bluuebunny/crossref_metadata_2025 huggingface dataset:
Ethanol‐Induced Conditioned Taste Aversion in BXD Recombinant Inbred Mice
Authors: Risinger Fred O. , Cunningham Christopher L.
Published: May 2006

Genetic differences in sensitivity to ethanol's aversive effects may play an important role in the development of alcohol‐seeking behavior and alcoholism. The present study examined the development of ethanol‐induced conditioned taste aversion in 20 BXD/Ty recombinant inbred strains of mice and their progenitor inbred strains, C57BL/6J (B6) and DBA/2J (D2). Adult male mice were given 1‐hr access to a saccharin‐flavored solution every 48 hr for 12 days. After all but the first and last saccharin access periods, they received ethanol injections (0, 2, or 4 g/kg, ip). Separate groups of unpaired control mice received 4 g/kg of ethanol 1 hr after water access. Saline control mice were also used for examining preference across a wide range of saccharin concentrations (0.019 to 4.864% w/v). As expected, saccharin consumption during taste conditioning declined over conditioning trials in a dose‐dependent manner, indicating development of ethanol‐induced conditioned taste aversion. Correlational analyses using strain means from recently published papers indicated no significant genetic correlation between taste conditioning and two phenotypes thought to reflect ethanol reinforcement or reward (ethanol drinking, conditioned place preference). However, there were significant genetic correlations between taste conditioning at the high dose and sensitivity to ethanol‐induced hypothermia, rotarod ataxia, and acute withdrawal. Quantitative trait locus (QTL) analyses of strain means indicated that taste aversion was associated (p0.01) with genetic markers on nine chromosomes (1, 2, 3, 4, 6, 7, 9,11, and 17). These QTLs were located near several candidate genes, including genes encoding several different acetylcholine receptor subunits, the 6 opioid receptor, and two serotonin receptors (lB and 1D). QTLs for saccharin preference were located on several of the same chromosomes (2,3,4,6, and 11). Two of these saccharin QTLs overlap candidate genes influencing sensitivity to sweet or bitter taste stimuli. In general, these findings support the conclusion that multiple genes influence ethanol‐induced conditioned taste aversion. Some of these genes appear to influence taste sensitivity, whereas others appear to mediate sensitivity to aversive pharmacological effects of ethanol.
Effect of Ethanol Drinking on the Gene Expression of Opioid Receptors, Enkephalinase, and Angiotensin‐Converting Enzyme in Two Inbred Mice Strains
Authors: Winkler Anett , Buzas Beata , Siems Wolf‐Eberhard , Heder Gottfried , Cox Brian M.
Published: May 2006

There is convincing evidence that genetic factors contribute to the predisposition to alcoholism. In this respect, alcohol‐preferring (like C57BL/6 mice) and alcohol‐avoiding lines (like DBA/2 mice) of animals served as models in the search for neurobiological substrates of excessive ethanol consumption. One of the systems that is thought to be associated with the incidence of alcoholism is the endogenous opioid system. In the first experiment, basal mRNA levels of μ‐ and §‐opioid receptors, and of opioid‐degrading enzymes enkephalinase (neutral endopeptidase 24.11; NEP) and angiotensin‐converting enzyme (ACE) in the brain regions of C57BL/6 and DBA/2 mice did not reveal genetically determined differences in these parameters between the two strains. Furthermore, in the brain regions studied, the corresponding enzyme activities of NEP and ACE did not differ significantly between the lines of mice, except for a higher NEP activity in the striatum and olfactoty bulb of DBA/2 mice (p< 0.01). In the second experiment, C57BL/6 and DBA/2 mice were offered a free choice between water and 10% ethanol solution for 4 weeks and were killed thereafter; from another group, ethanol was removed for 3 days and from a third group ethanol was removed for 3 weeks before killing. In the striatum, a highly significant increase in the ACE mRNA amount was detected after 3 weeks of removal of ethanol in C57BL/6 mice, whereas in DBA/2 mice the §‐opioid receptor mRNA level was increased at this time when compared with the corresponding ethanol treatment group. The most striking changes were seen in the hypothalamus, where μ‐opioid receptor, ACE, and NEP mRNA amounts markedly decreased after ethanol treatment in both strains. Thus, chronic ethanol intake caused significant changes in the gene expression of distinct components of the endogenous opioid system. These findings further underline an involvement of the opioid system in the effects of ethanol.
Acute Effects of Ethanol on Recombinant Kainate Receptors: Lack of Role of Protein Phosphorylation
Authors: Valenzuela C. Fernando , Cardoso Rita A. , Lickteig Ronald , Browning Michael D. , Nixon Kristin M.
Published: May 2006

This study examined the acute actions of ethanol on recombinant rat GIuR6 kainate receptors expressed in Xenopus oocytes and HEK 293 cells. Electrophysiological recordings showed that co‐application of ethanol with submaximal kainate concentrations resulted in similar inhibition of kainate‐gated currents in both expression systems. Manipulation of intracellular phosphorylation pathways by intracellular dialysis with a solution without ATP and GTP did not modify the inhibitory effects of ethanol. Moreover, co‐transfection of GIuR6 receptor subunits with PKA‐α catalytic subunit or the calcium/ calmodulin‐dependent protein kinase II (CamKII) catalytic fragment did not change the sensitivity of the receptor to ethanol. Treatment of Xenopus oocytes with specific inhibitors of PKC, PKA, CamKII, tyrosine kinases, and serine‐threonine protein phosphatases did not affect the 100 mM ethanol‐induced inhibition of GIuR6 receptor‐mediated currents. Biochemical experiments with transiently transfected HEK 293 cells confirmed published reports that GIuR6 receptors are minimally phosphorylated under basal conditions in these cells and also revealed that acute ethanol did not increase GIuR6 phosphorylation. These results suggest that, under our experimental conditions, ethanol inhibits recombinant GIuR6 receptor function by a direct effect on the receptor rather than an indirect action via protein phosphorylation.
What Makes Alcohol‐Dependent Individuals Early in Abstinence Crave for Alcohol: Exposure to the Drink, Images of Drinking, or Remembrance of Drinks Past?
Authors: Weinstein Aviv , Lingford‐Hughes Anne , Martinez‐Raga Jose , Marshall Jane
Published: May 2006

