bluuebunny/crossref_metadata_2025 huggingface dataset:
Hypothalamic Function in Response to 2‐Deoxy‐d‐Glucose in Long‐Term Abstinent Alcoholics
Authors: Umhau John C. , Petrulis Sarah G. , Diaz Rosalyn , Biddison James R. , George David T.
Published: April 2006

Background: The body adapts to diverse stressful stimuli with a response characterized by activation of the hypothalamic‐pituitary‐adrenal (HPA) axis. Chronic alcohol consumption can cause changes in the function of this neuroendocrine system. Although many studies have examined this phenomenon in drinking and recently sober alcoholics, few studies have examined HPA axis function in long‐term sober alcoholics.

Methods: To characterize HPA axis function in long‐term sober alcoholics, we used a challenge paradigm with 2‐deoxy‐d‐glucose (2‐DG). An infusion of 2‐DG (a nonmetabolizable glucose analog) induces a well‐characterized stress response. In a previous study, our laboratory found an exaggerated corticotropin and cortisol response in alcoholics abstinent 3 weeks; in this investigation we compared the effects of an infusion of 2‐DG on 19 healthy volunteers and 20 community‐living alcoholics who had been abstinent more than 6 months.

Results: In contrast to the previous study, long‐term sober alcoholics did not have an exaggerated corticotropin and cortisol response after 2‐DG.

Conclusions: Previously observed abnormalities in cortisol regulation in 3‐week‐sober alcoholics may be related to the acute effects of recent alcohol consumption and withdrawal. Future investigations into the metabolic function of alcoholics, particularly investigations involving the HPA system, should consider the possibility that normalization may not occur until long‐term abstinence has been achieved.
Comparison of Global Brain Gene Expression Profiles Between Inbred Long‐Sleep and Inbred Short‐Sleep Mice by High‐Density Gene Array Hybridization
Authors: Xu Yan , Ehringer Marissa , Yang Fan , Sikela James M.
Published: April 2006

Background: Inbred long‐sleep (ILS) and short‐sleep (ISS) mice show significant central nervous system–mediated differences in sleep time for sedative dose of ethanol and are frequently used as a rodent model for ethanol sensitivity. In this study, we have used complementary DNA (cDNA) array hybridization methodology to identify genes that are differentially expressed between the brains of ILS and ISS mice.

Methods: To carry out this analysis, we used both the gene discovery array (GDA) and the Mouse GEM 1 Microarray. GDA consists of 18,378 nonredundant mouse cDNA clones on a single nylon filter. Complex probes were prepared from total brain mRNA of ILS or ISS mice by using reverse transcription and 33P labeling. The labeled probes were hybridized in parallel to the gene array filters. Data from GDA experiments were analyzed with SQL‐Plus and Oracle 8. The GEM microarray includes 8,730 sequence‐verified clones on a glass chip. Two fluorescently labeled probes were used to hybridize a microarray simultaneously. Data from GEM experiments were analyzed by using the GEMTools software package (Incyte). Differentially expressed genes identified from each method were confirmed by relative quantitative reverse transcription–polymerase chain reaction (RT‐PCR).

Results: A total of 41 genes or expressed sequence tags (ESTs) display significant expression level differences between brains of ILS and ISS mice after GDA, GEM1 hybridization, and quantitative RT‐PCR confirmation. Among them, 18 clones were expressed higher in ILS mice, and 23 clones were expressed higher in ISS mice. The individual gene or EST's function and mapping information have been analyzed.

Conclusions: This study identified 41 genes that are differentially expressed between brains of ILS and ISS mice. Some of them may have biological relevance in mediation of phenotypic variation between ILS and ISS mice for ethanol sensitivity. This study also demonstrates that parallel gene expression comparison with high‐density cDNA arrays is a rapid and efficient way to discover potential genes and pathways involved in alcoholism and alcohol‐related physiologic processes.
Altered GABAA Receptor Subunit and Splice Variant Expression in Rats Treated With Chronic Intermittent Ethanol
Authors: Petrie Jennifer , Sapp Douglas W. , Tyndale Rachel F. , Park Maenghee Kang , Fanselow Michael , Olsen Richard W.
Published: April 2006

Background: Intermittent chronic administration of ethanol to rats has been shown previously to produce a hyperexcitable, kindling‐like state, accompanied by reduced inhibitory synaptic transmission in the hippocampus and changes in γ‐aminobutyric acid type A (GABAA) receptors. Further information is needed on the detailed changes in GABAA receptors and their time course and persistence, as is comparison to changes after chronic, continuous ethanol.

Methods: GABAA receptors were analyzed in the rat brain after chronic intermittent ethanol (CIE) by using radioligand binding, photoaffinity labeling of polypeptides, and estimates of messenger RNA (mRNA) levels of receptor subunits by reverse transcriptase–polymerase chain reaction (RT‐PCR) and in situ hybridization.

Results: CIE rats were confirmed to have increased GABAA receptor binding of the benzodiazepine partial inverse agonist and ethanol antidote ligand Ro15‐4513, due to increased expression of the α6 subunit polypeptide in the cerebellum, shown by photoaffinity labeling. Estimates of mRNA levels by use of RT‐PCR did not reveal any significant increase in α6 or in several other receptor subunits in several brain regions, but a decrease in the ratio of the long and short splice variants (L/S) of the γ2 subunit was detected in the hippocampus, especially the CA1 region.

