bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of the Production of Acetaldehyde by Astrocytes in Culture after Ethanol Exposure
Authors: Eysseric H. , Gonthier B. , Soubeyran A. , Bessard G. , Saxod R. , Barret L.
Published: May 2006

The nervous system is one of the main targets of ethanol toxicity. Astrocytes might play an important role in ethanol‐induced brain toxicity, because their integrity is essential for the normal growth and functioning of neurons. On the other hand, acetaldehyde has been implicated as a mediator in some of the biochemical, pharmacological, and behavioral effects of ethanol. The present study aimed at demonstrating the ability of astrocytes in culture to produce acetaldehyde from ethanol. Significant metabolization of ethanol with production of acetaldehyde was demonstrated in the primary culture of astrocytes. This production was quite low, compared with that usually observed in hepatocytes, but was in the same range as that measured in whole brain homogenates and corresponded to biologically active levels. Such a demonstration could bring new elements for understanding of ethanol neurotoxicity.
Immunohistochemical Determination of Hepatic Cytochrome P‐4502E1 in Formalin‐Fixed, Paraffin‐Embedded Sections
Authors: Cohen P. A. , Mak K. M. , Rosman A. S. , Kessova I. , Mishin V. M. , Koivisto T. , Lieber C. S.
Published: May 2006

Cytochrome P‐4502E1 (2E1) is inducible by chronic ethanol consumption that results in enhanced activation of anesthetics and commonly used drugs (such as acetaminophen) to hepatotoxins. Therefore, assessment of hepatic 2E1 is needed in prescribing these drugs for the management of alcoholic patients. Currently, measurement of 2E1 requires either immunohistochemistry on frozen sections or Western blot (WB) analysis of homogenized tissue in excess of that needed for pathology. To obtain a more widely applicable method, we developed a procedure to detect 2E1 by immunohistochemistry in formalin‐fixed, paraffin‐embedded liver biopsies obtained routinely for diagnosis. Data were collected from rats fed ethanol‐containing or control liquid diets for 3 weeks. lmmunostaining was performed using anti‐human rabbit 2E1 antibody as the primary antibody, and the immunoreaction was detected by the avidin‐biotin immunoperoxidase method after treating sections with target unmasking fluid, an antigen retrieval buffer that enhanced the staining of 2E1. In control rats, 2E1 staining was weak and perivenular. After ethanol feeding, it showed a lobular gradient, strongest perivenular and weakest periportal, similar to that seen in frozen sections. The staining intensity was scored as: 0 (no staining) to 3 (strong staining). The zonal staining was scored as follows: 1 = perivenular zonal staining, 2 = midzonal, and 3 = panlobular. With the product of the two scores, a significant difference was found between alcohol‐fed and control rats (5.1 ± 0.3 vs. 0.8 ± 0.2, p < 0.001). 2E1 assessments by WB were also significantly different for these rat pairs (68.5 ± 2.1 vs. 7.9 ± 0.8 arbitrary units/mg protein, p < 0.001), with a parallel increase of immunostaining scores and WB measurement of 2E1 content. This immunohistochemical method was then validated in 14 paraffin‐embedded percutaneous human liver biopsy samples. In livers of nonalcoholics, 2E1 staining was seen in the perivenular zone only, whereas in samples of alcoholics, the staining was perivenular to midzonal and sometimes periportal. A significant correlation between the zonal staining scores (rs= 0.67, p < 0.005) or intensity ± zonal staining scores (rs= 0.79, p < 0.001) and WB analysis was found. The immunohistochemical assessments of 2E1 expression in formalin‐fixed, paraffin‐embedded sections from livers of alcoholics was found to correlate with WB analysis, and lobular distribution was consistent with that seen in frozen sections. The proposed method should therefore be useful for the assessment of 2E1 content in paraffin‐embedded liver samples, thereby aiding in the management of heavy drinkers.
The Effect of Betaine in Reversing Alcoholic Steatosis
Authors: Barak Anthony J. , Beckenhauer Harriet C. , Badakhsh Shahrokh , Tuma Dean J.
Published: May 2006

The feeding of ethanol to experimental animals results in fatty infiltration of the liver. Recent findings have shown that ethanol‐induced steatosis is accompanied by a lowering in hepatic S‐adenosylmethionine (SAM) levels. It is known that SAM provides substrates for reduced glutathione formation and offers the cell protection from toxic metabolic oxidants. A recent study in this laboratory demonstrated that dietary supplementation with betaine generated increased SAM in the liver and protected against ethanol‐induced steatosis. The present study not only showed that betaine supplementation to rats protects the liver from alcoholic steatosis, but also demonstrated that once steatosis is established, treatment with betaine partially reversed the steatosis after cessation of ethanol feeding. Furthermore, this study indicated that betaine supplementation to the diet had the capacity to attenuate steatosis despite the continued feeding of ethanol.
Nerve Growth Factor and Basic Fibroblast Growth Factor Protect Rat Cerebellar Granule Cells in Culture against Ethanol‐Induced Cell Death
Authors: Luo Jia , West James R. , Pantazis Nicholas J.
Published: May 2006

