Long-term Ethanol Consumption Research Articles

TELOMERE LENGTH IN RHESUS MACAQUES WITH VOLUNTARY CHRONIC ETHANOL CONSUMPTION

Introduction. Alcohol abuse is associated with telomere shortening. There is no convincing evidence of a “safe” level of alcohol consumption in this regard. Long-term studies in rodents are not feasible, and clinical trials with the administration of alcohol to healthy individuals is not ethically acceptable. An approach based on a relevant model of voluntary alcohol consumption in monkeys under controlled conditions is a significant alternative. The aim of the study. To estimate the length of telomeres at long-term ethanol consumption by male rhesus macaques in under free choice with water Methods. The study was performed on fourteen mature male Rhesus macaques of groups with low (median 0.62 g/kg/day) and high (median 2.71 g/kg/day) ethanol consumption as 4% (v./v.) solution with condition of all-day access and free choice with drinking water. The duration of consumption was 920 days. The relative length of telomeres was determined by quantitative PCR according to Cawthon (2002) in blood leukocytes. Results. The relative average telomere length in the high-consumption group was 1.53±0.57 before the presentation of ethanol in the adaptation period (-32 day of the study), and at the consumption stage it was on 717 day 2.13±0.19 and on 917 day 4.61±0.7. In the low-consumption group, the average relative telomere length constituted 1.42±0.22, 1.55±0.15 and 3.3±0.47, respectively. The absolute count of leukocytes did not change significantly during the study. However, changes in the differential white cells count were revealed representing development of relative monocytosis by 917 day in both groups. Conclusion. The data obtained do not confirm the association of long-term alcohol consumption in moderate doses with telomere length. The completed study has limitations related to the lack of control without consumption and evaluation in one sex.

Effect of long-term ethanol consumption and a high-fat diet on mitochondrial respiration in rat pancreatic acinar cells and hepatocytes

Chronic alcohol consumption may cause pancreatitis and alcohol-related liver diseases. Both adaptation and damage of liver mitochondria in animals on chronic ethanol and high-fat diets were demonstrated. It is currently not clear if ethanol or its metabolites such as fatty acid ethyl esters can cause mitochondrial damage to the pancreas. The present study aimed to evaluate the effect of chronic ethanol administration in combination with a high-fat diet on mitochondrial respiration in both pancreatic acinar cells and hepatocytes of rats. Wistar male rats on a high-fat diet (35% calories) were administered ethanol (6 g/kg body weight) by oral gavage for 14 days. Pancreatic acini cells and hepatocytes were isolated with collagenase digestion. The respiration of isolated cells was studied with a Clark electrode. Ethanol administration to rats kept on a high-fat diet was followed by a rapid loss of animal weight during the first 5 days of the experiment and diminished secretory response of pancreatic acini to acetylcholine, however, no changes in acinar cells ultrastructure, basal, oligomycin-insensitive or FCCP-uncoupled respiration were found. Meanwhile ethanol caused a significant (~40%) increase in basal and maximal FCCP-uncoupled respiration rate of isolated hepatocytes. In conclusion, chronic ethanol administration to rats on a high-fat diet does not cause mitochondrial damage in the pancreas, while mitochondria of the liver adapt to ethanol by increasing respiration rate. Keywords: ethanol, hepatocytes, high fat diet, mitochondrial respiration, pancreatic acinar cells

Alterations of the Fatty Acid Profile and the Expression of Genes Related to FA Metabolism in Cirrhotic Liver Tissue.

In addition to direct damage to hepatocytes, long-term ethanol consumption leads to lipid accumulation and hepatic steatosis, as well as to the dysregulation of lipid metabolism. The final step in various liver diseases is cirrhosis. The aim of this study was to compare the FA (fatty acids) profile and expression levels of genes involved in lipid metabolism in cirrhotic liver tissue and normal liver tissue. Exploring the changes in the FA profile and expression of genes related to fatty acid metabolism in cirrhotic liver tissue reveals a molecular landscape that goes beyond the surface of traditional liver function assessments. Understanding the shifts in gene expression and fatty acid composition in liver tissue opens avenues for interventions that may aid in the treatment of cirrhosis in the future.

