bluuebunny/crossref_metadata_2025 huggingface dataset:
Adduction of Soluble Proteins with Malondialdehyde‐Acetaldehyde (MAA) Induces Antibody Production and Enhances T‐Cell Proliferation
Authors: Willis Monte S. , Klassen Lynell W. , Tuma Dean J. , Sorrell Michael F. , Thiele Geoffrey M.
Published: April 2006

Background: The alcohol metabolites malondialdehyde and acetaldehyde can combine to form stable adducts (MAA) which are found in the livers of humans and rats after significant alcohol ingestion. While adducted proteins induce antibody responses in the absence of adjuvants, the mechanisms by which these responses occur are unknown. Thus, it was the purpose of these studies to investigate how MAA modification stimulates antibody and T‐cell responses in the absence of adjuvants.

Methods: Hen egg lysozyme (HEL) was modified with increasing levels of MAA and was used as an immunogen, and antibody and T‐cell responses were determined. The role of scavenger receptors in the immunogenicity of MAA‐adducted proteins was also investigated.

Results: Maximum antibody response was induced after immunization with 1.8 nM MAA/nM HEL, and was primarily an IgG1 response to HEL as determined by inhibition ELISAs. T‐cell proliferative responses after immunization with HEL‐MAA were solely to HEL. Immunization with a scavenger receptor ligand in conjunction with HEL‐MAA increased the predominant IgG1 response and sharply decreased the IgG2a response by approximately 50%. Binding of HEL‐MAA by splenocytes was determined by flow cytometry to be approximately 15% greater than HEL alone, showing a doubling of the geometric mean fluorescence. Also, most of the cells that bound HEL‐MAA were class II positive, indicating that antigen‐presenting cells can bind the MAA‐adducted HEL, and potentially initiate immune responses.

Conclusions: MAA modification of proteins induces antibody and T‐cell proliferative responses in vivo. Initial studies suggest that these responses may be mediated by scavenger receptors that recognize MAA‐adducted proteins. This suggests a mechanism by which proteins modified with oxidative products associated with chronic ethanol consumption may alter immune responses that may play an active role in the development and/or progression of alcoholic liver disease.
Implicit and Explicit Alcohol‐Related Cognitions
Authors: Wiers Reinout W. , Stacy Alan W. , Ames Susan L. , Noll Jane A. , Sayette Michael A. , Zack Martin , Krank Marvin
Published: April 2006

This article presents the proceedings of a symposium at the 2001 RSA Meeting in Montreal, Canada organized by Reinout W. Wiers and Alan W. Stacy. The purpose of the symposium was to present recent applications of implicit cognitive processing theory to alcohol research. Basic cognitive research has demonstrated that implicit cognition influences memory and behavior without explicit recall or introspection. The presentations from this symposium show that implicit cognition approaches yield new insights into understanding drinking motivation. The presentations were: (1) An introduction by Alan W. Stacy; (2) Implicit cognition and alcohol use. Involvement of other variables? (Susan L. Ames); (3) Alcohol expectancies and the art of implicit priming (Jane A. Noll); (4) Parental alcoholism and the effects of alcohol on semantic priming (Michael A. Sayette); (5) Implicit arousal and explicit liking of alcohol in heavy drinkers (Reinout W. Wiers); and (6) Negative affective cues and associative cognition in problem drinkers (Martin Zack). Comments were provided by the discussant Marvin Krank. The presented studies demonstrated that: (1) implicit memories of alcohol associations are powerful predictors and cross‐sectional correlates of alcohol use; (2) implicit retrieval processes influence alcohol outcome expectancies and alcohol consumption; (3) alcohol consumption influences implicit memory processing; (4) heavy drinkers reveal different affective responses in implicit and explicit tasks; and (5) negative affect exerts an implicit priming effect for alcohol associations in problem drinkers. These findings illustrate the importance of implicit cognition in understanding alcohol abuse and demonstrate the potential of the theoretical framework for more widespread application across a variety of areas of alcohol research, including diagnostics for the risk of alcohol abuse, treatment, and prevention.
Acamprosate, MK‐801, and Ifenprodil Inhibit Neurotoxicity and Calcium Entry Induced by Ethanol Withdrawal in Organotypic Slice Cultures From Neonatal Rat Hippocampus
Authors: Mayer Sveta , Harris Barton R. , Gibson D. Alex , Blanchard John A. , Prendergast Mark A. , Holley Robert C. , Littleton John
Published: April 2006

Background The antirelapse drug acamprosate has previously been reported to inhibit activating effects of polyamines on N‐methyl‐D‐aspartic acid receptor (NMDAR) function. Because increased synthesis of polyamines has been suggested as a mechanism for potentiation of NMDAR function during ethanol withdrawal, we evaluated the effects of acamprosate, MK‐801, and ifenprodil in a cell culture model of ethanol withdrawal–induced neurotoxicity.

Methods Organotypic hippocampal cultures from 8‐day‐old neonatal rats were maintained in vitro for 23 days before experimental use. The ethanol withdrawal model consisted of exposing cultures to ethanol (70–100 mM) for 4 days before being “withdrawn” into Calcium‐Locke's buffer for 1 hr and then into minimal medium for 23 hr. Uptake of 45CaCl2 and propidium iodide by damaged cells was assessed 1 hr and 24 hr after the start of ethanol withdrawal, respectively. Additional studies examined effects of exposure to NMDA (50 μM) or spermidine (100 μM) on withdrawal‐induced hippocampal damage. Last, these studies examined the ability of the sodium salt of acamprosate (Na‐acamprosate, 200 μM), ifenprodil (50 μM), or MK‐801 (30 μM) to inhibit neurotoxicity and 45Ca2+ entry produced by these insults.

