Blood Ethanol Levels Research Articles

Maternal ethanol consumption before paternal fertilization: Stimulation of hypocretin neurogenesis and ethanol intake in zebrafish offspring

There are numerous clinical and pre-clinical studies showing that exposure of the embryo to ethanol markedly affects neuronal development and stimulates alcohol drinking and related behaviors. In rodents and zebrafish, our studies show that embryonic exposure to low-dose ethanol, in addition to increasing voluntary ethanol intake during adolescence, increases the density of hypothalamic hypocretin (hcrt) neurons, a neuropeptide known to regulate reward-related behaviors. The question addressed here in zebrafish is whether maternal ethanol intake before conception also affects neuronal and behavioral development, phenomena suggested by clinical reports but seldom investigated. To determine if preconception maternal ethanol consumption also affects these hcrt neurons and behavior in the offspring, we first standardized a method of measuring voluntary ethanol consumption in AB strain adult and larval zebrafish given gelatin meals containing 10% or 0.1% ethanol, respectively. We found the number of bites of gelatin to be an accurate measure of intake in adults and a strong predictor of blood ethanol levels, and also to be a reliable indicator of intake in larval zebrafish. We then used this feeding paradigm and live imaging to examine the effects of preconception maternal intake of 10% ethanol-gelatin compared to plain-gelatin for 14 days on neuronal development in the offspring. Whereas ethanol consumption by adult female HuC:GFP transgenic zebrafish had no impact on the number of differentiated HuC+ neurons at 28 h post-fertilization (hpf), preconception ethanol consumption by adult female hcrt:EGFP zebrafish significantly increased the number of hcrt neurons in the offspring, an effect observed at 28 hpf and confirmed at 6 and 12 days post-fertilization (dpf). This increase in hcrt neurons was primarily present on the left side of the brain, indicating asymmetry in ethanol's actions, and it was accompanied by behavioral changes in the offspring, including a significant increase in novelty-induced locomotor activity but not thigmotaxis measured at 6 dpf and also in voluntary consumption of 0.1% ethanol-gelatin at 12 dpf. Notably, these measures of ethanol intake and locomotor activity stimulated by preconception ethanol were strongly, positively correlated with the number of hcrt neurons. These findings demonstrate that preconception maternal ethanol consumption affects the brain and behavior of the offspring, producing effects similar to those caused by embryonic ethanol exposure, and they provide further evidence that the ethanol-induced increase in hcrt neurogenesis contributes to the behavioral disturbances caused by ethanol.

The Effect of Blood Ethanol Level on Mortality in Patients with Trauma

The Effect of Blood Ethanol Level on Mortality in Patients with Trauma

Chronic Intermittent Ethanol Exposure Selectively Increases Synaptic Excitability in the Ventral Domain of the Rat Hippocampus

Many studies have implicated hippocampal dysregulation in the pathophysiology of alcohol use disorder (AUD). However, over the past twenty years, a growing body of evidence has revealed distinct functional roles of the dorsal (dHC) and ventral (vHC) hippocampal subregions, with the dHC being primarily involved in spatial learning and memory and the vHC regulating anxiety- and depressive-like behaviors. Notably, to our knowledge, no rodent studies have examined the effects of chronic ethanol exposure on synaptic transmission along the dorsal/ventral axis. To that end, we examined the effects of the chronic intermittent ethanol vapor exposure (CIE) model of AUD on dHC and vHC synaptic excitability. Adult male Long–Evans rats were exposed to CIE or AIR for 10 days (12 h/day; targeting blood ethanol levels of 175–225 mg%) and recordings were made 24 h into withdrawal. As expected, this protocol increased anxiety-like behaviors on the elevated plus-maze and successive alleys test. Extracellular recordings revealed marked CIE-associated increases in synaptic excitation in the CA1 region that were exclusively restricted to the ventral domain of the hippocampus. Western blot analysis of synaptoneurosomal fractions revealed that the expression of two proteins that regulate synaptic strength, GluA2 and SK2, were dysregulated in the vHC, but not the dHC, following CIE. Together, these findings suggest that the ventral CA1 region may be particularly sensitive to the maladaptive effects of chronic ethanol exposure and provide new insight into some of the neural substrates that may contribute to the negative affective state that develops during withdrawal.

