bluuebunny/crossref_metadata_2025 huggingface dataset:
Personality and EEG β in Older Adults with Alcoholic Relatives
Authors: Pollock Vicki E. , Earleywine Mitchell , Gabrielli William F.
Published: April 2006

Research indicates that biological relatives of alcoholics are more likely to develop alcoholism than individuals without alcoholic relatives. Most research on these groups has focused on individuals who were relatively young (i.e., under 30 years old). In the present study, we evaluated middle‐aged and elderly nonalcoholic men and women who did and did not have alcoholic biological relatives to assess factors that might be involved in a later, rather than an earlier, onset of alcoholism. Psychological characteristics were assessed using the Tridimensional Personality Questionnaire. Biological characteristics were assessed using quantitative measures of the spontaneous electroencephalogram. The psychological measures did not distinguish the groups, but biological measures did. The results indicated that nonalcoholic individuals with alcoholic relatives showed elevated β as compared with sex‐ and age‐matched control subjects. Factors that might have mediated these findings are discussed, as are the implications of these outcomes.
Routine Activities and Alcohol Use: Constraints on Outlet Utilization
Authors: Gruenewald Paul J. , Treno Andrew J. , Nephew Thomas M. , Ponicki William R.
Published: April 2006

Studies of consumers' use of alcohol beverage outlets have provided a basis for understanding drinking behaviors in different drinking environments. These studies have shown that drinking environments are related to both demographic and drinking pattern measures. Absent from these studies has been a theoretical basis on which to make predictions regarding drinking patterns and choices of drinking environments under the various social, economic, and environmental constraints typically confronting alcohol consumers. This study presents one such theoretical approach.

The approach assumes that, in the context of individual preferences for alcohol, drinking choices are constrained by consumers' economic and time‐energy budgets for consumption. All other things being equal, it is suggested that greater budgets for consumption will be related to greater alcohol use, quality of beverages purchased, amenity values of purchase locations, or all three. Because on‐premise drinking entails greater economic costs, greater drinking levels will be related to lower utilization of on‐premise establishments.

The predictions of this approach were tested using data obtained from telephone surveys of consumers conducted in 1990 and 1991. The results showed that controlling for income, variables related to greater time‐energy budgets for consumption (i.e., marital status and household composition) were related to greater consumption levels and greater utilization of on‐premise establishments. Controlling for demographic measures, greater income was related to greater utilization of restaurants and increased beverage quality. Controlling for all other measures, frequencies of consumption were inversely related to consumption at on‐premise establishments, reflecting the expected moderation in costs for heavier consumers on a limited alcohol budget.
Anterior Hippocampal Volume Deficits in Nonamnesic, Aging Chronic Alcoholics
Authors: Sullivan Edith V. , Marsh Laura , Mathalon Daniel H. , Lim Kelvin O. , Pfefferbaum Adolf
Published: April 2006

Magnetic resonance imaging was used to quantify the volume of the hippocampus in 47 men with chronic alcoholism and 72 healthy male control subjects. The subjects ranged in age from 21 to 70 years, thus permitting a test of whether older alcoholics suffer greater brain tissue volume reduction than do younger ones. Comparison brain regions included temporal lobe gray matter, white matter, and cerebrospinal fluid, as well as measures of the lateral ventricles, third ventricle, and temporal horns. The results of this cross‐sectional study showed that the anterior, but not the posterior, portions of the hippocampus in both hemispheres were significantly smaller in the alcoholic than the healthy control group. Furthermore, the bilateral anterior hippocampal volume loss was greater in older than younger alcoholics. Despite the hippocampal volume deficit, these alcoholics did not demonstrate an explicit memory impairment; furthermore, memory test scores did not correlate significantly with hippocampal volumes. In the alcoholics, the age‐related volume loss, which was over and above that expected in normal aging, was also evident in the temporal cortex and white matter. Likewise, alcoholic ventricular enlargement was age‐related. Analysis of covariance revealed that the anterior hippocampal deficit persisted after accounting for the temporal lobe gray matter volume deficit. Multiple regression analysis revealed that the age‐related brain volume abnormalities observed in the alcoholics could not be attributed to duration of alcoholism or total lifetime consumption of alcohol.
Independent Associations among Maternal Alcohol Consumption and Infant Thyroxine Levels and Pregnancy Outcome
Authors: Hannigan John H. , Martier Susan S. , Naber John M.
Published: April 2006

Studies with animal models of alcohol‐related birth defects (ARBDs) suggest that reductions in circulating thyroid hormones, including thyroxine (T4), may be a persistent postnatal effect of fetal alcohol exposure. The few clinical reports of children with fetal alcohol syndrome (FAS) that address thyroid system function generally reported that FAS children have thyroid hormone levels within normal limits. For the current study, data bases from the Fetal Alcohol Research Center and the Michigan Department of Public Health Newborn Screening Program were assessed to correlate measures of maternal drug use during pregnancy and infant outcome (gestational age at birth, birthweight, “fetal growth”), with infant whole‐blood T4 levels. Multiple regression analyses accounted for demographic factors, infant age at testing, and variation in the T4 assay. As expected, alcohol intake and smoking each had a substantial negative impact on birthweight, gestational age at birth, and fetal growth, assessed as birthweight corrected for gestational age. Infant T4 levels were positively related to birthweight and gestational age and were more strongly related to fetal growth. Infant T4 levels were not influenced significantly by either maternal smoking or alcohol consumption. Smoking‐ and alcohol‐related reductions in birthweight, gestational age, or fetal growth were not associated significantly with variations in infant T4. Interesting questions remain regarding species differences and the influences of maternal alcohol consumption on T4 metabolism as a mechanism for ARBDs. However, the current data do not support the hypothesis that maternal alcohol consumption, or smoking, during pregnancy leads to compromised thyroid system function in newborn humans. Therefore, newborn screens for T4 are not likely to be effective in helping to detect infants with ARBDs or FAS.
Acute Prenatal Ethanol Exposure and Luteinizing Hormone‐Releasing Hormone Messenger RNA Expression in the Fetal Mouse Brain
Authors: Scott H. C. , Zoeller R. T. , Rudeen P. K.
Published: April 2006

