bluuebunny/crossref_metadata_2025 huggingface dataset:
Validity of the Obsessive Compulsive Drinking Scale (OCDS): Does Craving Predict Drinking Behavior?
Authors: Kranzler Henry R. , Mulgrew Christine L. , Modesto‐Lowe Vania , Burleson Joseph A.
Published: May 2006

Objective: The Obsessive Compulsive Drinking Scale (OCDS), a 14‐item, self‐report questionnaire, was developed to measure alcohol‐related craving. The OCDS may provide a measure of the state of illness among alcohol‐dependent individuals and may have value in predicting subsequent drinking behavior. The present study was conducted to evaluate the factor structure and the concurrent, construct, and predictive validity of the OCDS. Methods: Data on desire to drink and on drinking behavior were obtained from 127 alcohol‐dependent subjects who participated in a 12‐week outpatient pharmacotherapy trial and a 3‐month posttreatment follow‐up. Results: Principal components analysis of the OCDS indicated that three factors best described its structure: obsessions, drinking control and consequences, and alcohol consumption. Data also supported the concurrent and discriminant validity of the OCDS. However, the OCDS total score showed limited validity in predicting drinking during a posttreatment follow‐up period. Furthermore, the only empirically derived factor that predicted drinking during this period was the alcohol consumption factor. Conclusions: As might be expected, the OCDS questions on drinking behavior predict subsequent drinking behavior. However, the instrument does not appear to provide a general measure of alcohol‐related illness. The utility of the OCDS in studies of alcoholism treatment outcome requires clearer definition.
Do Patients with Alcohol Dependence Use More Services? A Comparative Analysis with other Chronic Disorders
Authors: Fortney John C. , Booth Brenda M. , Curran Geoffrey M.
Published: May 2006

Objective: The primary purpose of this research was to compare the service use of patients diagnosed with alcohol dependence to the service use of patients diagnosed with other chronic illnesses. The secondary purpose was to determine the impact of comorbid alcoholism on the service use of patients with chronic illnesses. Methods: The sample included 67,878 veterans diagnosed with alcohol dependence, depression, or diabetes who were treated by the Department of Veterans Affairs in 1993. The number of inpatient days and outpatient visits over a 4‐year period (1991 to 1995) were compared using regression models to control for differences in casemix. Results: Controlling for casemix, patients treated for alcohol dependence had significantly fewer outpatient visits than patients treated for either depression or diabetes. Patients treated for alcohol dependence also had significantly fewer inpatient days than patients treated for depression, but significantly more inpatient days than patients treated for diabetes. Comorbid alcoholism was prevalent among patients treated for depression and diabetes. Comorbid alcoholism increased the number of inpatient days for patients treated for depression or diabetes and increased the number outpatient visits for patients with depression. However, comorbid alcoholism decreased the number of outpatient visits for patients treated for diabetes. Conclusions: Results suggest that patients with alcohol use disorders should not be singled out as being more costly to treat than patients with other chronic illnesses. These findings are in stark contrast to those from studies comparing individuals with alcohol use disorders to relatively healthy individuals sampled from at‐risk populations. Key Words: Alcoholism, Comorbidity, Service Utilization.
Sleep‐Disordered Breathing in Alcoholics
Authors: Aldrich Michael S. , Brower Kirk J. , Hall Janette M.
Published: May 2006

Sleep apnea and related disorders contribute to disturbed sleep in abstinent alcoholics. In an earlier report from our group, sleep‐disordered breathing was common and increased with age in a cohort of 75 abstinent alcoholics. We now report an extension of the previous work that includes studies of an additional 103 abstinent alcoholics undergoing treatment for alcoholism (total sample = 188) and a comparison group of 87 normal subjects. The presence and severity of sleep‐disordered breathing was assessed with polysomnography. Among the alcoholics, sleep‐disordered breathing (defined as 10 or more apneas plus hypopneas per hour of sleep) was present in 3% of 91 subjects under age 40, 17% of 83 subjects age 40 to 59, and 50% of 14 subjects age 60 or over. Subjects with sleep‐disordered breathing were more likely to be male and had more severe sleep disruption and nocturnal hypoxemia and more complaints related to daytime sleepiness than subjects without sleep‐disordered breathing. In a multiple linear regression analysis, age and body mass index were significant predictors of the presence of sleep‐disordered breathing, whereas smoking history and duration of heavy drinking were not predictors after controlling for the effects of age and body mass index. Our findings suggest that sleep‐disordered breathing contributes significantly to sleep disturbance in a substantial proportion of older alcoholics and that symptomatic sleep‐disordered breathing increases with age in alcoholics. Sleep‐disordered breathing, when combined with existing cardiovascular risk factors, may contribute to adverse health consequences in alcoholics.
Implications of Depression on Outcome from Alcohol Dependence: A 3‐Year Prospective Follow‐up
Authors: Hodgins David C. , El‐Guebaly Nady , Armstrong Susan , Dufour Marie
Published: May 2006

The prognostic implications of comorbid depression for outcome from alcohol dependence are unclear. It has been suggested that drinking may represent self‐medication of depressive disorders and, alternatively, that the pharmacological properties of alcohol induce episodes of depression. In the present study, these questions were investigated by following 84 alcohol‐dependent individuals seeking treatment (34% women) for 3 years in a naturalistic, prospective design. During the follow‐up period, depressive and drinking outcomes were significantly related. However, there was no evidence that drinking reliably preceded depressive episodes or that depressive episodes precipitated heavy drinking. Despite their high co‐occurrence, this study did not find evidence of a strong, direct causal relationship between these phenomena.
Phases of Alcohol Problem Prevention Research
Authors: Holder Harold , Flay Brian , Howard Jan , Boyd Gayle , Voas Robert , Grossman Michael
Published: May 2006