Neuronal cell loss is one of the most debilitating effects of fetal ethanol exposure. Cultures of cerebellar granule cells are a useful model to investigate ethanol neurotoxicity, because ethanol depletes cell numbers in these cultures, which also occurs in vivo. The primary goal of the present study was to identify and characterize agents that can ameliorate the ethanol‐induced cell death that occurs in this culture system. Growth factors, such as nerve growth factor (NGF), basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and insulin‐like growth factor‐I (IGF‐I) can prevent neuronal degeneration after toxic insult in various experimental paradigms. These growth factors were investigated in the current study to determine whether or not they can mitigate ethanol‐induced death of cerebellar granule cells in culture. Results indicate that NGF and bFGF significantly reduced the ethanol‐induced cell loss. Both the NGF‐ and bFGF‐mediated neuroprotection required protein and RNA synthesis, because actinomycin D (RNA synthesis inhibitor) and cyclohex‐imide (protein synthesis inhibitor) blocked their neuroprotective effects. In addition to its neuroprotective effect, bFGF also had a neurotrophic effect and could enhance cell survival in the absence of ethanol exposure. NGF did not have a neurotrophic effect. Neither EGF nor IGF‐I was neuroprotective, although the latter did have a substantial neurotrophic effect. In conclusion, bFGF and NGF have long been recognized for their role in enhancing neuronal cell survival and differentiation. This study suggests that these growth factors can also provide neuroprotection against ethanol‐induced cell death.
Ethanol Consumption Suppresses Cell‐Mediated Inflammatory Responses and Increases T‐Helper Type 2 Cytokine Secretion in Trichinella spiralis‐Infected Rats
Authors: Na Hyung R. , Zhu Xi , Stewart George L. , Seelig Leonard L.
Published: May 2006

Alcohol consumption has been shown to be associated with immune suppression and immune modulation. In this study, the effects of ethanol ingestion on the host immune responses to Trichinella spiralis infection and the subsequent secretion of T‐helper cell‐associated cytokines were investigated in rats. At the early phase of T. spiralis infection, ethanol‐fed animals showed decreased numbers of blood neutrophils and eosinophils, and a decreased secretion of interferon‐γ (IFN‐γ) by mesenteric lymph node cells, compared with pair‐fed controls. Suppression of this early inflammatory response to infection in the ethanol‐treated groups resulted in a slower rate of expulsion of intestinal adult worms and a higher fecundity rate for female worms, compared with pair‐fed controls. A dramatic decrease in blood neutrophils in the ethanol‐treated groups was also manifested on day 9 postinfection. At that time, mesenteric lymph node cells from the ethanol‐treated groups secreted higher amounts of mast cell proliferation‐enhancing activity, which has been shown to contain T‐helper type 2‐associated cytokines. At the later phase of infection (day 12 to 20 day postinfection), ethanol‐treated animals contained higher numbers of blood eosinophils and secreted an increased amount of interleukin‐5 and mast cell proliferation‐enhancing activity, compared with pair‐fed controls. Although there was a slight rise with time after infection, the level of serum corticosterone was not significantly increased in the ethanol groups. Therefore, it is not likely that the observed immune modulations caused by ethanol consumption, especially in the early phase of infection, is the effect of elevated levels of corticosterone in the circulation. The present study found that ethanol consumption suppressed the initial amount of interferon‐γ secretion and inflammatory response, and may have directly or indirectly led to an enhancement of the secretion of T‐helper type 2‐type cytokines later in the primary immune response to T. spiralis infection.
Collagen Synthesis by Liver Stellate Cells Is Released from Its Normal Feedback Regulation by Acetaldehyde‐Induced Modification of the Carboxyl‐Terminal Propeptide of Procollagen
Authors: Ma Xiaoli , Svegliati‐Baroni Gianluca , Poniachik Jaime , Baraona Enrique , Lieber Charles S.
Published: May 2006

Acetaldehyde stimulates collagen synthesis in stellate cells and forms adducts with procollagen in the liver of alcoholics. To assess the possibility that modification of the carboxyl‐terminal propeptide by acetaldehyde affects its capacity to exert a feedback inhibition of collagen synthesis after splitting from procollagen, the propeptide was prepared by gel filtration of the bacterial collagenase digests of procollagen type I (obtained from 109 calvaria fibroblasts of newborn rats) and reacted with either 250 mM acetaldehyde and 100 mM CNBH3 or with 170;μM acetaldehyde without reducing agents, to mimick in vivo conditions. The unmodified propeptide produced a concentration‐dependent inhibition of collagen synthesis by Ito cells. By contrast, the acetaldehyde‐modified propeptide produced a lesser inhibition of procollagen synthesis in the cells, associated with a greater accumulation of collagen in the media. The incubation with 170 μM acetaldehyde and, to a lesser extent, 50 mM ethanol produced collagenase‐digestible adducts in stellate cells. Thus, the formation of acetaldehyde adducts with the carboxyl‐terminal propeptide of procollagen may account, at least in part, for the stimulatory effect of acetaldehyde on collagen synthesis by stellate cells and may lead to collagen accumulation through a decrease of the normal feedback regulation of collagen synthesis by the propeptide.
MK‐801 Administration During Ethanol Withdrawal in Neonatal Rat Pups Attenuates Ethanol‐Induced Behavioral Deficits
Authors: Thomas Jennifer D. , Weinert Stephen P. , Sharif Sharerah , Riley Edward P.
Published: May 2006