Biphasic effects of ethanol consumption on N,N-dimethylformamide-induced liver injury in mice

N,N-Dimethylformamide (DMF) is a well-documented occupational hazardous material, which can induce occupational liver injury. The current study was designed to investigate whether ethanol consumption can affect DMF-induced hepatotoxicity and the potential underlying mechanisms involved. We found that a single dose of ethanol (1.25, 2.5, or 5 g/kg bw by gavage) significantly repressed the increase in serum alanine transaminase (ALT) and aspartate transaminase (AST) activities and alleviated the liver histopathological changes in mice challenged with 3 g/kg DMF. In contrast, long-term moderate drinking (2.5 g/kg bw) significantly aggravated the repeated DMF (0.7 g/kg bw) exposure-induced increase in the serum ALT and AST activities. Mechanistically, acute ethanol consumption suppressed DMF-induced activation of the NLR family pyrin domain-containing protein 3 (NLRP3) inflammasome, while long-term moderate ethanol consumption promoted hepatocyte apoptosis in the mouse liver. Notably, cytochrome P4502E1 (CYP2E1) protein level and activity in mouse livers were not significantly affected by ethanol per se in the two models. These results confirm that regular drinking can increase the risk of DMF-induced hepatotoxicity, and suggest that DMF-handling workers should avoid consuming ethanol to reduce the risk of DMF-indued liver injury.

Psychoneuroimmunomodulating effect of lymphocytes with ortho-fluoro-benzonal modulated activity in syngeneic long-term alcoholized recipients

IntroductionLymphocytes are dysfunctional during long-term ethanol consumption and may contribute the progression from healthy to problem drinking. GABAA receptors are molecular targets of ethanol on lymphocytes, potentiating the effects of alcohol.ObjectivesWe first demonstrated that original compound ortho-fluoro-benzonal, artificial GABA receptor ligand, has immunostimulating properties and is able to restored long-term alcoholized mice lymphocytes activity in vitro through GABAA receptors. Based on the previous results we investigated effects of the ex vivo ortho-fluoro-benzonal modulated lymphocytes in recipients with experimental alcoholism.MethodsMale (CBAxC57Bl/6)F1 mice with 6-month 10% ethanol exposure were undergoing the transplantation of syngeneic long-term alcoholized mice lymphocytes, pretreated in vitro with ortho-fluoro-benzonal. Recipient’s ethanol consumption, parameters of the nervous and immune systems functional activities were estimated.ResultsIt was shown that lymphocytes modulated ex vivo with ortho-fluoro-benzonal after intravenous injection caused in syngeneic long-term alcoholized recipients ethanol consumption decrease and stimulation of behavioral activity in the “open field” test against the background of changes in the level of a number of cytokines in pathogenetically significant brain structures. Stimulation of humoral immune response, estimated by the relative number of antibody-forming spleen cells was also detected in recipients after lymphocytes transplantation. The injected immune cells were recorded in the parenchyma of the spleen and brain of recipients, which suggests, in particular, their direct influence on these functions.ConclusionsResults demonstrated that transplantation of ortho-fluoro-benzonal-modulated lymphocytes caused positive psychoneuroimmunomodulating effect in long-term alcoholized recipients, which makes it possible to consider adoptive immunotherapy as a promising method in the treatment of alcoholism.Disclosure of InterestNone Declared

Intensity of endoplasmic reticulum stress, autophagy, and apoptosis in the cerebral cortex of rats with chronic ethanol consumption under the influence of the complex compound of germanium with nicotinic acid