Conclusions: Changes in GABAA receptors were found in rats given CIE. Increased α6 subunit in the cerebellum was demonstrated by using both the binding to diazepam‐insensitive sites for [3H]Ro15‐4513 and increased levels of the 57‐kDa α6 polypeptide after photoaffinity labeling with this ligand. This increase appeared after 30 doses of ethanol and decayed to normal 1 week after ethanol was discontinued. The transient change in cerebellar α6 subunit‐containing receptors, also reportedly seen after chronic continuous ethanol, is thus unlikely to account for the persistently hyperexcitable, kindled, seizure‐susceptible state seen in CIE. However, the significant decrease in γ2 subunit L/S splice variant ratio in the hippocampus implies changes in GABAA receptor function, possibly involving protein phosphorylation by protein kinase C. Altered receptor trafficking and turnover associated with synaptic plasticity may contribute to the observed reduced inhibition in the hippocampus and other signs of alcohol dependence produced by CIE.
4‐Methylpyrazole Decreases Salivary Acetaldehyde Levels in ALDH2‐Deficient Subjects but Not in Subjects With Normal ALDH2
Authors: Väkeväinen Satu , Tillonen Jyrki , Salaspuro Mikko
Published: April 2006

Background: Carcinogenic acetaldehyde is produced from ethanol locally in the upper digestive tract via alcohol dehydrogenases (ADHs) of oral microbes, mucosal cells, and salivary glands. Acetaldehyde is further oxidized into less harmful acetate mainly by the aldehyde dehydrogenase‐2 (ALDH2) enzyme. ALDH2‐deficiency increases salivary acetaldehyde levels and the risk for upper digestive tract cancer in heavy alcohol drinkers. 4‐methylpyrazole (4‐MP) is an ADH‐inhibitor which could reduce the local production of acetaldehyde from ethanol in the saliva.

Methods: Five ALDH2‐deficient subjects and six subjects with normal ALDH2 ingested a moderate dose of alcohol (0.4 g/kg of body weight), whereafter their salivary acetaldehyde levels, heart rate, skin temperature, and blood pressure were followed for up to four hours. Blood acetaldehyde and ethanol levels were determined at 60 min. The experiment was repeated after a week. Two hours before the second study day, the volunteers received 4‐MP, 10–15 mg/kg of body weight orally.

Results: Total ethanol elimination rate decreased with 4‐MP by 38–46% in all subjects. 4‐MP also reduced blood acetaldehyde levels and suppressed the cardiocirculatory responses of the ALDH2‐deficient volunteers. In addition, salivary acetaldehyde production in ALDH2‐deficient subjects was significantly reduced when correlated with salivary ethanol levels. On the contrary, 4‐MP did not have any effect on salivary or blood acetaldehyde levels in subjects with normal ALDH2.

Conclusions: A single dose of 4‐MP before ethanol ingestion reduces ethanol elimination rate, the flushing reaction, and both blood and salivary acetaldehyde levels in ALDH2‐deficient subjects but not in subjects with the normal ALDH2 genotype. These results suggest that the role of oral mucosal and glandular ADHs in salivary acetaldehyde production is minimal and support earlier findings indicating that salivary acetaldehyde production is mainly of microbial origin in subjects with normal ALDH2.
Ethanol‐Induced Impairments in Spatial Working Memory Are Not Due to Deficits in Learning
Authors: Hoffmann Sarah E. , Matthews Douglas B.
Published: April 2006

Background: Acute ethanol administration impairs spatial reference memory and spatial working memory. However, the experimental designs previously used to test spatial working memory fail to make a distinction between the acquisition, or learning, of spatial information and the retention of this information. This study demonstrates that acute ethanol administration impairs spatial working memory, by using a novel experimental design that eliminates the confound between the learning of new spatial information and the testing of this information.

Methods: Long‐Evans male rats received three forced trials to the same place for food reward on an elevated radial arm maze. Subjects were then given six free‐choice trials in the first acquisition session, followed by a 30‐min consolidation period before an additional six free‐choice trials were administered—the retention session. Animals were trained to a criterion of five of six correct in both the acquisition and retention over 2 consecutive days. Once criteria were obtained, subjects received either saline or one of three ethanol doses immediately after the acquisition session to investigate whether ethanol alters retention of the learned spatial information.

Results: Acute ethanol administration impaired spatial working memory. Rats tested under saline and low‐dose ethanol (1.0 g/kg) made significantly more place choices than rats tested under moderate‐ or high‐dose ethanol (1.5 and 2.0 g/kg, respectively). Ethanol produced a temporary impairment in that no significant differences were found when subjects were retrained and retested 24 hr after initial testing.

Conclusions: These results demonstrate that acute ethanol administration impairs spatial working memory and that such a deficit is not contingent on a learning impairment. These results support earlier findings that acute ethanol administration impairs spatial working memory but provide a significant advance by validating a novel training procedure that allows for direct investigation of working memory. Ethanol's impairment of both spatial working memory and spatial reference memory strengthens the similarities between memory impairments due to ethanol administration and memory impairments due to hippocampal lesions.
In Utero Ethanol Exposure Causes Mitochondrial Dysfunction, Which Can Result in Apoptotic Cell Death in Fetal Brain: A Potential Role for 4‐Hydroxynonenal
Authors: Ramachandran Vinitha , Perez Antonio , Chen Juanjuan , Senthil Duraisamy , Schenker Steven , Henderson George I.
Published: April 2006

Background: In utero ethanol exposure causes abnormal fetal brain development that may partly be due to enhanced cell death. The mechanisms underlying this remain to be defined, but ethanol‐induced oxidative stress may play a role. The following studies investigated the effects of short‐term in utero ethanol exposure on fetal brain mitochondrial events that are known to elicit apoptotic cell death. Evidence is presented suggesting that 4‐hydroxynonenal (HNE), a toxic product of lipid oxidation, is a causal factor in the observed mitochondrial damage.