Alcohol exposure during development can produce central nervous system dysfunction, resulting in a wide range of behavioral alterations. The various mechanisms by which alcohol causes these behavioral changes, however, remain unknown. One mechanism that has been suggested is NMDA receptor‐mediated excitotoxic cell death produced by ethanol withdrawal. The present study examined whether MK‐801, an antagonist of the NMDA receptor that has been shown to protect against NMDA receptor‐mediated excitotoxicity, could block alcohol's adverse effects on behavior. Sprague‐Dawley rat pups were exposed to alcohol (6.0 g/kg) in a binge‐like manner on postnatal day 6 using an artificial rearing procedure. Subjects then received an injection of MK‐801 (0.1 mg/kg) or vehicle during withdrawal, 21 hr after ethanol exposure. At postnatal day 40, all subjects were tested on a serial spatial discrimination reversal task. Ethanolexposed subjects were impaired in both discrimination and reversal learning, and committed a significantly greater number of perseverative‐type errors, compared with controls. MK‐801 administration during ethanol withdrawal significantly attenuated ethanol‐induced deficits in reversal learning and increases in perseverative‐type errors, whereas MK‐801 exposure by itself had no significant effect on performance. Thus, exposure to MK‐801 during ethanol withdrawal partially protected against alcohol‐related disruptions in spatial reversal learning. These results support the suggestion that NMDA receptor‐mediated excitotoxicity may be one mechanism by which alcohol induces behavioral teratogenicity.
Polymorphism of ADH and ALDH Genes among Four Ethnic Groups in China and Effects upon the Risk for Alcoholism
Authors: Shen Yu‐Cun , Fan Jian‐Hua , Edenberg Howard J. , Li Ting‐Kai , Cui Yu‐Hua , Wang Yu‐Feng , Tian Cheng‐Hua , Zhou Chao‐Feng , Zhou Ru‐Lun , Wang Jing , Zhao Zhu‐Lin , Xia Guo‐Yan
Published: May 2006

The alcohol dehydrogenases (ADHs) and aldehyde dehydrogenases (ALDHs) that metabolize ethanol are polymorphic. Different alleles encode subunits of the enzymes that differ in their rate of metabolizing ethanol. These polymorphisms are distributed differently among populations and have been shown to influence the risk for alcoholism in some Asian populations. We have examined the allele frequencies at the ADH2, ADH3, and ALDH2 loci in four populations from China (Han, Mongolian, Korean, and Elunchun) and in alcoholics within each population. The four populations differ in allele frequencies, with the Elunchun having a much lower frequency of ADH2*2 alleles, and the Mongolian and Elunchun having a much lower frequency of ALDH2*2 alleles. Within each population, alleles at one or more of these three loci are protective against alcoholism, although the populations differ in which loci play significant roles. The protective allele at each locus (ALDH2*2, ADH2*2, and ADH3*1) encodes a subunit that either metabolizes ethanol to acetaldehyde more rapidly or slows the conversion of acetaldehyde to acetate. Taken as a whole, data demonstrate that genetic differences in the enzymes that metabolize alcohol can substantially affect the risk for alcoholism.
Effect of Pyridostigmine on the Thyroid‐Stimulating Hormone Response to Thyrotropin‐Releasing Hormone in Abstinent Alcoholics
Authors: Coiro V. , Vescovi P. P.
Published: May 2006

Alcoholism is sometimes associated with a blunted thyroid‐stimulating hormone (TSH) response to thyrotropin‐releasing hormone (TRH; peak minus baseline < 5 mlU/liter), despite basal TSH and thyroid hormone levels within the normal range. In light of the inhibitory effect of somatostatin on TSH secretion, we examined whether this condition is caused by an increased hypothalamic somatostatinergic tone in alcoholic subjects. To answer this question, 16 euthyroid male alcoholics (aged 38 to 50 years) with normal [n= 8; normal responder alcoholics (NRAs)] or blunted [n= 8; low responder alcoholics (LRAs)] TSH response to TRH were selected in a preliminary TRH test (200 μg in an intravenous bolus). In addition, 8 age‐ and weight‐matched normal men were tested with TRH and used as normal controls (NCs). NCs and alcoholic patients showed similar basal serum TSH, free triiodothyronine, and free thyroxine levels. All subjects were tested again with TRH, 60 min after treatment with 180 mg of pyridostigmine orally. According to selection criteria, NCs and NRA groups showed similar TSH responses to TRH, whereas TRH‐induced TSH rise was strikingly lower in LRAs than in NCs and the NRA group. Pyridostigmine did not change the basal levels of TSH in any group, whereas it enhanced in a similar manner the TRH‐induced TSH rise in the NC and NRA groups. No significant change in the TSH response to TRH was observed in LRA patients after pyridostigmine treatment. These data argue against the possibility that an enhanced somatostatinergic tone is responsible for the blunted TSH response to TRH observed in some alcoholic patients. This phenomenon might be attributed to alcohol‐related defects of stimulated pituitary thyrotroph secretory capacity in some individuals, possibly due to genetic vulnerability and/or the toxic effects of prolonged alcohol abuse.
Prodynorphin Allelic Distribution in Scandinavian Chronic Alcoholics
Authors: Geijer T. , Jönsson E. , Neiman J. , Gyllander A. , Sedvall G. , Rydberg U. , Terenius L.
Published: May 2006

Two regions of the human prodynorphin gene, the exon 4‐region coding for the opioid peptides and the putative promotor/exon 1‐region were analyzed for possible presence of polymorphisms. No polymorphism was detected in the exon 4‐region, whereas a GC/AT base‐pair exchange was observed 301 base pairs upstream of the exon 1/intron A boundary. This polymorphic marker was examined in Scandinavian chronic alcoholics (n= 70) and control subjects (n= 55). Prodynorphin allelic distributions were not significantly different in alcoholic patients and control subjects. The results suggest that no major influence on alcoholism is exerted through genes associated with this prodynorphin allelic marker.
Utility of Biological Markers during Outpatient Treatment of Alcohol‐Dependent Subjects: Carbohydrate‐Deficient Transferrin Responds to Moderate Changes in Alcohol Consumption
Authors: Huseby Nils‐Erik , Bjordal Einar , Nilssen Odd , Barth Tom
Published: May 2006