Results Ethanol withdrawal was associated with significantly greater toxicity and 45Ca2+ entry, relative to controls. Exposure to spermidine and NMDA during ethanol withdrawal further increased neurotoxicity and 45Ca2+ entry. Acamprosate, ifenprodil, and MK‐801 almost completely prevented ethanol withdrawal–induced toxicity and 45Ca2+ entry. Acamprosate also reduced spermidine‐induced neurotoxicity during ethanol withdrawal but was ineffective against NMDA‐induced toxicity or 45Ca2+ entry at this time.

Conclusions The results support the contention that acamprosate, like ifenprodil, interacts with polyamines and that these compounds may be effective in reducing consequences of ethanol withdrawal. NMDAR activation is also strongly implicated in ethanol withdrawal neurotoxicity, but whether acamprosate causes any of these effects in this preparation directly via the NMDAR remains uncertain.
The European NEAT Program: An Integrated Approach Using Acamprosate and Psychosocial Support for the Prevention of Relapse in Alcohol‐Dependent Patients With a Statistical Modeling of Therapy Success Prediction
Authors: Pelc Isidore , Ansoms Constant , Lehert Philippe , Fischer Felix , Fuchs Werner‐Joachim , Landron Frédéric , Pires Preto Anton José , Morgan Marsha Y.
Published: April 2006

Background A multicenter, prospective study was conducted in five European countries to observe outcome in alcohol misusers treated for 24 weeks with acamprosate and various psychosocial support techniques, within the setting of standard patient care.

Methods Patients diagnosed as alcohol dependent using DSM‐III‐R criteria were treated, for 24 weeks, with acamprosate and appropriate psychosocial support. Potential predictor variables were recorded at inclusion. Drinking behavior was monitored throughout; the proportion of cumulative abstinence days was the principal outcome measure. The influence of baseline clinical and demographic variables on outcome was assessed using multiple regression analysis. Adverse events were recorded systematically.

Results A total of 1289 patients were recruited; 1230 took at least one dose of the drug and provided at least one set of follow‐up data; 543 (42.1%)patients were observed for the full 24‐week period. The overall proportion of cumulative abstinence days was 0.48. Multiple physical and psychiatric comorbidities and a history of drug addiction were negatively correlated with outcome, as were, to a lesser extent, multiple previous episodes of detoxification, unemployment, and living alone. Older age and stable employment were positively associated with outcome. The difference in the unadjusted proportion of cumulative abstinence days between countries was significant (p < 0.001) but less so when adjusted for the predictive factors identified in the multivariate model (p < 0.019). Overall, outcome was not influenced by the nature of the psychosocial support provided. Adverse events were generally mild, with gastrointestinal disorders, which occurred in 21.5% of patients, being the most frequent.

Conclusions This open‐label study confirms the efficacy and safety of acamprosate in the treatment of alcohol dependence in the setting of standard patient care. Treatment benefit was observed irrespective of the nature of the psychosocial support provided. Predictors of the response to treatment were identified; their heterogeneous distribution within the study population explained, at least in part, the differences in outcome between countries.
Drinking During Pregnancy: Attitudes and Knowledge Among Pregnant Danish Women, 1998
Authors: Kesmodel Ulrik , Schiøler Kesmodel Pia
Published: April 2006

Background During the 1990s, most Western countries officially recommended that pregnant women abstain from alcohol. However, information about the potentially harmful effects of alcohol during pregnancy does not necessarily equate to understanding, and information and knowledge may not be associated with pregnant women's own attitudes toward drinking.

Methods From October to December 1998, we interviewed 439 Danish‐speaking pregnant women who were referred for routine antenatal care at their first visit at 15 to 16 weeks of gestation. The women were interviewed about their attitudes toward and beliefs and knowledge about drinking during pregnancy. Questions were also asked about information on alcohol provided to the women.

Results Seventy‐six percent of the women considered some alcohol intake during pregnancy to be acceptable, mostly on a weekly level. Binge drinking, however, was considered to be harmful by 85%. These attitudes were not associated with knowledge about the official recommendation or whether the woman had talked to her general practitioner or midwife about alcohol during pregnancy. Most of the women had received information on alcohol from the mass media or relatives, but most women believed that information about alcohol during pregnancy could best be communicated to them by health personnel. Only 21% were aware of the official recommendation from the Danish National Board of Health. One third had discussed alcohol with their general practitioner or midwife, but these women had mostly been advised that some alcohol intake was acceptable.

Conclusions Most of the women considered some alcohol intake during pregnancy to be acceptable, mostly on a weekly level, and their attitudes were independent of their knowledge about the subject. Most of the women had not been informed about alcohol during pregnancy.
Fine Mapping of Polymorphic Alcohol‐Related Quantitative Trait Loci Candidate Genes Using Interval‐Specific Congenic Recombinant Mice
Authors: Ehringer Marissa A. , Thompson Jessica , Conroy Otakuye , Yang Fan , Hink Raquel , Bennett Beth , Johnson Thomas E. , Sikela James M.
Published: April 2006

Background The inbred long‐sleep (ILS) and inbred short‐sleep (ISS) strains of mice are widely studied as a model of initial sensitivity to alcohol. Recently, a large comparative DNA sequencing study of candidate genes located within the four Lore quantitative trait loci (QTLs) associated with the ethanol‐induced loss of righting reflex in ILS and ISS mice has identified eight genes that contain coding region differences corresponding to amino acid changes. Here, recently developed interval‐specific congenic recombinant mice (ISCRs) have been used to map these genes in relationship to newly narrowed QTL regions.