Ethanol exposure during critical periods of development results in alterations of central nervous system morphology and function. In this study, the effects of acute ethanol exposure on the number of neurons expressing luteinizing hormone‐releasing hormone (LHRH) messenger RNA (mRNA) has been analyzed. Also, the expression of LHRH mRNA in the diagonal band of Broca/preoptic area (DBB/POA) was determined. Pregnant C57BL/6J mice were intubated with two doses of a 25% solution of ethanol or water (2.9 g/kg body weight) 4 hr apart on gestation day 7 (G7), G10, or G11. Animals were killed on G18, and in situ hybridization was utilized to detect neurons expressing LHRH mRNA. The number of neurons expressing LHRH mRNA was determined along their migration route from the rostrum into the forebrain. Ethanol exposure on G7 did not significantly change the number of neurons expressing LHRH mRNA on G18 compared with that in control animals. However, the number of neurons expressing LHRH mRNA in the nasal septum area in animals exposed to ethanol on G10 or G11 was significantly less than the number in control animals (p < 0.05). Prenatal ethanol exposure on any of the aforementioned treatment days did not alter the expression of LHRH mRNA at the level of the DBB/POA on G18 in ethanol‐treated animals compared with control animals. Also, neuron‐specific enolase mRNA expression at the level of the DBB/POA was not altered by prenatal ethanol exposure. Therefore, ethanol exposure on the aforementioned treatment days did not differentially affect LHRH mRNA expression compared with neuron‐specific enolase mRNA expression at the level of the DBB/POA. These data suggest that acute ethanol exposure may have specific effects on a subset of the LHRH neuron population, depending on the day of ethanol exposure during gestation and the neuroanatomical region in which the LHRH neurons are located.
Novel Synergistic Treatment of Ethanol Withdrawal Seizures in Rats with Dopamine and Serotonin Agonists
Authors: Mirovsky Yehudith , Yu Yi Lung , Wagner George C. , Sekowski Anna , Goldberg Melissa , Fisher Hans
Published: April 2006

A recent observation in this laboratory of a simultaneous increase in striatal dopamine and a decrease in serotonin in ethanol‐dependent rats during ethanol withdrawal prompted studies with combined dopaminergic + serotoninergic agonists to stop withdrawal seizures. Amphetamine (2 mg/kg) + fenfluramine (8 mg/kg) given jointly, but not separately, prevented ethanol withdrawal seizures as effectively as benzodiazepines (chlordiazepoxide), the current drugs of choice. The combination of amphetamine and fenfluramine, unlike chlordiazepoxide, significantly reduced intake of ethanol during and immediately following ethanol withdrawal.
Polymorphism at the P450IIE1 Locus Is Not Associated with Alcoholic Liver Disease in Caucasian Men
Authors: Carr Lucinda G. , Hartleroad Jeffrey Y. , Liang Yanbin , Mendenhall Charles , Moritz Thomas , Thomasson Holly
Published: April 2006

Because alcoholic hepatitis and cirrhosis, well‐known complications of alcohol abuse, do not occur in all alcoholics, genetic factors such as differences in alcohol‐metabolizing enzymes may play a role in the development of alcoholic liver disease. Cytochrome P450IIE1 catalyzes the oxidation of ethanol, producing acetaldehyde and free radicals capable of reacting with and peroxidizing cell membranes. Polymorphisms have been identified in the 51‐flanking region of the P450IIE1 gene that may alter the transcriptional activity of the gene. In this study, we analyzed the P450IIE1 genotypes at the polymorphic Pstl and Rsal restriction enzyme sites in 53 Caucasians with severe alcoholic liver disease to determine if there is an association between these polymorphisms and alcoholic liver disease. Subjects that tested positive for the hepatitis C virus were eliminated from the study. To identify the type A (homozygous for the d gene), type B (heterozygous for the d and c2 genes), and type C (homozygous for the c2 gene) genotypes at the P450IIE1 locus, DNA encompassing the polymorphisms was amplified by polymerase chain reaction, slot‐blotted, and probed with allele‐specific oligonucleotides. Allele frequencies for the d allele were 0.95 for alcoholics with severe liver disease, 0.95 for alcoholics without liver disease, and 0.98 for the general population. No differences in allele frequencies between alcoholic patients with severe liver disease and alcoholics without liver disease were observed.
Nitric Oxide‐Related Agents Alter Alcohol Withdrawal in Male Rats
Authors: Adams Michael L , Sewing Bryan N. , Chen Jingling , Meyer Edward R. , Cicero Theodore J.
Published: April 2006