A group of 25 alcohol‐dependent subjects in outpatient treatment were monitored for a period of 4 weeks. They were weekly interviewed for their alcohol consumption and their serum levels of carbohydrate‐deficient transferrin (CDT) and γ‐glutamyltransferase (GT) were analyzed. The majority of the patients reported an excessive and fairly constant alcohol intake during the observation period. When selecting those patients that reported periods of 1 or 2 weeks with moderate changes in alcohol consumption, corresponding changes in CDT were demonstrated. Thus, of 14 patients reporting an increased alcohol consumption for 2 weeks (mean values increased from 57 to 101 g/day), 11 showed an increase in CDT at the end of the period. The mean CDT value of all 14 increased from 5.5 to 6.7% (p < 0.05). Slight, but not significant, increases were noted in GT, indicating that CDT is more sensitive than GT in detecting increased alcohol consumption. Furthermore, of 17 patients that reported decreased alcohol consumption for one or several weeks, 14 showed decreased CDT and GT values. The mean values of all 17 were reduced from 5.1% to 4.5% (CDT) and from 126 units/liter to 97 units/liter (GT) (p < 0.05 for both parameters). The results indicate that CDT responds to moderate changes in alcohol consumption in alcohol‐dependent patients and may thus be useful as a corrective tool to self‐reports of alcohol consumption during outpatient treatments.
Differences in the Course of Alcohol Withdrawal in Women and Men: A Polish Sample
Authors: Wojnar Marcin , Wasilewski Dariusz , Matsumoto Halina , Cedro Artur
Published: May 2006

A retrospective study compared the course of alcohol withdrawal, including delirium tremens, in women and men hospitalized in the Nowowiejski Hospital in Warsaw from 1973 to 1987. Medical records pertaining to 1179 patients were analyzed; 13.8% of these patients were women and 86.2% were men. The study showed that women began intensive alcohol drinking later than men (p < 0.0001), but the period between the onset of alcohol abuse and the first occurrence of alcohol withdrawal was shorter in women than in men (p < 0.0001). In the period of heavy drinking before hospitalization, women consumed significantly less alcohol then men (p < 0.0001); moreover, women drank nonbeverage alcohol less frequently than men (p < 0.05). Women were hospitalized substantially longer than men (p < 0.0001), whereas the duration of alcohol withdrawal symptoms at the time of hospitalization was comparable in both groups. Withdrawal seizures were significantly more frequent among men than among women (p < 0.001). Significant differences in the patients’somatic conditions were not noted between the groups, with the exception of anemia and decreased potassium concentration, which were more frequently observed in women (both p < 0.0001), and of increased concentration of ALT and hypoproteinemia, which were more frequent in men (respectively, p < 0.05 and p < 0.01). Co‐existing personality disorders, depressive disorders, and anxiety disorders—as well as abuse of benzodiazepines and barbiturates—were more frequently observed in women (p < 0.0001). The period between the first hospitalization due to alcohol withdrawal and the time of death was significantly shorter in men than in women (p < 0.05). The results point to differences in the conditions and the course of alcohol dependence and alcohol withdrawal between women and men.
Effect of Treatment with Paromomycin on Endotoxemia in Patients with Alcoholic Liver Disease—A Double‐Blind, Placebo‐Controlled Trial
Authors: Bode Christiane , Schäfer Christian , Fukui Hiroshi , Bode J. Christian
Published: May 2006

The results of experimental and clinical studies support the hypothesis that gut‐derived endotoxins might be of relevance for the development and course of alcoholic liver disease. The aim of this study was to test the effect of a nonabsorbable, broad‐spectrum antibiotic on endotoxemia in patients with alcoholic liver disease. Fifty patients with alcoholic liver disease (27 with cirrhosis, 23 without cirrhosis) were randomly assigned to receive either paromomycin sulfate (3 × 1 g/day) or placebo in a double‐blind fashion for at least 3 weeks, and if possible 4 weeks. Endotoxin concentration, liver function tests, and other laboratory parameters were determined in weekly intervals. Endotoxin concentration was also determined in 15 healthy controls. Groups receiving paromomycin or placebo were similar for clinical and biological items collected initially. Mean initial endotoxin concentrations were significantly elevated in both groups (mean ± SEM; paromomycin, 16.7 ± 5.3 pg/ml; placebo, 17.5 ± 6.9 pg/ml; healthy controls, 2.3 ± 0.4 pg/ml). Although the mean endotoxin concentration was lower in the verum group after 1 week (paromomycin, 8.0 ± 1.9 pg/ml; placebo, 14.6 ± 3.5 pg/ml; p > 0.05), paromomycin treatment had no significant effect on endotoxin concentration or liver function tests during the 4‐week period. The beneficial effect of paromomycin treatment on endotoxemia in cirrhotics reported in earlier studies could not be reproduced under the conditions of this trial in patients with alcoholic liver disease.
Which Relapse Criteria Best Predict the Mortality Risk of Treated Alcoholics?
Authors: Yoshino Aihide , Kato Motoichiro , Yoshimasu Haruo , Tatsuzawa Yasutaka , Watanabe Tamotsu
Published: May 2006

We examined which relapse criteria best predict the mortality risk of treated male alcoholics. The subjects were 172 male alcoholics who had previously been hospitalized. Using three criteria which defined relapse as failure to maintain abstinence from alcohol, alcohol abuse, or dependence, the relapse of each subject had been evaluated during a previous 3‐year outcome study. Relative mortality risks in the next 3 years classified by the three relapse criteria were compared. The follow‐up rate was 93.6% and 31 subjects died. The age‐corrected relative mortality risk for subjects failing to maintain abstinence compared with abstainers was 5.32, while the relative mortality risks for the group abusing alcohol and for the group suffering alcohol dependence were 2.23 and 2.56, respectively. These results suggest that relapse defined as failure to maintain abstinence predicts a higher relative mortality risk than do criteria defining in terms of alcohol abuse and alcohol dependence.
Predicting Problem Drinking: A Test of an Interactive Social Learning Model
Authors: Curran Geoffrey M. , White Helene Raskin , Hansell Stephen
Published: May 2006