Methods Regions of candidate genes containing DNA differences corresponding to previously identified amino acid changes between ISS and ILS mice were amplified from either genomic DNA or complementary DNA from ISCR mice using polymerase chain reaction. The products were purified and directly sequenced to determine the genotypes for each polymorphism. On the basis of these genotypic data, each candidate gene was determined to be located either within or outside of recently narrowed Lore QTL intervals.

Results Of these eight candidates with protein‐coding differences, five are now excluded from their respective Lore intervals. The other three (Znf142, Ptprn, and Znf133) have been localized to the narrowed QTL intervals.

Conclusions These three central nervous system genes (Znf142, Ptprn, and Znf133) represent promising candidates for involvement in the differential sensitivity to alcohol exhibited between ILS and ISS mice. This study also demonstrates how the combination of high‐throughput comparative gene sequencing and concomitant genetic fine mapping of QTL regions with ISCRs can be an effective tool for accelerating the process of moving from QTL to gene.
Acute Alcohol Inhibits the Induction of Nuclear Regulatory Factor κB Activation Through CD14/Toll‐Like Receptor 4, Interleukin‐1, and Tumor Necrosis Factor Receptors: A Common Mechanism Independent of Inhibitory κBα Degradation?
Authors: Mandrekar Pranoti , Dolganiuc Angela , Bellerose Gary , Kodys Karen , Romics Laszlo , Nizamani Rabia , Szabo Gyongyi
Published: April 2006

Background Nuclear translocation and DNA binding of the nuclear factor κB (NF‐κB) is an early event in inflammatory cell activation in response to stimulation with bacterial components or cytokines. Cell activation via different receptors culminates in a common pathway leading to NF‐κB activation and proinflammatory cytokine induction. We have previously shown that acute alcohol inhibits NF‐κB activation by lipopolysaccharide (LPS) in human monocytes. Here we investigated whether acute alcohol treatment of human monocytes also inhibits NF‐κB when induced through activation of the interleukin (IL)‐1 or tumor necrosis factor (TNF) receptors.

Methods Human peripheral blood monocytes were treated with LPS, TNFα, and IL‐1β in the presence or absence of 25mM alcohol for 1 hr. NF‐κB activation was determined by electrophoretic mobility shift assays using nuclear extracts. Inhibitory κBα (IκBα) was estimated by Western blotting in cytoplasmic extracts. Chinese hamster ovary cells expressing human CD14 were treated with LPS in the presence or absence of alcohol to study NF‐κB and IκBα regulation.

Results Our results indicate that acute alcohol inhibits IL‐1β‐ and TNFα‐induced NF‐κB activation. We further show in CD14/toll‐like receptor 4–expressing Chinese hamster ovary cells the specificity of alcohol‐mediated inhibition of NF‐κB via the toll‐like receptor 4/CD14 receptors. Inhibition of NF‐κB by acute alcohol was concomitant with decreased levels of the IκBα molecule in the cytoplasm of LPS, IL‐1, and TNFα‐activated monocytes.

Conclusions These data suggest a unique, IκBα‐independent pathway for the inhibition of NF‐κB activation by acute alcohol in monocytes. Universal inhibition of NF‐κB by acute alcohol via these various receptor systems suggests a target for the effects of alcohol in the NF‐κB activation cascade that is downstream from IκBα degradation. Further, these results demonstrate that acute alcohol is a potent inhibitor of NF‐κB activation by mediators of early (LPS) or late (IL‐1, TNFα) stages of inflammation in monocytes.
Effects of Ethanol Exposure on Subsequent Acquisition and Extinction of Ethanol Self‐Administration and Expression of Alcohol‐Seeking Behavior in Adult Alcohol‐Preferring (P) Rats: II. Adult Exposure
Authors: Rodd‐Henricks Zachary A. , Bell Richard L. , Kuc Kelly A. , Murphy James M. , McBride William J. , Lumeng Lawrence , Li Ting‐Kai
Published: April 2006

Background In a preceding study, we reported that ethanol (EtOH) consumption during periadolescence in alcohol‐preferring (P) rats produced significant effects on the acquisition, extinction, Pavlovian spontaneous recovery (PSR), and reacquisition of operant self‐administration of EtOH. The objective of the present study was to determine if EtOH consumption during adulthood produced similar effects on subsequent operant behaviors.

Methods Adult female P rats (>135 days of age) were given 24 hr free‐choice access to 15% EtOH for 30 days or were similarly housed and received water only. After a 15 day period of no EtOH access and without any prior training, adult alcohol drinking and adult alcohol‐naïve rats were placed in standard two‐lever (15% EtOH and water) chambers to examine acquisition of EtOH self‐administration. After stable responding was established on a concurrent fixed ratio (FR) 5 FR1 schedule for EtOH versus water, the P rats underwent extinction training for nine sessions. After extinction and a 2 week home cage period (with no operant sessions or access to EtOH), rats were returned to the operant chambers in the absence of reward for seven consecutive sessions to test for PSR. After PSR testing, animals were maintained in their home cage for a week, before being reintroduced to the operant chambers and allowed to respond for EtOH and water.