Evidence has been reported supporting the hypothesis that nitric oxide (NO) partially mediates the expression of morphine dependence. To examine whether NO‐related agents also affect the expression of alcohol dependence, adult male rats were treated chronically with alcohol. Upon withdrawal of alcohol administration, abstinence signs were observed after treatment with a NO synthase (NOS) inhibitor, NG‐nitro‐l‐arginine methyl ester (NAME), or a NO donor, isosorbide dinitrate (ISDN). Withdrawal severity was based primarily on the presence and intensity of tremors, rigidity, hyperactivity, and spontaneous and audiogenic convulsions. The NOS inhibitor, NAME (10–100 mg/kg), injected during alcohol withdrawal significantly inhibited withdrawal severity decreasing the intensity of signs of hyperactivity, tremors, and rigidity, but not affecting the occurrence of convulsions. The NO donor, ISDN (30 mg/kg), administered during alcohol withdrawal significantly increased the severity of most withdrawal signs. These results suggest that NO mediates some aspects of the expression of alcohol dependence.
Regulation of Intracellular pH in Periportal and Perivenular Hepatocytes Isolated from Ethanol‐Treated Rats
Authors: Benedetti Antonio , Baroni Gianluca Svegliati , Marucci Luca , Mancini Raniero , Bassotti Cristina , Macarri Giampiero
Published: April 2006

The aim of this study was to gain information on intracellular pH (pHi) regulation in periportal (PP) and perivenular (PV) hepatocytes isolated from rats pair‐fed liquid diets with either ethanol (T rats) or isocaloric carbohydrates (C rats).

pHi was analyzed by the pH‐sensitive dye BCECF in perfused sub‐confluent hepatocyte monolayers. Cells were acid‐loaded by pulse exposure to NH4Cl and were alkali‐loaded by suddenly reducing external C02 and HC03‐(from 10% and 50 mm, respectively, to 5% and 25 mm) at constant pHout.

In cells from C rats: (a) steady‐state pHi was higher in PP than in PV hepatocytes in the presence, but not in the absence, of bicarbonate; (b) pHi recovery from an acid load was 35% higher in PP than in PV cells in the presence of HC03‐, whereas it was similar in HC03‐‐free experiments; and, on the contrary, (c) pHi recovery from an alkaline load was 30% higher in PV than in PP cells.

In cells from T rats: (a) steady‐state pHi was always lower than in cells isolated from pair‐fed animals; (b) steady‐state pHi was similar in PP and PV hepatocytes either in the presence or absence of bicarbonate in the perfusate; (c) pHi recovery from an acid load was not significantly different in PP and PV cells either in the presence of HC03‐ or in HC03‐‐free experiments; and (d) pHi recovery from an alkaline load was similar in PP and PV cells.

Our data suggest that chronic ethanol treatment selectively modifies pH, by affecting the activity of ion transport mechanisms regulating pHi in PP and PV hepatocytes isolated from rat liver.
Effects of Prenatal Exposure to Alcohol Plus Caffeine in Rats: Pregnancy Outcome and Early Offspring Development
Authors: Hannigan John H.
Published: April 2006

The factors determining susceptibility to fetal alcohol syndrome (FAS) are not fully understood. We used an animal model of alcohol‐related birth defects to assess the coteratogenic potential of caffeine as a risk factor in FAS. Rats were exposed prenatally to alcohol (˜15 g/kg/day) with or without caffeine (˜84 mg/kg/day) from gestation days 6 through 20 via liquid diet. All control groups were pair‐fed to the alcohol‐exposed groups. In addition, some controls had free access to lab chow and water. Prenatal exposure to alcohol or caffeine reduced both maternal weight gain during pregnancy and birth‐weight of offspring. The combination of alcohol plus caffeine produced an additive effect in reducing birthweight and synergistic effects in increasing postnatal offspring mortality. Prenatal alcohol exposure had a significant negative impact on several developmental indices, including grip strength and negative geotaxis. Prenatal caffeine exposure did not affect maturational measures and did reduce offspring serum levels of the zinc‐dependent enzyme alkaline phosphatase. This study in rats demonstrated that caffeine can exacerbate some of the effects of alcohol on prenatal development, specifically reduced birthweight, litter size, and postnatal survival, but that caffeine does not appear to alter prenatal alcohol‐induced delays in early postnatal maturation of survivors. The relative impact of intralitter birthweight rank on developmental outcome was also assessed. Birthweight influenced development, but did not interact significantly with either prenatal alcohol or caffeine. The results imply that high levels of maternal caffeine intake in humans could increase the likelihood that a child exposed prenatally to alcohol would be born with significantly lowered birthweight, one of the cardinal diagnostic criteria for FAS.
Molecular Biological Aspects of Alcohol‐Induced Liver Disease
Authors: Arnon Ron , Esposti Silvia Degli , Zern Mark A.
Published: April 2006

Molecular biological investigations have become a predominant methodology applied to the study of alcohol‐induced liver disease. The enzymatic pathways responsible for ethanol metabolism, and their genetic as well as environmental control, have become the focus of detailed investigation. More recently, the significance of cytokines in the pathogenesis of alcohol‐induced liver disase has also become a major area of speculation. This review focuses on the advances made in studies of two important enzymes responsible for alcohol metabolism, alcohol dehydrogenase and aldehyde dehydrogenase, as well as the investigation of the proinflammatory and profibrogenic cytokines involved in the process of hepatic fibrogenesis. The quality and quantity of new discoveries made in the field of alcohol‐induced liver disease is impressive, especially when one realizes that molecular biological approaches have been employed in this area for only 15 years. However, in most cases the studies have been predominantly descriptive, with little direct relevance to the therapeutics of alcoholism and alcohol‐induced organ injury. Because the groundwork has been laid, one hopes that the next 15 years will rectify this failure.
Ethanol Inhibition of Inducible Nitric Oxide Synthase Activity in C6 Glioma Cells
Authors: Syapin Peter J.
Published: April 2006