This study tested a social learning model and explored the direct and interactive relationships between personality and environment in predicting problem alcohol use. We used longitudinal data from a nonclinical sample of males and females first tested in adolescence and followed into young adulthood. Hierarchial regression analyses were used to test main effects and interaction models. The cross‐sectional data supported an interactive social learning model. Both personality and environment variables significantly predicted problem drinking. Two interactions between heavy drinking peer groups and personality variables were significant. Contrary to our hypothesis, the direction of the interaction was negative. In contrast, the longitudinal analyses did not provide strong support for our interactive model. Personality variables were significant predictors longitudinally, but in only one analysis did an environment variable significantly predict problem drinking. Furthermore, none of the interactions was significant predictors over time. Overall, the findings suggest that social learning models based on the interaction of personality and environmental influences may be more appropriate for predicting concurrent, as opposed to future problems, and that future research should include person‐environment interactions. In addition, cultural tolerance of heavy drinking may be an important determinant of the role of psychological vulnerability in the development of problem drinking.
Combination of Naltrexone and Fluoxetine on Rats’Propensity to Take Alcoholic Beverage
Authors: Gardell Luis R. , Whalen Christine A. , Chattophadyay Soma , Cavallaro Colleen A. , Hubbell Christopher L. , Reid Larry D.
Published: May 2006

Naltrexone (NTX) and fluoxetine (FLU) are useful for treating alcoholism and depression, respectively. Furthermore, these afflictions co‐vary. Given the possibility that people might be prescribed NTX and FLU concurrently, we assessed the effects of these two agents on rats’propensity to drink an alcoholic beverage. Rats were given 66 days of access to a sweetened alcoholic beverage and water for 2 hr daily. At first, they took little ethanol, but after 20 days, they took on average 2.0 to 2.5 g/kg of ethanol, daily during the 2‐hr session. They also took sufficient water to maintain their health. After 30 days, they were divided into four groups to receive, 30 min before the drinking session, 1 of 4 different kinds of injections. For the next 20 days, one group received placebo daily. Another group received 5 mg/kg of NTX dairy and another 5 mg/kg of FLU dairy. The fourth group received both 5 mg/kg of NTX and 5 mg/kg of FLU dairy. After 20 days, the doses of NTX and FLU were doubled across an additional 10 days. Both NTX and FLU reduced rats’intake of alcoholic beverage. The combinations of NTX and FLU, however, were no more effective in reducing rats’intake of alcoholic beverage than either alone. Also, the small dose of NTX seemed to lose its effectiveness with repeated administrations. A second experiment con‐firmed the conclusion that small doses of NTX lose their effectiveness in suppressing intake of alcoholic beverage across repeated administrations. In summary, data provide no support for the idea that FLU and NTX would act synergistically to reduce propensity to take alcoholic beverages.
Effects of Chronic Ethanol on the Mobilization of Arachidonate and Docosahexaenoate Stimulated by the Type 2A Serotonin Receptor Agonist (±)‐2,5‐Dimethoxy‐4‐iodoamphetamine Hydrochloride in C6 Glioma Cells
Authors: Garcia Martha C. , Kim Kun‐Yang , Hough Christopher , Kim Hee‐Yong
Published: May 2006

We studied the effects of chronic ethanol exposure on the mobilization of polyunsaturated fatty acids stimulated by activation of the type 2A serotonin receptor in C6 glioma cells. In our in vitro model, we prelabeled cells with [3H]arachidonate and [14C]docosahexaenoate and subsequently stimulated with the type 2A serotonin receptor agonist (±)‐2,5‐dimethoxy‐4‐iodoamphetamine hydrochloride. In as early as 10 days of exposure to 20 or 50 mM ethanol, the (±)‐2,5‐dimethoxy‐4‐iodoamphetamine hydrochloride‐stimulated mobilization of [3H]arachidonic acid ([3H]AA) and [14C]docosahexaenoic acid ([14C]DHA) was significantly inhibited, and this inhibition was accompanied by decreased mobilization of intracellular [Ca2+]i. Exposure to ethanol did not alter significantly the release of [3H]AA and [14C]DHA stimulated by the calcium ionophore A23187 nor the incorporation of [3H]AA and [14C]DHA into cellular lipids. Decreased mobilization of polyunsaturated fatty acids and calcium in astroglia may contribute to neurotoxicity caused by chronic ethanol exposure.
Ethanol Increases Surface‐Localized Fibrinolytic Activity in Cultured Endothelial Cells
Authors: Aikens Michael L. , Benza Raymond L. , Grenett Hernan E. , Tabengwa Edlue M. , Davis Glenda C. , Demissie Sossiena , Booyse Francois M.
Published: May 2006