Results Both the adult alcohol‐drinking and adult alcohol‐naïve groups rapidly acquired EtOH self‐administration, expressed a pronounced PSR, which was augmented by EtOH priming and the presence of a discriminative stimulus (odor cue), and increased responding when EtOH was reinstated. Adult pre‐exposure to EtOH did not alter any of the operant measures.

Conclusions The results of this study suggest that, unlike the results with EtOH pre‐exposure during periadolescence, chronic alcohol drinking by P rats in adulthood did not produce sufficient long‐lasting changes in neuronal function to alter subsequent operant acquisition of alcohol self‐administration, alcohol relapse, or alcohol‐seeking behavior.
Neurophysiological Profiles of Replicate Line 2 High‐Alcohol‐Drinking (HAD‐2) and Low‐Alcohol‐Drinking (LAD‐2) Rats
Authors: Katner S. N. , Slawecki C. J. , Ehlers C. L.
Published: April 2006

Rationale A select number of electrophysiological findings have been demonstrated to differentiate rat lines selectively bred for high and low ethanol preference.

Objective In the present study, EEGs and event‐related potentials (ERPs) of high‐alcohol‐drinking (HAD) and low‐alcohol‐drinking (LAD) rats from replicate line 2 (HAD‐2 and LAD‐2) were assessed to determine if their neurophysiological profiles are similar to selected lines previously evaluated.

Methods Rats obtained from Indiana University were implanted with cortical and amygdalar recording electrodes. Baseline EEG and ERPs were assessed in ethanol‐naïve HAD‐2 and LAD‐2 rats. Animals subsequently were trained to self‐administer ethanol by using a sucrose‐substitution procedure.

Results Compared with LAD‐2 rats, HAD‐2 rats displayed greater parietal cortical power in the 6 to 32 Hz frequency range of the EEG. Greater parietal cortical peak frequency in the 2 to 4 Hz range and decreased frontal, parietal, and amygdalar peak frequencies in the 16 to 32 Hz frequency range were also seen. Compared with LAD‐2 rats, HAD‐2 rats had decreased P2 latency of ERPs recorded in the parietal cortex. HAD‐2 rats also had greater frontal, parietal, and amygdalar P2 amplitudes, greater frontal and parietal cortical P1 amplitudes, and greater parietal cortical P3 amplitudes compared with LAD‐2 rats. As anticipated, HAD‐2 rats consumed significantly greater levels of sucrose, sucrose‐ethanol, and ethanol over the course of the sucrose‐substitution procedure compared with LAD‐2 rats.

Conclusions These data suggest that increased cortical power is associated with high ethanol preference in a number of selectively bred rat lines. However, unique electrophysiological characteristics may index alcohol preference in each line.
Operant Self‐Administration of Ethanol in Sardinian Alcohol‐Preferring Rats
Authors: Vacca Giovanni , Serra Salvatore , Brunetti Giuliana , Carai Mauro A. M. , Samson Herman H. , Gessa Gian Luigi , Colombo Giancarlo
Published: April 2006

Background “Work” for ethanol, that is, the ability of a laboratory animal to press a lever to gain access to ethanol, has been proposed as (a) a requirement for definition of an animal model of alcoholism and (b) a measure of ethanol‐reinforcing properties. The present study evaluated oral self‐administration of ethanol under an operant (lever pressing) procedure in selectively bred Sardinian alcohol‐preferring (sP) and alcohol‐nonpreferring (sNP) rats.

Methods Rats from both lines were initiated to self‐administer 10% ethanol, on a fixed ratio 1 schedule and in daily 30 min sessions, by using the Samson sucrose fading procedure. Subsequently, rats were exposed to increasing concentrations of ethanol up to 30% on a fixed ratio 4 schedule. Finally, the extinction responding for ethanol, defined as the maximal number of lever responses reached by each rat in the absence of ethanol reinforcement, was determined.

Results The results indicated that sP rats acquired and maintained lever pressing for ethanol, self‐administering mean amounts of ethanol in the range of 0.6 to 1.1 g/kg/session, which gave rise to mean blood ethanol levels in the 30 to 45 mg% range. Extinction responding for ethanol in sP rats averaged 73. In contrast, once sucrose was faded out, sNP rats displayed minimal levels of responding for ethanol, and extinction responding averaged 6.

Conclusions The results of the present study extend to the sP/sNP rat lines the finding that ethanol can be established as a reinforcer in selectively bred alcohol‐preferring rats, whereas it has modest, if any, reinforcing properties in alcohol‐nonpreferring rats.
Serum Ubiquitin‐Protein Conjugates in Normal Subjects and Patients With Alcoholic Liver Diseases: Immunoaffinity Isolation and Electrophoretic Mobility
Authors: Takagi Masaru , Yamauchi Masayoshi , Takada Koji , Ohkawa Kiyoshi
Published: April 2006

Background We previously reported that levels of multiubiquitin chains, representing ubiquitin‐protein conjugates, were significantly higher in sera of patients with alcoholic liver cirrhosis than in normal subjects and patients with other types of alcoholic liver disease.

Methods To characterize them, ubiquitin‐immunoreactive proteins were purified from sera of healthy human volunteers and patients with alcoholic liver diseases by using affinity chromatography on a Sepharose column containing an immobilized monoclonal antibody recognizing conjugated ubiquitin.