Craving is a major factor in addiction, predicting poorer outcome to treatment. To improve our understanding of craving for alcohol, we have compared in the laboratory the effects of inducting craving for alcohol by exposure to the sight and the smell of an alcoholic beverage, imagery of craving scripts, and recall of autobiographical memories of craving. We used subjective measures of craving, together with autonomic measures, in 14 abstinent alcohol‐dependent individuals in the first month after detoxification. All subjects reported a significant increase in ratings of urges after exposure to alcoholic drinks, following the imagery of craving and after recalling autobiographical memories of craving. Physiological measures have shown that craving imagery as well as memory induction were equally effective as exposure to alcoholic drinks in modestly increasing autonomic arousal (indicated by systolic blood pressure). Our preliminary findings support the existing evidence in nicotine and opiate dependence that images and memories are as effective as in vivo exposure in eliciting craving for drugs.
Nalmefene Causes Greater Hypothalamic‐Pituitary‐Adrenal Axis Activation than Naloxone in Normal Volunteers: Implications for the Treatment of Alcoholism
Authors: Schluger James H. , Ho Ann , Borg Lisa , Porter Margaret , Maniar Swapnil , Gunduz Mithat , Perret Guillaurne , King Andrea , Kreek Mary Jeanne
Published: May 2006

Among other actions, opioid antagonists modulate the control endogenous opioids exert on the hypothalamic‐pituitary‐adrenal (HPA) axis. Naloxone, nalmefene, and naltrexone are the opioid antagonists approved for use in man and are primarily μ‐opioid selective. Naltrexone and nalmefene have been demonstrated to be useful in the treatment of alcoholism. Compared with naloxone, nalmefene has a longer half‐life, is more potent at the preceptor, and has a higher affinity for K‐ and §‐opioid receptors. We conducted an inpatient study comparing the effects of 10 and 30 mg doses of intravenous naloxone and nalmefene in normal, nonsubstance nor alcohol‐abusing, volunteers. Significant increases in ACTH and cortisol were observed after both antagonists, without an apparent dose‐response relationship; however, both doses of nalmefene resulted in greater HPA axis activation than either dose of naloxone (ACTH: p <0.005). These results indicate that K‐ and 8‐opioids may play important roles in the regulation of the HPA axis; nalmefene may be useful as both a probe to explore the HPA axis physiology and as a pharmacotherapeutic agent.
Alcoholic Beverage Preference and Risks of Alcohol‐Related Medical Consequences: A Preliminary Report from the National Longitudinal Alcohol Epidemiologic Survey
Authors: Chou S. Patricia , Grant Bridget F. , Dawson Deborah A.
Published: May 2006

In studying the alcohol‐morbidity association, a substantial amount of attention and efforts has been focused on volume of alcohol intake. Considerably less is known about the differential health effects of beverage types. The present study used a most recent national household survey of the U.S. general population on drinking practices, alcohol use disorders, and their associated disabilities. The prevalence of a broad range of alcohol‐related diseases was examined with respect to preferred beverage type, as well as consumption level. Our findings showed a reduced health risk associated with beer and wine drinking for a number of physical disorders, and a somewhat favorable cardiovascular effect of these two beverage types in relation to abstention. Among preferrers of beer, wine, and liquor, the results indicate that liquor preference is associated with elevated morbidity for several medical consequences. However, interpretation of results and the public health implications of these findings need to be taken cautiously, because sociodemographic and other behavioral characteristics were not considered in this preliminary report.
The Effect of Drinking Intensity and Frequency on Serum Carbohydrate‐Deficient Transferrin and γ‐ Glutamyl Transferase Levels in Outpatient Alcoholics
Authors: Anton Raymond F. , Stout Robert L. , Roberts James S. , Allen John P.
Published: May 2006

Whereas heavy alcohol consumption is known to elevate serum carbohydrate‐deficient transferrin (CDT) and γ‐glutamyl transferase (GGT) levels, the contribution of drinking pattern to these effects is not completely understood. We present data on 423 men and 146 women evaluated 1 year after treatment in a large‐scale alcoholism treatment study (Project MATCH). Relationships between drinking frequency (number of days drinking), intensity (drinks per drinking day), and blood levels of CDT and GGT were analyzed by using response surface regression models and thin‐plate spline‐smoothing techniques. Both models indicated differences between CDT‐ and GGT‐drinking pattern relationships in men and, also, a difference between men and women in CDT drinking‐pattern relationships. For men, CDT levels appeared to respond primarily to frequency of drinking, whereas GGT was influenced primarily by drinking intensity. For women, both CDT and GGT were influenced more by drinks per drinking day (intensity) than by number of days drinking (frequency). The data confirm both the independent nature of these biological markers of alcohol consumption and gender differences in alcohol‐induced CDT response reported previously.
Ethanol Modulation of Excitatory and Inhibitory Synaptic Interactions in Cultured Cortical Neurons
Authors: Marszalec William , Aistrup Gary L. , Narahashi Toshio
Published: May 2006