We build on precedents from other health research to present a phases model of research for alcohol problem prevention that accommodates the special characteristics of this research. We propose a five‐level model, in which research moves along a series of relevant continua: from basic to more and more applied research; from descriptive hypothesis‐generating pilot studies to full‐fledged, methodologically sophisticated, hypothesis‐testing studies; from smaller to larger samples for testing; from greater to lesser control of experimental conditions; from more artificial “laboratory” environments to real‐world geographically defined communities; from testing the effects of single prevention strategies to more complex studies of multiple strategies integrated into intervention systems; and from research‐driven outcome studies to “demonstration” projects that evaluate the capacity of various types of communities to implement prevention programs based on prior evaluations. The five phases of research are: (1) foundational research to define and determine the prevalence of specific alcohol‐involved problems, establish causal factors and processes that yield the specific problems or increase the risk of a problem, and provide the foundations for the development of effective prevention interventions; (2) developmental (preliminary effectiveness) studies to develop and test the likely effectiveness, safety, and costs of new interventions or to assess the effectiveness, safety, and costs of an existing intervention; (3) efficacy studies to determine the effects, safety, and costs of an intervention under optimal conditions of implementation (or availability or enforcement) and acceptance (or adoption at the community, organizational, or group level; or participation, compliance, or adherence at the individual level); (4) effectiveness studies of the real‐world effectiveness of preventive interventions with purposeful or natural variation in implementation and acceptance;, and (5) demonstration studies of the effects of interventions when widely disseminated. The proposed phases model for alcohol problem prevention research presented herein differs in significant ways from the models established by other National Institutes of Health agencies. Greater emphasis is placed on natural experiments, on methods development along the whole research continuum, on collapsing or combining research phases when appropriate, on recognizing the critical importance of behavioral parameters early as well as late in the research sequence, and on extending the research continuum to embrace diffusion and dissemination (i.e., technology transfer) studies. We also include examples of phased research in existing alcohol studies and a discussion of relevant issues, including cost, special populations, methods, and dissemination. If systematically followed, this model has the potential to contribute to wider testing and dissemination of prevention interventions of known effectiveness.
Size of Mamillary Bodies in Health and Disease: Useful Measurements in Neuroradiological Diagnosis of Wernicke's Encephalopathy
Authors: Sheedy Donna , Lara Ana , Garrick Therese , Harper Clive
Published: May 2006

Background: As the resolution of noninvasive neuroimaging techniques improves, small structures such as the mamillary bodies can be visualized and measured. The mamillary bodies are pathologically small in a number of neurological disorders, the most common and important is chronic Wernicke's encephalopathy (WE), or the Wernicke‐Korsakoff syndrome (WKS), as it is often called. This disorder is caused by vitamin B1 deficiency (thiamin) and is seen most commonly in people who drink excessive amounts of alcohol. The disorder is easily preventable by using oral or parenteral thiamin. The aim of this study was to establish a range for the volume of the mamillary bodies in normal and in various disease states, particularly WE.

Methods: Brains were taken from 2212 sequential autopsies performed at the New South Wales Institute of Forensic Medicine from 1996 through 1997. After fixation in 10% formalin, the brains were sectioned coronally and a block containing the mamillary bodies was dissected. The maximum vertical and transverse diameters of the mamillary bodies were measured using Mitutoyo vernier callipers and the volume calculated (V = 413 (a2b), where a and b are the vertical and transverse radii, respectively).

Results: There were 164 cases with significant pathological changes in the mamillary bodies. These included cases with WE (25), Alzheimer's disease (10), infarction (11), and trauma (55). All but two of the WE cases were chronic or acute on chronic. The mean volume of the mamillary bodies was reduced by 60% in cases with chronic WE and by 25% in cases with Alzheimer's disease. In normal cases, there was a significant age‐related reduction in volume, and males had larger mamillary bodies than females. Cases with alcoholic cirrhosis of the liver had normal mamillary body volumes.

Conclusions: There is a gross shrinkage of the mamillary bodies in cases of chronic WE, but clinicians need to consider other diagnoses, such as Alzheimer's disease, which can also result in shrinkage. These quantitative data will be helpful in the neuroradiological diagnosis of some of these disorders, particularly WE.
In Vivo Mammillary Body Volume Deficits in Amnesic and Nonamnesic Alcoholics
Authors: Sullivan E. V. , Lane B. , Deshmukh A. , Rosenbloom M. J. , Desmond J. E. , Lim K. O. , Pfefferbaum A.
Published: May 2006

Background: Neuropathological studies use the presence of mammillary body (MB) pathology as a cardi nal, diagnostic feature of Wernicke's encephalopathy (WE) in neuropsychiatric diseases, most notably alcoholism. Although Korsakoff's Syndrome (KS), which is marked behaviorally by dense global amnesia, is a typical sequela of WE, it remains controversial whether these two conditions necessarily co‐occur and whether MB pathology is therefore a diagnostic requisite for KS.

Methods: We investigated these issues by examining, in vivo, 24 nonamnesic alcoholics (ALC), 5 amnesic alcoholics (KS), and 51 normal controls with three‐dimensional MRI and memory testing. MB volume was dctermined from successive, 1 mm thick slices.