Methods: Mitochondria were isolated from control and ethanol‐exposed fetal brains (days 17 and 18 of gestation). Permeability transition was determined spectrophotometrically, and cytochrome c and apoptosis‐inducing factor (AIF) release were assessed by Western blotting. Caspase‐3 activity and DNA fragmentation were determined both as markers for mitochondrially mediated apoptosis and as consequences of cytochrome c and AIF release.

Results: Maternal ethanol intake caused an increase in mitochondrial permeability transition, and this was accompanied by cytochrome c and AIF release from fetal brain mitochondria that exceeded control values by 62 and 25%, respectively (p < 0.05). In utero ethanol exposure resulted in a 30% increase in caspase‐3 activity and a 25% increase in DNA fragmentation (p < 0.05) in the fetal brain. HNE levels were increased by 23% (p < 0.05) in mitochondria by in vivo ethanol exposure. In vitro treatment of fetal brain mitochondria with HNE (25–100 μM) also caused increases in mitochondrial permeability transition, as well as dose‐dependent releases of cytochrome c and AIF.

Conclusions: These studies illustrate that in utero ethanol exposure can elicit a cascade of events in the fetal brain that are consistent with mitochondrially mediated apoptotic cell death. Additionally, the increase in mitochondrial content of HNE after ethanol intake and the ability of HNE added to fetal brain mitochondria to mimic these effects of in vivo ethanol exposure support a potential role for HNE in the proapoptotic responses to ethanol.
Effect of Alcohol Consumption on Host Release of Interleukin‐17 During Pulmonary Infection With Klebsiella pneumoniae
Authors: Shellito Judd E. , Zheng Min Quan , Ye Peng , Ruan Sanbao , Shean Mary K. , Kolls Jay
Published: April 2006

Background: A link between alcohol abuse and bacterial pneumonia has been recognized for centuries, but mechanisms to explain this relationship are unclarified. Interleukin‐17 (IL‐17) is a lymphocyte‐derived cytokine that is part of the inflammatory cytokine cascade. Previous studies from our laboratory indicated that IL‐17 is released in lung tissue in a murine model of bacterial pneumonia caused by Klebsiella pneumoniae. The effects of alcohol consumption on pulmonary release of IL‐17 are unknown.

Methods: Mice were maintained on 20% ethanol in drinking water or on a control diet without alcohol. After 2 weeks, alcohol and control mice were challenged with intratracheal K. pneumoniae. Mice were followed for survival after bacterial challenge, neutrophil recruitment was assayed as myeloperoxidase, and IL‐17 was measured in lung lavage fluid by enzyme‐linked immunosorbent assay. In additional experiments, splenocytes from control mice were incubated with ethanol in vitro, and release of IL‐17 was measured in culture supernatants. Finally, control and alcohol mice received intrapulmonary gene transfer of E‐1–deleted adenovirus containing the murine IL‐17 gene. These mice were then challenged with K. pneumoniae and followed for survival and neutrophil recruitment.

Results: In these studies, we demonstrate that a 2‐week history of ethanol consumption in mice suppresses release of IL‐17 into lung tissue, decreases neutrophil recruitment, and increases mortality from experimental K. pneumonia. In vitro experiments confirm a direct suppressive effect of ethanol on the release of IL‐17 from splenocytes. In vivo administration of the IL‐17 gene in an adenoviral vector to alcohol‐consuming mice results in release of IL‐17 into lavage fluid and normalizes neutrophil recruitment and mortality after bacterial challenge.

Conclusions: The results of these experiments strongly implicate IL‐17 as an important pathway for the immunosuppression associated with alcohol abuse and support gene therapeutic approaches to augment immune function in the alcoholic host or to treat infections associated with alcoholism.
Cardioprotective Effect of Propranolol From Alcohol‐Induced Heart Muscle Damage as Assessed by Plasma Cardiac Troponin‐T
Authors: Patel Vinood B. , Ajmal Raheela , Sherwood Roy A. , Sullivan Andrew , Richardson Peter J. , Preedy Victor R.
Published: April 2006

Background: Heavy alcohol consumption from either long‐term misuse or binge drinking is associated with poor cardiac contractility, mitochondrial dysfunction, and ventricular arrhythmias. The aim of this study was to measure circulating cardiac troponin‐T as a marker for myocardial damage following acute and chronic alcohol administration.

Methods: In acute studies, male Wistar rats were treated with alcohol (75 mmol/kg body weight, intraperitoneal) and plasma was collected 2.5 hr after alcohol administration for analysis of rat cardiac troponin‐T. In addition, rats were pretreated with cyanamide (an inhibitor of acetaldehyde dehydrogenase), various beta‐blockers, xanthine oxidase inhibitors, or lisinopril before acute alcohol dosing. In chronic studies, rats were fed alcohol (as 35% of total dietary calories) for 6 weeks.