Nitric oxide (NO), a free radical gas, has been implicated in the CNS actions of ethanol. The brain contains several cell types that can produce NO, including neurons and glia. This study examined the effect of acute and chronic ethanol exposure on the activity of the inducible isoform of nitric oxide synthase (iNOS) found in neuroglia. Experiments were performed using intact rat C6 glioma cells, and NO production was assessed by nitrite accumulation after iNOS induction by coadministration of phorbol 12‐myristate 13‐acetate (PMA) and lipopolysaccharide (LPS). Ethanol was inhibitory at high concentrations (IC50= 150 mm) when acutely present during the 24‐hr period subsequent to initiation of enzyme induction. In contrast, cells exposed to ethanol were inhibited chronically at clinically relevant lower concentrations (IC50= 30 mm with 10 days exposure). Chronic inhibition was both time‐ and concentration‐dependent. Inhibition by ethanol seems to be a consequence of interference with LPS signal transduction. Acutely, ethanol did not affect the ability of PMA to synergize with LPS to induce activity, but it attenuated the ability of LPS to synergize with the PMA. Ten days exposure to 50 mm ethanol decreased the LPS potency by 4‐fold in the presence of a maximally activating concentration of PMA, although not significantly changing PMA potency. Inhibition by chronic ethanol exposure was long‐lasting, being retained over 24 hr in cells returned to control conditions. Thus, chronic ethanol may downregulate key components needed for iNOS expression. These results suggest that acute toxic and chronic ethanol exposure can inhibit NO production in the brain, and considering the ability of NO to protect neurons from free radical injury, this action could contribute to ethanol‐in‐duced brain damage.
Effects of Acute and Repeated Ethanol Exposures on the Locomotor Activity of BXD Recombinant Inbred Mice
Authors: Phillips Tamara J. , Huson Mary , Gwiazdon Courtney , Burkhart‐Kasch Sue , Shen Elaine H.
Published: April 2006

Investigations of ethanol's (EtOH's) complex response profile, including locomotor and other effects, are likely to lead to a more in‐depth understanding of the constituents of alcohol addiction. Locomotor activity responses to acute and repeated EtOH (2 g/kg, ip) exposures were measured in BXD recombinant inbred (RI) mice and their C57BL/6J (B6) and DBA/2J (D2) progenitors. Both the acute response and the change in initial EtOH response with repeated treatments were strain‐dependent. The coefficient of genetic determination was 0.38–0.49 for initial locomotor response to EtOH, and 0.29 for change in response. Changes in response were largely attributable to sensitization of locomotor stimulation. Quantitative trait loci (QTL) analyses identified significant marker associations with basal activity, acute locomotor response, and change in response. Markers were for QTL on several chromosomes, and there was only one case of overlap in marker associations among phenotypes. Acute locomotor response and locomotor sensitization were negatively correlated with 3% EtOH preference drinking data collected in BXD Rl strains. Overall, these results demonstrate locomotor sensitization induced by EtOH, suggest independence of genetic determination of locomotor responses to acute and repeated EtOH exposure, and partially support a relationship between reduced sensitivity to the locomotor stimulant/sensitizing effects of EtOH and EtOH consumption.
Liver Sinusoid during Chronic Alcohol Consumption in the Rat: An Electron Microscopic Study
Authors: Sarphie Theodore G. , Deaciuc Ion V. , Spitzer John J. , D'Souza Nympha B.
Published: April 2006

Transmission and scanning electron microscopic studies were performed on the liver sinusoid, with emphasis on sinusoidal endothelial cells, in rats fed a liquid diet containing either alcohol or dextrin (control) for 14 weeks. Animals were also treated with either Gram‐negative bacterial lipopolysaccharide (LPS; 100 μg/100 g body weight, intravenously) or sterile saline (control). All specimens were prepared after perfusion fixation of the liver. Livers of rats fed dextrin‐containing liquid diet displayed the ultrastructural features typical of the sinusoid and its endothelial cells. Livers from alcohol‐fed animals, however, were characterized by massive loss of sieve‐plate architecture of the sinusosidal endothelium, which was virtually replaced with a meshwork of enlarged openings with diameters frequently exceeding 1 urn. Morphological evidence of Kupffer cell activation could also be seen along with significant fatty infiltration of the hepatocyte. Conversely, LPS administration to dextrin‐fed animals induced an apparent decrease in fenestration of the sinusoidal endothelial cell, accompanied by morphological evidence of enhanced endocytotic activity and cytoplasmic swelling. The changes seen 3 hr after LPS administration were markedly advanced at 24 hr. LPS administration to alcohol‐fed rats accentuated the alterations observed after alcohol treatment alone. Additionally, the presence of platelets in the sinusoid as well as adhering to the hepatocyte microvilli in the space of Disse, along with the presence of Ito and Kupffer cell activation, greater than that observed in the alcohol‐treated rats, is morphological evidence consistent with the disruption of vascular integrity in the liver. Interestingly, LPS treatment did not reverse the effects of alcohol on the sinusoidal endothelial cell fenestration. The possible functional consequences of alcohol‐ and LPS‐induced ultrastructural alterations of the sinusoidal endothelial cell, and of the hepatic sinusoid in general, are evaluated in light of the available data on scavenging function of the sinusoidal endothelial cell.
Structural Analysis of Peptide‐Acetaldehyde Adducts by Mass Spectrometry and Production of Antibodies Directed Against Nonreduced Protein‐Acetaldehyde Adducts
Authors: Lin Renee C. , Smith Jean B. , Radtke David B. , Lumeng Lawrence
Published: April 2006