Results: The ALC group had significantly smaller MB volumes bilaterally (mean = 54.5 ± 22.0 mm3) than controls (mean = 66.3 ± 17.1 mm3), and the KS group had even smaller MB volumes than the ALC group (mean = 20.7 ± 14.8 mm3). Only 2 ALC patients met historical clinical criteria for past WE, and their MB volumes were well within range of the remaining 22 ALC patients. Although all five KS patients met historical clinical criteria for WE, three KS did not have accompanying dementia and had the same degree of MB volume loss as the ALC; the remaining two KS had accompanying dementia and MB volumes half the volume of the ALC group and of KS patients without dementia.

Conclusions: These findings provide volumetric in vivo evidence that: (1) MB volume deficits do occur in alcoholics without amnesia, although these deficits are not present in all such alcoholics; (2) greater MB volume deficits are present in alcoholics with clinically detectable amnesia or dementia; (3) MB shrinkage is related to severity of cognitive and memory dysfunction, which suggests a continuum of MB pathology in chronic alcoholism to KS; and (4) the presence of WE in all of the KS patients and in the two ALC patients with the greatest long‐term declarative memory deficit supports the possibility of an additional and unique pathology distinguishing nonamnesic and amnesic alcoholism.
Purkinje Cell Vulnerability to Developmental Ethanol Exposure in the Rat Cerebellum
Authors: Pierce Dwight R. , Williams D. Keith , Light Kim Edward
Published: May 2006

Background: Ethanol cxposure is a consistent and reliable producer of neuronal toxicity, especially during periods of enhanced neuronal vulnerability. For rat cerebellar Purkinje cells, the postnatal period during the time of the brain growth spurt exhibits the greatest vulnerability to ethanol. Analyses of studies completed over more than 20 years provides sufficient detail to allow for the determination of the specific vulnerable window for ethanol‐induced loss of Purkinje cells.

Methods: Data reporting Purkinje cell counts after ethanol exposure were compiled from 18 studies published since 1975. We conducted linear regression analysis between peak blood ethanol concentration (BEC) and percent reduction in Purkinje cells for the following individual postnatal (PN) days: PN4, PN5, PN6, PN7, and PN8 or beyond (+). The slope of the regression and the coefficients of determination (r2) were the primary factors of interest. Analysis of variance of the regressions was conducted to identify whether the slopes were significantly different from zero, or from each other.

Results: Exposures involving the PN4‐6 period demonstrated the greatest significance in the relationship between BEC and reduction of Purkinje cell number. No significant differences were identified between different ethanol exposure techniques or for different Purkinje cell counting techniques. In addition, the initial day of exposure and the duration of exposure were not identified as critical variables.

Conclusions: The literature database, developed over the past 20 years is clear in its direction that studies designed to identify the ethanol‐specific mechanisms of Purkinje cell death are best designed to involve ethanol exposure during the vulnerable window of postnatal days 4‐6.
Ethanol Feeding Selectively Impairs the Spreading of Rat Perivenous Hepatocytes on Extracellular Matrix Substrates
Authors: Tuma Dean J. , Smith Teresa E. , Schaffert Courtney S. , Kharbanda Kusum K. , Sorrell Michael F.
Published: May 2006

Background: Hepatocytes require attachment and subsequent spreading on an extracellular matrix for their proper growth, function and survival. Our previous studies have shown that ethanol feeding selectively impairs perivenule hepatocyte attachment to various extracellular matrices. This study was undertaken to determine whether zonal differences in hepatocyte spreading in response to ethanol feeding occurs and to ascertain the influence of ethanol consumption on the zonal expression of the β1, subunit of integrins, which are the major surface receptors responsible for matrix binding and subsequent interactions.

Methods: Hepatocytes from the perivenous and periportal regions of the liver were isolated by digitonin/collagenase perfusion from rats that were pair‐fed for 2 to 3 weeks with a liquid diet containing either ethanol or isocaloric carbohydrate. The ability of perivenous and periportal hepatocytes to spread on plates coated with either type IV collagen, laminin, fibronectin or polylysine was determined. In addition, the isolated cells were used for the analysis of total cellular and surface β1integrin expression.

Results: With all of the matrix substrates tested, the spreading of perivenous hepatocytes isolated from the ethanol‐fed animals was markedly impaired, while the spreading of periportal hepatocytes was essentially unaffected by ethanol feeding. Both the total cellular as well as the surface expression of the β1integrin subunit in perivenous cells from the ethanol‐fed rats were significantly higher than from the perivenous control cells, whereas the total and surface expression of the β1integrin in periportal cells isolated from ethanol‐fed and control rats were not significantly different.

Conclusions: The results indicated that in addition to impairing hepatocyte attachment, ethanol feeding also impairs another critical step of the adhesion process, that of hepatocyte spreading on extracellular matrix substrates. This defect occurred preferentially in perivenous cells and not periportal cells and was associated with an increase in β1integrin expression, suggesting that a compensatory mechanism occurs as an attempt by the perivenous cells to overcome impaired cell‐matrix interactions caused by ethanol. Overall, these alterations in extracellular matrix‐hepatocyte interactions could lead to alterations of hepatocyte structure and function and potentially play a role in alcoholic liver injury.
Effects of Benzodiazepines on Acute and Chronic Ethanol‐Induced Nociception in Rats
Authors: Gatch Michael B.
Published: May 2006

Background

: Acute and chronic ethanol produces antinociception, and ethanol withdrawal induces hyperalgesia.