Epidemiological studies demonstrated a positive association between moderate alcohol consumption and reduced cardiovascular mortality that may be mediated, in part, through increased fibrinolysis. These studies were conducted to determine whether low concentrations of alcohol (0.025 to 0.1%, v/v) directly affected the surface‐localized versus secreted/solution phase fibrinolytic activity in live cultured endothelial cell (EC) types. Confluent live cultured ECs [human umbilical vein ECs (HUVECs), human saphenous vein ECs (HSVECs), and porcine aortic ECs (PAECs)] were preincubated (0 to 20 min, 4°C) in the absence or presence of varying concentrations of alcohol (0 to 0.1%, v/v), in the presence of saturating levels of 125I‐labeled Glu‐plasminogen (2 μM) and 125I‐plasmin Mr 20‐kDa light‐chain formation quantitated by phosphorimaging autoradiography analysis. Endogenous plasminogen activator (PA)‐mediated fibrinolytic activity was time‐ and dose‐dependent; reached a maximum ˜5‐ to 10‐fold increase at 0.05% alcohol in HUVECs, HSVECs, and PAECs; was completely inhibited by anti‐t‐PA IgG in HUVECs; and partially inhibited by both anti‐t‐PA (˜40%) and anti‐u‐PA IgG (˜60%) in HSVECs. Complete inhibition of alcohol‐induced (0.05%) fibrinolytic activity in cultured HUVECs by 2 mM tranexamic acid (an antagonist of plasminogen binding) indicated that the increased fibrinolytic activity was receptor‐bound and localized to the EC surface, rather than present in or secreted into the medium (solution phase). Finally, the alcohol‐induced increased fibrinolytic activity in cultured HUVECs returned to essentially normal control levels in ˜1 hr. These studies have demonstrated a direct effect of low alcohol on EC fibrinolytic activity that may contribute, in part, to the decreased risk for thrombosis, coronary artery disease, and myocardial infarction associated with moderate alcohol consumption.
Chronic Ethanol Exposure Alters Leukocyte Subsets in Repopulating Spleens, But Does Not Alter Negative Selection in Thymuses of Sublethally Irradiated Mice
Authors: Livant Emily J. , Welles Elizabeth G. , Ewald Sandra J.
Published: May 2006

Results from previous in vitro experiments in this laboratory suggested that ethanol may affect selection processes in the thymus. To determine whether ethanol allows escape of potentially autoreactive T‐cell clones from negative selection, we fed ethanol to sublethally irradiated, young, adult C57BR mice during the time of thymic and splenic repopulation as a new model of human third trimester fetal alcohol exposure. The mice received a whole‐body, sublethal dose (6 Gy) of gamma irradiation at 5 to 6 weeks of age. Feeding of a liquid diet providing 25% of calories as ethanol (EDC) or an isocaloric control liquid diet was begun 3 days after irradiation and was continued for 5 weeks. Each EDC mouse had 2 weight‐ and age‐matched controls, 1 pair‐fed (PF), and 1 fed ad libitum (AD LIB). Average blood alcohol concentrations (90 to 440 mg/100 ml) were higher than those reported previously for neonatal mice exposed to ethanol through lactation. At 5 weeks after irradiation, the EDC mice had lower total thymocyte numbers (p < 0.05) and a higher proportion of CD4‐ CD8‐ thymocytes than either the PF or AD LIB mice (p < 0.05), which is consistent with findings using in utero models of ethanol exposure. Ethanol exposure also altered the proportion of leukocyte subsets in repopulating spleens. B cells were the most sensitive to the detrimental effects of ethanol and, as a percentage of total nucleated cells in the spleen, B cells were decreased in the EDC group, compared with both the PF and AD LIB groups (p < 0.05). C57BR mice normally delete by negative selection thymocytes bearing vβ17+ T‐cell receptors. There was no discernible effect of ethanol exposure during thymic and splenic repopulation on the expression of Vβ17a on thymocytes and splenic T lymphocytes, indicating that ethanol does not affect negative selection.
Alcohol‐Induced Expression of the CD14 Endotoxin Receptor Protein in Rat Kupffer Cells
Authors: Järveläinen Harri A. , Oinonen Teija , Lindros Kai O.
Published: May 2006

Gut‐derived endotoxins (lipopolysaccharide, LPS) are believed to contribute to alcohol‐induced liver disease (ALD) by stimulating Kupffer cells, the resident liver macrophages, to release proinflammatory cytokines. This activation is largely mediated by CD14, a high‐affinity membrane‐anchored receptor for LPS. We observed, by chemiluminescence‐enhanced detection, an increase in immunoreactive CD14 protein in Kupffer cells isolated from rats treated with ethanol for 2 weeks. Immunocytofluorescence experiments confirmed that this increase was confined to the membranes of Kupffer cells from the alcohol‐treated rats. The increase was regulated pretranslationally: a 3‐fold elevation (p< 0.01) in the hepatic level of CD14 mRNA was observed. The marked increase in CD14 expression suggests a new mechanism by which alcohol increases the LPS‐mediated cytokine signaling by the liver macrophages, thus promoting the interaction between alcohol and endotoxins in the development of liver damage.
Performance by Adult Rats in Sensory‐Mediated Radial Arm Maze Tasks Is Not Impaired and May Be Transiently Enhanced by Chronic Exposure to Ethanol
Authors: Steigerwald Elizabeth S. , Miller Michael W.
Published: May 2006

The effect of chronic exposure to ethanol on learning and memory was assessed in rats. Adult rats were fed a liquid diet containing ethanol (Et) ad libitum, pair‐fed an isocaloric/isonutritious control diet (Ct), or fed lab chow and water (Ch) ad libitum. These diets were provided for as long as 28 weeks prior to testing and during the period of behavioral testing. Rats were required to learn a radial arm maze task that depended upon extramaze cues (visual and spatial) or intramaze cues (various odors). Each rat was tested twice daily for 26 days. The number of reference and working memory errors and the time required to successfully navigate the maze (latency) were recorded. Et‐fed rats consistently performed the same or better than the Ct‐fed or Ch‐fed rats. The enhanced behavior was task‐specific and transient. The enhancement was observed in rats after 14 or 20 weeks of ethanol exposure, but not in rats exposed to ethanol for 28 weeks. Et‐fed rats performed significantly better on the reference memory task in the odor condition of the maze task, whereas their performance in the spatial task was similar to that for the two control groups. Thus, after an extended period of exposure (e.g., 28 weeks), ethanol had no effect on learning and memory. On the other hand, under certain conditions (14 or 20 weeks), ethanol transiently enhanced memory.
Effect of Prenatal Ethanol Exposure on Fetal Calcium Metabolism
Authors: Keiver K. , Ellis L. , Anzarut A. , Weinberg J.
Published: May 2006