Acetaldehyde can form protein‐acetaldehyde adducts (AAs) in vivo and may play a role in the genesis of alcoholic liver disease. The nature of the chemical modification of proteins by acetaldehyde in vivo has not been elucidated. In vitro, acetaldehyde can form reversible adducts including a Schiff's base with lysine (K) and imidazolidinone with terminal amino groups of proteins such as human hemoglobin (Hb). In this study, we used FAB/MS to analyze the products of peptide‐AAs (pep‐AAs) formed by incubating acetaldehyde with Hb peptides. We then used an octabranched multiple antigen peptide (MAP) system containing Hb peptide‐AAs to raise antibodies. Three Hb peptides [i.e., 8‐pep consisting of 8 residues (V1HLTPVEK8) at the N‐terminus of β‐chain of human sickle‐cell Hb, 11‐pep‐gly consisting of 11 residues (G56NPKVKAHGKK66) in a segment of β‐chain rich in lysine, and 11‐pep‐pro that consists of the same sequence as 11‐pep‐gly, except G56 was replaced by proline (P)] were incubated with 1 mM acetaldehyde at 4°C for 7d without NaCNBH3 (nonreduced conditions). Analysis by FAB/MS showed that 8‐pep formed an imidazolidinone at the N‐terminal valine, 11‐pep‐gly formed a Schiff's base and imidazolidinone at the N‐terminus, whereas 11‐pep‐pro that lacks a free α‐amino group formed only a Schiff's base at K59. By contrast, incubation of these Hb peptides with 250 mM acetaldehyde and NaCNBH3 at 37°C for 1 hr (reduced conditions) produced mono‐and diethylated modifications of all available K residues, as well as the N‐terminal amino group. MAP‐11‐pep‐gly and MAP‐11‐pep‐pro were prepared and reacted with 1 mM acetaldehyde under nonreduced conditions. Antibodies against these nonreduced MAP‐11 ‐pep‐AAs were raised. By ELISA assay, antibodies against nonreduced MAP‐11‐pep showed reactivity with Hb‐AA prepared under nonreduced conditions. Unexpectedly, these antibodies also reacted with Hb‐AA prepared under reduced conditions as well. Our studies therefore showed: (1) acetaldehyde can modify peptides with or without NaCNBH3 producing different chemical modifications; (2) lysine residues, although located in close vicinity in a peptide, are not equally reactive in forming a stable AA; and (3) antibodies raised by nonreduced MAP‐11 ‐pep‐AA immunogens can recognize both reduced and nonreduced epitopes in Hb‐AA.
Ethanol‐Induced Depletion of Lymphocytes from the Mesenteric Lymph Nodes of C57B1/6 Mice Is Associated with RNA But Not DNA Degradation
Authors: Sibley Don A. , Fuseler John , Slukvin Igor , Jerrells Thomas R.
Published: April 2006

The effects of ethanol (EtOH) consumption by adult female C57BI/6 mice on lymphocyte populations of the mesenteric lymph nodes (MLNs) were determined by feeding mice with the Lieber‐DeCarli liquid diet by a pair‐feeding paradigm. Histological analysis of the MLNs of EtOH‐fed mice showed a progressive loss of lymphocytes from the medullary regions at 3, 5, and 7 days after initiation of the EtOH diet. The stromal cells in the medullary region also demonstrated a progressive alteration in stellate morphological features at times corresponding to those of loss of lymphocytes from this region. Microscopic evaluation of the follicle regions of MLNs obtained from mice fed an EtOH‐containing diet showed no appreciable alterations in morphological characteristics. The number of tingible body macrophages in the germinal centers of the follicles, however, was increased after 3 days of EtOH diet feeding and declined progressively after this time. Flow cytometric analysis of isolated lymphocytes showed a depletion of both T and B cell populations from the MLNs. In contrast to B cells, however, T cells were depleted through 7 days of EtOH diet feeding. Total RNA isolated from the MLNs of mice consuming the EtOH‐containing diet was progressively degraded. No degradation of DNA was observed. These study results establish that continuous consumption of dietary EtOH adversely affects the cellularity of MLN, resulting in a progressive loss of lymphocytes that is associated with degradation of total RNA.
The Neurosteroid, 3α‐Hydroxy‐5α‐pregnan‐20‐one, Protects against Bicuculline‐Induced Seizures during Ethanol Withdrawal in Rats
Authors: Devaud Leslie L. , Purdy Robert H. , Morrow A. Leslie
Published: April 2006

Prolonged alcohol consumption leads to the development of tolerance to and dependence on ethanol, resulting in a decreased response to the sedative/hypnotic effects of ethanol, and by negative symptomatology following abrupt termination of use. One symptom associated with ethanol withdrawal in humans, as well as laboratory animals, is enhanced susceptibility to seizures. This study investigated the effects of the neurosteroid, 3α‐hydroxy‐5α‐pregnan‐20‐one (3α‐5α‐THP), on alterations in seizure sensitivity associated with ethanol withdrawal. 3α‐5α‐THP is a potent anxiolytic and anticonvulsant agent that acts via selective interactions with GABAA receptors. Extensive evidence suggests that some aspects of ethanol dependence and withdrawal are mediated by alterations in GABAA receptor function. Withdrawal from chronic ethanol exposure elicited dramatic increases in seizure susceptibility in male and female rats. Administration of 3α‐5α‐THP just before seizure threshold determinations blocked the increased seizure susceptibility induced by ethanol withdrawal. Ethanol‐withdrawn animals were protected by 3α‐5α‐THP at a dose that had no effect on control animal seizure thresholds. Moreover, male and female rats displayed differential responses to the seizure‐threshold lowering effects of ethanol withdrawal, as well as the protection by 3α‐5α‐THP pretreatment. These findings suggest that there are gender differences associated both with ethanol withdrawal as well as the protection by 3α‐5α‐THP in ethanol‐dependent rats.
Alcohol Acceptance, Preference, and Sensitivity in Mice. II. Quantitative Trait Loci Mapping Analysis Using BXD Recombinant Inbred Strains
Authors: Rodriguez Lawrence A. , Plomin Robert , Blizard David A. , Jones Byron C. , McClearn Gerald E.
Published: April 2006

