Daily Ethanol Research Articles

Daily Ethanol Drinking Followed by an Abstinence Period Impairs Bone Marrow Niche and Mitochondrial Function of Hematopoietic Stem/Progenitor Cells in Rhesus Macaques.

Unhealthy consumption of alcohol is a major public health crisis with strong associations between immunological dysfunctions, high vulnerability to infectious disease, anemia, and an increase in the risk of hematological malignancies. However, there is a lack of studies addressing alcohol-induced changes in bone marrow (BM) and hematopoiesis as fundamental aspects of immune system function. To address the effect of chronic alcohol consumption on hematopoietic stem and progenitor cells (HSPCs) and the BM niche, we used an established rhesus macaque model of voluntary alcohol drinking. A cohort of young adult male rhesus macaques underwent a standard ethanol self-administration protocol that allowed a choice of drinking alcohol or water 22hours/day with periods of forced abstinence that elevated subsequent intakes when alcohol availability resumed. Following the last month of forced abstinence, the monkeys were euthanized. HSPCs and bone samples were collected and analyzed in functional assays and by confocal microscopy. HSPCs from alcohol animals exhibited reduced ability to formgranulocyte-monocyte and erythroidcolonies in vitro. HSPCs also displayed a decrease in mitochondrial oxygen consumption linked to ATP production and basal respiratory capacity. Chronic alcohol use led to vascular remodeling of the BM niche, a reduction in the number of primitive HSPCs, and a shift in localization of HSPCs from an adipose to a perivascular niche. Our study demonstrates, for the first time, that chronic voluntary alcohol drinking in rhesus macaque monkeys leads to the long-term impairment of HSPC function, a reduction in mitochondrial respiratory activity, and alterations in the BM microenvironment. Further studies are needed to determine whether these changes in hematopoiesis are persistent or adaptive during the abstinent period and whether an initial imprinting to alcohol primes BM to become more vulnerable to future exposure to alcohol.

Freeze-Dried Immobilized Kefir Culture in Low Alcohol Winemaking.

Low alcohol wines represent a rising trend in the global market. Since for ethanol removal, certain physicochemical methods that negatively affect wine quality are applied, the aim of this present study was to evaluate the efficiency of freeze-dried, immobilized kefir culture on natural supports (apple pieces, grape skins and delignified cellulosic material) in low alcohol winemaking at various temperatures (5–30 °C). Initially, genetic analysis of kefir culture was performed by Next Generation Sequencing. There was an immobilization of kefir culture on grape skins-enhanced cell survival during freeze-drying in most cases, even when no cryoprotectant was used. Simultaneous alcoholic and malolactic fermentations were performed in repeated batch fermentations for >12 months, using freeze-dried free or immobilized cells produced with no cryoprotectant, suggesting the high operational stability of the systems. Values of great industrial interest for daily ethanol productivity and malic acid conversion [up to 39.5 g/(Ld) and 67.3%, respectively] were recorded. Principal Component Analysis (PCA) showed that freeze-drying rather than the fermentation temperature affected significantly minor volatiles. All low alcohol wines produced were accepted during the preliminary sensory evaluation.

Influence of daily beer or ethanol consumption on physical fitness in response to a high-intensity interval training program. The BEER-HIIT study