Alcohol consumption has adverse effects on both adult and developing bone. The mechanisms by which alcohol affects bone, however, are unknown. This study examined the possibility that maternal alcohol consumption may affect fetal bone development by altering fetal levels of parathyroid hormone (PTH), 1,25(OH)2D, or calcitonin (hormones that regulate calcium (Ca) and bone metabolism in the adult animal). Female Sprague‐Dawley rats were bred and divided into three groups: 1 group was fed lab chow ad libitum (Control; C) and the other 2 groups received a liquid diet with (Ethanol; E) or without (Pair‐fed; PF) ethanol. Blood from dams and fetuses was collected on day 21 of gestation, and selected fetuses were stained for determination of the degree of bone ossification. Mean fetal body weight and fetal skeletal ossification were reduced in the E compared with PF and C groups. Total Ca levels in fetal serum, however, showed a trend to be increased in E compared with PF and C fetuses, and no significant group differences were found in fetal serum levels of albumin, PTH, or calcitonin. Serum levels of 25‐OH‐D and 1,25(OH)2D were significantly decreased in E and PF, compared with C fetuses. Total Ca levels in maternal serum did not vary with the group; however, serum albumin levels were higher in E, compared with PF and C dams, suggesting that serum ionic Ca levels may have been reduced in the E dams. Serum levels of 25‐OH‐D were reduced in the E, compared with PF and C dams, whereas levels of 1,25(OH)2D were elevated. PTH levels did not vary among groups. Interestingly, serum calcitonin levels were elevated in the E, compared with PF and C, dams. These results indicate that the effects of ethanol on fetal bone development do not appear to be related to alterations in fetal serum levels of PTH, 1,25(OH)2D, or calcitonin. Maternal ethanol consumption, however, results in reduced appetite and a decrease in dietary Ca intake. Despite the reduced Ca intake, the ability of the dam to maintain Ca homeostasis appeared intact, although this may be dependent on the duration of ethanol consumption.
Zinc Administration Improves Gastric Alcohol Dehydrogenase Activity and First‐Pass Metabolism of Ethanol in Alcohol‐Fed Rats
Authors: Caballería Joan , Giménez América , Andreu Hernán , Deulofeu Ramón , Parés Albert , Caballería Llorenç , Ballesta Antonio M. , Rodés Joan
Published: May 2006

The effects of zinc on first‐pass metabolism (FPM) of ethanol and gastric and hepatic alcohol dehydrogenase (ADH) activities have been investigated in two groups of male Wistar rats fed a liquid ethanol diet with normal zinc content (7.6 mg/liter), or zinc supplemented (76 mg/liter), for 21 days, and in two pair‐fed groups receiving the same diets without ethanol. Alcoholic rats with normal dietary zinc had lower FPM (1.64 ± 0.25 vs. 2.43 ± 0.20 mM ± hr, p < 0.05) and gastric ADH activity (184 ± 7 vs. 335 ± 41 μmol/min/mg protein, p < 0.01) than control rats. Zinc supplementation did not produce any change in FPM or in gastric ADH activity in control rats. By contrast, in alcoholic rats, the zinc supplement increased gastric ADH activity (247 ± 31 vs. 184 ± 7 μmol/min/mg protein, p < 0.05) and decreased the areas under the curve of blood ethanol concentrations after the intragastric administration of 0.25 g/kg of body weight of ethanol (0.78 ± 0.07 vs. 1.71 ± 0.24 mM ± hr, p < 0.05), thereby increasing the FPM. In conclusion, in alcohol‐fed rats, the administration of zinc supplements restores gastric ADH activity and improves the FPM of ethanol. These effects may be one of the mechanisms in which zinc has a beneficial role in preventing the development of alcoholic hepatic lesions.
Involvement of Dopamine D2 Receptors in the Suppressive Effect of the Thyrotropin‐Releasing Hormone Analog TA‐0910 on Alcohol Intake in Alcohol‐Preferring Rats
Authors: Mason George A. , Rezvani Amir H. , Overstreet David H. , Hamedi Mani , Walker Cheryl H. , Yang Ying , Garbutt James C.
Published: May 2006

Pharmacological experiments were conducted to determine the neuronal mechanisms involved in the suppressive effects of the thyrotropin‐releasing hormone analog TA‐0910 on alcohol intake in alcohol‐preferring (P) rats. We previously reported that single intraperitoneal injections of TA‐0910 dose‐dependently reduced alcohol intake in P rats without altering fluid or total calorie intake; however, after several consecutive, once‐daily injections, P rats developed tolerance to the suppressive effects of TA‐0910 on alcohol intake and cross‐tolerance to like effects of the dopamine D2 agonist bromocriptine, but not to like effects of the serotonin uptake inhibitor fluoxetine. In the present study, rats were injected with vehicle or different doses of the D2 antagonist s (–)‐eticlopride (0.01 to 0.05 mg/kg) or the D1 antagonist R(+)‐SCH23390 (0.1 to 0.5 mg/kg) and 20 min later with TA‐0910 (0.75 mg/kg). Alcohol and water intakes were measured at 2,4,6, and 24 hr, and food was measured every 24 hr. Both s(–)‐eticlopride and R(+)‐SCH23390 produced modest reductions in alcohol intake alone; however, only s(–)‐eticlopride antagonized the suppressive effect of TA‐0910 on alcohol intake. In related experiments, it was confirmed that the dopamine D3 agonist 7‐hydroxy‐N,N‐di‐n‐propyl‐2‐aminotetralin reduced alcohol intake in P rats, and it was found that tolerance to this effect did not develop during or after seven consecutive once‐daily injections. Furthermore, this effect of 7‐hydroxy‐N,N‐di‐n‐propyl‐2‐aminotetralin was not diminished in rats made tolerant to the effect of TA‐0910 on alcohol intake. These data, those of previous studies, and recent preliminary findings support involvement of dopamine D2, but not D1 or D3 receptors in mediating the suppressive effect of TA‐0910 on alcohol intake of P rats.
A Study Examining Intravenous Ethanol‐Conditioned Place Preference in C57BL/6J Mice
Authors: Kelley Brian M. , Bandy Angela‐Leigh E. , Middaugh Lawrence D.
Published: May 2006