Quantitative trait loci (QTL) mapping of complex phenotypes has emerged as an important feature of the recombinant inbred (Rl) strain methodology. In this second study of our series on alcohol‐related behaviors in mice, we examine alcohol acceptance, preference, and hypnotic dose sensitivity (HDS) to a standard dose of alcohol measured in BXD RI strains to identify candidate QTL regions responsible for their heritability. We detected highly significant marker associations for acceptance on chromosome 12 (Eif4e), for preference on chromosome 1 (D1Rti2) and chromosome 7 (D7Mit7), and for HDS on chromosome 7 (Mpmv1). These are the strongest QTL associations that we detected, but several other candidate QTL regions are reported. Given the limited number of BXD RI strains available, the large number of markers used herein, and the consequent chance of identifying false marker associations, these RI QTL mapping results must be seen as tentative, but an important first step toward identifying QTL for alcohol‐related behaviors.
Turnover of Ethyl‐Linoleate in Rat Plasma and Its Distribution in Various Organs
Authors: Hungund Basalingappa L. , Zheng Zhihong , Barkai Amiram I.
Published: April 2006

The fate of [14C]ethyl‐linoleate (EthLin) after its intravenous administration was investigated in pentobarbital‐anesthetized rats. The disappearance of [14C]EthLin from the plasma was very rapid and followed quite closely a biexponential function of time. Fitting of the experimental data to a two‐compartmental mammillary model revealed that the labeled compounds are eliminated from the plasma with a half‐life of <1 min during the early time following the intravenous injection and that a large portion of the EthLin is hydrolyzed instantly to linoleic acid and ethanol. About 9–11 % of the plasma [14C]EthLin or its breakdown products are irreversibly cleared from the plasma compartment each minute. Most of the 14C‐labeled compounds that originated in the plasma were recovered in the rat liver and lungs and to a lesser extent in the heart, spleen, and kidneys. Two hr after the [14C]EthLin administration, ‐2.5–5.5% of the total radioactivity in the various organs was still associated with EthLin. Such accumulations, although relatively small, indicate that fatty acid ethyl esters (FAEEs) may be taken up from the plasma. Thus, some of the FAEEs that are formed in certain organs may spillover to the circulating blood where much of it would be hydrolyzed to free fatty acids, but reuptake from the plasma may still account, to some extent, to FAEE‐induced damage in chronic alcohol abusers.
Chronic Alcohol Administration Stimulates Nitric Oxide Formation in the Rat Liver With or Without Pretreatment by Lipopolysaccharide
Authors: Wang Ji‐Feng , Greenberg Stanley S. , Spitzer John J.
Published: April 2006

This study examines the effect of chronic alcohol consumption on nitric oxide release from the liver of rats with or without lipopolysaccharide (LPS) (Escherichia coli) treatment. Reactive nitrogen intermediates (RNIs) in plasma were monitored with an NOx Analyzer, and nitric oxide (NO) production was measured as nitrite or nitrite + nitrate accumulation in perfusates of the perfused liver, and in supematants of the freshly isolated hepatic cells after incubation for 3 hr in Hank's balanced salt solution buffer containing 1 Mm l‐arginine. RNI concentration in plasma of control rats was 32.0 ± 3.4 μm (mean ×se). Livers from diet‐fed control rats produced RNIs at the barely detectable rate of 7.8 ± 1.5 nmol/hr × g wet liver. Six hr after administration of LPS (1 mg/kg, iv), plasma RNI levels in diet‐fed control rats increased to 426.9 ± 29.4 μm, and RNI release from the perfused liver was also markedly elevated to 97.7 ± 7.7 nmol/hr × wet g liver, indicating hepatic NO release as a potentially important source for the increased RNI in plasma. The presence of NG‐monomethyl‐l‐arginine (0.5–1 mm) or the absence of l‐arginine in the perfusate inhibited LPS‐induced stimulation of RNI release. EGTA (1 mm) had little effect, indicating that the increased RNI release was likely to be due to inducible NO synthase activity. The release of RNIs by freshly isolated Kupffer cells increased 13‐fold, and this small cell mass contributed almost half of the hepatic RNI production under these conditions. Plasma ALT concentration was elevated after LPS administration, indicating incipient liver damage. Chronic alcohol administration resulted in increased RNI levels in plasma (44.6 ± 4.2) that were accompanied by increased spontaneous release in the perfused liver (16.4 ± 1.5 nmol/hr wet g). Enhanced activity of Kupffer cells was responsible for this increase. After administration of LPS, the increase in plasma RNIs (565.72 ± 49.6 μm) was slightly higher in chronic alcoholic rats than in control animals. This was also accompanied by a somewhat higher RNI release from the perfused liver (125.40 ± 12.2 nmol/hr × wet g) in the same group. Hepatocytes were responsible for the post‐LPS increase in alcoholic rats. Plasma ALT concentration was higher in alcoholic than control, diet‐fed rats after LPS, indicating more liver damage in that group. The exact role of the elevated hepatic RNI release in affecting hepatic function in chronic alcoholic animals remains to be clarified.
Neuroactive Steroid Sensitivity in Withdrawal Seizure‐Prone and ‐Resistant Mice
Authors: Finn Deborah A. , Roberts Amanda J. , Crabbe John C.
Published: April 2006