Methods

: A radiant heat tail‐flick assay was used to assess the effects of benzodiazepine ligands on ethanol‐induced changes in nociception in rats. Acute activity of cumulative doses of ethanol (0.5–2.0 g/kg) and diazepam (0.1–10 mgkg), a benzodiazepine agonist, was tested alone and after pretreatment with flumazenil (1.0–10 mg/kg), a benzodiazepine antagonist. Chronic effects of ethanol were tested in three groups of rats that received a liquid diet for 10 days. One group received ethanol alone; one group received ethanol and twice‐daily injections of flumazenil (10 mg/kg); and one received a dextrin control diet. Acute withdrawal was tested at 12 hr after removal of the liquid diet. Effects of cumulative doses of diazepam (1.0–10 mg/kg) were tested during withdrawal (12 hr) in the ethanol‐alone group.

Results

: Acute doses of ethanol produced a small but significant degree of antinociception, which was fully suppressed by flumazenil. Acute doses of diazepam did not produce antinociception. Chronic exposure to ethanol produced antinociception on days 2 through 8. Tolerance developed by day 10, and hyperalgesia was seen 12 hr after removal of ethanol. Administration of diazepam or ethanol during withdrawal reversed the hyperalgesia induced by ethanol withdrawal. However, flumazenil (10–50 mg/kg) failed to reverse the antihyperalgesic effect of either diazepam or ethanol. No antinociception was seen in either the ethanoliflumazenil or dcxtrin control groups.

Conclusions

:These results suggest that the antinociceptive effects of both acute and chronic ethanol are at least partially mediated by GABA receptors, and that diazepam's antihyperalgesic effects may not be mediated by the GABA acid receptor.
Mismatch Negativity and Auditory Sensory Memory in Chronic Alcoholics
Authors: Polo M. Dolores , Escera Carles , Gual Antoni , Grau Carles
Published: May 2006

Background & Methods

: Preattentive auditory processing and sensory memory were investigated by means of mismatch negativity (MMN) in a sample of 22 middle‐aged abstinent chronic alcoholics and 25 age‐matched healthy controls. Stimuli were presented at two inter‐stimulus intervals (ISIs, 0.75 sec and 2.0 sec) in separate blocks.

Results

: No significant differences in amplitude or latency of MMN were found between aicoholic and control subjects in either of the two ISI conditions. However, when age was included as a factor in the analysis, MMN amplitude was attenuated in chronic alcoholics who were older than 40 years of age.

Conclusions

: These results indicate that the automatic stimulus‐change detector mechanism associated with MMN gencration is impaired in chronic alcoholics over the age of 40, suggesting that the neurotoxic effects of chronic consumption of alcohol are more prone to appear after a critical age.
Suppression of Alcohol Intake by Chronic Naloxone Treatment in P Rats: Tolerance Development and Elevation of Opiate Receptor Binding
Authors: Overstreet David H. , Kampov‐Polevoy Alexey B. , Rezvani Amir H. , Braun Chris , Bartus Raymond T. , Crews Fulton T.
Published: May 2006

Background

: This study was planned to determine the feasibility of using a slow release naloxone preparation to treat alcoholism, because compliance with medication is a significant problem in alcoholics.

Methods

: Experiments were performed in alcohol‐preferring P rats maintained either on continuous access or on limited access (1 hr/day) to alcohol with water and food provided ad libitum. Naloxone (Nx) was administered either by twice daily subcutaneous injections or by slow release (1.1 mg/kg/hr) osmotic minipump. In limited access experiments, Nx was injected immediately before access to alcohol.

Results

: An initial experiment estimated the dose‐effect curve for Nx subcutaneous suppression on alcohol intake. Nx (2.5–20 mgkg) had a stronger effect during the first 2 hr after injection (ED50= 2.1 mgkg); however, the effect was more modest on 24‐hr consumption. Similar results were found with chronic Nx treatment. Low doses of Nx (0.5 and 2.0 mgkg) injected immediately before limited access to alcohol produced almost complete suppression of alcohol intake for at least 14 consecutive days. However, 14 days of treatment with 26 mg/kg/day by minipump or injection produced an initial 50% suppression of 24‐hr alcohol intake with the gradual development of tolerance. An acute challenge with Nx immediately after the pumps were scheduled to be empty provided additional evidence of tolerance development in chronically Nx‐treated rats. Brain μ‐opiate receptors, estimated autoradiographically by using the ligand [3H][D‐Ala2,N‐Me‐Phe4, Gly‐015][tyrosyl‐3,5‐3H]‐enkephalin, showed that rats chronically exposed to Nx and showing tolerance to Nx suppression of drinking exhibited 17% to 250% increases in [3H][D‐Ala2, N‐Me‐Phe4, Gly‐ol5][tyrosyl‐3,5‐3H]‐enkephalin binding.

Conclusions

: High doses of Nx are required to suppress continuous access alcohol consumption in P rats, and tolerance develops to the ethanol consumption‐suppressing effect of Nx that may be related to increases in μ‐opiate receptors.
Ethanol Consumption, Amino Acid and Glutathione Blood Levels in Patients With and Without Chronic Liver Disease
Authors: Loguercio C. , Del Vecchio Blanco F. , De Girolamo V. , Disalvo D. , Nardi G. , Parente A. , Del Vecchio Blanco C.
Published: May 2006

Background

: Ethanol abuse and liver cirrhosis cause a reduction of glutathione blood levels; liver cirrhosis induces an alteration of the plasma amino acid pattern. We evaluated whether or not ethanol abuse affects amino acid levels, particularly those that are involved in metabolizing glutathione in the plasma and erythrocytes of chronic alcohol abusers with or without liver cirrhosis.