BackgroundHigh-intensity interval training (HIIT) is an effective approach to improve physical fitness, but consuming beer, which is a regular practice in many physically active individuals, may interfere with these effects. The purposes of this study were to investigate the effects of a 10-week (2 days/week) HIIT program on cardiorespiratory fitness, muscle strength and power parameters, and also to assess the possible influence on them of a moderate consumption of beer (at least from Monday to Friday) or its alcohol equivalent.Methods: Young (24 ± 6 years old) healthy adults (n = 73, 35 females) were allocated to five groups. Four groups participated in the HIIT intervention program while the fifth group was a control Non-Training group (n = 15). Participants in the training groups chose whether they preferred receiving alcohol or alcohol-free beverages. Those choosing alcohol were randomized to either beer or ethanol intake: (i) T-Beer group (alcohol beer, 5.4%; n = 13) or (ii) T-Ethanol (sparkling water with vodka, 5.4%; n = 14). Those choosing alcohol-free intake were randomized to (iii) T-Water group (sparkling water, 0.0%; n = 16), or (iv) T-0.0Beer group (alcohol-free beer, 0.0%; n = 15). Men ingested 330 ml of the beverage at lunch and 330 ml at dinner; women ingested 330 ml at dinner. Before and after the intervention, maximal oxygen uptake in absolute and relative terms (VO2max.), maximal heart rate, total test duration, hand grip strength and four types of vertical jumps were measured.ResultsHIIT induced significant improvements in absolute and relative values of VO2max, and total test duration (all p < 0.05) in all the training groups; also, clinical improvements were found in hand grip strength. These positive effects were not influenced by the regular intake of beer or alcohol. No changes in the vertical jumps occurred in any of the groups.ConclusionsA moderate beer or alcohol intake does not mitigate the positive effect of a 10-week HIIT on physical fitness in young healthy adults.Trial registrationClinicalTrials.gov ID: NCT03660579. Registered 20 September 2018. Retrospectively registered.

Long-term exposure to daily ethanol injections in DBA/2J and Swiss mice: Lessons for the interpretation of ethanol sensitization.

Most mice ethanol sensitization studies focused on neurobiology at the expense of its behavioral characterization. Furthermore, relatively short ethanol exposures (10 to 20 injections) were used in these studies. The first aim of the present study is to better characterize the development and expression of ethanol sensitization after an extended exposure of 45 daily injections. In some previous studies, mice were classified as “respondent” and “resistant” to ethanol sensitization. The second aim of the present study is to test the long-term reliability of such categorizations and the consequences of their use on the interpretation of the ethanol sensitization results. Swiss and DBA/2J female mice received 45 consecutive daily ethanol administrations (respectively 2.5 and 2.0 g/kg) and their locomotor activity was daily recorded to test the development of ethanol sensitization. At the end of the procedure, a challenge test assessed the inter-group ethanol sensitization.The results of the present study show that ethanol sensitization continues to develop beyond 20 days to reach maximal levels after about 25 injections in DBA/2J mice and 40 injections in Swiss mice, although the core phase of the development of ethanol sensitization occurred in both strains during the first 20 days. Remarkably, ethanol sensitization after such a long daily ethanol treatment resulted in both an upward shift of the magnitude of ethanol stimulant effects and a prolongation of these effects in time (up to 30 minutes). Mice classified as “resistant to ethanol sensitization” according to previous studies developed very significant levels of ethanol sensitization when tested after 45 ethanol injections and are best described as showing a delayed development of ethanol sensitization. Furthermore, mice classified as respondent or resistant to ethanol sensitization also differ in their acute response to ethanol, such that it is difficult to ascertain whether these classifications are specifically related to the sensitization process.

Re-examining the relationship between alcohol consumption and coronary heart disease with a new lens

Moderate alcohol consumption has been related to lower risk of coronary heart disease (CHD) in the literature. To examine whether alcohol drinking during the past 12 months and heaviest drinking period were differentially associated with the risk of CHD, we designed a case-control study using a population-based health survey of U.S. adults conducted from 2012 to 2013. Respondents who reported to have doctor-ascertained CHD served as cases (n = 1671), and those free of CHD and other alcohol-related health conditions served as controls (n = 17,629) in logistic regressions. Sex-specific quartiles of average daily ethanol intake were ascertained and calculated for the past 12 months and during the period of heaviest lifetime drinking. We further split current drinkers into reducers and non-reducers (past 12 months relative to the heaviest drinking period) to examine CHD risk profiles in association with the 12-month drinking level. Current-drinker reducers (AOR, 95% CI = 1.57 [1.10–2.27] for men; AOR, 95% CI = 1.33 [1.02–1.72] for women) and former drinkers (AOR, 95% CI = 2.06 [1.43–2.97] for men; AOR, 95% CI = 1.51 [1.19–1.92] for women) more often had CHD than lifetime abstainers. Male heavy drinkers during the heaviest drinking period (AOR, 95% CI = 2.25 [1.52–3.32]) were more likely to manifest CHD than lifetime abstainers. In addition, individuals with diagnosed CHD were significantly more likely to have reduced drinking in the past. A change in alcohol consumption over the life course among former and current drinkers may distort the true alcohol-CHD relationship.