Results: The results of the time course study showed that acute alcohol administration significantly raised plasma cardiac troponin‐T levels after 2.5 hr and 6 hr, but not after 24 hr. The effects of alcohol on cardiac troponin‐T were potentiated with cyanamide pretreatment. Acute ethanol, alone or with cyanamide pretreatment, decreased systolic blood pressure and increased heart rates. Beta‐blocker pretreatment with propranolol reduced the alcohol‐induced increase in plasma troponin‐T, whereas lisinopril potentiated this effect. The beta‐blockers, atenolol and metoprolol, and the xanthine oxidase inhibitors, allopurinol and oxypurinol, were unable to reduce elevated troponin‐T. However, pretreatment with the beta‐blocker timolol moderated the acute alcohol‐induced increase in troponin‐T. In the chronic alcohol rat model, no differences were observed between alcohol and control pair‐fed rats, suggesting the inducement of tolerance.

Conclusions: In conditions of acute exposure, ethanol‐induced lesions are characterized by raised plasma cardiac troponin‐T possibly due to β1 and/or β2 adrenergic activation.
Effects of Prenatal Ethanol Exposure on Hypothalamic‐Pituitary‐Adrenal Regulation After Adrenalectomy and Corticosterone Replacement
Authors: Glavas Maria M. , Hofmann Candace E. , Yu Wayne K. , Weinberg Joanne
Published: April 2006

Background: Previous studies have demonstrated that rats prenatally exposed to ethanol (E) exhibit hypothalamic‐pituitary‐adrenal (HPA) hyperresponsiveness, demonstrated by increased and/or prolonged elevations of adrenocorticotropin (ACTH) and/or corticosterone (CORT) after stress. This study investigated possible mechanisms of HPA hyperresponsiveness in E rats by manipulating CORT feedback regulation of HPA activity via adrenalectomy (ADX) with or without CORT replacement.

Methods: Male Sprague‐Dawley rat offspring from prenatal E, pair‐fed (PF) and ad libitum–fed control (C) groups were tested at 90 to 120 days of age. Rats were either sham‐operated or underwent ADX, with or without CORT replacement. CORT (25 μg/ml) was replaced via the drinking water to achieve basal plasma CORT levels and maintain a phasic CORT signal. Seven days after surgery, animals were decapitated at the diurnal peak either under basal conditions or after a 15‐min restraint stress, and trunk blood was collected.

Results: After ADX, loss of the CORT feedback signal resulted in increased plasma ACTH in all groups compared with those in sham animals. In addition, under basal conditions, ADX E rats had significantly greater plasma ACTH levels than both PF and C rats. However, no differences were seen in ADX rats after stress. CORT replacement after ADX was partially effective in normalizing ACTH levels under both basal and stress conditions, with no differences among E, PF, and C animals.

Conclusions: These results suggest that E males may exhibit enhanced stimulatory inputs to the hypothalamus, increased pituitary sensitivity to secretagogues, or both, which may be revealed after ADX. In contrast, E animals seem similar to controls in their ability to use an exogenous CORT signal to regulate HPA activity.
Mitochondrial DNA Damage and Impaired Mitochondrial Function Contribute to Apoptosis of Insulin‐Stimulated Ethanol‐Exposed Neuronal Cells
Authors: de la Monte Suzanne M. , Wands Jack R.
Published: April 2006

Background: Ethanol inhibition of insulin signaling may contribute to impaired central nervous system development in fetal alcohol syndrome. An important consequence of ethanol inhibition of insulin signaling is increased apoptosis due to reduced levels of insulin‐stimulated phosphoinositol‐3‐kinase activity.

Methods: We used viability assays, end‐labeling, Western blot analysis, and MitoTracker (Molecular Probes, Eugene, OR) fluorescence labeling to determine whether ethanol‐induced central nervous system neuronal cell death was mediated in part by increased mitochondrial (Mt) DNA damage and impaired Mt function.

Results: In ethanol‐exposed, insulin‐stimulated PNET2 central nervous system–derived human neuronal cells, reduced viability was associated with increased Mt DNA damage, reduced Mt mass (manifested by reduced Mt protein expression and MitoTracker Green fluorescent labeling), and impaired Mt function (manifested by reduced levels of 3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyltetrazolium bromide activity, cytochrome oxidase‐Complex IV, Subunit II expression, and MitoTracker Red fluorescence). The adverse effects of ethanol on Mt function were reduced by pretreating the cells with broad‐spectrum caspase inhibitors and nearly abolished by nerve growth factor stimulation, with or without concomitant treatment with global caspase inhibitors.

Conclusions: These results suggest that ethanol‐induced death of insulin‐stimulated immature neuronal cells is mediated in part by impaired Mt function associated with Mt DNA damage and reduced Mt mass, and therefore it is likely to contribute to neuronal loss associated with fetal alcohol syndrome. The findings also suggest that the adverse effects of ethanol on insulin‐stimulated survival and metabolic function could be overcome by stimulating with growth factors that support Mt function through insulin‐independent pathways.
Relationship of Brain Ethanol Metabolism to the Hypnotic Effect of Ethanol. II: Studies in Selectively Bred Rats and Mice
Authors: Zimatkin Sergey M. , Liopo Anton V. , Satanovskaya Valentina I. , Bardina Lubov R. , Deitrich Richard A.
Published: April 2006

Background : To clarify the role of brain acetaldehyde in the hypnotic effect of ethanol, we compared the ethanol‐oxidizing capacity (rate of acetaldehyde accumulation) and catalase and aldehyde dehydrogenase activity in the brains of animals genetically selected for different sensitivities to the hypnotic effect of ethanol.