The effects of traditional fermented beverages on ethanol, ethyl glucuronide and ethyl sulphate levels

BackgroundTraditional beverages such as şalgam, boza, kımız and kefir are frequently being consumed in Turkey. During the production of these beverages fermentation provides long time bio-preservation due to protective effects of ethanol, a product of microorganisms’ metabolism. Especially in traffic controls and in rehabilitation process of alcoholic patients, fermented beverages may cause confusion since it may be linked with false positivity of ethanol and ethanol metabolite tests. The present study aims to assess blood ethanol and urine ethanol metabolite levels after consumption of traditional fermented beverages.MethodTwelve participants consumed 300 mL standardized homemade traditional şalgam, boza, kımız and kefir. Blood samples were collected before and after (at 45th min) consumption and urine samples were collected before and after (at 4th hour) consumption from the participants. The samples of these four beverages and the blood samples were analyzed using headspace gas chromatography for ethanol levels, and urine samples were analyzed with liquid chromatography tandem mass spectrometry for ethyl-glucuronide and ethyl-sulphate levels.ResultsEthanol levels in şalgam, boza, kımız, and kefir samples, used at the present study, were 80.02 mg/dl, 30.02 mg/dl, 57.83 mg/dl and 6.07 mg/dl, respectively. After consumption of beverages, blood ethanol levels were negative for all analyzed samples. No statistically significant correlation was found between initial and post-consumption ethyl-glucuronide and ethyl-sulphate levels.

Pre-offense alcohol intake in homicide offenders and victims: A forensic-toxicological case-control study

Background and aimAlcohol is associated with violent behavior, although little is known regarding to what extent alcohol increases homicide risk. We aimed to estimate risks of homicide offending and victimization conferred by the presence of ethanol in blood by using toxicological data from homicide victims and offenders and from controls who had died in vehicle-related accidents. MethodsFrom nationwide governmental registries and databases, forensic-toxicological results were retrieved for victims (n = 200) and offenders (n = 105) of homicides committed during the years 2007–2009 and individuals killed in vehicle-related accidents (n = 1629) during the years 2006–2014. Ethanol levels in blood exceeding 0.01 g/100 ml were considered positive. ResultsUsing logistic regression, we found that the presence of ethanol in blood conferred a significantly increased risk of homicide offending (age-adjusted odds ratio [aOR] = 3.6, 95% confidence interval [95% CI] = 2.3–5.6) and homicide victimization (aOR = 2.1, 95% CI = 1.4–3.0). After stratification by sex, risk estimates in females were about 3-fold greater than in males for both homicide offending ([aOR = 11.0, 95% CI = 2.4–49.8] versus [aOR = 3.1, 95% CI = 1.9–4.9]) and victimization ([aOR = 5.4, 95% CI = 2.4–12.2] versus [aOR = 1.7, 95% CI = 1.1–2.8]). Sensitivity analyses yielded similar estimates. ConclusionsThe results of the present study are consistent with prior findings suggesting alcohol to be an important risk factor for homicide offending and victimization. Surprisingly, however, associations were more pronounced in females, although additional studies that control for potential confounders are warranted to facilitate speculations about causality.

Differential effects of acute versus chronic stress on ethanol sensitivity: Evidence for interactions on both behavioral and neuroimmune outcomes

