased the RSV infection and subsequent acute lung injury.

Biomarkers of Affective Dysregulation Associated with In Utero Exposure to EtOH.

Children with fetal alcohol spectrum disorders (FASD) exhibit behavioral and affective dysregulation, including hyperactivity and depression. The mechanisms are not known, but they could conceivably be due to postnatal social or environmental factors. However, we postulate that, more likely, the affective dysregulation is associated with the effects of EtOH exposure on the development of fetal serotonergic (5-HT) and/or dopaminergic (DA) pathways, i.e., pathways that in postnatal life are believed to regulate mood. Many women who use alcohol (ethanol, EtOH) during pregnancy suffer from depression and take selective serotonin reuptake inhibitors (SSRIs), which might influence these monoaminergic pathways in the fetus. Alternatively, monoaminergic pathway abnormalities might reflect a direct effect of EtOH on the fetal brain. To distinguish between these possibilities, we measured their expressions in fetal brains and in fetal brain-derived exosomes (FB-Es) isolated from the mothers' blood. We hypothesized that maternal use of EtOH and/or SSRIs during pregnancy would be associated with impaired fetal neural development, detectable as abnormal levels of monoaminergic and apoptotic biomarkers in FB-Es. Fetal brain tissues and maternal blood were collected at 9-23 weeks of pregnancy. EtOH groups were compared with unexposed controls matched for gestational age (GA). The expression of 84 genes associated with the DA and 5-HT pathways was analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) on microarrays. FB-Es also were assayed for serotonin transporter protein (SERT) and brain-derived neurotrophic factor (BDNF) by enzyme-linked immunosorbent assay (ELISA). Six EtOH-exposed human fetal brain samples were compared to SSRI- or polydrug-exposed samples and to unexposed controls. EtOH exposure was associated with significant upregulation of DA receptor D3 and 5-HT receptor HTR2C, while HTR3A was downregulated. Monoamine oxidase A (MAOA), MAOB, the serine/threonine kinase AKT3, and caspase-3 were upregulated, while mitogen-activated protein kinase 1 (MAPK1) and AKT2 were downregulated. ETOH was associated with significant upregulation of the DA transporter gene, while SERT was downregulated. There were significant correlations between EtOH exposure and (a) caspase-3 activation, (b) reduced SERT protein levels, and (c) reduced BDNF levels. SSRI exposure independently increased caspase-3 activity and downregulated SERT and BDNF. Early exposure to EtOH and SSRI together was associated synergistically with a significant upregulation of caspase-3 and a significant downregulation of SERT and BDNF. Reduced SERT and BDNF levels were strongly correlated with a reduction in eye diameter, a somatic manifestation of FASD. Maternal use of EtOH and SSRI during pregnancy each was associated with changes in fetal brain monoamine pathways, consistent with potential mechanisms for the affective dysregulation associated with FASD.

Abstract 14241: Alcohol Exposure Among Patients With Dilated Cardiomyopathy: A Report From the DCM Precision Medicine Study

Introduction: Alcohol-related cardiomyopathy is characterized by cardiac dysfunction and dilatation due to prolonged and excessive alcohol (ETOH) consumption. This study aims to determine the prevalence, use patterns and genetic associations of ETOH exposure in patient with dilated cardiomyopathy (DCM) and their first-degree relatives (FDR). Hypothesis: 1. There is increased prevalence of EOTH use among patients who develop DCM. 2. Non-genetic co-morbidities are more common in patients who use ETOH and developed DCM. 3. Probands and FDR who use ETOH and develop DCM are more likely to have genetic variants. Methods: We analyzed clinical and genetic data on individuals aged 18 and older enrolled in the US DCM Precision Medicine Study. Partial and full DCM phenotypic expression were included in the analysis. The AUDIT-C score was used to evaluate the intensity of ETOH use. ETOH use and presence of DCM-related rare genetic variants were examined in FDRs using GEE type logistic mixed models adjusted for intra-family correlation and site heterogeneity. Results: A total of 2,595 subjects (1,188 probands and 1,407 FDRs) were included. Prior and current ETOH users were 270 (23%) and 546 (46%) for probands, and 30 (10%) and 168 (54%) for FDRs with DCM/partial DCM. Avg. years of drinking were 25.2 for probands and 24.0 for FDRs with DCM/partial DCM; the percent with AUDIT-C positive scores were 31.5 for probands and 32.7 for FDRs with DCM/partial DCM. In probands, moderate or high/severe ETOH use was more likely to be in men, white, Hispanic and those with </=12 yrs. of education (p<0.001). Patients drinking heavily were more likely to have hypertension, diabetes, hypercholesterolemia, and history of arrhythmia/conduction disease. Among FDRs, DCM/partial DCM was associated with the presence of DCM-related variants but not with moderate-severe drinking years in quartile after controlling for demographics and comorbidities. ETOH use did not modify the genetic effect on DCM/partial DCM. Conclusions: There is a high prevalence of prior or current ETOH use among probands and FDRs with DCM/partial DCM with an average ETOH use between 24-25 yrs. ETOH use was not found to associate with DCM or to modify the association between the presence of genetic variants and development of DCM.