Methods

:. We studied 10 chronic alcohol abusers without liver cirrhosis, 10 with alcoholic cirrhosis, 10 affected by hepatitis C virus‐related cirrhosis, and 10 healthy subjects. Glutathione, γ‐glutamyl‐cysteine, and cysteine were determined by fluorescent HPLC, glutamic acid, glycine, and other free amino acids by cation exchange chromatography both in the plasma and erythrocytes of all studied subjects.

Results and Conclusions

: In both alcoholics and cirrhotics, we found a significant increase of plasma‐aromatic amino acid and methionine levels, whereas glutathione was significantly reduced. The erythrocytes of these patients showed a significant increase of cysteine, glutamic acid, and glycine; γ‐glutamyl‐cysteine was normal; and glutathione and other free amino acids were significantly decreased. Data suggest that, independent of liver cirrhosis, ethanol abuse affects the metabolism of amino acids and glutathione in both the plasma and the erythrocytes.
Prenatal Ethanol Reduces the Activity of Adult Midbrain Dopamine Neurons
Authors: Shen Roh‐Yu , Hannigan John H. , Kapatos Gregory
Published: May 2006

Background

: Prenatal ethanol exposure has been demonstrated to reduce dopamine (DA) neurotransmission in the forebrain area, which could be contributed by altered electrical activity in midbrain DA neurons. This hypothesis was tested in the present study.

Methods

: The effects of prenatal ethanol exposure on the spontaneous activity of DA neurons in the substantia nigra and ventral tegmental area were investigated with extracellular single‐unit recording techniques in adult male rats. Pregnant rats were administered single daily doses of 0, 3, or 5 g/kg ethanol via intragastric intubation from gestation day 8 through 20. An additional control group did not receive the intubation procedure.

Results

: Prenatal ethanol treatment significantly reduced the number of spontaneously active DA neurons in the substantia nigra and ventral tegrnental area in 3‐ to 5‐month‐old male offspring. The firing rate and firing pattern of the remaining spontaneously active DA neurons were not altered. There were no differences in the spontaneous activity of DA neurons between the nonintubated and 0 g/kg control groups, indicating possible intubation‐induced stress did not influence the activity of DA neurons in adult offspring. Similar prenatal ethanol effects were also determined from older animals (14–16 months old), suggesting that the reduction in the spontaneous activity of DA neurons is a persistent phenomenon in adulthood after prenatal ethanol exposure. Furthermore, the reduction in the number of spontaneously active DA neurons was not the result of a loss in DA neurons per se, as revealed by the results of tyrosine hydroxylase immunohistochemistry. The prenatal ethanol exposure‐induced reduction in DA neuronal activity may result from depolarization inactivation, because systemically administered apomorphine (20 μg) increased the spontaneous activity of DA neurons.

Conclusions

: Prenatal ethanol exposure induced a long‐lasting reduction in the activity of midbrain DA neurons in adult animals. The effect was not the result of cell loss but possible changes in the electrical properties of DA neurons. The decreased electrical activity in midbrain DA neurons could contribute to the prenatal ethanol exposure‐induced reduction in DA content and metabolites observed in previous studies and the attention/hyperactivity problems reported in children with fetal alcohol effects/fetal alcohol syndrome.
Ethanol Directly Excites Dopaminergic Ventral Tegmental Area Reward Neurons
Authors: Brodie Mark S. , Pesold Christine , Appel Sarah B.
Published: May 2006

Background

: The mesolimbic/mesoeortical dopamine pathway mediates the rewarding effects of ethanol and other drugs of abuse like cocaine and opiates. Dopaminergic neurons of the ventral tegmental area (VTA) arc the cells of origin of the mesolimbic/mesocortical dopamine pathway. Ethanol's rewarding properties result from its ability to excite dopaminergic cell bodies in the VTA which results in increased dopamine release in the nucleus accumbens. Many recent papers have speculated that ethanol excitation of dopaminergic VTA neurons is indirect, either that ethanol acts on GABAergic or other interneurons, which in turn modulate the activity of dopaminergic VTA neurons, or that ethanol modulates the action of neurotransmitter‐gated ion channels in the VTA.

Methods

: VTA neurons were acutely dissociated and plated onto a cover slip in an electrophysiological recording chamber. These neurons generated spontaneous action potentials which could be measured with cell attached loose patch recording. The dissociation procedure truncated the dendritic trees, severed synaptic contacts and widely dispersed these neurons. Dopamine (10–50 nM) and ethanol (20–120 mM) were bath applied and their effects on firing rate were measured. After some experiments, plated cells were fixed and processed for immunostaining of tyrosine hydroxylase, the rate‐limiting enzyme in dopamine synthesis.

Results

: All neurons met electrophysiological criteria previously established for dopaminergic VTA neurons. Dopamine inhibited all VTA neurons tested, indicating the presence of dopamine autoreceptors. All neurons identified as dopaminergic by these electrophysiological and pharmacological criteria, and that were processed for immunohistochemistry, stained positive for tyrosine hydroxylase immunoreactivity. All acutely dissociated VTA neurons, identified as dopaminergic by electrophysiological, pharmacological and immunohistochemical criteria, were robustly excited by behaviorally relevant concentrations of ethanol. The ethanol‐induced excitation was concentration‐dependent.