Acute alcohol intoxication induces significant alterations in brain cytokines. Since stress challenges also profoundly impact central cytokine expression, these experiments examined the influence of acute and chronic stress on ethanol-induced brain cytokine responses. In Experiment 1, adult male rats were exposed to acute footshock. After a post-stress recovery interval of 0, 2, 4, or 24 h, rats were administered ethanol (4 g/kg; intragastric), with trunk blood and brains collected 3 h later. In non-stressed controls, acute ethanol increased expression of Il-6 and IκBα in the hippocampus. In contrast, rats exposed to footshock 24 h prior to ethanol demonstrated potentiation of hippocampal Il-6 and IκBα expression relative to ethanol-exposed non-stressed controls. Experiment 2 subsequently examined the effects of chronic stress on ethanol-related cytokine expression. Following a novel chronic escalating stress procedure, rats were intubated with ethanol. As expected, acute ethanol increased Il-6 expression in all structures examined, yet the Il-6 response was attenuated exclusively in the hippocampus in chronically stressed rats. Later experiments determined that neither acute nor chronic stress affected ethanol pharmacokinetics. When ethanol hypnosis was examined, however, rats exposed to chronic stress awoke at significantly lower blood ethanol levels compared to acutely stressed rats, despite similar durations of ethanol-induced sedation. These data indicate that chronic stress may increase sensitivity to ethanol hypnosis. Together, these experiments demonstrate an intriguing interaction between recent stress history and ethanol-induced increases in hippocampal Il-6, and may provide insight into novel pharmacotherapeutic targets for prevention and treatment of alcohol-related health outcomes based on stress susceptibility.

Activated mesenchymal stem cell administration inhibits chronic alcohol drinking and suppresses relapse-like drinking in high-alcohol drinker rats.

Neuroinflammation has been reported to follow chronic ethanol intake and may perpetuate alcohol consumption. Present studies determined the effect of human mesenchymal stem cells (hMSCs), known for their anti-inflammatory action, on chronic ethanol intake and relapse-like ethanol intake in a post-deprivation condition. Rats were allowed 12-17weeks of chronic voluntary ethanol (10% and 20% v/v) intake, after which a single dose of activated hMSCs (5×105 ) was injected into a brain lateral ventricle. Control animals were administered vehicle. After assessing the effect of hMSCs on chronic ethanol intake for 1week, animals were deprived of ethanol for 2weeks and thereafter an ethanol re-access of 60min was allowed to determine relapse-like intake. A single administration of activated hMSCs inhibited chronic alcohol consumption by 70% (P<0.001), an effect seen within the first 24hours of hMSCs administration, and reduced relapse-like drinking by 80% (P<0.001). In the relapse-like condition, control animals attain blood ethanol ('binge-like') levels >80mg/dl. The single hMSC administration reduced relapse-like blood ethanol levels to 20mg/dl. Chronic ethanol intake increased by 250% (P<0.001) the levels of reactive oxygen species in hippocampus, which were markedly reduced by hMSC administration. Astrocyte glial acidic fibrillary protein immunoreactivity, a hallmark of neuroinflammation, was increased by 60-80% (P<0.001) by chronic ethanol intake, an effect that was fully abolished by the administration of hMSCs. This study supports the neuroinflammation-chronic ethanol intake hypothesis and suggest that mesenchymal stem cell administration may be considered in the treatment of alcohol use disorders.

Effect of alcohol dehydrogenase-1B and -7 polymorphisms on blood ethanol and acetaldehyde concentrations in healthy subjects with a history of moderate alcohol consumption.

This study aims to evaluate the effect of ADH1B and ADH7 genotypes on blood acetaldehyde and ethanol levels after alcohol ingestion, and to measure the genotoxic effect of smoking and ethanol on the buccal cells, also controlling for ADH variants. We recruited healthy Italian subjects with at least a moderate history of alcohol consumption. All subjects were given an alcoholic drink of 0.4 g ethanol /kg of body weight. Blood venous samples were collected at baseline, and 30, 60, 90, and 120 minutes after ingestion. Buccal cells were collected before ethanol ingestion. Sixty subjects were enrolled in the study. Individuals with the ADH1B GG genotype had median ethanol levels of 5.0mM (IQR 3.4-7.2), and those with the ADH1B GT/TT genotype had 4.7mM (IQR 4.2-4.8). Corresponding acetaldehyde levels were 1.5μM (IQR 0.7-2.6) for ADH1B GG genotype and 1.6μM (IQR 1.5-1.7) for ADH1B CG/GG genotype. Individuals with the ADH7 CC genotype had median ethanol levels of 5.0mM (IQR 3.3-7.2), while 5.0mM (IQR 4.7-5.6) was in those with the ADH7 CG/GG genotype. Corresponding acetaldehyde levels were 1.5 μM (IQR 0.7-2.6) for ADH7 CC genotype and 1.5 μM (IQR 1.4-1.6) for ADH7 CG/GG genotypes. A non-significant increase in the frequency of karyolitic and pyknotic cells was found in the group of heavy drinkers and current smokers, when compared to the moderate drinkers and the non-smokers. Our study does not support the hypothesis that ADH1B and ADH7 genotypes affect blood ethanol and acetaldehyde concentration.