Methods: We used high, low, or control alcohol‐sensitive rats (HAS, LAS, and CAS) and short‐ and long‐sleep mice (SS and LS), as well as SS × LS recombinant inbred mice with known strain differences in mean duration of ethanol‐induced sleep. We studied the rate of accumulation of acetaldehyde from ethanol in brain homogenates of these animals and correlated those values with their hypnotic sensitivity to ethanol.

Results: Acetaldehyde accumulation from ethanol was significantly higher in the brain homogenates from HAS rats and LS mice with high sensitivity to the hypnotic effect of ethanol in vivo, compared with LAS rats and SS mice with low sensitivity to ethanol. A correlation was found between the duration of ethanol‐induced sleep and the in vitro rate of accumulation of ethanol‐derived acetaldehyde in the brains of recombinant SS × LS mice strains. There was no correlation of sleep time with brain catalase levels. There were no line differences in brain catalase or aldehyde dehydrogenase or in alcohol or aldehyde dehydrogenase activity in livers of LAS, CAS, and HAS rats or in SS and LS mice.

Conclusions: A correlation between the brain acetaldehyde accumulation, but not catalase levels, and the central effect of ethanol was demonstrated in animals genetically differing in initial sensitivity to the hypnotic effect of ethanol.
Fetal and Infantile Alcohol‐Mediated Associative Learning in the Rat
Authors: Abate Paula , Spear Norman E. , Molina Juan Carlos
Published: April 2006

Background : Infant rats express conditioned responses to an odor experienced prenatally as a chemosensory cue associated with moderate alcohol intoxication. This study examined postnatal intake of a chemosensory cue (cineole) that had been paired with alcohol's unconditioned effects. It also tested the interaction between prenatal association and postnatal conditioning with cineole and alcohol.

Methods: Pregnant female rats intubated with cineole were given ethanol (EtOH) .25 or 4.0 hr later. Other groups received only water or water paired with ethanol. During postnatal day 15 (PD15), infant consumption of cineole solution was assessed. After the cineole drinking test, pups were intubated with EtOH or water to assess infant conditioning. On PD16, all pups were tested for mouthing to milk alone or to a milk‐cineole solution.

Results: Statistical analysis confirmed fetal associative conditioning attributable to the unconditioned effects of prenatal alcohol. Fetuses given explicit pairings of cineole and alcohol ingested less cineole on PD15 than control fetuses given a 4‐hr interval between cineole and alcohol.

On PD16, consumption of cineole was significantly increased by prenatal exposure to cineole. Teratogenic effects of this dose of prenatal alcohol did not affect postnatal associative or nonassociative behavior.

Conclusions: Prenatal associative learning can be established through temporal contiguity between fetal chemosensory stimulation and alcohol's unconditioned properties. This associative memory survives to infancy and modulates intake patterns and behavioral reactivity to substances that were prenatally paired with alcohol intoxication.
Chronic Daily Ethanol and Withdrawal: 2. Behavioral Changes During Prolonged Abstinence
Authors: Rasmussen Dennis D. , Mitton Dennis R. , Green Jillian , Puchalski Stephaney
Published: April 2006

Background : Our previous investigations have suggested that alcohol abuse may induce persistently compromised hypothalamo‐pituitary‐adrenal (HPA) function, which could increase risk for subsequent alcohol abuse. The apparent similarity of chronic alcohol abuse–induced HPA defects to the compromised HPA functions associated with posttraumatic stress disorder and atypical depression also suggest potential common mechanisms shared with varied neurobehavioral disorders. Accordingly, we have investigated persistent behavioral effects of previous repetitive daily ethanol consumption and withdrawal.

Methods: Male Sprague‐Dawley rats received either daily ad libitum chow, ad libitum liquid diet containing ethanol, or pair‐fed isocaloric control liquid diet. The ethanol was gradually introduced over 4 weeks, maintained at 5% w/v for four subsequent weeks, and then gradually removed over 1 week.

Results: Four weeks after removal of ethanol from the diet, the previously ethanol‐consuming rats exhibited greater (p < 0.05) evidence of anxiety in the elevated plus‐maze test and in the novel cork‐gnawing test, as well as greater (p < 0.05) locomotor response to a novel environment, compared with controls.

Conclusions: These results suggest that repetitive daily alcohol consumption and withdrawal can induce not only persistent defects in HPA function, but also persistent increases in anxiety and behavioral responsiveness to novelty, all consistent with characteristics of abstinent alcoholics as well as with rats that self‐administer increased amounts of alcohol and other drugs of abuse. This suggests that alcohol abuse can induce persistent common or interacting changes in neuroendocrine and behavioral responses that may increase risk for subsequent abuse.
Comparison of the Adaptive Functioning of Children Prenatally Exposed to Alcohol to a Nonexposed Clinical Sample
Authors: Whaley Shannon E. , O'Connor Mary J. , Gunderson Brent
Published: April 2006

Background : Several studies show impairments in the social and adaptive behaviors of children prenatally exposed to alcohol. However, there remains limited consensus on whether the alcohol exposure directly affects social functioning or whether its effect is mediated by deficits in IQ. In addition, no studies have investigated whether deficits in social functioning are significantly more pronounced in children prenatally exposed to alcohol than in children referred to psychiatric treatment who were not prenatally exposed. We explored the effect of alcohol exposure on social and adaptive functioning and explored whether or not social and adaptive functioning are significantly more impaired in children prenatally exposed to alcohol than in a clinical sample of children.