The reinforcing effects of intravenous (IV) ethanol were examined in C57BL/6J (C57) mice with a conditioned‐place‐preference (CPP) paradigm. Before CPP testing, adult mice underwent jugular catheterization. On the following day, subjects were acclimated to a two‐compartment CPP chamber. A 15‐min nondrug pretest was conducted to determine compartment preference. For the treatment group, IV ethanol [30% (v/v), 3.4 μl/min, 25 min] was paired with the nonpreferred compartment, whereas IV saline was paired with the preferred compartment. The control group received IV saline in both compartments. Two conditioning sessions were conducted per day (0900 and 1500), and the order of the infusions was counterbalanced across subjects. The drug‐free posttest was identical to the pretest, except that it occurred on the day after the final drug/compartment pairing. The entire procedure required 6 days. After just two pairings with ethanol, with a cumulative ethanol dose of only 0.82 g/kg/day, significant CPP was noted in the treatment group, whereas no change in compartment preference was noted for the control group. A separate group of C57 mice were trained to discriminate intraperitoneal ethanol (1.5 g/kg) from saline using a two‐lever drug discrimination paradigm. After training was complete, these mice also underwent jugular catheterization. Substitution testing was conducted with IV ethanol [30% (v/v), 6.4 μl/min, 12 min] and saline. The results indicate that the subjective effects of ethanol did not differ according to the route of administration. Together, these experiments provide evidence that ethanol is rewarding for C57 mice, as indexed by ethanol CPP, and that the subjective effects of intravenously and intraperitoneally administered ethanol are similar.
Alcoholic Myopathy: Evaluation with Magnetic Resonance Imaging—A Case Study
Authors: Kishi Toshiro , Kitani Mitsuhiro , Uegaki Jun , Naganuma Rokuichi
Published: May 2006

Magnetic resonance imaging (MRI) of thigh and leg muscles was performed in a patient with alcoholic myopathy showing myalgia, hypercreatine kinasemia, and hypermyoglobinemia. High signal intensities in both T1‐ and T2‐weighted images were widely distributed in the affected muscle groups, which most likely reflected lipid accumulation. Although he had hypermyoglobinemia, MRI and muscle biopsy did not show findings of rhabdomyolysis, such as necrosis, regeneration, and edema. We suggest that the high signal intensities in this case may have indicated „prerhabdomyolysis” related to alcohol abuse and that muscle MRI is useful in the evaluation of alcoholic myopathy, mainly predicting the onset of rhabdomyolysis.
Ethanol Action on Neural Networks Studied with Multineuron Recording in Freely Moving Animals
Authors: Woodward Donald J. , Janak Patricia H. , Chang Jing‐Yu
Published: May 2006

The advent of new chronic multineuron recording techniques for examining neural activity in behaving animals has initiated a new phase in the analysis of the neuronal mechanisms that underlie ethanol and other drug self‐administration. The technique allows for the simultaneous recording of groups of individual neurons in one or more brain regions during ongoing behavior; therefore, the spatiotemporal patterns of neuronal activity during specific behavioral events can be determined. We have successfully applied this technique to rat models of cocaine and heroin self‐administration. Recently, using rats, we have been able to record from neurons in areas of the mesocorticolimbic circuit during ethanol‐reinforced operant responding. In this review, we describe the current and future application of this new behavioral neurophysiology to the investigation of the neurobiology of alcohol addiction.
Monitoring Relapse Drinking During Disulfiram Therapy by Assay of Urinary 5‐Hydroxytryptophol
Authors: Helander Anders
Published: May 2006

Screening for recent alcohol use by testing urine for the ratio of 5‐hydroxytryptophol (5HTOL) to 5‐hydroxyindole‐3‐acetic acid (5HIAA) was performed in 10 methadone patients on disulfiram (Antabuse) maintenance therapy in an outpatient setting. Apart from alcohol ingestion, treatment with aldehyde dehydrogenase inhibitors such as disulfiram is the only known cause of an abnormally high 5HTOL/5HIAA ratio. After introduction of drug therapy, increased ratios were observed in all patients. The new higher level reached was relatively stable over time within the same patient but variable between patients. Four patients continued to consume alcohol, as evidenced by 5HTOL/5HIAA ratios well above the new individual plateau, while still taking 400 mg disulfiram 3 times per week under strict supervision. To try to achieve sobriety in two patients who drank frequently while on therapy, the disulfiram dose was doubled. Continued testing demonstrated this to increase the 5HTOL/5HIAA steady‐state level further, and the absence of extreme values above this new baseline level indicated adherence to abstinence with possibly one single relapse. When disulfiram administration was discontinued, as planned, by five of the patients, four of them returned to drinking very soon. The present results show that during disulfiram maintenance the continuous inhibition of aldehyde dehydrogenase produces a new higher and dose‐related 5HTOL to 5HIAA steady state level in urine, but relapse to drinking will still lead to further increased 5HTOL/5HIAA ratios. It is also suggested that an individual dose‐titration regimen, whereby the disulfiram dose is raised gradually until 5HTOL/5HIAA testing indicates sobriety, will improve therapeutic effectiveness.