Results SDS‐PAGE analysis followed by Western blotting revealed that the immunoaffinity‐purified proteins mainly contained multiple components with molecular masses greater than 60 kDa, almost all of which were immunostained with the ubiquitin antibody. This size heterogeneity was in agreement with the property of ubiquitin‐protein conjugates in all cells examined. These results indicate that the immunoaffinity‐purified serum proteins are polyubiquitinated proteins presumably derived from some somatic cells. These ubiquitinated proteins obtained from the alcoholic cirrhosis patients were stained more strongly than those from the normal subjects and patients with other types of alcoholic liver disease, although equal amounts of multiubiquitin chains were analyzed simultaneously. In addition, marked differences were observed in the two‐dimensional PAGE pattern of the ubiquitin‐protein conjugates purified from the alcoholic cirrhosis patient serum compared with those from the normal human serum: four distinct broad spots (presumably polyubiquitin‐protein complexes) were observed only in the former.

Conclusions This is the first report on isolation of ubiquitin‐protein conjugates from human serum, and it indicates that not only their levels, but also their molecular compositions, were greatly affected by alcoholic cirrhosis.
Acute Ethanol Administration and Acute Allopregnanolone Administration Impair Spatial Memory in the Morris Water Task
Authors: Matthews Douglas B. , Morrow A. Leslie , Tokunaga Sayaka , McDaniel Janelle R.
Published: April 2006

Background Acute ethanol administration degrades performance on many learning and memory tasks, including tasks that are dependent on spatial information. One common test of spatial learning and memory is the Morris water task, a task that requires subjects to learn the spatial location of a submerged escape platform located in a pool of cloudy water. However, although some studies report that acute ethanol administration degrades spatial memory performance in the Morris task, other studies report no significant performance impairment. Acute ethanol administration also produces a dose‐ and time‐dependent increase in the concentration of the endogenous neuroactive steroid 3α‐hydroxy‐5α‐pregnan‐20‐one (allopregnanolone) in rat brain. Given that ethanol and allopregnanolone are both γ‐aminobutyric acid type A receptor modulators, both drugs should produce similar degradations in spatial learning and memory.

Methods Adult male rats were trained with either the spatial version or the nonspatial version of the Morris water task. After 4 days of training, the spatial or nonspatial memory performance of subjects was assessed after either an ethanol (1.0, 1.5, or 2.0 g/kg) or allopregnanolone (12.5, 17.0, or 20.0 mg/kg) challenge.

Results Acute ethanol administration and acute allopregnanolone administration impaired spatial memory performance in a dose‐dependent manner in the Morris water task. In addition, the impairment was selective in that neither acute ethanol nor acute allopregnanolone administration impaired nonspatial memory performance in the Morris water task.

Conclusions Acute ethanol administration and acute allopregnanolone administration impaired spatial memory performance but did not impair nonspatial memory performance in the Morris water task. These results demonstrate that both ethanol and allopregnanolone produce selective cognitive deficits that are not due to general sensory or motor deficits.
Acetylcholine and Alcohol Sensitivity of Neuronal Nicotinic Acetylcholine Receptors: Mutations in Transmembrane Domains
Authors: Borghese Cecilia M. , Ali Deeba N. , Bleck Virginia , Harris R. Adron
Published: April 2006

Background The effect of n‐alcohols on glycine and γ‐aminobutyric acid type A receptors depends on two specific amino acids (AAs) located in the transmembrane domains (TM) 2 and 3. Our aim was to assess whether the corresponding AAs in the neuronal nicotinic acetylcholine receptor (nAChR) also formed a binding pocket for alcohols.

Methods We made single AA substitutions in the homologous sites in rat neuronal nAChR α2 and α4 (αL261 and αL283) and expressed them in Xenopus laevis oocytes in combination with β4 wild type. The effect of different n‐alcohols was studied in α4(L261A)β4 and α4(L283A)β4 nAChRs. The effect of ethanol, propanol, and octanol on acetylcholine (ACh) responses was studied in α2(L261X)β4 and α2(L283X)β4 nAChRs.

Results Most of the mutations in the α2 subunit, in either the 261 or the 283 position, induced changes in ACh sensitivity and increased alcohol action, but none was able to reduce ethanol potentiation. In α4(L283A)β4, enhancement of potentiation by short‐chain alcohols was observed, as well as a change from inhibition to potentiation for long‐chain alcohols. The exposure of the AAs was assessed through the action of a charged thiol‐specific reagent on α2(L261C)β4 and α2(L283C)β4, and these experiments suggest that the AA in TM2 is located in a water‐accessible position, whereas the AA in TM3 is inaccessible. However, a noncharged thiol‐specific reagent did not affect either ACh responses or ethanol effect on α2(L261C)β4.

Conclusions The AAs located at positions 261 and 283 of the α2 and α4 nAChR subunits do not seem to form a binding pocket for alcohols. Additional studies are required to determine whether alcohols act on a site near these AAs or on sites unrelated to the TM2‐TM3 site found in glycine and γ‐aminobutyric acid type A receptors.
Short of Complete Abstinence: An Analysis Exploration of Multiple Drinking Episodes in Alcoholism Treatment Trials
Authors: Wang Sue‐Jane , Winchell Cellia J. , McCormick Cynthia G. , Nevius S. Edward , O'Neill Robert T.
Published: April 2006

Background In alcoholism treatment clinical trials, conventional analysis of efficacy outcomes often focuses on the time to a first event, where the event may be “any drinking”, “safe (or low risk) drinking”, “moderate drinking” or “heavy drinking,” in addition to multiple outcomes such as frequency of drinking days, percent abstinence days, etc.