The effects of ethanol on spontaneous excitatory and inhibitory postsynaptic currents (EPSCs and IPSCs) were studied in a culture of embryonic rat cortical neurons. In these experiments, EPSCs and IPSCs were recorded concurrently as inward and outward currents, respectively. These spontaneous currents were dominated by a slow (<1 Hz) repetitive pattern of prolonged N‐methyl D‐aspartate (NMDA)‐EPSCs and co‐occurring IPSCs when Mg2+ was left out of the perfusate. A 3‐ to 5‐min bath perfusion of 100 mM ethanol reduced the average integrated EPSC by 65%, while simultaneously potentiating IPSCs by about 3‐fold. EPSC frequency was also reduced by about one‐third. NMDA‐mediated EPSCs were inhibited more than non‐NMDA currents. A perfusion of 30 mM ethanol was less effective and probably represents a threshold concentration for these effects. The ethanol inhibition of currents evoked by directly applied glutamate or NMDA to these cells was much less than that observed for spontaneous EPSCs. Currents evoked by exogenous γ‐aminobutyric acid (GABA) application were never potentiated by ethanol. When spontaneous NMDA‐EPSCs were blocked with an NMDA antagonist, ethanol no longer potentiated the IPSCs. However, benrodiazepine treatment increased these IPSCs 2‐fold. In other experiments, spontaneous IPSCs were blocked by a GABAA antagonist. Here, the EPSCs occurred as groups of repetitive bursts. Ethanol decreased the total number of EPSCs per burst but did not decrease their overall amplitude, as in the control recordings. Thus, the way in which ethanol affects concurrently recorded spontaneous EPSCs and IPSCs appears different from the way in which it affects isolated GABA‐ and NMDA‐evoked currents. In addition, the antagonist studies show that concurrently activated NMDA and GABA channels each tend to limit the responses of the other. Thus, the overall effect of ethanol on spontaneous activity may result, in part, by a modification of this synaptic interaction.
Development of Alcohol Drinking Behavior in Rat Lines Selectively Bred for Divergent Alcohol Preference
Authors: McKinzie D. L. , Nowak K. L. , Murphy J. M. , Li T.‐K. , Lumeng L. , McBride W. J.
Published: May 2006

A characteristic of heritable alcoholism is an early onset of alcohol abuse, which may begin at or before the age of adolescence. The objective of the present study was to determine the ontogeny of alcohol drinking behavior before and during puberty in the selectively bred alcohol‐preferring (P), alcohol‐nonpreferring (NP), high alcohol drinking (HAD), and low alcohol drinking (LAD) lines of rats. In addition, the effects of postweaning housing conditions (single‐ or pair‐housed) and initiation procedure (4 days forced ethanol or free‐choice) were evaluated in male and female P rats. Results indicate that high alcohol drinking in P and HAD (replicate line 2) rats, as well as low alcohol drinking behavior in NP and LAD (replicate line 2) rats, is present as early as 3 to 4 weeks of age. Ethanol intakes in juvenile P and HAD rats reached levels of ∼4 to 5 g/kg/day by 38 to 41 days of age and were comparable with levels observed in adults. Neither housing conditions nor ethanol initiation procedure significantly altered the acquisition or magnitude of alcohol intake levels in juvenile male and female P rats. These results suggest that the neural substrates underlying divergent ethanol drinking behavior in P/NP and HAD/LAD lines of rats are present early in life.
Effects of Ethanol on Gene Expression in Rat Bone: Transient Dose‐Dependent Changes in mRNA Levels for Matrix Proteins, Skeletal Growth Factors, and Cytokines Are Followed by Reductions in Bone Formation
Authors: Turner Russell T. , Wronski Thomas J. , Zhang Minzhi , Kidder Louis S. , Bloomfield Susan A. , Sibonga Jean D.
Published: May 2006

Several studies were performed in female rats to determine dose and time course changes in mRNA levels for matrix proteins in bone after a single administration of ethanol. As expected, dose‐dependent transient increases in blood ethanol were measured. Additionally, there was mild hypocalcemia with no change in immunoreactive parathyroid hormone. Coordinated dose‐dependent increases in mRNA for type 1 collagen, osteonectin, and osteocalcin were noted in the proximal tibial metaphysis 6 hr after ethanol was given, with the peak values occurring at a dose of 1.2 g/kg (0.4 ml). Similar increases in mRNA levels for matrix proteins were noted in lumbar vertebrae after ethanol treatment. The changes were specific for bone; ethanol had no effect on mRNA levels for matrix proteins in the uterus or liver, although the mRNA concentrations tended to be reduced in uterus. Message levels for several cytokines implicated in the regulation of bone turnover were also assayed; mRNA levels for transforming growth factor‐β1, transforming growth factor‐β2, interferon‐γ, and interleukin‐6 were unchanged at doses ranging from 0.14 to 1.7 g/kg. At the highest dose of ethanol, the mRNA level for tumor necrosis factor‐α was elevated while the level for insulin‐like growth factor‐1 was reduced. The time course effects of ethanol (0.4 ml dose) were determined in a separate experiment. Ethanol resulted in a transient increase in mRNA levels for the three bone matrix proteins assayed. However, matrix protein synthesis, as determined by incorporation of 3H‐proline into the proximal tibial metaphysis, was not changed after 6 hr. The changes in mRNA levels for the matrix proteins were preceded by brief, transient decreases in mRNA levels for interleukin‐1 β, interferon‐γ, and migration inhibitory factor, and followed by a more prolonged decrease in the mRNA level for insulin‐like growth factor‐1. A subsequent study was performed to determine the effects of repetitive daily treatment with ethanol on rat bone. After 7 days, there were highly significant decreases in the mRNA level for type 1 collagen, as well as decreased bone formation. These results suggest that ethanol may alter bone metabolism by disturbing signal transduction pathways that regulate the expression of genes for bone matrix proteins, skeletal growth factors, and cytokines.
Distribution and Kinetics of Ethanol Metabolism in Rat Brain
Authors: Zimatkin Sergey M. , Liopo Anton V. , Deitrich Richard A.
Published: May 2006