Non-Competitive NMDA Receptor Antagonist Hemantane Reduces Ethanol Consumption in Long-Term Alcohol Experienced Rats.

Activity of hemantane, an amino adamantane derivative, exhibiting the properties of lowaffinity non-competitive NMDA receptor antagonist, was evaluated in experimental in vivo models of alcoholism. Hemantane had no effects on the formation and manifestation of behavioral sensitization to ethanol in DBA/2 mice. Under conditions of free choice between 10% ethanol and water, hemantane (20 mg/kg/day for 14 days, intraperitoneally) significantly reduced the daily ethanol intake in random-bred male rats with formed alcohol motivation (>4 g/kg of ethanol). During modelling of withdrawal syndrome, hemantane administered intraperitoneally in doses of 5-20 mg/kg dose-dependently attenuated alcohol-deprivation effect after acute withdrawal with no effects on protracted abstinence. It was found that hemantane suppressed alcohol drinking behavior in long-term ethanol experienced rats and attenuated alcohol-seeking behavior after acute withdrawal.

ProBDNF/p75NTR/sortilin pathway is activated in peripheral blood of patients with alcohol dependence

Alcohol dependence is a worldwide problem with a great social and economic burden in many countries. A number of studies have suggested that BDNF (mature BDNF) and its precursor (proBDNF) play important roles in the alcohol dependence. However, what roles of the mBDNF/proBDNF pathways play during the pathological process of alcohol dependence are not clearly understood. In our clinical study, peripheral blood was sampled from 30 male patients with alcohol dependence and 50 healthy males (as control). The protein levels of proBDNF, p75NTR, sortilin, mBDNF, TrkB and mRNA levels of BDNF, p75NTR, sortilin, and TrkB were detected in the peripheral blood in our study. We found that the protein levels of proBDNF and p75NTR were increased, but not the sortilin protein level; while the TrkB protein level was decreased in the alcohol dependence patients compared with healthy controls. Moreover, the mRNA levels of p75NTR and sortilin from the lymphocytes were slightly increased; while BDNF and TrkB were significantly decreased. The ELISA results of mBDNF and TrkB were declined in the alcohol dependence group. The levels of mBDNF and TrkB were negatively correlated with the average amount of daily ethanol consumption, and the levels of proBDNF, p75NTR and sortilin were positively correlated with the average amount of ethanol consumption per day. The ratio of proBDNF to mBDNF was altered in alcohol dependence patients. The balance between the proBDNF/p75NTR and mBDNF/TrkB signalling pathways appeared dysregulated in alcohol dependence. Our results suggested that both pathways may participate in the complex processes of alcohol dependence.

Alcoholic Cardiomyopathy: Translation Model.

We developed a translation model of alcoholic cardiomyopathy in rats. By the end of forced alcoholization (the rats received 10% ethanol solution as the only source of fluid for 24 weeks; mean daily ethanol consumption was 5.0-6.5 g/kg), the rats developed dilated heart failure. Echocardiography and morphometric study of the myocardium revealed a decrease in inotropic function of the heart and dilatation of the right and left ventricles. Fatty degeneration of the myocardium (pathognomonic sign of alcoholic cardiomyopathy) and decrease in electrical stability of cardiomyocytes reliably reproduce the clinical pattern of alcoholic cardiomyopathy.

Behavioral Sensitization to the Disinhibition Effect of Ethanol Requires the Dopamine/Ecdysone Receptor in Drosophila.