The aim of the research – to determine the level of BAX, caspase-3, GRP78, IRE1 and Beclin-1 in the cerebral cortex of rats with chronic ethanol consumption and under conditions of exposure to the germanium complex with nicotinic acid (MIGU-1). Materials and methods. Female rats had free access to 20 % C2H5OH as the only source of fluid for 110 days. Starting from the 90th day, the animals were injected with MIGU-1 (10 mg/kg/day, IP). The expression level of BAX, caspase-3, GRP78, IRE1 and Beclin-1 in the tissue was determined by Western blot analysis. Results. In rats with chronic ethanol consumption, the level of BAX-dimer increased by 2.06 times (p˂0.001). The introduction of MIGU-1 caused a decrease in the level of BAX-dimer by 1.42 times (p˂0.05). In rats with chronic ethanol consumption, the level of caspase-3 increased by 2.12 times (p˂0.05), cleaved caspase-3 increased by 6.37 times (p˂0.05). When MIGU-1 was administered, the level of caspase-3 decreased by 1.73 times (p˂0.05). Under the conditions of MIGU-1 administration, protein bands of cleaved caspase-3 were reduced to an undetectable level. In rats with chronic ethanol consumption, the level of GRP78 increased by 1.72 times (p˂0.05). After administration of MIGU-1, no changes in the level of GRP78 were recorded. Long-term ethanol consumption increased the levels of IRE1 by 1.74 times (p˂0.05) and p-IRE1 by 2.7 times (p˂0.001). In the presence of MIGU-1, the levels of IRE1 and p-IRE1 did not change. Under the conditions of chronic ethanol consumption, an increase in the levels of Beclin-1 by 2.33 times (p˂0.001) and p-Beclin-1 by 4.69 times (p˂0.001) was observed. Administration of MIGU-1 did not affect the level of Beclin-1, while the level of p-Beclin-1 decreased by 3.09 times (p˂0.001). Conclusions. Long-term ethanol consumption triggers metabolic changes in the cerebral cortex, resulting in ER stress, UPR activation, autophagy, and apoptosis. Administration of MIGU-1 alleviates ER stress by selectively inhibiting specific branches of apoptosis through effects on Beclin-1 levels, suggesting an effect of MIGU-1 on neuronal survival under chronic ethanol consumption

Myrrh Attenuates Gonadal Toxicity and DNA Fragmentation Induced by Ethanol in Male Rats

Background: Long-term ethanol consumption causes pathological alterations to the male reproductive system, possibly resulting in infertility. Myrrh is a resin with antioxidant properties that is produced from the bark of Commiphora tree species. This research aimed to observe the protective impact of myrrh extract supplementation on male reproductive injury and toxicity induced by ethanol treatment. Methods: Adult male Sprague Dawley rats were orally treated with 40% ethanol (3 g/kg) in combination with myrrh extract (500 mg/kg) for four weeks to investigate the curative efficiency of myrrh extract against ethanol toxicity from physiological, histopathological and genetic aspects. Result: The findings revealed that the sperm count, testosterone level and DNA concentration diminished significantly in the ethanol-treated rats compared to control rats, while the contents of malondialdehyde (MDA), heat shock protein 70 (HSP70) and proinflammatory mediators, including tumor necrosis factor-α (TNF-α) and nitric oxide (NO), were elevated significantly; severe degenerative changes in the testicular structure and strong testicular DNA fragmentation were also observed. Co-treatment with myrrh attenuated lipid peroxidation and improved testicular histomorphology, testosterone levels and DNA concentrations and pattern, while at the same time, the levels of both HSP70 and TNF-α levels were increased. Our results provide insight into the curative impacts of myrrh on ethanol toxicity resulting from the rich content of curzerene in myrrh, which has antioxidant activities.

Development of an LC-MS/MS assay with automated sample preparation for phosphatidylethanol (PEth)– Not your typical clinical marker

Phosphatidylethanol (PEth) is a group of phospholipids formed exclusively in the presence of ethanol on the erythrocyte membrane, making it a direct biomarker for long-term ethanol consumption for which a clinical reference interval has been established. Here, we describe an assay for quantitation for two most abundant PEth homologues, PEth 16:0/18:1 and PEth 16:0/18:2, from human whole blood, and present challenges overcome throughout the development process. Since PEth is localized within erythrocyte membranes, a reliable sample preparation technique is an important aspect of PEth analysis. Therefore, various erythrocyte lysing agents for recovery of exogenously spiked standards and controls were evaluated to identify one that performed comparably to the recovery of endogenous analytes found in authentic samples. A supported liquid extraction (SLE) technique was employed for sample cleanup and enrichment which together with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis enabled automated sample preparation, appropriate chromatographic resolution, and minimal system carryover. This resulted in a laboratory developed test with an analytical measurement range (AMR) of 10–1000 ng/mL (slope = 0.9902–1.0138, R2 = 0.9958–0.9972), that was precise (intra-day precision: 3.4–4.1%; inter-day precision: 4.4–8.2% over the AMR), accurate when compared with an available external laboratory test (slope = 0.9943–1.0206, R2 = 0.9635–0.9678, no lower decision point interpretation changes), with effective analyte recovery (77.2–83.5%), and established stability characteristics, while chromatographically separating the analytes to ensure no additive effects due to the isotopic distribution of the opposing analyte.

Effects of ethanol and sex on propionate metabolism evaluated via a faster 13C-propionate breath test in rats.