Methods We consider the multivariate failure time analytic methods. In alcoholism treatment trials, the naturalistic course of drinking behavior during treatment intervention often presents with a gradual change in drinking before the emergence of a more stable drinking or abstinence pattern. Thus, for each subject, evaluation of all drinking events, and incorporating the event times over a defined duration, may give a more comprehensive description of his/her drinking pattern. As a consequence, the efficacy of a new treatment for alcoholism may be elevated with greater statistical sensitivity.

Results The utility of the multiple failure time method is demonstrated via a real case study for evaluation of alcoholism treatments. The multiple event time analyses showed that the risk of having “any drinking days” or “heavy drinking days” during the entire duration of the study was significantly lower with experimental treatment than with placebo. Further explorations showed that the treatment effect was primarily observed in the later relapse events and not the first event with respect to relapse to any drinking episodes. Such effect would have missed using the traditional time to first event analysis approach. The observed effect of treatment with respect to relapse to multiple heavy drinking episodes was shown not only in the first event but also in the later events.

Conclusion The multiple failure time approach may be applicable when ‘drinking failure’ is variously defined as a single drink, one at‐risk drinking day, one heavy drinking day, or one alcohol‐related social, occupational or medical problem. If “a drinking episode” is properly defined and the design gains statistical efficiency, the multiple event analytic strategy should provide improved statistical power to detect treatment effects.
Drinks of the Father: Father's Maximum Number of Drinks Consumed Predicts Externalizing Disorders, Substance Use, and Substance Use Disorders in Preadolescent and Adolescent Offspring
Authors: Malone Stephen M. , Iacono William G. , McGue Matt
Published: April 2006

Background The maximum number of drinks consumed in 24 hr seems to be an interesting phenotype related to alcoholism. The goal of the present study was to determine in an epidemiologic sample whether this measure of drinking history in fathers predicted externalizing behavioral disorders, substance use, and substance abuse in preadolescent and adolescent offspring and whether any such associations would be independent of paternal alcohol dependence diagnoses.

Methods Subjects were male and female twins from both age cohorts of the Minnesota Twin Family Study, a population‐based longitudinal study, and were approximately 11 or 17 years of age, respectively, upon study enrollment. In both age cohorts, diagnoses of conduct disorder, oppositional defiant disorder, and attention‐deficit/hyperactivity disorder served as outcome measures. In addition, measures of lifetime substance use and of the presence of symptoms of substance abuse were derived for the 11‐year‐old cohort when subjects were approximately 14 years old and diagnoses of substance abuse were derived for the older cohort at age 17. An extension of logistic regression using generalized estimating equations served to assess whether paternal maximum alcohol consumption predicted filial outcome measures.

Results Paternal maximum alcohol consumption was consistently associated with conduct disorder, substance use, and substance abuse or dependence in male and female offspring. These associations were not mediated by a primary effect of paternal alcoholism.

Conclusions Paternal maximum alcohol consumption was uniquely associated with those offspring characteristics most reliably found in adolescent children of alcoholic parents. This phenotype might supplement DSM diagnoses of alcohol dependence to reduce the number of false positives in genetic research.
Effects of Chronic Hepatic Dysfunction on Pulmonary Glutathione Homeostasis
Authors: Foreman Marilyn G. , Hoor Terri Ten , Brown Lou Ann S. , Moss Marc
Published: April 2006

Background The development of acute respiratory distress syndrome (ARDS) in patients with pre‐existing cirrhosis of the liver is associated with very high mortality. One possible cause may be alteration of pulmonary antioxidant capacity as a result of chronic hepatic dysfunction. Glutathione (GSH) is the most substantial nonprotein thiol in living organisms and likely plays a key role in neutralizing the oxidants and reactive oxygen species that are increased in ARDS. The lung is unable to synthesize GSH and is dependent on the liver. During periods of oxidant stress, individuals may exhibit relative deficiencies of GSH. With cirrhosis, the end result of chronic alcohol ingestion, this deficiency is more profound.

Methods Sixteen stable subjects with cirrhosis primarily due to alcohol consumption and 15 healthy controls underwent bronchoscopy and bronchoalveolar lavage with concurrent measurement of GSH in the plasma and the alveolar epithelial lining fluid (ELF).

Results For standardizing for saline dilution of the epithelial lining fluid as a result of bronchoalveolar lavage, GSH values are expressed in relation to immunoglobulin A (IgA). GSH in the ELF was profoundly reduced in the cirrhotic group [12.5 μg of GSH per μg of IgA (5.3–16.9 μg)] compared with the control group [64.0 μg of GSH per μg of IgA (55.1–242.5 μg);p < 0.001]. The ratio of oxidized GSH to total GSH in the ELF was also significantly increased in the cirrhotic group [9.2% (5.1–16.4%) vs. 3.4% (1.7–5.7%);p < 0.003]

Conclusions Despite a total reduction in GSH concentrations in the alveolar epithelial lining fluid of individuals with cirrhosis, the amount of oxidized GSH is increased. There is increased utilization of GSH despite the low supply in stable individuals with cirrhosis during steady state. These perturbations in GSH homeostasis in the alveolar epithelial lining fluid may be a factor in the poor outcomes seen in these individuals with ARDS.
Alcohol Unmasks Simian Immunodeficiency Virus‐Induced Cognitive Impairments in Rhesus Monkeys
Authors: Winsauer Peter J. , Moerschbaecher Joseph M. , Brauner Ian N. , Purcell Jeanette E. , Lancaster Jack R. , Bagby Gregory J. , Nelson Steve
Published: April 2006

Background Alcohol and human immunodeficiency virus (HIV) produce similar neuropathological profiles, including loss of neurons in the frontal cortex. Additionally, HIV‐positive patients with a history of alcohol abuse have greater neurologic deficits, and chronic alcohol abuse produces electrophysiological deficits earlier in the HIV disease process. Few studies, preclinical or clinical, have examined whether alcohol administration exacerbates the neuropsychological deficits in subjects with lentiviruses such as HIV.