Male flies under the influence of ethanol display disinhibited courtship, which is augmented with repeated ethanol exposures. We have previously shown that dopamine is important for this type of ethanol-induced behavioral sensitization but the underlying mechanism is unknown. Here we report that DopEcR, an insect G-protein coupled receptor that binds to dopamine and steroid hormone ecdysone, is a major receptor mediating courtship sensitization. Upon daily ethanol administration, dumb and damb mutant males defective in D1 (dDA1/DopR1) and D5 (DAMB/DopR2) dopamine receptors, respectively, showed normal courtship sensitization; however, the DopEcR-deficient der males exhibited greatly diminished sensitization. der mutant males nevertheless developed normal tolerance to the sedative effect of ethanol, indicating a selective function of DopEcR in chronic ethanol-associated behavioral plasticity. DopEcR plays a physiological role in behavioral sensitization since courtship sensitization in der males was reinstated when DopEcR expression was induced during adulthood but not during development. When examined for the DopEcR’s functional site, the der mutant’s sensitization phenotype was fully rescued by restored DopEcR expression in the mushroom body (MB) αβ and γ neurons. Consistently, we observed DopEcR immunoreactivity in the MB calyx and lobes in the wild-type Canton-S brain, which was barely detectable in the der brain. Behavioral sensitization to the locomotor-stimulant effect has been serving as a model for ethanol abuse and addiction. This is the first report elucidating the mechanism underlying behavioral sensitization to another stimulant effect of ethanol.

Lessons and Challenges from a 6-Month Randomized Pilot Study of Daily Ethanol Consumption

BackgroundObservational studies and crossover feeding studies suggest that moderate alcohol use may benefit cardiovascular risk, but we know of no long-term randomized trials that have tested this hypothesis. ObjectiveWe evaluated the feasibility of an efficacy study of daily ethanol use in a 6-mo randomized pilot study in adults at higher cardiovascular risk. MethodsIn a double-blind, randomized, controlled parallel-design trial, we screened 67 adults aged ≥55 y and randomly assigned 45 participants to consume 150 mL of an artificially sweetened beverage with or without 10% grain alcohol daily for 6 mo. Participants were asked to consume no other alcohol and returned monthly to receive the beverage and undergo measurement of HDL cholesterol, liver function tests, and complete blood counts. ResultsOf the 45 randomly assigned participants, 39 completed the trial; the primary reason cited for attrition was inconvenience. None of the participants reported problem drinking or developed any serious adverse events or abnormal biochemical findings. However, we observed no differences in concentrations of HDL cholesterol, HDL lipoprotein subclasses, aspartate aminotransferase, alanine aminotransferase, γ-glutamyltransferase, mean corpuscular volume, or adiponectin between the alcohol and control arms, suggesting that adherence was poor. Every participant accurately identified their assigned beverage, most with great certainty. ConclusionsIn this parallel-design pilot study of daily alcohol use, we observed none of the expected changes in markers of alcohol intake, which suggests poor adherence to this pure alcohol intervention. Our results suggest that long-term trials of alcohol consumption, if they are conducted in light drinkers similar to these, must use pragmatic designs for maximal feasibility. This study was registered at clinicaltrials.gov as NCT01377727.

THE RELATION OF ALCOHOL CONSUMPTION WITH CARDIOVASCULAR DISEASES AND RISK FACTORS (ESSE-RF TRIAL IN KEMEROVSKAYA OBLAST)