It was found that the accumulation of acetaldehyde produced from 50 mM ethanol in rat brain homogenates takes place in all major brain regions. The velocity varied between 3.5 to 7.1 nmol/mg of protein/hr. The rate increased in the following orderbrain hemispheres, striatum, brainstem, hypothalamus, and cerebellum. Significant regional differences in this process were found in the initial period of incubation (5 min), acetaldehyde accumulation was maximal in the brain hemispheres; but, in the 30‐ to 60‐min period, it became significantly higher in the cerebellum. Inhibition of this process by the catalase inhibitor, 3‐amino‐1,2,4‐triazole (8 mM), was minimal in the brainstem (27%) and maximal (57%) in the cerebellum, despite nearly complete inhibition of catalase. This would indicate that processes other than catalase activity must contribute to acetaldehyde accumulation.
Ethanol Inhibits the Potentiation of Endotoxin by Dibutyryl cAMP and 2‐Methylthio ATP In Vivo
Authors: Zhao Xinfang , Jie Ouyang , Li Hua , Xie Jianming , Greenberg Stan S.
Published: May 2006

Ethanol (ETOH) selectively suppressedEscherichia coliendotoxin lipopolysaccharide (LPS)‐stimulated, but not dibutyryl cAMP (db‐cAMP)‐stimulated upregulation of inducible nitric oxide synthase (iNOS) in rat alveolar macrophages (AMs) in vivo (Zhao et al.,Alcohol. Clin. Exp. Res.21:1062–1074,1997). LPS‐induced stimulation of iNOS is inhibited in vitro by db‐cAMP and purine‐2‐Y (PPY) receptor‐mediated agonists. We examined the effect of ETOH on this interaction in rat lung AMs in vivo. Two hours after co‐administration of LPS [0.6 mg/kg, intratracheal (i.t.)] with db‐cAMP (0.1 mg/kg, i.t) or 2‐methylthio‐adenosine‐triphosphate (2‐mes‐ATP) to Sprague‐Dawley rats (225 to 250 g) (n= 10 to 24/gp) iNOS messenger ribonucleic acid (mRNA), iNOS protein, and nitrate and nitrite anions [reactive nitrogen intermediates (RNIs)] in bronchoalveolar lavage fluid (BALf), and the ex vivo incubates of AMs were increased more than when these compounds were given individually to the rats. Co‐administration of LPS with the autacoids did not affect LPS‐stimulated increases of tumor necrosis factor‐α (TNFα) mRNA, but inhibited LPS‐stimulated BALf TNFα protein and attenuated LPS‐mediated decreases of cell‐associated TNFα. Pretreatment of rats with ETOH (4.5 g/kg, ip) or diethyldithiocarbamate (DETC;5 mg/kg, i.t.), an inhibitor of the nuclear transcription factor NF kappa B (NF‐κB), 30 min before co‐administration of LPS with the autacoids restored iNOS mRNA levels to that obtained with the autacoid alone. Pretreatment of rats with ETOH decreased iNOS protein and RNI below that produced by either compound given individually to the rats. Although pretreatment of rats with DETC also decreased iNOS protein and RNI levels produced by LPS, the levels of both substances were elevated above that of the autacoid alone, LPS‐induced upregulation of iNOS mRNA was associated with elevated levels of p65/p50 NF‐κB in the nucleus of AMs. Neither db‐CAMP or 2‐mes‐ATP affected LPS‐stimulated NF‐κB‐DNA binding. Moreover, ETOH and DETC inhibited LPS‐stimulated NF‐κB‐DNA binding. We conclude that in rat AMs in vivo: (1) both db‐CAMP and P2Y‐receptor stimulation summate with, rather than inhibit, LPS‐induced upregulation of the iNOS mRNA, protein, and RNI; (2) ETOH and DETC inhibit LPS‐induced upregulation of iNOS mRNA only when stimulated through the NF‐κB pathway; and (3) ETOH inhibits both autacoid and LPS‐stimulated formation of iNOS protein by a mechanism independent of its ability to suppress iNOS transcription.
Alcohol and Heightened Aggression in Individual Mice
Authors: Miczek Klaus A. , Barros Helena M. , Sakoda Leigh , Weerts Elise M.
Published: May 2006