PP-2 Ethanol content in registered paediatric preparations: address the excess

Background Ethanol is commonly used in many pe-diatric liquid formulations as a solvent or preservative, with concentrations varying widely between products per formulation design. Therefore, acute or chronic use of certain products in paediatric patients may expose them to excessive amounts of ethanol. Both the U.S. Food and Drug Administration (FDA) and the Europe-an Medicines Agency (EMA) recommend to exclude ethanol from medicinal products intended for use in children whenever possible. If deemed necessary, however, the FDA has set (1995) a maximum quantity of ethanol content in over-the-counter (OTC) pediat-ric formulations for different ages. Recently (2014), the EMA suggested to lower the ethanol content limit set previously (2006). The use of ethanol in paediatric for-mulations is being evaluated these days by the Israeli Drug Registration Department and new regulations regarding limitations of ethanol content in liquid pediatric formulations (both OTC and prescription drugs) are being finalised. The objective of our study was to com-pile a list of oral liquid paediatric formulations registered in Israel that contain ethanol, and identify products whose ethanol concentration may produce dangerous ethanol blood levels in infants, either by taking the recommended dosage or by accidental consumption of large amounts of the preparation. Methods The Israeli Drug Registry was searched using the following keywords: elixir, solution, syrup, suspension, drops, tincture and ethanol, and the results were sent to the Drug Registration Department to provide the exact amounts of ethanol in each product per manufacturer’s data. Thirty-nine registered products were identified, of which 29 are currently registered for paediatric use. The majority of formulations are indicated for the treatment of cold and cough symptoms (n=14). Other formulations include antihistaminic, antiemetic and sedative prepara-tions (n=6); analgesics and antipyretics (n=3); antibiotics and anti-epileptics. Results The preparations were found to differ widely in ethanol content, ranging from Conclusion Ethanol consumption is strongly discour-aged during pregnancy and lactation, whereas children are constantly at risk for exposure to ethanol through routine use of registered medications. Many paediatricians and parents acknowledge the harmful effect of ethanol in this young population, but are unaware of the high potential for exposure. More than half (n=26) of the available products that contain alcohol do not meet the require-ments set by the new Israeli regulations and would be subject to changes accordingly (either in total volume or alcohol content of the formulation).

Methanol Poisoning with Fatality- Case Series in Dhaka Medical College Hospital in Bangladesh

Background: Adulterated methanol is highly toxic and leads to severe metabolic acidosis and blindness and mortality is very high if not treated with specific antidote. The time of ingestion and severity of illness is very short and comprehensive quick assessment and care is crucial for survival of patient. The selective antidote Fomepizole is not available in Bangladesh and pure intravenous ethanol is also out of reach in health care facility. The oral ethanol has its legislative regulation. Occasional clusters of severe methanol poisoning leading to fatality has been observed in recent years in Bangladesh. Here is a case series of 8 cases of adulterated methanol poisoning in a tertiary care hospital with lethality. Results and discussion: From November 2012 to January, 2013, in sphere of 3 months medicine units of Dhaka Medical College Hospital (DMCH) experienced 8 cases of methanol poisoning with fatality. Six patient presented in unconscious states within 3 to 7 h of consuming methanol while two patient presented within 48 h. All of them had gastrointestinal toxicity with variable episodes of vomiting. Three patients presented with visual impairment while only two out of eight had normal ophthalmoscopy. Respiratory distress was uniformly found in all patients before unstable profound shock. Blood ethanol level was not performed in any patient due to lack of available facility. Seven patients received only supportive measures ranging from steroids to sodium bi carbonate while one patient presented in severe toxicity and died quickly before any supportive measures. Antidote was not prescribed in any patient in the form of fomepizole or intravenous or oral ethanol. Conclusion: The judicious use of antidote even in the form of oral ethanol and folinic acid can save the precious life. A national guideline should be uniformly practiced by the physicians to combat the catastrophic methanol poisoning in Bangladesh.