Aim. To analyze the relation of alcohol consumption level with cardiovascular diseases and risk factors in the population of Kemerovskaya Oblast. Material and methods. The study was performed under the framework of multicenter epidemiological trial ESSE-RF in Kemerovskaya Oblast, with 1628 participants with the age 25-64 y.o. The data was analyzed concerning the prevalence, volume and type of alcohol beverages by the respondents, as the prevalence of a range of cardiovascular diseases and risk factors. By original formulae, total volume was calculated of the consumed alcohol in units of the “safe” ethanol dose (24 grams) with further grouping to “non consuming” and consuming “moderately”, “in-between” and “over”. Results. Overconsumption of alcohol, comparing to non-consumers, is associated with high risk of hypercholestrolemia (OR 1,76; 95% CI: 1,12-2,75), hypertriglyceridemia (OR 2,69: 1,52-4,77), body overweight (OR 1,68: 1,04-2,71), smoking (OR 2,24: 1,48-3,41). The relation of low physical activity with alcohol consumption is found in all grades. Increase of the mean daily ethanol dosage is positively linearly related with the risk of arterial hypertension (OR 1,04: 1,11-2,75), hypercholesterolemia (OR 1,06: 1,01-1,12), increased level of low density lipoproteins (OR 1,09: 1,02-1,16), smoking (OR 1,17: 1,10-1,24), low physical activity (OR 1,09: 1,02-1,16). There is U-shaped and upside-down J-shaped relation for the level of psychological risk factors (stress, anxiety, depression) and volumes of consumed alcohol. By the considered cardiovascular diseases it can be noted only, the existence of statistically non-significant (0,1>р>0,05) tendency to decrease myocardial infarction risk and stroke in anamnesis, with the increase of the consumed alcohol volumes comparing to non-drinkers. Conclusion. The study shows that overconsumption of alcohol, as the increase of mean daily dosage of ethanol are associated with the range of cardiovascular risk factors. There is U-shaped and upside-down J-shaped relation of psychoemotional risk factors with the volumes of consumed alcohol. By the considered cardiovascular dseases, there are no significant relations for the level of consumed alcohol, however, there is non-significant tendency for myocardial infarction and stroke risk factors diminishing in the anamnesis, with the increase of consumed alcohol volumes.

Validating a Hazardous Drinking Index in a Sample of Sexual Minority Women: Reliability, Validity, and Predictive Accuracy

ABSTRACTBackground: Although sexual minority women (SMW) are at increased risk of hazardous drinking (HD), efforts to validate HD measures have yet to focus on this population. Objectives: Validation of a 13-item Hazardous Drinking Index (HDI) in a large sample of SMW. Methods: Data were from 700 adult SMW (age 18-82) enrolled in the Chicago Health and Life Experiences of Women study. Criterion measures included counts of depressive symptoms and post-traumatic stress disorder (PTSD) symptoms, average daily and 30-day ethanol consumption, risky sexual behavior, and Diagnostic and Statistical Manual (DSM-IV) measures of alcohol abuse/dependence. Analyses included assessment of internal consistency, construction of receiver operating characteristic (ROC) curves to predict alcohol abuse/dependence, and correlations between HDI and criterion measures. We compared the psychometric properties (diagnostic accuracy and correlates of hazardous drinking) of the HDI to the commonly used CAGE instrument. Results: KR-20 reliability for the HDI was 0.80, compared to 0.74 for the CAGE. Predictive accuracy, as measured by the area under the receiver operating characteristic curve for alcohol abuse/dependence, was HDI: 0.89; CAGE: 0.84. The HDI evidenced the best predictive efficacy and tradeoff between sensitivity and specificity. Results supported the concurrent validity of the HDI measure. Conclusions: The Hazardous Drinking Index is a reliable and valid measure of hazardous drinking for sexual minority women.

N-acetylcysteine treatment blocks the development of ethanol-induced behavioural sensitization and related ΔFosB alterations