Alcoholism is regarded as a risk factor for vitamin B12 (VB12) deficiency. Because VB12 serves as a coenzyme of methylmalonyl-CoA mutase, a key enzyme in propionate metabolism, the 13C-propionate breath test (PBT) has been studied as a non-invasive diagnostic modality for VB12 deficiency. However, the conventional PBT requires 2 h, which is inconvenient in clinical practice. We hypothesized that a faster PBT can be used to evaluate propionate metabolism and is more easily adaptable for clinical practice. To evaluate a faster PBT for assessing the effects of long-term ethanol consumption on propionate metabolism in ethanol-fed rats (ERs). ERs were obtained by replacing standard drinking water (for control rats, CRs) with 16% ethanol solution in descendants of F344/DuCrj rats. Faster PBT was performed by administering 13C-propionate aqueous solution to male and female ERs and CRs by inserting a metal tubule from the mouth to the stomach; exhaled gas was collected in a bag to measure its 13CO2/12CO2 isotope ratio via infrared isotope spectrometry. Serum VB12 and alanine transaminase (ALT) levels were measured via chemiluminescence immunoassay and the lactate dehydrogenase-ultraviolet method, respectively. We evaluated statistical differences in mean body weight, change in 13CO2 (Δ13CO2‰), peak Δ13CO2‰, and serum VB12 and ALT, between males and females and between ERs and CRs using the t-test and Mann-Whitney U test for normally and non-normally distributed variables, respectively. Males weighed significantly more than females (P < 0.001); CRs weighed significantly more than ERs (P < 0.008). Δ13CO2 reached a peak (Cmax) at 20 min and 30 min in females and males, respectively, decreasing after 20-30 min without rebound in all groups. Males had significantly higher Cmax and Δ13CO2 at 15-45 min than females (P < 0.05; for all pairs). Propionate metabolism was enhanced in male ERs relative to male CRs, whereas metabolism did not differ markedly between ERs and CRs for females. Males had higher serum VB12 levels than females, without prominent differences between the ER and CR groups. Male CRs had notably higher ALT levels than male ERs. Thus, chronic ethanol consumption may trigger fatty acid production via intestinal bacteria and changes in gut microbiome composition. Faster PBT shows that 16% ethanol consumption promotes propionate metabolism without inducing liver injury. This PBT may be used clinically to evaluate gut flora status.

Tauopathy and alcohol consumption interact to alter locus coeruleus excitatory transmission and excitability in male and female mice.

Alcohol use disorder is a major public health concern in the United States. Recent work has suggested a link between chronic alcohol consumption and the development of tauopathy disorders, such as Alzheimer's disease and frontotemporal dementia. However, relatively little work has investigated changes in neural circuitry involved in both tauopathy disorders and alcohol use disorder. The locus coeruleus (LC) is the major noradrenergic nucleus in the brain and is one of the earliest sites to be affected by tau lesions. The LC is also implicated in the rewarding effects of ethanol and alcohol withdrawal. In this study we assessed effects of long-term ethanol consumption and tauopathy on the physiology of LC neurons. Male and female P301S mice, a humanized transgenic mouse model of tauopathy, underwent 16 weeks of intermittent access to 20% ethanol from 3 to 7 months of age. We observed higher total alcohol consumption in female mice regardless of genotype. Male P301S mice consumed more ethanol and had a greater preference for ethanol than wild-type (WT) males. At the end of the drinking study, LC function was assessed using exvivo whole cell electrophysiology. We found significant changes in excitatory inputs to the LC due to both ethanol and genotype. We found significantly increased excitability of the LC due to ethanol with greater effects in female P301S mice than in female WT mice. Our study identifies significant changes in the LC due to interactions between tauopathy and long-term ethanol use. These findings could have important implications regarding LC activity and changes in behavior due to both ethanol- and tauopathy-related dementia.