Methods To examine the combined effects of alcohol and immunodeficiency viruses (IVs) on neuropsychological functioning, four groups of young adult rhesus monkeys trained to respond under two complex behavioral tasks were administered ethanol 4 days per week via an intragastric catheter for 3 months and then infected with simian immunodeficiency virus (SIV). Behavioral testing after SIV inoculation was conducted in each group (−ethanol [EtOH]/−SIV, −EtOH/+SIV, +EtOH/−SIV, and +EtOH/+SIV) on days when alcohol was not administered to avoid a direct confound and on several occasions when ethanol or sucrose was administered as a probe of the effect of alcohol alone and the effect of caloric supplementation on the food‐maintained tasks, respectively.

Results During the days of the week when ethanol was not administered, little or no disruption was observed in either response rate or the percentage of errors (accuracy) across the different treatment groups. In contrast, behavioral testing during alcohol administration revealed that subjects in the various treatment groups had different susceptibilities to ethanol administration. As expected, a two‐way ANOVA (ethanol condition, SIV condition) indicated there were significant main effects of ethanol on both response rate and percent errors in both behavioral tasks, but it also indicated there was a significant interaction between ethanol administration and SIV infection on the accuracy of responding in the acquisition (learning) task. In addition, the main effect of SIV on percent errors was in the performance task.

Conclusions The fact that alcohol administration in SIV‐infected monkeys produced greater behavioral deficits than either alcohol or SIV alone further strengthens the supposition that IVs adversely affect neural substrates involved in cognition and that the adverse effects of many central nervous system drugs may be enhanced in IV‐infected individuals.
Muscle Antioxidant Status in Chronic Alcoholism
Authors: Fernández‐Solà Joaquim , García Gloria , Elena Montserrat , Tobías Esther , Sacanella Emilio , Estruch Ramón , Nicolás José‐María
Published: April 2006

Background Chronic myopathy due to excessive ethanol intake is one of the most frequent causes of acquired skeletal myopathy in developed countries. Its pathogenesis is multi‐factorial, only partially clarified, and antioxidant imbalance has been suggested to influence its development, being a type II glucolytic, fast‐twitch fiber subset more sensitive to this effect.

Methods We assessed superoxide dismutase, glutathione peroxidase, and glutathione reductase enzyme activities as well as the total antioxidant status capacity in muscle samples obtained from 41 chronic alcoholic males and 12 age‐matched controls. Alcoholic skeletal myopathy was defined according to standard histologic criteria. We evaluated the influence of ethanol consumption, caloric and protein nutritional status, and the presence of skeletal myopathy with the tissue activities of these antioxidant enzymes.

Results Chronic alcoholics showed a 16% reduction in glutathione peroxidase and a 13% increase of superoxide dismutase in the skeletal muscle, compared with controls (p < 0.05, both). Muscle antioxidant changes in chronic alcoholics were not related to the presence of skeletal myopathy, parameters of alcohol consumption, or conventional nutritional parameters.

Conclusions Antioxidant muscle enzyme activities are partially disturbed in chronic alcoholism, although not related to the presence of myopathy, amount of ethanol consumed, or the nutritional status of the patients. Further studies should assess other aspects not included in the present study such as muscle site‐specific changes in antioxidant status/oxidative damage, specific fiber‐type sensitivity to alcohol, and type and quantity of antioxidant content of the diet or in the alcohol beverages.
Alcohol Potentiates HIV‐1 Infection of Human Blood Mononuclear Phagocytes
Authors: Wang Xu , Douglas Steven D. , Metzger David S. , Guo Chang‐Jiang , Li Yuan , O'Brien Charles P. , Song Li , Davis‐Vogal Annet , Ho Wen‐Zhe
Published: April 2006

Background Acute and chronic alcohol abuse impairs various functions of the immune system and thus has been implicated as a cofactor in HIV infection. The mechanisms by which alcohol affects the function of human immune cells that are the targets for HIV are unknown.

Methods Human blood monocyte‐derived macrophages (MDM) were incubated with or without alcohol (10–40 mM) for 24 hr and then infected with HIV for 24 hr. Culture supernatants were harvested for HIV reverse transcription assay. HIV entry receptor (CCR5, CD4, and CXCR4) expression was determined by reverse transcription‐polymerase chain reaction and flow cytometry assays. β‐Chemokines were analyzed using enzyme‐linked immunosorbent assay. Different HIV strains (Bal, SF‐162, 89.6, and UG024) were used for infection experiments. In addition, ADA (macrophage‐tropic strain) and murine leukemia virus envelope‐pseudotyped HIV infection was carried out.

Results Although alcohol had little effect on HIV T‐lymphocyte‐tropic strain infection, it significantly enhanced HIV R5 strain infection in MDM. The enhancing effect of alcohol on the HIV R5 strain was further evidenced by the observation that the R5 (ADA) strain envelope‐pseudotyped HIV infection is markedly increased by alcohol, whereas murine leukemia virus envelope‐pseudotyped HIV infection was not affected. Alcohol significantly up‐regulated CCR5 receptor expression and inhibited the endogenous production of β‐chemokines by MDM.