Methods: A sample of 33 alcohol‐exposed children was compared with a sample of 33 clinic‐referred nonexposed children. The groups were compared on measures of communication, daily living skills, and socialization. The groups were matched on sex, age, IQ, and outpatient or inpatient status.

Results: Analyses revealed that the prenatally alcohol‐exposed children did not differ significantly from the nonexposed children in any of the domains of adaptive functioning. However, with age, exposed children showed a more rapid decline in socialization standard scores compared with the nonexposed clinical sample.

Conclusions: Young children who were exposed to alcohol prenatally show deficits in all domains of adaptive functioning. Although these deficits do not seem to differ from those exhibited by young children with psychiatric problems but no prenatal exposure, deficits in socialization behavior of prenatally exposed children may become more significant with age.
Electroencephalogram Spectral Characteristics After Alcohol Ingestion in Japanese Men With Aldehyde Dehydrogenase‐2 Genetic Variations: Comparison With Peripheral Changes
Authors: Nishimura Fusae T. , Fukunaga Tatsushige , Nishijo Hisao , Ono Taketoshi , Kajiura Hideaki , Yokomukai Yoshiko
Published: April 2006

Background : Electroencephalographic (EEG) activity induced by alcohol ingestion may be influenced by individual differences in alcohol metabolism. Specifically, an absence of the low Km isozyme of aldehyde dehydrogenase (ALDH), found in many Asians, may be related to alterations of EEGs.

Methods: EEG power spectral changes induced by 0.4 ml/kg of alcohol ingestion were compared in two groups of Japanese subjects: NN (ALDH2*1/1, n= 19) and ND (ALDH2*2/1, n= 12). Peripheral changes in heart rate and facial skin temperature were evaluated after the same treatment. Blood ethanol, acetaldehyde, and catecholamine levels were determined to evaluate mediation of the ethanol metabolite, acetaldehyde, and its indirect action on EEGs through the effects on peripheral systems.

Results: As expected, blood acetaldehyde was about 10‐fold higher in the ND subjects compared with the NN subjects during the postingestion period. Ethanol produced characteristic EEG changes during and after ingestion. The ND subjects, however, displayed brief periods of decrease in slow α immediately after alcohol ingestion and showed no long‐term EEG changes. The EEG changes were parallel to peripheral changes.

Conclusions: The lack of increased EEG power in the ND subjects may be partly mediated by a direct action of the high concentration of acetaldehyde in the central nervous system that penetrates from the peripheral blood. In addition to this direct action of acetaldehyde, an indirect action, such as feedback from peripheral changes, might prevent the characteristic increase in α and β power observed in the NN subjects.
Effects of Alcohol Consumption on Indices of Iron Stores and of Iron Stores on Alcohol Intake Markers
Authors: Whitfield J. B. , Zhu G. , Heath A. C. , Powell L. W. , Martin N. G.
Published: April 2006

Background : Alcohol increases body iron stores. Alcohol and iron may increase oxidative stress and the risk of alcohol‐related liver disease. The relationship between low or “safe” levels of alcohol use and indices of body iron stores, and the factors that affect the alcohol‐iron relationship, have not been fully characterized. Other aspects of the biological response to alcohol use have been reported to depend on iron status.

Methods: We have measured serum iron, transferrin, and ferritin as indices of iron stores in 3375 adult twin subjects recruited through the Australian Twin Registry. Information on alcohol use and dependence and smoking was obtained from questionnaires and interviews.

Results: Serum iron and ferritin increased progressively across classes of alcohol intake. The effects of beer consumption were greater than those of wine or spirits. Ferritin concentration was significantly higher in subjects who had ever been alcohol dependent. There was no evidence of interactions between HFE genotype or body mass index and alcohol. Alcohol intake–adjusted carbohydrate‐deficient transferrin was increased in women in the lowest quartile of ferritin results, whereas adjusted γ‐glutamyltransferase, aspartate aminotransferase, and alanine aminotransferase values were increased in subjects with high ferritin.

Conclusions: Alcohol intake at low level increases ferritin and, by inference, body iron stores. This may be either beneficial or harmful, depending on circumstances. The response of biological markers of alcohol intake can be affected by body iron stores; this has implications for test sensitivity and specificity and for variation in biological responses to alcohol use.
Influence of Alcohol Intake on Risk for Increased Low‐Density Lipoprotein Cholesterol in Middle‐Aged Japanese Men
Authors: Nakanishi Noriyuki , Yoshida Hiroshi , Nakamura Koji , Kawashimo Haruhito , Tatara Kozo
Published: April 2006

Background : Decreased low‐density lipoprotein (LDL) cholesterol in chronic alcoholics is well known. However, the importance of light to moderate alcohol consumption is less certain.

Methods: We investigated the association of alcohol intake with risk for increased LDL cholesterol over 5 years in a cohort of 933 Japanese male office workers aged 35 to 54 years who had LDL cholesterol levels less than 140 mg/dl and were not taking medication for dyslipidemia, hypertension, diabetes, liver disease, or hyperuricemia at study entry. Incident‐increased LDL cholesterol was defined by an LDL cholesterol level of 140 mg/dl or more or use of medication for dyslipidemia. Each individual's slope for LDL cholesterol was also calculated with a simple linear regression model.