Accidental Ethanol Ingestion in a 32 Day Old Infant

Alcohol exposure can occur in all age groups but occurs much less commonly in infants. Prior studies have shown that no level of alcohol exposure is safe, and can affect the brain and other areas of development. We report a 32 day old previously healthy male who presented to an outside hospital early morning for not acting normal. Earlier that morning, mom had fed him 3 ounces of formula mistakenly prepared with a clear liquid thought to be water, but later confirmed to be gin. The baby was mildly agitated on arrival, and the blood alcohol level 4 hrs post-ingestion was 230 mg/dl. This is one of the youngest documented cases of alcohol ingestion, and was remarkable for having a mild course with normal vital signs and electrolytes, and a non-focal physical exam. This case highlights the importance of having a high level of suspicion for alcohol exposure and a low threshold for checking blood ethanol levels in infants presenting with altered mental status, as there may be no hallmark signs, symptoms, or electrolyte abnormalities.

Hanging Fatalities in Central Bangkok, Thailand: A 13-Year Retrospective Study.

Hanging is violent asphyxial death. The objective of this study is to assess the data of hanging cases. A descriptive-retrospective study was conducted. We studied 244 hanging cases autopsied in Forensic Division, Department of Pathology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand, between January 2001 and December 2013. The study included 197 men (80.7%) and 47 women (19.2%). Their age ranged from 14 to 93 years. Most of these cases were incomplete hanging (83.6%). Features of hanging victims, such as tongue protrusion; congestion of face; petechial hemorrhage of face, conjunctiva, and internal organs; and neck injuries, significantly correlated with complete hanging. The predominant occupation of hanging victims was in the service industry (63.1%). Suicides usually occurred in private homes or apartments (84.8%). A suicide note was found in 6.1% of cases. The most common ligature material used was nylon rope, found in 61.1% of cases. The most underlying diseases of the victims in hanging cases were tuberculosis and human immunodeficiency virus infection, 9 cases each. Blood ethanol levels of 29 cases (11.8%) were detected to be higher than 150 mg%. Methamphetamine and benzodiazepine were detected in 5.3% and 3.3% of cases, respectively. This study provides comprehensive baseline data of hanging cases in central Bangkok.

Preliminary study of the effects of orengedokuto on breath alcohol concentration

ABSTRACTAimOrengedokuto (coptis detoxification decoction) is a medicinal extract widely used in Japan for the alleviation of symptoms of alcohol hangover and flush reaction. There have been no studies, however, on the effects of this medication on the concentration of alcohol in humans. The aim of this preliminary experiment was therefore to determine the effects of orengedokuto given before alcohol consumption, on breath alcohol concentration.MethodsThree healthy adults participated in the experiment. Breath alcohol concentration was measured with a breathalyzer at multiple intervals under two conditions: one with only the alcoholic beverage and one with orengedokuto extract (Matsuura orengedokuto fine powdered extract, 1.45 g) followed by an alcoholic beverage.ResultsThere was a lower peak and a faster decrease in breath alcohol concentration when the extract was given prior to alcohol consumption, suggesting that the extract accelerated the metabolism and/or inhibited the absorption of alcohol.ConclusionsThis preliminary study was limited by small sample size and lack of blood sampling. Additional trials using a cross‐over design with a larger sample size to further investigate the effects of orengedokuto on blood ethanol and acetaldehyde levels after consumption of alcohol, are currently in progress.