The objective of the current research was to study the large individual differences in alcohol effects on aggressive behavior under systematically varied conditions in experimental protocols with mice. Three experiments were conducted with outbred Swiss‐Webster derived mice that identified those individuals whose aggressive behavior was reliably heightened by low acute alcohol doses. In all experimental protocols, low alcohol doses were orally administered to a “resident” male mouse that subsequently confronted an “intruder” opponent for 5 min while all salient elements of aggressive behavior and motor activities were quantified. In all three experiments, alcohol (1.0 g/kg) heightened aggressive behavior by at least two standard deviations of the individual's water vehicle control mean in 27% of the mice. In 64% of mice, no reliable change in aggressive behavior was detected after the identical alcohol treatment, and in 9% of the mice alcohol decreased aggressive behavior. Experiments differed in protocol indicating that these aggression‐heightening effects were evident in resident mice that were either maintained at restricted or unlimited amounts of food, housed singly or in breeding pairs with a female partner, and conditioned to perform daily a food‐reinforced task or remained undisturbed. The first experiment found the aggression‐heightening effects to persist during weekly challenges for at least 2 months (n= 8 of 30). The second experiment showed these effects at intervals from 5 to 60 min after alcohol administration. Blood alcohol concentrations reached peak level within 5 to 10 min after oral administration in mice that had confronted an intruder. Those mice in whom alcohol heightened aggressive behavior (n= 21) did not differ from those that showed suppressed levels (n= 9) in terms of blood alcohol concentrations (79.6 vs. 82.4 mg%), suggesting that the intensity and frequency of aggressive behavior after alcohol were not directly dependent on the amount of alcohol in the circulation. The third experiment revealed that alcohol's (0.1 to 5.6 g/kg) effects on heightened aggressive behavior (n= 11) are dissociated from those on concurrently measured high‐ or low‐rate operant performance as engendered by a multiple FR 30‐FI 600 sec schedule of reinforcement. Current results indicate that this alcohol effect is relatively specific to aggressive behavior in individual animals, offering the opportunity for neuropharmacological and molecular characterization.
Effects of In Utero Alcohol Exposure on B‐Cell Development in the Murine Fetal Liver
Authors: Biber Kristen L. , Moscatello Kim M. , Dempsey Deborah C. , Chervenak Robert , Wolcott R. Michael
Published: May 2006

Fetal alcohol syndrome is one of the leading causes of birth defects in this country. Children exposed to alcohol in utero suffer from growth and mental retardation, physical abnormalities, and immune dysfunction. Previous work from this laboratory demonstrated that B lymphopoiesis is delayed in mice exposed to alcohol in utero. The deficit in B‐cell development was apparent shortly after birth and extended to well after weaning. Because lymphopoiesis begins in the fetal liver, the current study was done to determine if fetal B‐cell development was affected as well by in utero exposure to alcohol. We now show that the effects of in utero alcohol exposure on B lymphopoiesis do not become apparent until late in gestation. Flow cytometry was used to enumerate several intermediates in the B‐cell developmental pathway. These phenotypic analyses showed that before day 17 of gestation, B‐lineage intermediates developed normally when compared with control animals. However, between days 17 and 18 of gestation, an abnormality in the population dynamics of B‐lineage intermediates became apparent in the fetal liver of alcohol‐exposed mice. Early intermediates in the B‐cell developmental pathway were present in normal numbers; however, the more mature progenitors as well as B cells were decreased in number by gestational day 18. These data suggest that in utero alcohol exposure disrupts the ability of B‐lineage intermediates to progress along the developmental pathway to maturity, thereby leaving the animal immunocompromised at birth.
Acetaldehyde‐Induced Increase in Paracellular Permeability in Caco‐2 Cell Monolayer
Authors: Rao R. K.
Published: May 2006

Evidence indicates that endotoxin‐mediated liver injury plays an important role in the pathogenesis of alcoholic liver disease. Elevated plasma endotoxin level in alcoholics is suggested to be caused by enteric bacterial overgrowth and/or increased intestinal permeability to endotoxin. In this study, the effect of ethanol and acetaldehyde on the paracellular permeability was evaluated in Caco‐2 cell monolayers. Ethanol was administered into the incubation medium, whereas acetaldehyde was administered by exposing cell monolayers to vapor phase acetaldehyde, or by direct administration of an acetaldehyde generating system (AGS), ethanol + NAD++ alcohol dehydrogenase. Paracellular permeability was assessed by measuring transepithelial electrical resistance (TER), sodium chloride dilution potential, and unidirectional flux of d‐[2‐3H]mannitol. Administration of ethanol up to 900 mM produced no significant effect on paracellular permeability. Vapor phase acetaldehyde, generated from 5 to 167 mM acetaldehyde solutions in neighboring wells, resulted in a time‐ and dose‐dependent increase in acetaldehyde concentration (99 to 760 μM) in the buffer bathing cell monolayer. Acetaldehyde induced a reduction of TER and dilution potential, and an elevation of mannitol flux in a time and concentration‐related manner, without affecting the ability of cells to exclude trypan blue. Removal of acetaldehyde after 1, 2, or 4 hr treatment and subsequent incubation in the absence of acetaldehyde resulted in a time‐dependent reversal of TER to baseline values. Administration of AGS also reduced TER and dilution potential, associated with an increase in mannitol flux. This effect of AGS was prevented by 4‐methylpyrazole, an alcohol dehydrogenase inhibitor. These results show that acetaldehyde, but not ethanol, reversibly increases the paracellular permeability of Caco‐2 cell monolayer.
Soluble Proteins Modified with Acetaldehyde and Malondialdehyde Are Immunogenic in the Absence of Adjuvant
Authors: Thiele Geoffrey M. , Tuma Dean J. , Willis Monte S. , Miller Jacqueline A. , McDonald Thomas L. , Sorrell Michael F. , Klassen Lynell W.
Published: May 2006