Conclusions

: These data provide strong evidence that ethanol directly excites dopaminergic VTA neurons, 21s this excitation still occurs in the absence of input from surrounding neurons.
In Vivo Modulation of 73 kDa Heat Shock Cognate and 78 kDa Glucose‐Regulating Protein Gene Expression in Rat Liver and Brain by Ethanol
Authors: Tunici Patrizia , Schiaffonati Luisa , Rabellotti Elisa , Tiberio Laura , Perin Antonio , Sessa Angela
Published: May 2006

Background: In cultured cells of various origin, ethanol induces the synthesis of 70 kDa family heat shock proteins (hsp70 family), which play a role in the protection of protein traffic and secretion, as well as in cytoskeleton organization. To assess whether ethanol also can induce such genes in vivo, we studied the behavior of hsp70, hsc73, and grp78 messenger ribonucleic acids (mRNAs) and related proteins in the liver and brain of rats acutely treated with ethanol.

Methods: Overnight fasted Sprague‐Dawley rats (220‐250 g) were acutely treated with a low (2 g/kg body weight) or a high (5 g/kg body weight) dose of ethanol as a 30% solution in saline or an equal volume of saline (controls) by gastric intubation. Animals were killed at various times after treatments (3‐72 hr). Messenger RNA levels for different members of hsp70 family (hsp70; 73 kDa heat shock cognate, or hsc73; and 78 kDa glucose‐regulating protein, or grp78) were determined by Northern blot analysis and hybridization with specific complementary deoxyribonuclcic acid (cDNA) probes. The amounts of related proteins were assayed by Western blot analysis with specific antibodies. Autoradiograms and fluorograms were subjected to densitometric scanning.

Results: Ethanol (2 g/kg) caused a slight increase in hsc73 and grp78 mRNA levels only in the liver, without enhancing the amount of proteins. Ethanol (5 g/kg) increased the level of hsc73 and grp78 mRNAs and related proteins in the liver. In the brain, the amount of hsc73 mRNA was enhanced, but this did not change hsc73 protein. In addition, we observed an increase in cerebral grp78 transcript and related protein. Hsp70 gene was not induced in the examined tissues by either dose of ethanol.

Conclusions: Hepatic and cerebral hsc73 and grp78 genes are responsive to ethanol in vivo, and their activation may signal the cell's effort to counteract the harmful action of ethanol.
Disruption of Operant Oral Self‐Administration of Ethanol, Sucrose, and Saccharin by the AMP/Kainate Antagonist, NBQX, but Not the AMPA Antagonist, GYKI 52466
Authors: Stephens D.N. , Brown G.
Published: May 2006

Background: AMPAergic transmission has been suggested to be important in mediating the rewarding effects of drugs. We therefore examined the role of non‐NMDA glutamate‐receptors in mediating the reinforcing properties of ethanol in an oral self‐administration paradigm.

Methods: The competitive antagonist NBQX (2,3‐dihydroxy‐6‐nitro‐7‐sulfamoylbenzo(f)quinoxaline) and the non‐competitive antagonist, GYKI 52466 (1‐(4‐aminophenyl)‐4‐methyl‐7,8‐methylene‐dioxy‐5H‐2,3‐benzodiazepine), were administered to animals previously trained to self‐administer ethanol, sucrose, or saccharin, on a progressive ratio schedule.

Results: GYKI 52466 (5 & 10 mg/kg) had no effect on operant responding for ethanol, but increased spontaneous locomotor activity, whereas the highest doses of NBQX (3 & 6 mg/kg) significantly decreased operant responding and correspondingly decreased the breaking point in responding for ethanol on a progressive ratio schedule. NBQX (but not GYKI 52466) also significantly decreased operant responding for a food reinforcer (sucrose) as well as for a sweet tasting reinforcer (saccharin). Doses of NBQX that disrupted operant performance caused a significant reduction in locomotor activity, indicating that NBQX decreased ethanol self‐administration only at doses which disrupt motor activity.

Conclusions: These results suggest non‐NMDA glutamate‐receptors may not have a specific role in the maintenance of responding for an ethanol reinforcer, but may generally disrupt operant performance. Because, unlike GYKI 52466, NBQX possesses an action at central kainate receptors, these results suggest that kainate, but not AMPA (α‐amino 3‐hydroxy‐5‐methyl‐4‐isoxazole propionate) receptors may be important in mediating the reinforcing effects of ethanol.
Development of a Composite Measure for Assessing Alcohol Treatment Outcome: Operationalization and Validation
Authors: Cisler Ron A. , Zweben Allen
Published: May 2006

This article operationally describes and empirically validates a composite outcome measure developed for use in a multisite alcohol treatment matching study. Using empirically based clinical guidelines to establish alcohol consumption and alcohol‐related problems criteria, 1,726 subjects were classified as abstinent, moderate drinking without problems, heavy drinking or problems, or heavy drinking and problems at intake and 3, 6, 9, 12, and 15 months postintake. Subjects with poorer composite outcome also had poorer outcomes related to quantity and frequency of alcohol consumption, alcohol‐related problems, serum γ‐glutamyltranspeptidase and other nonalcohol‐related measures assessing psychiatric dysfunction, psychosocial functioning, and purpose or meaning in life. Differences in the present composite measure relative to other categorical measures used in research to date and the potential for incorporating nonalcohol‐specific variables into composite measures are discussed.
Acetaldehyde Enhances Murine α2(I) Collagen Promoter Activity by Ca2+‐Independent Protein Kinase C Activation in Cultured Rat Hepatic Stellate Cells
Authors: Anania Frank A. , Womack Lawrence , Potter James J. , Mezey Esteban
Published: May 2006