Effects of aqueous extracts from Panax ginseng and Hippophae rhamnoides on acute alcohol intoxication: An experimental study using mouse model

Ethnopharmacological relevanceAcute alcohol intoxication (AAI) is a frequent emergency, but therapeutic drugs with superior efficacy and safety are lacking. Panax ginseng (PG) and Hippophae rhamnoides (HR) respectively has a wide application as a complementary therapeutic agent in China for the treatment of AAI and liver injury induced by alcohol. We investigated the effects of aqueous extracts from PG and HR (AEPH) on AAI mice and identified its underlying mechanisms. Materials and methodsModels of AAI were induced by intragastric administration of ethanol (8g/kg). Seventy-two Specific pathogen-free (SPF) male Kunming mice were randomly divided into six groups: normal group, positive control group, AEPH of low dosage (100mg/kg) group, AEPH of medium dose (200mg/kg) group, AEPH of high dosage (400mg/kg) group and model group. The mice were treated with metadoxine (MTD, 500mg/kg) and AEPH. Thirty minutes later, the normal group was given normal saline, while the other groups were given ethanol (i.g., 8g/kg). The impact of AEPH was observed. In the same way, another seventy-two Kunming mice were randomly divided into six groups equally. The blood ethanol concentration at 0.5, 1, 1.5, 2, 3 and 6h after ethanol intake was determined by way of gas chromatography. The activity of alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) and microsomal ethanol oxidase (EO) in liver, and the concentration of β-endorphin (β-EP), leucine-enkephalin (LENK) in the brain were determined by enzyme-linked-immunosorbent serologic assay (ELISA). ResultsAEPH markedly prolonged alcohol tolerance time and shortened sober-up time after acute ethanol administration. AEPH decreased blood ethanol levels in six tests after ethanol intake. The 7-day survival rate of AEPH group was obviously superior to model group. AEPH increased the activities of ADH, ALDH, and decreased EO activity in liver. The crucial find was that AEPH markedly decreased β-EP and LENK concentration in the brain. ConclusionsAEPH can markedly increase the levels of ADH, ALDH, decrease EO activity in liver and decrease the concentration of β-EP and LENK in the brain to against acute alcohol intoxication in mice.

Auto-brewery Syndrome in the Setting of Long-standing Crohn’s Disease: A Case Report and Review of the Literature

A 71-year-old male with 50-year history of Crohn's disease was evaluated for acute onset of dizziness and slurred speech. Blood ethanol levels were elevated despite abstinence from alcohol for over 30 years. CT enterography demonstrated massive dilation of the small bowel with anastomotic stricture. Auto-brewery syndrome may be considered in a patient with chronic obstruction or hypomotility presenting with elevated serum ethanol levels in the setting of high carbohydrate intake. Although treatment algorithms lack validation, judicious use of antibiotic therapy, carbohydrate control, and short courses of antifungal therapy have all been reported in the literature. Importantly, clinical consideration of 'auto-brewery' should be undertaken with substantial caution, given the lack of validated mechanisms linking endogenous ethanol production to peripheral blood ethanol.

Selective alterations of NMDAR function and plasticity in D1 and D2 medium spiny neurons in the nucleus accumbens shell following chronic intermittent ethanol exposure