Recent studies have shown that the alcohol metabolites malondialdehyde and acetaldehyde can combine to form a stable adduct (MAA) on proteins. This adduct has been detected in the livers of rats chronically consuming ethanol, and serum antibodies to MAA have been observed at significantly higher concentrations in ethanol‐fed when compared with pair‐fed or chow‐fed control rats. More recently, preliminary studies have strongly suggested that the MAA adduct is capable of stimulating antibody responses to soluble proteins in the absence of adjuvants. The antibodies produced recognize either the MAA epitope or the carrier protein itself. Therefore, it was the purpose of this study to examine the potential immunogenicity of MAA‐modified exogenous proteins in the absence of adjuvants. Balb/c mice were immunized in the presence or absence of adjuvant with different concentrations of unmodified or MAA‐modified proteins. The antibody response to both the MAA epitope and unmodified protein epitopes were determined by ELISA. In the absence of adjuvant, significant antibody responses were induced to both the MAA epitope and nonmodified protein epitopes. Smaller immunizing doses of MAA‐protein conjugate favored the production of antibodies to nonmodified proteins, whereas larger doses induced a strong anti‐MAA response. In studies to begin determining a mechanism for the specificity of the response in the absence of adjuvants, peritoneal macrophages were found to bind and degrade MAA‐adducted proteins through the use of a scavenger receptor. This indicated that MAA‐adducted proteins may be specifically taken up and epitopes presented to the humoral immune system in the absence of adjuvants. Importantly, these are the first data showing that an alcohol‐related metabolite can induce an antibody response in the absence of adjuvant and suggesting a mechanism by which antibody to the MAA adduct or its carrier (exogenous or endogenous) proteins may be generated in vivo.
Appetitive and Consummatory Behaviors in the Control of Ethanol Consumption: A Measure of Ethanol Seeking Behavior
Authors: Samson Herman H. , Slawecki Craig J. , Sharpe Amanda L. , Chappell Ann
Published: May 2006

Models of ethanol self‐administration in animals have demonstrated that ethanol can reinforce a variety of behaviors, independent of ethanol's caloric or fluid properties. However, the processes that control self‐administration remain unclear. Determining factors related to ethanol seeking behavior, independent of consumption, Is central to the concepts of intake regulation. The model described in this article proposes a method to separate the initial appetitive (seeking) behavior from the following consummatory (drinking) behavior to assess each behavior type. Rats were trained to lever press to gain access to a drinking tube connected to a fluid bottle containing either 10Y0 ethanol or 3% sucrose for 20 min. When the response requirement to obtain access to the tube was increased, it was found that both solutions supported the same amount of responding (breakpoint was at approximately a fixed ratio 32 requirement), indicating equal reinforcer strength. However, regardless of the response requirement, if access to the fluids occurred, intakes were not changed. This suggests that factors besides those of reinforcer efficacy are important in controlling the size of the consummatory bout. Based on these findings, we believe that this model will be useful in determining factors related to seeking behaviors and the control of drinking bout size.
Fetal Alcohol Effects in Alcoholic Veteran Patients
Authors: Tishler Peter V. , Henschel Carroll E. , Ngo Tu A. , Walters Ellen E. , Worobec Thomas G.
Published: May 2006

Fetal alcohol syndrome is often associated with severe physical and neuropsychiatric maldevelopment. On the other hand, some offspring of women who drank during pregnancy appear to be affected in minimal ways and function relatively well within society. We questioned whether this effect of prenatal alcohol in the adult is generally minimal. To bear on this, we determined whether we could distinguish alcohol‐exposed from nonexposed individuals in a population of male veterans, selected because of both their accepted level of function within society (e.g., honorable discharge from the military) and their admission to an alcohol treatment unit (thus, a greater likelihood of parental alcoholism, because of its familial aggregation). Consecutively admitted alcoholics (cases; n= 77) with likely maternal alcohol ingestion during their pregnancy or the first 10 years of life were matched with alcoholics with no maternal alcohol exposure during these periods (controls; n= 161). Each subject completed questionnaires regarding personal birthweight, alcohol, drug, educational and work histories, and family (including parental) alcohol and drug histories. We measured height, weight, and head circumference; checked for facial and hand anomalies; and took a frontal facial photograph, from which measurements of features were made. Data were analyzed by univariate statistics and stepwise logistic regression. No case had bona fide fetal alcohol syndrome. With univariate statistical analyses, the cases differed from the controls in 10 variables, including duration of drinking, width of alae nasae, being hyperactive or having a short attention span, and being small at birth. By stepwise logistic regression, the variables marital status, small size at birth, duration of drinking, and the presence of a smooth philtrum were marginally (the first two) or definitely (the last two) significant predictors of case status. Analysis of only the 37 cases in whom maternal prenatal drinking was the most likely yielded a marginal association for small size at birth (odds ratio = 3.1, p= 0.08) and a significant association for the presence of a smooth philtrum (odds ratio = 11.9, p= 0.005). Predictability was poor in all regression models. Based on the presence of this single physical finding (smooth philtrum), we estimate that the prevalence of manifestations of fetal alcohol exposure (fetal alcohol effects) is 6 to 13% in adult male veteran children (not necessarily nonveteran offspring) of women who drank alcohol during pregnancy. Thus, in our study of adult veterans, most individuals who were born to women who drank during pregnancy could not be differentiated from normal individuals, and those who were affected were distinguished by a single, nonspecific physical finding.
The Potential Mechanism of Induction of Inducible Nitric Oxide Synthase mRNA in Alveolar Macrophages by Lipopolysaccharide and Its Suppression by Ethanol, In Vivo
Authors: Greenberg Stan S. , Jie Ouyang , Zhao Xinfang , Wang Ji‐feng , Giles Thomas D.
Published: May 2006