Withdrawal Seizure‐Prone (WSP) and ‐Resistant (WSR) mice, which were selectively bred for severe (WSP) or mild (WSR) handling‐induced convulsions (HICs) following chronic ethanol inhalation, were found to differ in sensitivity to the anticonvulsant effects of the neuroactive steroid 3α‐hydroxy‐5α‐pregnan‐20‐one (3α,5α‐P). 3α,5α‐P (5 or 10 mg/kg, ip) significantly increased seizure thresholds to pentylenetetrazol in ethanol‐naive males of both the WSP and WSR lines. In general, WSP mice were more sensitive than WSR mice to the anticonvulsant effect of 3α,5α‐P. Subsequent studies in male WSP mice exposed to ethanol vapor or air for 24 hr demonstrated enhanced sensitivity to the anticonvulsant effect of 3α,5α‐P (0.5–20 mg/kg, ip) during ethanol withdrawal. Only the highest dose affected HICs in air‐exposed animals, whereas both the two highest doses significantly reduced HICs in ethanol‐exposed mice. These results provide the first demonstration that 3α,5α‐P attenuates ethanol withdrawal convulsions and indicate enhanced sensitivity to the anticonvulsant effect of 3α,5α‐P in animals withdrawing from ethanol dependence.
Staurosporine, a Protein Kinase Inhibitor, Increases the Intoxicating Potencies of Ethanol and Other n‐Alkanols in Rana pipiens Tadpoles
Authors: Firestone Susan , Firestone Leonard L.
Published: April 2006

Central nervous system protein kinases are the intracellular effectors for many of the signal transduction pathways essential to neurotransmission. Although the in vitro activity of at least one of these important enzymes, protein kinase C, is diminished by therapeutic concentrations of ethanol and other central depressants, the relationship of this effect to intoxication in vivo is not known. If intoxication by ethanol involves central protein kinase inhibition, then other inhibitors of these enzymes should enhance ethanol's intoxicating potency. To test this hypothesis, we compared the median effective concentrations of ethanol and two other n‐alkanols for loss‐of‐righting reflex in Rana pipiens tadpoles pretreated with staurosporine and in untreated controls. Alkanol concentrations were confirmed by gas chromatography and staurosporine concentrations by ultraviolet absorbance spectrophotometry. Results obtained with 650 animals demonstrate that pretreatment with staurosporine concentrations in the nanomolar range significantly decrease the median effective concentration for ethanol (56% of control; p < 0.001), butanol (38% of control; p < 0.001), and octanol (59% of control; p < 0.001). This finding supports that central protein kinase inhibition may be involved in the acute intoxicating effects of ethanol and other n‐alkanols.
Polymorphism of a Class 3 Aldehyde Dehydrogenase Present in Human Saliva and in Hair Roots
Authors: Dyck Lillian E.
Published: April 2006

The types of aldehyde dehydrogenases (ALDH) present in human hair roots and in saliva were investigated. ALDH was detected by activity staining following separation of crude extracts by isoelectric focusing. Hair roots were found to express ALDH1, ALDH2, ALDH3, and ALDH4, whereas saliva expressed ALDH3. Two different patterns of ALDH3 were detected in hair roots collected from 42 donors, 40 expressed one pattern (variant I) and two another pattern (variant II) of activity staining. The variant I pattern of hair root ALDH3 changed with repetitive freezing and thawing of the sample, whereas the variant II pattern was stable. In contrast to hair root ALDH3, all patterns of ALDH3 activity in saliva were stable. The patterns of ALDH3 activity present in human hair roots that had been frozen and thawed twice matched those present in saliva collected from the same individual. Three polymorphisms of ALDH3 (variants I, II, and III) were detected in the 33 saliva samples analyzed. Variants I and II were inherited in each of three generations of a 10‐member family.
Mechanisms of Acetaldehyde‐Mediated Growth Inhibition: Delayed Cell Cycle Progression and Induction of Apoptosis
Authors: Zimmerman Barbara T. , Crawford Garrett D. , Dahl Rolf , Simon Francis R. , Mapoles John E.
Published: April 2006

Chronic ethanol exposure has been associated with pleiotropic effects on cellular function in vivo and in vitro, including inhibition of growth. To date, it has been difficult to dissociate the primary effects of ethanol from the effects of ethanol metabolism, generation of acetaldehyde, and reducing equivalents. We have previously described the development of a Chinese hamster ovary cell line, A‐10, which expresses a transfected murine‐liver alcohol dehydrogenase. Cultures of these cells accumulate acetaldehyde due to the low level of aldehyde dehydrogenase. One noticeable effect of chronic acetaldehyde exposure, but not ethanol exposure, is the inhibition of cell growth. This study focuses on the mechanisms that underlie this growth inhibition. Our studies with the A‐10 cell on the rates of [3H]thymidine incorporation and flow cytometry of asynchronous cultures indicated that acetaldehyde did not lead to arrest of the cell cycle in the G1 phase as has been found in other models of ethanol exposure. Rather, we observed a generalized delay in cell cycle progression. However, the slower cell cycle did not account exclusively for the slower rates of cell accumulation. Chronic exposure to acetaldehyde also increased the rate of cell death. The increased rate of cell death was both cumulative and dose‐dependent. The dead cells accumulated in the medium and were apoptotic. Apoptosis was confirmed using morphological criteria and quantitation of DNA fragmentation. These data lend additional support to the idea that chronic acetaldehyde exposure can affect the mechanisms that regulate cell division and the apoptotic program.
Effect of Ethanol on p36 Protein Kinase Substrate and Insulin Receptor Substrate 1 Expression and Tyrosyl Phosphorylation in Human Hepatocellular Carcinoma Cells
Authors: Bhavani K. , de la Monte S. , Brown N. V. , Xu Y.‐Y. , Sasaki Y. , Wands J. R.
Published: April 2006