Results: Three hundred twenty‐one men developed increased LDL cholesterol during 3785 person‐years of follow‐up. After controlling for potential predictors of increased LDL cholesterol, the relative risk for increased LDL cholesterol compared with nondrinkers was 0.89 for those who drank 0.1 to 22.9 g/day of ethanol, 0.74 for those who drank 23.0 to 45.9 g/day of ethanol, 0.64 for those who drank 46.0 to 59.9 g/day of ethanol, and 0.54 for those who drank 69.0 g/day or more of ethanol (p < 0.001). Slopes of LDL cholesterol level decreased significantly as alcohol intake increased. From multiple linear regression analyses, alcohol intake remained as an independent negative factor for slopes of LDL cholesterol level.

Conclusions: Alcohol intake is negatively associated with development of increased LDL cholesterol in middle‐aged Japanese men.
Fetal and Maternal Sheep Hypothalamus Pituitary Adrenal Axis Responses to Chronic Binge Ethanol Exposure During the Third Trimester Equivalent
Authors: Cudd Timothy A. , Chen Wei‐Jung A. , West James R.
Published: April 2006

Background : We tested the hypothesis that in utero ethanol exposure results in changes in fetal and maternal adrenocorticotropin (ACTH) and cortisol during the third trimester equivalent, by using a chronically instrumented fetal sheep model.

Methods: Pregnant ewes received saline or ethanol intravenously 3 consecutive days per week from day 109 to day 132 of gestation. Fetal and maternal blood samples were collected on days 118 and 132.

Results: Maternal and fetal ACTH and cortisol values increased on days 118 and 132 of gestation in response to ethanol infusions that created blood ethanol concentrations (BECs) that are easily achievable by human drinkers. Peak ACTH and cortisol values were detected 30 to 60 min after peak BECs were achieved.

Conclusions: Chronic ethanol exposure during the third trimester equivalent in sheep resulted in repeated activation of the hypothalamus‐pituitary‐adrenal axis in both the mother and fetus. Temporally, the patterns of maternal and fetal responses to ethanol infusion were similar. We conclude that ovine maternal ethanol exposure during the third trimester equivalent increases fetal ACTH and cortisol concentrations, hormonal responses that may play a role in mediating alcohol‐related birth defects.
Direct Evidence for a Cause‐Effect Link Between Ethanol Potentiation of GABAA Receptor Function and Intoxication From Hyperbaric Studies in C57, LS, and SS Mice
Authors: Davies Daryl L. , Alkana Ronald L.
Published: April 2006

Background: This article uses a direct ethanol antagonist, increased atmospheric pressure, to further test the causative link between ethanol potentiation of γ‐aminobutyric acid (GABA) type A receptor function and ethanol's behavioral effects. This was done by determining whether initial biochemical findings in long‐sleep (LS) mice extended to other genotypes and whether the previously reported insensitivity of short‐sleep (SS) mice to pressure antagonism of ethanol‐induced loss of righting reflex extended to a nonselected ethanol‐induced behavior.

Methods: The effects of 12 times normal atmospheric pressure of helium‐oxygen gas (heliox) versus ethanol (25–200 mM) potentiation of GABA‐activated Cl− uptake in brain membranes (microsacs) from C57, LS, and SS mice were tested by using a 36Cl− flux assay. The effects of pressure versus ethanol's (2 g/kg) anticonvulsant effect in SS mice were tested by using time to onset of isoniazid‐induced myoclonic seizures.

Results: Exposure to 12 times normal atmospheric pressure heliox antagonized ethanol potentiation of GABA‐activated Cl− uptake in all three genotypes across a range of ethanol concentrations that cause ethanol's behavioral and anesthetic effects. Pressure did not affect baseline receptor function. The threshold for initiating ethanol potentiation differed between genotypes in accordance with their behavioral sensitivities to ethanol (C57 and LS, ≤25 mM; SS, >50 mM). Pressure antagonized ethanol's anticonvulsant effect in SS mice.

Conclusions: The results add important direct evidence supporting the hypothesis that ethanol potentiation of GABAA receptor function is an initial action of ethanol causing its behavioral effects. These findings also provide insight into possible effects of selective breeding on GABAA receptor function.
Examination of a CYP2E1 Repeat Polymorphism in a Monkey Model of Alcohol Abuse
Authors: Walker Stephen J. , Grant Kathleen A. , Vrana Kent E.
Published: April 2006

Background: Cytochrome P‐450 2E1 (CYP2E1) is involved in alcohol metabolism, and the expression of this enzyme displays wide phenotypic variability in humans. It has been proposed that some of this variability in expression may be a consequence of the size of a repeat polymorphism in the 5″ regulatory region of the gene and that the polymorphism may segregate with alcoholism. This study examined whether the repeat polymorphism exists in macaque monkeys and whether it associates with excessive alcohol consumption in this animal model.

Methods: Ten outbred cynomolgus monkeys (Macaca fascicularis) that displayed a voluntary alcohol consumption ranging from 1.0 to 3.6 g/kg/day were genotyped for a CYP2E1 repeat polymorphism. This polymorphism has been documented in the region from −2519 base pair (bp) to −1953 bp of the human CYP2E1 gene 5″ distal promoter.