Ethanol addiction is a serious public health problem that still needs more effective pharmacological treatment. A key factor in the development and maintenance of this disease is the advent of neuroadaptations in the mesocorticolimbic brain pathway upon chronic ethanol abuse. In general, these neuroadaptations are maladaptive and affect numerous neurotransmitter systems and intracellular molecules. One of these molecules is ΔFosB, a transcription factor that is altered after chronic drug use. Behavioural sensitization is a useful model for the study of the neuroadaptations related to addiction. Recent works have shown a role for the imbalance of glutamatergic neurotransmission in the symptoms found in addicted people. In this sense, the treatment with N-acetylcysteine, a l-cysteine prodrug that acts by restoring extrasynaptic concentrations of glutamate through the activation of cystine-glutamate antiporter, has shown promising results in the treatment of addiction. Thus, an animal model of behavioural sensitization was used to evaluate the effects of N-acetylcysteine treatment in the behavioural and molecular alterations induced by chronic ethanol administration. Swiss mice were subject to 13 days of daily ethanol administration to induce behavioural sensitization. Two hours before each ethanol administration and locomotor activity evaluation, the animals received intraperitoneally N-acetylcysteine injections. Immediately after the last test session, their brains were removed for ΔFosB and cystine-glutamate antiporter quantification. It was found that N-acetylcysteine treatment blocked ethanol-induced behavioural sensitization, the increase of ΔFosB content in the prefrontal cortex, and its reduction in the nucleus accumbens. The results suggest a possible use of N-acetylcysteine in ethanol-related disorders.

Ethanol Induced Brain Lipid Changes in Mice Assessed by Mass Spectrometry.

Alcohol abuse is a chronic disease characterized by the consumption of alcohol at a level that interferes with physical and mental health and causes serious and persistent changes in the brain. Lipid metabolism is of particular interest due to its high concentration in the brain. Lipids are the main component of cell membranes, are involved in cell signaling, signal transduction, and energy storage. In this study, we analyzed lipid composition of chronically ethanol exposed mouse brains. Juvenile (JUV) and adult (ADU) mice were placed on a daily limited-access ethanol intake model for 52 days. After euthanasia, brains were harvested, and total lipids were extracted from brain homogenates. Samples were analyzed using high resolution mass spectrometry and processed by multivariate and univariate statistical analysis. Significant lipid changes were observed in different classes including sphingolipids, fatty acids, lysophosphatidylcholines, and other glycerophospholipids.

Alcohol consumption and corresponding factors: A novel perspective on the risk factors of esophageal cancer

Esophageal cancer is the eighth most common type of cancer in the world, and the sixth most common cause of mortality from cancer. Alcohol consumption is the major risk factor for esophageal cancer, due to the worldwide prevalence and high carcinogenicity of the ethanol metabolite. In epidemiological studies, the efficiency of alcohol intake to enhance the risk of esophageal cancer is altered by daily ethanol consumption, type of alcoholic beverages ingested, time since quitting drinking, age of drinking initiation, differences in population and subtypes of esophageal cancer. Corresponding factors, including gene polymorphisms, tobacco smoking, oral microorganisms and folate deficiency, reveal a synergistic effect in concurrent alcohol users that may lead to an increased risk of developing esophageal cancer. Consequently, esophageal cancer prevention involves multiple aspects, including quitting drinking and smoking, maintaining an adequate oral health and ingesting adequate quantities of folate, particularly in genetically high-risk populations.

A U-shaped relationship between alcohol consumption and cardiometabolic index in middle-aged men.

BackgroundCardiometabolic index (CMI) is a new index for discriminating diabetes. The purpose of this study was to determine whether CMI is affected by habitual alcohol drinking.MethodsThe subjects were 21572 men (35-60 years) receiving annual health checkups. They were divided by average daily ethanol consumption into non-, light (<22 g), moderate (≥22 and < 44 g), heavy (≥44 and < 66 g) and very heavy (≥66 g) drinkers. Relationship between alcohol intake and CMI was investigated with adjustment for age and histories of smoking and regular exercise.ResultsLog-transformed CMI was significantly lower in light, moderate and heavy drinkers than in nondrinkers and was lowest in light drinkers, while there was no significant difference in log-transformed CMI of nondrinkers and very heavy drinkers. Odds ratio vs. nondrinkers for high CMI was significantly lower than the reference level of 1.00 in light, moderate and heavy drinkers and was lowest in light drinkers but was not significantly different from the reference level in very heavy drinkers. Odds ratio of subjects with vs. those without high CMI for hyperglycemia was significantly higher than the reference level in all of the alcohol groups and was significantly lower in moderate drinkers but was not significantly different in the other drinker groups when compared with the nondrinker group.ConclusionThere is a U-shaped relationship between alcohol consumption and CMI, and moderate drinking but not excessive drinking attenuates the association between CMI and hyperglycemia.