This study reviews the putative mechanism of ethanol (ETOH)‐mediated downregulation of inducible nitric oxide synthase (iNOS) messenger RNA (mRNA) and protein and upregulation of constitutive NOS activity (ecNOS) in immunocompetent cells and endothelium, in vivo. Current evidence supports the hypothesis that ETOH inhibits the phospholipased‐tyrosine kinase pathway involved in the phosphorylation and activation of NADPH oxidase and myeloperoxidase, which upregulates the formation of reactive oxygen intermediates and mitogen‐activated protein kinase cascade, including the extracellular receptor‐linked kinase 1 and 2 (erkl and erk2). This decreases reactive oxygen intermediate formation, tyrosine kinase‐induced phosphorylation, and activation of transcription factors that, in turn, decreases the expression of iNOS mRNA. Also, ETOH‐mediated attenuation of endotoxin‐induced downregulation of nuclear protein kinase C activity appears to decrease the stability of expressed iNOS mRNA. ETOH‐mediated inhibition of tyrosine kinase activity may also explain the ability of ETOH to upregulate ecNOS enzymatic activity, because tyrosine kinase activity suppresses ecNOS enzymatic activity.
Differential Sensitivity to Alcohol Reinforcement in Groups of Men at Risk for Distinct Alcoholism Subtypes
Authors: Conrod Patricia J. , Pihl Robert O. , Vassileva Jasmin
Published: May 2006

The present study examines the relationship of familial and personality risk factors for alcoholism to individual differences in sensitivity to the positively and negatively reinforcing properties of alcohol. Sixteen sons of male alcoholics with multigenerational family histories of alcoholism (MFH) and 11 men who self‐report heightened sensitivity to anxiety (HAS) were compared with 13 age‐matched family history negative, low anxiety sensitive men (FH‐LAS) on sober and alcohol‐intoxicated response patterns. We were interested in the effects of alcohol on specific psychophysiological indices of “stimulus reactivity,” anxiety, and incentive reward. Alcohol significantly dampened heart rate reactivity to aversive stimulation for the MFH and HAS men equally, yet did not for the FH‐LAS group. HAS men evidenced idiosyncrasies with respect to alcohol‐induced changes in electrodermal reactivity to avenive stimulation (an index of anxiety/fear‐dampening), and MFH men demonstrated elevated alcohol‐intoxicated resting heart rates (an index of psychostimulation) relative to the FH‐LAS men. The results are interpreted as reflecting a sensitivity to the “stimulus reactivity‐dampening” effects of alcohol in both high‐risk groups, yet population‐specific sensitivities to the fear‐dampening and psychostimulant properties of alcohol in the HAS and MFH groups, respectively.
Selective Acceleration of Auditory Processing in Chronic Alcoholics during Abstinence
Authors: Pekkonen Eero , Ahveninen Jyrki , Jääskeläinen Iiro P. , Seppä Kaija , Näätänen Risto , Sillanaukee Pekka
Published: May 2006

Simultaneous auditory processing between the hemispheres was studied with a whole‐head magnetometer in 13 abstinent chronic alcoholics and 10 healthy control subjects. Auditory stimuli were presented monaurally with interstimulus intervals of 0.5 and 2.5 sec in different blocks. The N100m response, which contributes to stimulus detection, was significantly accelerated in the hemisphere ipsilateral to the ear stimulated in abstinent alcoholics. The MMNm response reflecting automatic stimulus‐change detection peaked earlier in alcoholics, and the ipsilateral N100m latency correlated significantly with the abstinence duration. These results suggest that auditory processing is accelerated in the auditory cortex ipsilateral to the stimulated ear in chronic abstinent alcoholics and that the accelerated processing is at least partly reversible. This may be caused by the hyperexcitation in the brain related to the ethanol withdrawal.
Effect of Pyrazole and Dexamethasone Administration on Cytochrome P450 2E1 and 3A Isoforms in Rat Liver and Kidney: Lack of Specificity of p‐Nitrophenol as a Substrate of P450 2E1
Authors: Zerilli Alain , Lucas Danièle , Dreano Yvonne , Picart Daniel , Berthou François
Published: May 2006

The induction effects of pyrazole and dexamethasone (known to be specific to P450 2E1 and 3A enzymes, respectively), given alone or simultaneously, were studied in rat liver and kidney microsomes. Pyrarole treatment induced the catalytic activity and the amount of P450 2E1 enzyme in both organs. Immunoreactive P450 2E1 and 4‐nitrophenol 2‐hydroxylation increased 8‐ and 13‐fold, respectively (versus control), in the kidney, but only 2.4‐ and 2.7‐fold (versus control) in the liver after pyrazole treatment. As assessed by nifedipine oxidation activity, dexamethasone treatment increased the P450 3A catalytic activity approximately 4‐fold (versus control) in the liver, but not in the kidney, suggesting that P450 3A was not inducible in the kidney. Pyrazole decreased P450 3A activity in the liver but did not modify it in the kidney. A combination of both chemicals induced both enzymes, but to a lesser extent than treatment with each single chemical compound. Furthermore, the 2‐hydroxylation of p‐nitro‐phenol, considered one of the most specific substrates for monitoring the level of P450 2E1, was mediated also by P450 3A, at least in dexamethasone‐treated rats. Finally, this experimental work demonstrated that P450 3A induction is organ‐specific, and it also demonstrated the lack of specificity of p‐nitrophenol as a P450 2E1 substrate.
More Vasopressin mRNA in the Paraventricular Hypothalamic Nucleus of Alcohol‐Preferring Rats and High Alcohol‐Drinking Rats Selectively Bred for High Alcohol Preference
Authors: Hwang Bang H. , Froehlich Janice C. , Hwang Wayne S. , Lumeng Lawrence , Li Ting‐Kai
Published: May 2006