Results: Individual polymerase chain reaction amplification of genomic DNA from each of the 10 monkey samples revealed a single band of approximately 400 bp in the region corresponding to the human CYP2E1 polymorphism. Polymerase chain reaction amplicons from the 10 individuals were sequenced, and each one generated a 370 bp sequence that is 90% identical to the human gene sequence. However, unlike human alleles that contain five to eight repeats, the cynomolgus monkey is homozygous for a single copy of the repeat most closely resembling repeat 8 (88% identical) in the human gene.

Conclusions: These data demonstrate that the CYP2E1 distal promoter region in monkeys is very similar to the human sequence yet lacks the extensive repeated DNA found in humans. This includes the rare repeats 3 and 4, which have been postulated to play a role in transcription regulation and to associate with alcohol abuse liability in humans. These data suggest that the CYP2E1 polymorphism arose late in evolution and that the regulation of the gene by this genetic region is not associated with a heavy alcohol drinking phenotype in the cynomolgus monkey.
The Role of Biomarkers in Alcoholism Medication Trials
Authors: Allen John P. , Litten Raye Z. , Strid Nuria , Sillanaukee Pekka
Published: April 2006

Background: Increasingly, biomarkers are being incorporated into the research design of clinical trials on medications to reduce drinking in alcoholics. To date, however, there has been little analysis of the unique roles that biomarkers can play in such investigations or of the practical and conceptual considerations that surround their best use in this context.

Methods: Clinical trials of alcoholism medications published between 1985 and the present were abstracted to determine how biomarkers were used and how changes in them related to self‐report measures of drinking.

Results: Six uses of biomarkers were identified: determination of subjects to be included or excluded in the trial; description of baseline sample characteristics; primary and secondary outcome assessment; corroboration of self‐reports of drinking status; specification of patients likely to respond to the medication; and evaluation of drug safety.

Conclusion: Use of biomarkers in such studies appears warranted, particularly as an objective source of information on treatment efficacy that can be considered with patient self‐report measures of drinking status. Biomarkers related to liver functioning also can assist in determination of drug safety for medications metabolized by the liver.
Confirmation That Offspring From Families With Alcohol‐Dependent Individuals Have Greater Hypothalamic‐Pituitary‐Adrenal Axis Activation Induced by Naloxone Compared With Offspring Without a Family History of Alcohol Dependence
Authors: Wand Gary , McCaul Mary E. , Gotjen Deidre , Reynolds Joanna , Lee Shing
Published: April 2006

Background: This study was designed to confirm our previous findings that nonalcoholic offspring from families with alcohol‐dependent individuals have greater hypothalamic‐pituitary‐adrenal axis activation induced by opioid blockade compared with nonalcoholic subjects without a family history of alcohol dependence.

Methods: Sixty‐four nonalcoholic subjects aged 18 to 25 years were enrolled in the protocol. Twenty‐seven subjects were offspring from families with alcohol dependence and were designated as family history–positive subjects (FHP). Thirty‐seven subjects were biological offspring of non–alcohol‐dependent parents and were designated as family history–negative subjects (FHN). Subjects received naloxone hydrochloride (0, 50, 125, 375, and 500 μg/kg) in double‐blind, randomized order; adrenocorticotropin (ACTH) and cortisol were monitored over 120 min.

Results: No hormone differences at baseline or during placebo administration were identified between FHP and FHN subjects. FHP subjects had greater ACTH and cortisol response to opioid receptor blockade induced by naloxone hydrochloride compared with FHN subjects.

Conclusions: These observations confirm previous findings that differences in ACTH and cortisol dynamics between FHP and FHN subjects can be unmasked by opioid receptor blockade.
Origins and Consequences of Age at First Drink. II. Familial Risk and Heritability
Authors: McGue Matt , Iacono William G. , Legrand Lisa N. , Elkins Irene
Published: April 2006

Background: The association of age at first drink (AFD) and alcoholism may reflect a common inherited vulnerability to disinhibitory behavior and psychopathology rather than a direct influence of the former on the latter. We tested the common‐inherited‐vulnerability hypothesis by determining whether AFD is familial and heritable.

Methods: A sample of 1,232 14‐year‐old twins was classified according to their biological parents’ AFD.

Results: Lifetime symptoms of externalizing disorders (i.e., conduct disorder and oppositional defiant disorder) were significantly higher in the sons but not the daughters of parents whose AFD came before age 15 years. Offspring symptoms of internalizing disorders (i.e., major depressive disorder, separation anxiety disorder, and overanxious disorder) were not significantly associated with parental early use of alcohol in either sons or daughters. Early use of alcohol in mothers but not fathers was associated with a significant increase in both sons’ and daughters’ risk of alcohol use by age 14. The contribution of genetic and environmental factors to risk of early alcohol use was investigated in a sample of 416 monozygotic (MZ) and 225 like‐sex dizygotic (DZ) 14‐year‐old twin pairs. Twin similarity for early alcohol use was substantially greater in MZ than DZ twins in boys but about equally similar in MZ and DZ twins in girls. Estimated heritability of early alcohol use was significantly greater in boys (55%) than girls (11%), in part because the genetic factors underlying symptoms of disinhibitory psychopathology contributed more to risk of early alcohol use in boys than girls.

Conclusions: Early use of alcohol is familial and, at least in males, heritable. The familial transmission of early alcohol use is caused in part by genetic risk for disinhibitory psychopathology in males and to shared environmental factors in girls. These results provide some support for a common‐inherited‐vulnerability hypothesis, and suggest that the processes underlying early alcohol use may differ in boys and girls.