Increased presynaptic regulation of dopamine neurotransmission in the nucleus accumbens core following chronic ethanol self-administration in female macaques.

Hypofunction of striatal dopamine neurotransmission, or hypodopaminergia, is a consequence of excessive ethanol use and is hypothesized to be a critical component of alcoholism, driving alcohol intake in an attempt to restore dopamine levels; however, the neurochemical mechanisms involved in these dopaminergic deficiencies are not fully understood. Here we examined the specific dopaminergic adaptations that produce hypodopaminergia and contribute to alcohol use disorders using direct, sub-second measurements of dopamine signaling in nonhuman primates following chronic ethanol self-administration. Female rhesus macaques completed 1 year of daily (22 h/day) ethanol self-administration. Subsequently, fast-scan cyclic voltammetry was used in nucleus accumbens core brain slices to determine alterations in dopamine terminal function, including release and uptake kinetics, and sensitivity to quinpirole (D2/D3 dopamine receptor agonist) and U50,488 (kappa opioid receptor agonist) induced inhibition of dopamine release. Ethanol drinking greatly increased uptake rates, which were positively correlated with lifetime ethanol intake. Furthermore, the sensitivity of dopamine D2/D3 autoreceptors and kappa opioid receptors, which both act as negative regulators of presynaptic dopamine release, was moderately and robustly enhanced in ethanol drinkers. Greater uptake rates and sensitivity to D2-type autoreceptor and kappa opioid receptor agonists could converge to drive a hypodopaminergic state, characterized by reduced basal dopamine and an inability to mount appropriate dopaminergic responses to salient stimuli. Together, we outline the specific alterations to dopamine signaling that may drive ethanol-induced hypofunction of the dopamine system and suggest that the dopamine and dynorphin/kappa opioid receptor systems may be efficacious pharmacotherapeutic targets in the treatment of alcohol use disorders.

Effects of Alcohol Consumption on Hepatocarcinogenesis in Japanese Patients With Fatty Liver Disease.

The effect of ethanol consumption on hepatocarcinogenesis in patients with fatty liver disease (FLD) is not clear. We aimed to investigate the influence of alcohol consumption on hepatocarcinogenesis and determine the risk factors for hepatocellular carcinoma (HCC) in a large number of Japanese patients with FLD without viral hepatitis. This multicenter, retrospective cohort study was conducted at a specialized center for hepatology in Japan and included 9959 patients with FLD without viral hepatitis, diagnosed by ultrasonography from January 1997 through December 2011. The patients' level of ethanol consumption was divided into 4 categories: <20 g/day (n= 6671), 20-39 g/day (n= 753), 40-69 g/day (n= 1589), and ≥70 g/day (n= 946). The primary endpoint was the onset of HCC. Statistical analyses performed included the Kaplan-Meier method and Cox proportional hazard analysis. The median follow-up period was 5.4 years. Of the study cohort, 49 cases (0.49%) developed HCC during the follow-up period. The annual incidence rate of HCC was 0.05% in patients with FLD and a daily ethanol consumption <20 g/day. Increasing levels of ethanol consumption were associated with increased annual incidence rates of HCC: 0.06% for patients with 20-39 g/day ethanol consumption (hazard ratio [HR], 1.54; 95% confidence interval [CI], 0.34-7.04), 0.16% for patients with 40-69 g/day ethanol consumption (HR, 3.49; 95% CI, 1.50-8.12), and 0.22% for patients with ≥70 g/day ethanol consumption (HR, 10.58; 95% CI, 5.06-22.13), compared with patients with ethanol consumption <20 g/day. Multivariate analysis showed that ethanol consumption ≥40 g/day was an independent risk factor for HCC: for 40-69 g/day the HR was 2.48 (95% CI, 1.01-6.05; P<.047) and for ≥70 g/day the HR was 12.61 (95% CI, 5.68-28.00; P < .001). Based on a multicenter, retrospective analysis of almost 10,000 patients with FLD, ethanol consumption ≥40 g/day is an independent risk factor for HCC.