Protein kinase C (PKC) inhibitors decrease α1(I) collagen mRNA in stellate cells exposed to 200 μmol/liter of acetaldehyde. The purpose of these studies was to determine whether PKC activation plays a role in transcriptional activation of the α2(I) collagen gene. Cultured stellate cells were exposed to 200 μmol/liter of acetaldehyde. PKC, inositol triphosphate, diacylglycerol (DAG), and intracellular free calcium (Ca2+,) were measured. α1(I) and α2(I) collagen messages were determined by reverse transcriptase‐polymerase chain reaction. Activation of the α2(I) collagen promoter was determined in transiently transfected stellate cells. Acetaldehyde exposure enhanced PKC activity translocation to the particulate fraction at 20 min. Acetaldehyde did not increase Ca2+i, or inositol triphosphate but increased DAG levels at 20 min and 3 hr. Acetaldehyde increased both the α1(I) and α2(I) collagen messages in stellate cells. Calphostin C, a specific PKC inhibitor, which blocks DAG binding, eliminated both activation of the α2(I) collagen promoter by acetaldehyde and mRNA production by reverse transcriptase‐polymerase chain reaction analysis. Similarly, D609, an inhibitor of DAG production, also inhibited α2(I) collagen gene expression. This study shows that collagen production by acetaldehyde is mediated by a calcium‐independent PKC mechanism.
Effects of Prenatal Ethanol Exposure on Hypothalamic‐Pituitary‐Adrenal Responses to Chronic Cold Stress in Rats
Authors: Kim C. Kwon , Giberson Pam K. , Yu Wayne , Zoeller R. Thomas , Weinberg Joanne
Published: May 2006

Animals prenatally exposed to ethanol typically exhibit hypothalamic‐pituitary‐adrenal (HPA) hyperresponsiveness to stressors. In contrast to previous studies that have investigated effects of prenatal ethanol exposure on HPA responses to acute or intermittent stressors, our study investigated HPA responses to a chronic continuous stressor, cold stress (4°C for 0,1, or 3 days). We tested the hypothesis that prenatal ethanol exposure would result in increased plasma corticosterone (CORT) and adrenocorticotropin (ACTH) responses and increased peptide [corticotropin‐releasing factor and vasopressin] mRNA levels in the paraventricular nucleus (PVN) of the hypothalamus compared to that in control animals. In addition, CORT and ACTH responses were measured after exposure to an acute stressor (ip isotonic saline injection), superimposed during chronic cold exposure, to examine possible sensitization of the HPA response to the acute stress. Thus, blood samples were collected at the end of each of the three periods of cold exposure, either before (0 min) or 15 min after acute stress. The subjects were adult male and female Sprague‐Dawley rat offspring from prenatal ethanol (E), pair‐fed (PF), and ad libitum‐fed control (C) treatment groups. Exposure to cold stress resulted in significant body weight loss in E males at 1 day and in both males and females of all prenatal treatment groups by 3 days of cold stress. Males in all prenatal groups also exhibited significant increases in adrenal weight:body weight ratios. Cold stress alone (0 min condition) increased CORT levels in E males and overall ACTH levels in E males and females compared to controls. ACTH levels were also higher overall in E compared to control males after acute stress (15 min condition). Sensitization of the CORT response to acute stress was observed in males but not females across all prenatal treatment groups. Corticotropin‐releasing factor and vasopressin mRNA levels in the PVN were not significantly affected by prenatal treatment or chronic cold stress in either males or females. In contrast, both males and females displayed increases in PVN thyrotropin‐releasing hormone (TRH) mRNA levels after cold stress. These data support and extend previous work demonstrating differential effects of prenatal ethanol exposure on HPA responsiveness of male and female offspring, and suggest that E males may be more vulnerable to the effects of chronic cold stress than E females.
Chronic Ethanol‐Initiated Apoptosis in Hepatocytes Is Induced by Changes in Membrane Biogenesis and Intracellular Transport
Authors: Slomiany Amalia , Piotrowski Elizabeth , Grabska Maria , Piotrowski Jerzy , Slomiany Bronislaw L.
Published: May 2006

The outcome of chronic ethanol consumption recorded in liver by in situ staining of the genomic DNA in fragmented nuclei indicates the course of cellular events that has been coined as apoptosis or programmed cell death. Hence, we designed the study to determine which ethanol‐induced modification of the cellular make‐up is responsible for the hastening the cell damage. The in vitro assays were performed with cellular organelles and cytosol prepared from hepatocytes derived from rats subjected to 9 weeks of chronic alcohol consumption. The results were compared with the pair‐fed controls receiving isocaloric liquid diet. In the initial phase of the studies, we established that the process of apoptosis was not triggered by the aberrant activity of neutral or acidic sphingomyelinase. The hepatocytes derived from alcohol and control diets manifested equal enzymatic activity. The de novo synthesis of sphingoid bases and ceramides in the alcohol‐derived sample of endoplasmic reticulum was reduced, but the in situ apoptosis was up to 36‐fold higher than in the control. Also, the isolated hepatocytes contained a 2‐ to 4‐fold higher amount of nucleosomal fragments in the cytosolic extracts. The endosomes from liver hepatocytes of ethanol‐consuming rats, in the presence of the cytosol and mitochondria from pair‐fed controls, disclosed 2 to 3 times higher apoptotic potential than sample consisting of ethanol‐derived fractions only, and 3 to 5 times higher than the control‐derived fractions. The substantial increase in apoptosis, as recorded in the amount of DNA fragments released to the cytosol from the fresh nuclei, was also recorded when the microsomal membranes of endoplasmic reticulum and Golgi were incubated in the conditions with preserved intracellular transport. The maximal 20‐fold increase of apoptotic activity was recorded in the incubation mixtures of ethanol‐derived endoplasmic reticulum‐Golgi membranes with control‐derived cytosol in the presence of the ATP generating system. Results infer that the intracellular transport vesicles, generated from ethanol‐modified membranes in the presence of the substrates that are available in the cytosol of the control hepatocytes, activate the apoptotic activity in the in vitro system. This interpretation is supported by the results of analysis of the clathrin‐coated transport vesicles that, in contrast to nonclathrin transport vesicles, contain a sizable accumulation of ceramides that are known to induce apoptosis.
Modulation of Caspase‐3 Activity and Fas Ligand mRNA Expression in Rat Liver Cells In Vivo by Alcohol and Lipopolysaccharide
Authors: Deaciuc Ion V. , Fortunato Franco , D'Souza Nympha B. , Hill Daniell B. , Schmidt Jack , Lee Eun Y. , McClain Craig J.
Published: May 2006