A major mouse model widely adopted in recent years to induce pronounced ethanol intake is the ethanol vapor model known as “CIE” or “Chronic Intermittent Ethanol.” One critical question concerning this model is whether the rapid induction of high blood ethanol levels for such short time periods is sufficient to induce alterations in N-methyl-d-aspartate receptor (NMDAR) function which may contribute to excessive ethanol intake. In this study, we determined whether such short term intermittent ethanol exposure modulates NMDAR function as well as other prominent electrophysiological properties and the expression of plasticity in both D1 (D1+) and D2 (D1−) dopamine receptor expressing medium spiny neurons (MSNs) in the nucleus accumbens (NAc) shell. To distinguish between the two subtypes of MSNs in the NAc we treated Drd1a-TdTomato transgenic mice with CIE vapor and electrophysiological recordings were conducted 24 h after the last vapor exposure. To investigate CIE induced alterations in plasticity, long-term depression (LTD) was induced by pairing low frequency stimulation (LFS) with post synaptic depolarization. In ethanol naïve mice, LFS induced synaptic depression (LTD) was apparent exclusively in D1+ MSNs. Whereas in slices prepared from CIE treated mice, LFS induced synaptic potentiation (LTP) in D1+ MSNs. Furthermore, following CIE exposure, LFS now produced LTD in D1− MSNs. We found that CIE exposure induced an increase in excitability in D1+ MSNs with no change in D1− MSNs. After CIE, we found a significant increase in spontaneous EPSCs (sEPSCs) frequency in D1+ but not D1− MSNs suggesting alterations in baseline α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) mediated signaling. CIE induced changes in NMDAR function were measured using the NMDA/AMPA ratio and input-output curves of isolated NMDAR currents. We observed a significant increase in NMDAR function in D1+ MSNs and a decrease in D1− MSNs after ethanol vapor exposure. The reversal of NMDAR function may account for the CIE induced alterations in the expression of plasticity. The cell type specific alterations in excitatory signaling in the NAc shell may constitute an important neuroadaptation necessary for the expression of increased ethanol consumption induced by intermittent ethanol vapor exposure.This article is part of the Special Issue entitled ‘Ionotropic glutamate receptors’.

Oleoylethanolamide prevents neuroimmune HMGB1/TLR4/NF-kB danger signaling in rat frontal cortex and depressive-like behavior induced by ethanol binge administration.

Alcohol abuse is frequently characterized by a specific pattern of intake in binge drinking episodes, inducing neuroinflammation and brain damage. Here, we characterized the temporal profile of neuroinflammation in rats exposed to intragastric binge ethanol administrations (3 times/day × 4 days) and tested the anti-inflammatory/neuroprotective properties of the satiety factor oleoylethanolamide (OEA). Pre-treatment with OEA (5 mg/kg, i.p.) previous each alcohol gavage blocked the expression of high mobility group box 1 (HMGB1) danger signal and the innate immunity Toll-like receptors 4 (TLR4) in frontal cortex, and inhibited the nuclear factor-kappa B (NF-kB) proinflammatory cascade induced by alcohol binge administration. OEA reduced the levels of interleukin-1beta (IL-1β), the monocyte chemoattractant protein-1 (MCP-1), and the enzymes cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in ethanol binged animals. Elevations in plasma tumor necrosis factor alpha (TNF-α) and IL-1β after ethanol were also inhibited by OEA. OEA also prevented ethanol-induced lipid peroxidation, caspase-8 and pro-apoptotic caspase-3 activation in frontal cortex. Additionally, OEA blocked the rise in blood corticosterone levels after ethanol with no alteration in blood ethanol levels and may affect ethanol-induced gut permeability for endotoxin. Finally, OEA, administered as a pre-treatment during the ethanol binge, exerted antidepressant-like effects during acute withdrawal. Altogether, results highlight a beneficial profile of OEA as a potent anti-inflammatory, antioxidant, neuroprotective and antidepressant-like compound to treat alcohol abuse.

Ethanol-induced conditioned taste aversion in Warsaw Alcohol High-Preferring (WHP) and Warsaw Alcohol Low-Preferring (WLP) rats

The aversive action of the pharmacological properties of ethanol was studied in selectively bred Warsaw Alcohol High-Preferring (WHP) and Warsaw Alcohol Low-Preferring (WLP) rats. For this study, a conditioned-taste aversion test was used. Male WHP and WLP rats were submitted to daily 20-min sessions for 5 days, in which a saccharin solution (1.0g/L) was available (pre-conditioning phase). Next, this drinking was paired with the injection of ethanol (0, 0.5, 1.0g/kg), intraperitoneally [i.p.] immediately after removal of the saccharin bottle (conditioning phase). Afterward, the choice between the saccharin solution and water was extended for 18 subsequent days for 20-min daily sessions (post-conditioning phase). Both doses of ethanol did not produce an aversion to saccharin in WLP and WHP rats in the conditioning phase. However, injection of the 1.0g/kg dose of ethanol produced an aversion in WLP rats that was detected by a decrease in saccharin intake at days 1, 3, 7, and 10 of the post-conditioning phase, with a decrease in saccharin preference for 16 days of the post-conditioning phase. Conditioned taste aversion, measured as a decrease in saccharin intake and saccharin preference, was only visible in WHP rats at day 1 and day 3 of the post-conditioning phase. This difference between WLP and WHP rats was apparent despite similar blood ethanol levels in both rat lines following injection of 0.5 and 1.0g/kg of ethanol. These results may suggest differing levels of aversion to the post-ingestional effects of ethanol between WLP and WHP rats. These differing levels of aversion may contribute to the selected line difference in ethanol preference in WHP and WLP rats.