Voluntary ethanol consumption reduces GABAergic neuroactive steroid (3α,5α)3-hydroxypregnan-20-one (3α,5α-THP) in the amygdala of the cynomolgus monkey.

Neuroactive steroids such as (3α,5α)3-hydroxypregnan-20-one (3α,5α-THP, allopregnanolone) enhance the gamma-aminobutyric acid (GABA)-ergic effects of ethanol and modulate excessive drinking in rodents. Moreover, chronic ethanol consumption reduces 3α,5α-THP levels in human plasma, rat hippocampus and mouse limbic regions. We explored the relationship between 3α,5α-THP levels in limbic brain areas and voluntary ethanol consumption in the cynomolgus monkey following daily self-administration of ethanol for 12 months and further examined the relationship to hypothalamic-pituitary-adrenal (HPA) axis function prior to ethanol exposure. Monkeys were subjected to scheduled induction of ethanol consumption followed by free access to ethanol or water for 22 h/day over 12 months. Immunohistochemistry was performed using an anti-3α,5α-THP antibody. Prolonged voluntary drinking resulted in individual differences in ethanol consumption that ranged from 1.2 to 4.2 g/kg/day over 12 months. Prolonged ethanol consumption reduced cellular 3α,5α-THP immunoreactivity by 13 ± 2 percent (P < 0.05) in the lateral amygdala and 17 ± 2 percent (P < 0.05) in the basolateral amygdala. The effect of ethanol was most pronounced in heavy drinkers that consumed ≥3 g/kg ≥ 20 percent of days. Consequently, 3α,5α-THP immunoreactivity in both the lateral and basolateral amygdala was inversely correlated with average daily ethanol intake (Spearman r = -0.87 and -0.72, respectively, P < 0.05). However, no effect of ethanol and no correlation between drinking and 3α,5α-THP immunoreactivity were observed in the basomedial amygdala. 3α,5α-THP immunoreactivity following ethanol exposure was also correlated with HPA axis function prior to ethanol exposure. These data indicate that voluntary ethanol drinking reduces amygdala levels of 3α,5α-THP in non-human primates and that amygdala 3α,5α-THP levels may be linked to HPA axis function.

Chronic ethanol self-administration in macaques shifts dopamine feedback inhibition to predominantly D2 receptors in nucleus accumbens core

BackgroundGiven the high level of homology between nonhuman primates and humans in regard to anatomy, physiology and ethanol drinking patterns, nonhuman primates represent an unparalleled preclinical model for examining the neurobiological basis of ethanol abuse. MethodsHere we examined the neurochemical consequences of chronic daily ethanol use using fast-scan cyclic voltammetry in brain slices containing the nucleus accumbens core or dorsolateral caudate taken from male cynomolgus macaques following ethanol drinking. ResultsWe found that in both regions the ability of ethanol to decrease dopamine release was unchanged, indicating that ethanol self-administration does not produce tolerance or sensitization to ethanol effects on dopamine release at the dopamine terminal at this time point. We also found that in the nucleus accumbens core, autoregulation of dopamine release was shifted from equal D2 and D3 receptor involvement in control animals to primarily D2 receptor-mediated in drinkers. Specifically, the effect quinpirole, a D2/D3 receptor agonist, on dopamine release was equal across groups; however, dopamine signals were reversed to a greater extent by the selective D3 receptor antagonist SB-277,011A in control animals, indicating a greater contribution of D2 receptors in quinpirole-induced inhibition following ethanol self-administration. In the dorsolateral caudate, the effects of quinpirole and reversal with SB-277,011A was not different between ethanol and control slices. ConclusionsThis work provides novel insight into the dopaminergic adaptations resulting from chronic ethanol use in nonhuman primates and indicates that alterations in D2/D3 dopamine autoreceptor signaling may be an important neurochemical adaptation to ethanol consumption during early use.