Effects of Long-Term Ethanol Consumption on Learning-Memory Functions in Mice and the Mechanisms Involved

To investigate the effect of long-term ethanol consumption on learning-memory functions in mice and the mechanisms involved. Thirty male C57BL6/J mice were randomly assigned to 3 groups, with 10 mice in each group. The three groups included a control group in which the mice were given water ad libitum for 30 days, a long-term ethanol consumption group, or the EtOH group, in which the mice were given 6% (volume fraction) ethanol ad libitum for 30 days, and a long-term alcoholism group, or the EtOH+G group, in which the mice were given 5% (volume fraction) ethanol ad libitum for 30 days plus intermittent intragastric gavage of 20% ethanol at 3.5 g per kilogram body mass once every three days. After 30 days, the learning-memory functions of the mice were evaluated. At the conclusion of the experiment, the brain tissue of the mice was collected in order to examine the oxygen consumption rate (OCR) of mitochondria, the levels of pan-acetylation and protein oxidative stress in the hippocampal tissue, and the expression of sirtuin-3 (SIRT3) in hippocampus. Morris water maze test showed that, compared with those of the control group, the times of crossing the platform and the percentage of platform time in the EtOH group and the EtOH+G group were both lower, and the EtOH+G group had the lowest results ( P<0.05). Western blot results showed that long-term ethanol intake increased the levels of protein oxidative stress and pan-acetylation in the hippocampal tissue and down-regulated SIRT3 expression of hippocampal mitochondria. The results of mitochondrial complex Ⅱ respiration showed that the brain mitochondrial 3-state respiration in the EtOH group and the EtOH+G group was lower than that in the control group ( P<0.05). Compared that with the control group, the mitochondrial maximum respiration in EtOH+G group was decreased ( P<0.05). Both long-term ethanol consumption and long-term alcoholism can reduce learning-memory functions and long-term alcoholism has the greater impact of the two. The potential mechanism may involve the down-regulation of the expression of SIRT3 protein in the hippocampus, which results in an increased level of pan-acetylation and enhanced expression of oxidative stress protein in the hippocampus, affects the mitochondrial functions of the brain, inhibits the oxidative phosphorylation capacity of mitochondrial complex Ⅱ, reduces the ATP energy supply of the brain tissue, and thus affects the learning-memory function.

Neural serotonergic circuits for controlling long-term voluntary alcohol consumption in mice

Alcohol-use-disorders are chronic relapsing illnesses, often co-morbid with anxiety. We have previously shown using the “drinking-in-the-dark” model in mice that the stimulation of the serotonin receptor 1A (5-HT1A) reduces ethanol binge-drinking behaviour and withdrawal-induced anxiety. The 5-HT1A receptor is located either on Raphe neurons as autoreceptors, or on target neurons as heteroreceptors. By combining a pharmacological approach with biased agonists targeting the 5-HT1A auto- or heteroreceptor and a chemogenetic approach (DREADDs), here we identified that ethanol-binge drinking behaviour is dependent on 5-HT1A autoreceptors and 5-HT neuronal function, with a transition from DRN-dependent regulation of short-term (6 weeks) ethanol intake, to MRN-dependent regulation after longer ethanol exposure (12 weeks). We further identified a serotonergic microcircuit (5-HTMRN→DG) originating from the MRN and projecting to the dentate gyrus (DG) of the hippocampus, that is specifically affected by, and modulates long-term ethanol consumption. The present study indicates that targeting Raphe nuclei 5-HT1A autoreceptors with agonists might represent an innovative pharmacotherapeutic strategy to combat alcohol abuse.

Comparative Study of the Behavior of Wistar Rats, Dopamine Transporter Heterozygous Rats and Rats after Long-Term Ethanol Consumption in the Morris Water Maze

Comparative Study of the Behavior of Wistar Rats, Dopamine Transporter Heterozygous Rats and Rats after Long-Term Ethanol Consumption in the Morris Water Maze

UFR2709, an Antagonist of Nicotinic Acetylcholine Receptors, Delays the Acquisition and Reduces Long-Term Ethanol Intake in Alcohol-Preferring UChB Bibulous Rats.

Alcoholism is a worldwide public health problem with high economic cost and which affects health and social behavior. It is estimated that alcoholism kills 3 million people globally, while in Chile it is responsible for around 9 thousand deaths per year. Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels expressed in the central nervous system, and they were suggested to modulate the ethanol mechanism involved in abuse and dependence. Previous work demonstrated a short-term treatment with UFR2709, a nAChRs antagonist, which reduced ethanol intake using a two-bottle free-choice paradigm in University of Chile bibulous (UChB) rats. Here, we present evidence of the UFR2709 efficacy in reducing the acquisition and long-term ethanol consumption. Our results show that UFR2709 (2.5 mg/kg i.p.) reduces the seek behavior and ethanol intake, even when the drug administration was stopped, and induced a reduction in the overall ethanol intake by around 55%. Using naïve UChB bibulous rats, we demonstrate that UFR2709 could delay and reduce the genetically adaptive impulse to seek and drink ethanol and prevent its excessive intake.