Group 1 mGlu-family proteins promote neuroadaptation to ethanol and withdrawal-associated hippocampal damage

BackgroundGroup 1 mGlu-family proteins (i.e., mGlu) consist of mGlu1 and mGlu5 and their activity may influence voluntary ethanol intake. The present studies sought to examine the influence of these receptors on the development of ethanol dependence using in vitro and in vivo models of chronic, intermittent ethanol (CIE). MethodsRat hippocampal explants were exposed to CIE with or without the addition of mGlu1 antagonist (7-hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester (CPCCOEt; 0.5, 1, and 3μM) or mGlu5 antagonist (E)-2-methyl-6-styryl-pyridine (SIB-1893; 20, 100, and 200μM) to assess sparing of withdrawal-induced cytotoxicity. In a separate study, adult male rats were administered CIE with or without the addition of oral administration of group 1 mGlu antagonist 2-methyl-6-(phenylethynyl)-pyridine (MPEP; 3mg/kg). Blood ethanol levels (BELs) were determined at 0930h on Day 2 of Weeks 1, 2, and 3. Withdrawal behavior was monitored during Day 6 of the third consecutive withdrawal. ResultsCIE produced significant hippocampal cytotoxicity. These effects were attenuated by co-exposure to CPCCOEt (3μM) with ethanol in the CA3. By contrast, these effects were blocked by SIB-1893 (20μM) in each primary cell layer. Oral administration of MPEP with ethanol significantly attenuated behavioral effects of subsequent withdrawal and reduced BELs. ConclusionsThese data demonstrate that ethanol activates group 1 mGlu-family proteins to promote withdrawal-associated cytotoxicity in vitro and physical dependence in vivo. These findings suggest that group 1 mGlu-family proteins may be therapeutic targets for treatment of alcohol use disorders.

Ethanol Reversal of Tolerance to the Respiratory Depressant Effects of Morphine.

Opioids are the most common drugs associated with unintentional drug overdose. Death results from respiratory depression. Prolonged use of opioids results in the development of tolerance but the degree of tolerance is thought to vary between different effects of the drugs. Many opioid addicts regularly consume alcohol (ethanol), and post-mortem analyses of opioid overdose deaths have revealed an inverse correlation between blood morphine and ethanol levels. In the present study, we determined whether ethanol reduced tolerance to the respiratory depressant effects of opioids. Mice were treated with opioids (morphine, methadone, or buprenorphine) for up to 6 days. Respiration was measured in freely moving animals breathing 5% CO2 in air in plethysmograph chambers. Antinociception (analgesia) was measured as the latency to remove the tail from a thermal stimulus. Opioid tolerance was assessed by measuring the response to a challenge dose of morphine (10 mg/kg i.p.). Tolerance developed to the respiratory depressant effect of morphine but at a slower rate than tolerance to its antinociceptive effect. A low dose of ethanol (0.3 mg/kg) alone did not depress respiration but in prolonged morphine-treated animals respiratory depression was observed when ethanol was co-administered with the morphine challenge. Ethanol did not alter the brain levels of morphine. In contrast, in methadone- or buprenorphine-treated animals no respiratory depression was observed when ethanol was co-administered along with the morphine challenge. As heroin is converted to morphine in man, selective reversal of morphine tolerance by ethanol may be a contributory factor in heroin overdose deaths.