The purpose of this study was to determine if exacerbation of apoptosis precedes liver injury during chronic exposure of rats to alcohol. After 7 weeks of feeding an alcohol‐ or dextrin‐containing liquid diet, the animals were treated with Gram‐negative bacterial lipopolysaccharide (1 mg kg‐1 body weight, intravenously) or sterile saline and sacrificed 3 hr after the treatment. Alanine:2‐oxoglutarate aminotransferase (ALT) and 1actate:NAD oxidoreductase [lactate dehydrogenase (LDH)] were measured in plasma. The caudate lobe of the liver was resected for histology, while the rest of the organ was perfused with collagenase to isolate hepatocytes, Kupffer cells (KCs), and sinusoidal endothelial cells (SECs) by centrifugal elutriation. Hepatocyte mitochondria were isolated by differential centrifugation of the cell homogenate. Reduced and oxidized glutathione (GSH and GSSG) in isolated hepatocytes and hepatocyte mitochondria, and malondialdehyde in hepatocytes were assayed. Caspase‐3 activity and Fas ligand mRNA expression were determined in hepatocytes, KCs, and SECs. Plasma ALT and LDH activity, liver histology, GSH, GSSG and their ratio, and malondialdehyde content were not affected by alcohol treatment. Caspase‐3 activity was significantly increased in alcohol‐treated rats in all three cell types, with the lowest response observed in hepatocytes and the highest in KCs. Fas ligand mRNA expression, which had the highest level in SECs, followed by KCs and hepatocytes, was not affected by alcohol administration. Lipopolysaccharide had the following effects: an increase in ALT in both pair‐ and alcohol‐fed rats, and LDH only in alcohol‐fed rats, a decrease in GSH + GSSG levels in both mitochondria and hepatocytes, an elevation of malondialdehyde content in hepatocytes, a raise in caspase‐3 activity in all groups and cell types, and an augmentation of Fas ligand expression in hepatocytes and KCs, but not in SECs. These data suggest that, during chronic alcohol consumption, an exacerbated apoptosis precedes alcohol‐induced liver injury.
Effects of c‐Src Tyrosine Kinase on Ethanol Sensitivity of Recombinant NMDA Receptors Expressed in HEK 293 Cells
Authors: Anders Douglas L. , Blevins Tana , Sutton Greg , Chandler L. Judson , Woodward John J.
Published: May 2006

N‐Methyl‐D‐aspartate (NMDA) receptors are ligand‐gated ion channels that are important mediators of the actions of the excitatory amino acid neurotransmitter glutamate. Previous studies have shown that ethanol inhibits the function of both wild‐type receptors found in neurons and recombinant NMDA receptors expressed in heterologous cells, such as oocytes and transfected mammalian cells. Although some studies have reported that certain subunit combinations display an enhanced sensitivity to ethanol, this effect is not observed in all experimental systems. This discrepancy may be due to varying levels of endogenous modulators, such as kinases, between different cell preparations. In this study, we investigated the effects of tyrosine phosphorylation on the ethanol sensitivity of NMDA receptor function using a recombinant cell system where levels of both NMDA subunits and protein kinases can be more carefully controlled. Human embryonic kidney (HEK 293) cells were transfected with different NMDA receptor subunits and a c‐Src‐green fluorescent protein (GFP) fusion protein that could be directly visualized in living cells. Agonist‐stimulated calcium flux was measured in single cells using fura‐2 video imaging. As expected, cells transfected with the NR1/NR2B subunits were more sensitive to inhibition by the NR2 selective antagonist ifenprodil than those transfected with NR1/NR2A or NRl/NR2A/NR2B subunits. All receptor combinations were inhibited by ethanol (25 and 100 mM), with the NRl/NR2B combination being slightly more sensitive than NRl/NR2A or NRl/NR2A/NR2B. Control and NMDA‐receptor transfected HEK 293 cells displayed a low degree of tyrosine phosphorylation as measured by immunofluorescence and Western immunoblotting using an anti‐phosphotyrosine antibody. Phosphorylation was markedly enhanced in cells transfected with the c‐Src‐GFP fusion protein. The sensitivity of NMDA receptors to either 25 or 100 mM ethanol, or 10 μM ifenprodil, was not significantly altered by co‐transfection with c‐Src‐GFP. These results indicate that, although NMDA receptors can be a target of c‐Src tyrosine kinase, tyrosine phosphorylation by this enzyme does not modulate the inhibitory effects of ethanol on NMDA‐activated currents.