Both the selectively bred alcohol‐preferring (P) and high alcohol‐drinking (HAD) rats exhibit alcohol preference, and develop tolerance to alcohol more quickly than their counterparts, the alcohol‐nonpreferring (NP) and low alcohol‐drinking (LAD) rats, respectively. It has been shown that the P rats retain developed tolerance longer than do NP rats, and alcohol drinking increases concurrently with the development of tolerance. Although alcohol preference and tolerance are fundamental elements of alcoholism, the exact mechanisms underlying these two phenotypes in P and HAD rats are not well understood. Recent studies have suggested that arginine vasopressin (AVP) may be involved in modulation of alcohol tolerance. Accordingly, this study was designed to examine whether the AVP mRNA level in the hypothalamus differs in rats that have been selectively bred for alcohol preference and nonpreference. A 35S‐AVP antisense oligodeoxynucleotide probe was used for in situ hybridization to localize AVP mRNA in the paraventricular hypothalamic nucleus (PVN) and supraoptic nucleus (SON), two major sites for AVP synthesis in the hypothalamus. Quantitative autoradiography demonstrated that P rats had higher levels of AVP mRNA in the PVN than NP rats. Similarly, higher levels of AVP mRNA were also found in the PVN of HAD rats, compared with LAD rats. The AVP mRNA levels in the SON were similar in the alcohol‐preferring and alcohol‐nonpreferring rat lines. Basal plasma AVP levels were higher in NP rats than in P rats as determined by radioimmunoassay, whereas plasma AVP levels were not significantly different between HAD and LAD rats. The results suggest that increased AVP gene expression in the PVN may contribute to alcohol preference and the development of alcohol tolerance.
Effect of Ethanol Administration on the Level of Dolichol in Rat Liver Microsomes and Golgi Apparatus
Authors: Cottalasso Damiano , Bellocchio Antonella , Pronzato Maria Adelaide , Domenicotti Cinzia , Traverso Nicola , Gianelli Maria Vittoria , Marinari Umberto Maria , Nanni Giorgio
Published: May 2006

Data obtained in our laboratory had suggested that acute ethanol administration (6 g/kg body weight) selectively and rapidly affects the intracellular system of protein glycosylation at the level of the Golgi apparatus. Dolichols are important membrane components, and dolichyl phosphate is a glycosyl sugar carrier for N‐glycosylation of proteins in endoplasmic reticulum and is considered rate‐limiting for this process. In this study, modifications in the concentration and distribution of liver microsomal dolichols after acute ethanol administration were investigated. Between 3 and 24 hr after ethanol administration, the microsomal dolichyl phosphate concentration was significantly lower than in control animals. The highest reduction was observed at 12 hr (–52%). An earlier and more marked reduction of total dolichol was observed in the Golgi apparatus, and, in particular, in the secretory fraction F1 (–70% at 6 hr). Ethanol treatment of isolated hepatocytes led to a significant reduction of the de novo synthesis of both dolichyl phosphate and free dolichol. Moreover, in vitro experiments have demonstrated that pro‐oxidant agents lead to a significant decrease of both free dolichol and dolichyl phosphate. Our results suggest that acute ethanol administration induces a marked decrease of dolichols, probably by increasing the degradation and impairing the biosynthetic pathway of these molecules.
Cultural and Subgroup Issues in Measuring Consumption
Authors: Caetano Raul
Published: May 2006

In May 1997, the National Institute on Alcohol Abuse and Alcoholism sponsored an International Workshop on Consumption Measures and Models for Use in Policy Development and Evaluation in Bethesda, MD. This paper is based on the “Cultural and Subgroup Issues in Measuring Consumption” session chaired by Raul Caetano; discussants were Raul Caetano, Salme Ahlström, Fe Caces, Paul Lemmens, Robin Room, Kathryn Graham, and Richard Wilsnack.
Special Issues of Measurement in Longitudinal Studies
Authors: Fillmore Kaye Middleton
Published: May 2006

This paper is based on the session discussing special issues of “Measurement in Longitudinal Studies” at the International Conference on Consumption Measures and Models for Use in Policy Development and Evaluation, May 12‐14, 1997 in Bethesda, MD. The session chair was Kaye M. Fillmore; and presenters were Thomas Harford, Bryan Johnstone, Paul Lemmens, Robin Room, Anders Romelsjo, and John Searles.
Evaluating Competing Models of Alcohol‐Related Harm
Authors: Greenfield Thomas K.
Published: May 2006

This paper summarizes “Evaluating Competing Models of Alcohol‐Related Harm” of the International Workshop on Consumption Measures and Models for Use in Policy Development and Evaluation, May 12‐14, 1997, Bethesda, MD. The session chair was Thomas Greenfield; presenters were Gerald Williams, Thomas Greenfield, Lorraine Midanik, Kathryn Graham, Jürgen Rehm, Anders Romelsjö, Deborah Dawson, Klaus Mäkelä, Tim Stockwell, and Paul Gruenewald; and discussants were Tim Stockwell, Timo Alanko, and Eric Single.
Alcohol as a Risk Factor for Brain Damage: Neurologic Aspects
Authors: Neiman Jack
Published: May 2006

Alcohol consumption in excess affects the brain negatively, both immediately and in the long‐term. Brain lesions in alcohol abusers are multiple and are multifactorial in origin. The toxic effect of ethanol, withdrawal from alcohol, nutritional deficits, and electrolyte disturbances, as well as liver damage, may contribute to the ethiopathogenesis of brain injury. The susceptibility of the brain to the negative effects of alcohol may be influenced by sex and age. The role of genetic factors and interactions of several licit and illicit drugs with alcohol needs further investigation.