Bone impairment in adolescent female rats chronically exposed to ethanol

Ethanol consumption has increased among teenagers worldwide considerably,including females. Long-term ethanol consumption in women has been reported to cause bone metabolism imbalance. However, only few studies are available on the impact of long-term ethanol consumptionon bone morphology during adolescence. Here, we report the effects of chronic ethanol consumption on bone structure in adolescent female rats. Twenty female Wistar rats (35 days old) received, by gavage, distilled water (control) or ethanol (6.5 g/kg/day, 22.5% w/v) once daily for 55 days. After ethanol administration, animals were perfused, and the femora were collected. Morphometric evaluations were performed by electron microscopy scanning. Femora length, cortical bone thickness and medullar bone diameter was measured. The results demonstrated that ethanol exposure during adolescence reduced the length of femurs, with a decrease of the anterior thickness, posterior thickness, and mid-lateral diameter (P&lt;0.05). Thus, long-term ethanol intake may lead to alterations on bone morphometry, reducing the thickness of compact bone and femur length in adolescent females.

On the Use of Heterogeneous Stock Mice to Map Transcriptomes Associated With Excessive Ethanol Consumption.

We and many others have noted the advantages of using heterogeneous (HS) animals to map genes and gene networks associated with both behavioral and non-behavioral phenotypes. Importantly, genetically complex Mus musculus crosses provide substantially increased resolution to examine old and new relationships between gene expression and behavior. Here we report on data obtained from two HS populations: the HS/NPT derived from eight inbred laboratory mouse strains and the HS-CC derived from the eight collaborative cross inbred mouse strains that includes three wild-derived strains. Our work has focused on the genes and gene networks associated with risk for excessive ethanol consumption, individual variation in ethanol consumption and the consequences, including escalation, of long-term ethanol consumption. Background data on the development of HS mice is provided, including advantages for the detection of expression quantitative trait loci. Examples are also provided of using HS animals to probe the genes associated with ethanol preference and binge ethanol consumption.

Changes in glucocorticoid receptor sensitivity following long‐term ethanol self‐administration in male and female rhesus macaques

Stress can alter ethanol consumption and long-term use results in adaptation of the stress response, highlighting a bidirectional relationship that remains poorly understood. Non-human primates are an excellent model to investigate the interaction of stress and long-term ethanol use due to their similar neuroendocrine system to humans and propensity to chronically drink intoxicating amounts of alcohol. Here, circulating stress hormones (adrenocorticotropin hormone (ACTH) and cortisol) were measured in response to a low (17 mg/kg) and high (130 mg/kg) dose of dexamethasone, a potent glucocorticoid receptor (GR) agonist. The dexamethasone response was compared before (baseline) and after 7 months of daily 22 h/d ethanol self-administration in young adult male (n=6) and female (n=6) rhesus macaques. At both timepoints, the high dose dexamethasone challenge resulted in 90-100% suppression of both ACTH and cortisol with no difference between sexes. At baseline, low dose dexamethasone reduced both ACTH and cortisol, but to a lesser degree and with greater between-subject variability, 0 – 60%. Following seven-months of daily ethanol consumption (average intakes ranging from 0.6 – 4.0 g/kg/day) the response to low dose dexamethasone was greater. This effect was more robust in females compared to males. Additionally, the change in suppression between baseline and following long-term daily ethanol consumption correlated with daily intake, such that the magnitude of change over time was greater in heavier drinkers. Because stress increases reliance on putamen-dependent or habitual behavior and heavy ethanol drinking leads to excitatory-inhibitory neurotransmission imbalance within the putamen, we tested the hypothesis that long-term ethanol self-administration will alter striatal transmission via the GR. Therefore, we used whole cell patch clamp electrophysiology to investigate the effect of bath application of dexamethasone onto putamen medium spiny neurons (MSNs). Dexamethasone rapidly decreased the area and amplitude of spontaneous inhibitory postsynaptic currents among binge drinkers. These data show that acute dexamethasone, presumably via activation of the membrane bound GR, activated fewer GABAergic receptors among binge drinkers. These data highlight a unique role of the GR within the putamen that may mediate stress dependent changes contributing to addiction vulnerability, particularly among binge drinkers.