Conclusion Alcohol, through the down‐regulation of β‐chemokine production and the up‐regulation of CCR5 receptor expression, enhances HIV R5 strain infection of MDM and may have an important role as a cofactor in the progression of HIV disease.
Modulation of Immunity and Viral‐Host Interactions by Alcohol
Authors: Thiele Geoffrey M. , Szabo Gyongyi , Kovacs Elizabeth J. , Bautista Abraham P. , Sosa Laura , Jerrells Thomas R.
Published: April 2006

This manuscript represents the proceedings of a symposium at the 2001 RSA Meeting in Montreal, Canada. The organizers/chairs were Gyongyi Szabo and Geoffrey M. Thiele. The presentations were (1) Introduction, by Gyongyi Szabo; (2) Chemokine dysregulation after acute ethanol exposure, by Elizabeth J. Kovacs; (3) Chemokine production and innate immunity in the livers of simian immunodeficiency virus‐infected Macaca mulatta following chronic alcohol administration, by Abraham P. Bautista; (4) Influence of ethanol consumption on the severity and progression of hepatitis associated with cytomegalovirus infection, by Laura Sosa and Thomas R. Jerrells; (5) Scavenger receptor involvement in the immune response to the metabolites of chronic ethanol ingestion, by Geoffrey M. Thiele; and (6) Mechanisms of impaired accessory cell functions due to alcohol exposure and hepatitis C infection, by Gyongyi Szabo.
Early‐Onset Alcoholism With Conduct Disorder: Go/No Go Learning Deficits, Working Memory Capacity, and Personality
Authors: Finn Peter R. , Mazas Carlos A. , Justus Alicia N. , Steinmetz Joseph
Published: April 2006

Background: Two studies were conducted to investigate the disinhibitory mechanisms that (1) discriminate early‐onset alcoholism (EOA) with conduct disorder (CD; antisocial EOA) from a nonantisocial subtype of EOA and (2) are associated with novelty‐seeking and low harm avoidance.

Methods: Young adults with antisocial EOA (n= 96), with nonantisocial EOA (without CD;n= 80), with CD alone (n= 50), and controls (n= 125) were given two go/no go tasks (one with monetary loss and the other with shock punishment), the Digit Span test (working memory capacity), and personality measures of harm avoidance, novelty‐seeking/impulsivity, excitement‐seeking, and negative affectivity.

Results: Study 1 revealed that antisocial EOA subjects had poor behavioral inhibition compared with nonantisocial EOAs and controls on both go/no go tasks and with the CD‐alone group on the monetary‐loss task. Low Digit Span scores accentuated poor inhibition in antisocial EOAs on the monetary loss, but not the shock task. EOA with low Digit Span was associated with higher hit rates on the shock task. Study 2 revealed that antisocial EOAs had high novelty‐seeking/impulsivity and low harm avoidance compared with both nonantisocial EOAs and controls. Low harm avoidance was associated with poor inhibition with shock punishment, and this association was mediated by CD. For subjects with low Digit Span scores, novelty‐seeking/impulsivity was associated with poor inhibition to monetary‐loss punishment and higher hit rates to shock punishment.

Conclusions: The results suggest two disinhibitory mechanisms that distinguish antisocial from nonantisocial EOA: an increased sensitivity to reward in nonaversive contexts associated with novelty‐seeking/impulsivity and a decreased sensitivity to punishment in aversive contexts associated with low harm avoidance. Results also suggest that EOA and novelty‐seeking/impulsivity are associated with a greater response to rewards in those with low working memory capacity.
Chronic Ethanol Consumption Increases Plasma Leptin Levels and Alters Leptin Receptors in the Hypothalamus and the Perigonadal Fat of C57BL/6 Mice
Authors: Obradovic Tanja , Meadows Gary G.
Published: April 2006

Background: Leptin, a protein secreted from adipocytes, and leptin receptors, expressed in the hypothalamus and fat tissue, regulate energy balance and adiposity. Leptin receptors exist as several forms that are delineated by the length of their cytoplasmic domains. Only the longest form is physiologically active through stimulation of the signal transducers and activators of transcription (STAT) second messenger system. Previous studies demonstrated that chronic ethanol consumption increases body adiposity and circulating leptin levels. This study examined the effect of chronic alcohol intake on the expression of leptin receptors and associated STAT second messengers in the hypothalamus and fat tissue.

Methods: Female C57BL/6 mice were given water or 20% w/v ethanol and Purina 5001 laboratory chow ad libitum for 2 or 5 weeks. Plasma leptin levels were determined by radioimmunoassay and body adiposity was evaluated by measuring the amount of perigonadal fat. Expression of leptin receptors as well as STAT molecules in whole‐cell lysates of the hypothalamus and the perigonadal fat were assayed by Western blot analysis.

Results: Plasma leptin and perigonadal fat were increased after 5 weeks. The overall expression of leptin receptors was increased while the expression of the physiologically active long form of leptin receptor was decreased in the hypothalamus and the perigonadal fat of ethanol‐consuming mice. Ethanol consumption also decreased hypothalamic expression of STAT3, a protein associated with leptin receptor activation in this region of the brain. In contrast, STAT1, a protein associated with leptin receptor activation in adipose tissue, was significantly elevated in the perigonadal fat of ethanol‐consuming mice.

Conclusions: Chronic ethanol consumption increases the circulating level of leptin, and this is accompanied by altered expression of leptin‐sensing molecules in the hypothalamus and peripheral adipose tissue. These results suggest that chronic ethanol intake affects metabolism by altering the leptin system that regulates energy balance.