Ethanol inhibits insulin (IN) and epidermal growth factor (EGF)‐induced hepatocyte DNA synthesis. Growth factor receptor kinases, such as IN and EGF, phosphorylate insulin receptor substrate (IRS‐1) and p36 protein kinase substrate, respectively, on tyrosine residues. IRS‐1 and p36 are thought to be important intracellular signal transduction molecules involved in the regulation of cell growth. These investigations explored the effect of ethanol additions on the expression and tyrosyl phosphorylation (TP) of p36 and IRS‐1 in a human hepatocellular carcinoma cell line (FOCUS) in relationship to cell proliferation induced by IN and serum growth factor stimulation. It was found that p36 was constitutively and highly expressed in serum‐starved cells and protein, and mRNA levels did not change with cell proliferation induced by growth factors. However, exposure of FOCUS cells to ethanol additions substantially inhibited TP of p36. The early TP of IRS‐1 induced by IN stimulation was also reduced by ethanol additions. Finally, there was a parallel decrease of FOCUS cell proliferation in ethanol‐ex‐posed cultures. These studies suggest that one possible mechanism of ethanol inhibitory effect on cell proliferation is through reduced TP of putative intracellular signal transduction molecules, such as p36 and IRS‐1.
Rat Mitochondrial Aldehyde Dehydrogenase Polymorphism and Major Histocompatibility Complex RT1.A Phenotypes Are Not Associated with Alcohol Drinking in P and NP Rats
Authors: Carr Lucinda G. , Kirchner Jeffrey , Magnes Les , Lumeng Lawrence , Li Ting‐Kai
Published: April 2006

Previous studies revealed polymorphisms in mitochondrial aldehyde dehydrogenase (ALDH2) and the major histocompatibility complex RT1.A that appeared in association with alcohol drinking preference in the alcohol‐preferring (P) and alcohol‐nonpreferring (NP) rat lines. To determine the strength of these associations, the P and NP lines were crossed, and cosegregation studies were performed in the F2 progeny. The ALDH2Q allele and the ALDH2R allele, found in the P and NP lines, respectively, were found not to be associated with the high and low drinking animals in the F2 progeny. Kinetic studies with ALDH2Q and ALDH2R isozymes purified from the mitochondria of P and NP rat livers, respectively, showed that the polymorphism would not be expected to be associated with altered aldehyde metabolism. The association between RT1.A and alcohol preference (RT1.Auu with high and RT1.A with low alcohol intake) also could not be confirmed in the segregating F2 progeny of the P × NP intercross.
Increases and Time‐Course Variations in β‐Hexosaminidase Isoenzyme B and Carbohydrate‐Deficient Transferrin in Serum from Alcoholics Are Similar
Authors: Hultberg B. , Isaksson A. , Berglund M. , Ailing C.
Published: April 2006

β‐Hexosaminidase B‐isoforms (β‐hexosaminidase B, P, and intermediate forms; abbreviated herein as “Hex B”) and serum carbohydrate‐deficient transferrin (CDT) are two markers of alcohol abuse. In the present study, we have compared “Hex B” with CDT as markers of alcohol abuse in a group of alcoholics hospitalized for detoxification after a period of heavy alcohol abuse. We have also followed the disappearance rate of these two markers from circulation. “Hex B” was elevated in 38 of 42 patients hospitalized for detoxication, whereas CDT was elevated in 35 of 42 patients. A highly significant correlation was noted between “Hex B” and CDT in these patients (p= 0.52, p < 0.001). Neither “Hex B” nor CDT correlated with γ‐glutamyltransferase or AST. The disappearance rates from serum of “Hex B” and CDT were determined in 21 hospitalized patients followed for up to 15 days. “Hex B” and CDT showed similar time‐course variation and half‐lives, 6.5 ± 3.7 (mean ×sd) and 8.6 ± 4.1 days, respectively. The possible reasons for a relation between these two markers are discussed, and it is concluded that more experience of both “Hex B” and CDT in unselected populations is needed to establish the diagnostic potential of these tests as markers of alcohol abuse.
Antagonism of Ethanol by Pretreatment or Posttreatment with RO 15–4513 and Indomethacin Alone or in Combination
Authors: Elmer Gregory I. , George Frank R.
Published: April 2006

RO 15–4513, an inverse agonist at the GABA/benzodiazepine receptor Cl‐ channel complex, antagonizes multiple effects of ethanol. Prostaglandin synthesis inhibitors, such as indomethacin, also antagonize several effects of alcohols. However, prostaglandin synthesis inhibitors and RO15–4513 each provide only partial antagonism of ethanol, typically seen as a dose‐related effect with a maximum efficacy of ˜50%. The purpose of this study was to: (1) compare the relative efficacy of these compounds for antagonizing ethanol; (2) compare the effectiveness of preethanol treatment versus postethanol treatment with each drug; and (3) compare the effect of RO 15–4513 and indomethacin in combination with the effects of each drug alone. The results show that indomethacin significantly decreased duration of loss of the righting reflex when administered either pre‐or postethanol. Conversely, RO 15–4513 decreased duration of loss of the righting reflex only when given preethanol. When coadministered, RO 15–4513 and indomethacin did not show additive or synergistic effects. Compounds from these two drugs classes should continue to prove useful in elucidating ethanol's mechanisms of action.