Modulated in vitro lymphocytes in the treatment of alcoholism: Experimental study

IntroductionImmune cells are dysfunctional during long-term ethanol consumption and may contribute to the progression from healthy to problem drinking. Lymphocytes from mice with chronic ethanol exposure characterized by impaired functional activity, manifested in the combination of increased spontaneous proliferation against the background of low sensitivity to T- cell mitogens.ObjectivesWe first demonstrated that original compound meta-chloro-benzhydryl-urea (m-ch-BHU) in vitro restored long-term alcoholized mice lymphocytes activity through GABA(A) receptors. We also revealed the possibility of animal’s behavioral regulation by the transplantation of immune cells with definite functional characteristics, also modulated by psychoactive drugs. Based on the previous results we investigated effects of m-ch-BHU modulated lymphocytes transplantation in recipients with experimental alcoholism.MethodsMale (CBAxC57Bl/6)F1 mice with 6-month 10% ethanol exposure were undergoing the transplantation of syngeneic long-term alcoholized mice lymphocytes, pretreated in vitro with m-ch-BHU. Recipient’s ethanol consumption, nervous and immune systems functional activities were estimated.ResultsIt was shown that transplantation of lymphocytes with in vitro m-ch-BHU modulated functional activity caused in syngeneic recipients with chronic alcohol exposure essential ethanol consumption decrease and stimulation of motor and exploratory activities in the “open field” against the background of cytokines modulation in brain. The significant stimulation of humoral immune response, estimated by the relative number of antibody-forming spleen cells, and stimulation of DTH reaction were also detected in recipients after lymphocytes transplantation.ConclusionsResults demonstrated that transplantation of m-ch-BHU modulated lymphocytes caused positive psychoneuroimmunomodulating effect in animals with chronic alcohol exposure, so, it may be considered as a promising method in the treatment of alcoholismDisclosureNo significant relationships.

P.002 Effects of long-term ethanol consumption: study the possible interaction between brain-derived neurotrophic factor and serotonin brain systems

The results of a long-term study of the Campanula altaica phenotypic variation in the Western Siberian forest steppe (Novosibirsk) are described. Two ecotypes (subalpine and meadow) and three populations (Altai, Mountain Altai, and Tomsk) were assessed for the variation in phenological and several structural traits. The PAleontological STatistics (PAST) tool was used for statistical data processing. The adaptive changes in plant seasonal rhythms during their cultivation mainly appeared as a high lability of the flowering phase, whereas most phenological traits did not significantly differ at an intrapopulation level, reflecting an endogenous character of seasonal development. The intraspecific variation of structural traits is more pronounced: significant differences in the flowering shoot height, seed productivity per capsule, and several other characteristics are observed. The same traits of the accessions with different origins display different degrees and directions of changes. The least variable among the examined traits are the duration of vegetation, corolla diameter, and mass of 1000 seeds. High individual variation is a factor determining C. altaica as culturally sustainable species.

Distinct Region- and Time-Dependent Functional Cortical Adaptations in C57BL/6J Mice after Short and Prolonged Alcohol Drinking.

Alcohol (ethanol) use disorder is associated with changes in frontal cortical areas including the anterior cingulate cortex (ACC) and orbitofrontal cortex (OFC) that contribute to cognitive deficits, uncontrolled drinking, and relapse. Acute ethanol exposure reduces intrinsic excitability of lateral OFC (lOFC) neurons, while chronic exposure and long-term drinking influence plasticity of intrinsic excitability and function of glutamatergic synapses. However, the time course that these adaptations occur across a history of ethanol drinking is unknown. The current study examined whether short-term and long-term voluntary ethanol consumption using an intermittent access paradigm would alter the biophysical properties of deep-layer pyramidal neurons in the ACC and lOFC. Neuronal spiking varied in the ACC with an initial increase in evoked firing after 1 d of drinking followed by a decrease in firing in mice that consumed ethanol for one week. No difference in lOFC spike number was observed between water controls and 1-d ethanol drinking mice, but mice that consumed ethanol for one week or more showed a significant increase in evoked firing. Voluntary ethanol drinking for 4 weeks also produced a total loss of ethanol inhibition of lOFC neurons. There was no effect of drinking on excitatory or inhibitory synaptic events in ACC or lOFC neurons across all time points in this model. Overall, these results demonstrate that voluntary drinking alters neuronal excitability in the ACC and lOFC in distinct ways and on a different time scale that may contribute to the impairment of prefrontal cortex-dependent behaviors observed in individuals with alcohol use disorder (AUD).