bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Hepatic Alcohol Dehydrogenase and Human Erythrocyte Catalase Do Not Metabolize the Cytochrome P‐4502E1 Substrate, Chlorzoxazone
Authors: McCarver‐May D. Gail , Durisin Lubica
Published: April 2006

Studies of cytochrome P‐4502E1 (CYP2E1)‐mediated oxidation of ethanol have been hampered by the lack of a suitable probe for in vivo human studies. Chlorzoxazone, a prescribed skeletal muscle relaxant, is metabolized to 6‐hydroxychlorzoxazone by CYP2E1 and has been advocated as a specific probe of this enzyme on the basis of microsomal studies. The applications of this probe may include delineating the contribution of CYP2E1 to in vivo human ethanol metabolism. However, the activity of nonmicrosomal enzymes in metabolizing chlorzoxazone is unknown. Alcohol dehydrogenase (ADH), predominantly a hepatic cytosolic enzyme, may be more important than CYP2E1 in the oxidation of ethanol to acetaldehyde. The contribution of catalase in the in vivo oxidation of ethanol to acetaldehyde is controversial. To determine if either of these enzymes metabolizes chlorzoxazone and whether ethanol oxidation by either enzyme is inhibited by chlorzoxazone or its metabolite, multiple in vitro studies were performed. ADH enzyme kinetics were performed with human recombinant β1β1 and β3β3 ADH with ethanol and chlorzoxazone (0.5 to 2.5 mM). Neither ADH isoenzyme exhibited NAD+‐dependent oxidation of chlorzoxazone, but displayed Michaelis‐Menten kinetics for ethanol with Km values of 89 μM and 34 mM, for β1β1, and β3β3, respectively. Typical in vivo concentrations of chlorzoxazone and its metabolite, 6‐hydroxychlorzoxazone, did not alter β1β1, or β3β3 ADH‐mediated oxidation of ethanol to acetaldehyde. Studies of human hepatic nonmicrosomal enzyme activity were expanded to include all nonmicrosomal NAD+‐dependent hepatic enzymes by starch gel electrophoresis assessment. Human hepatic enzymatic activity in the presence of chlorzoxazone was similar to that observed in the control sample (no added substrate), suggesting a lack of metabolism by NAD+‐dependent enzymes. Similarly, human erythrocyte catalase, in the presence of a hydrogen peroxide generating system, did not metabolize chlorzoxazone. Furthermore, neither chlorzoxazone nor 6‐hydroxychlorzoxazone altered the catalase‐induced formation of acetaldehyde from ethanol. These data are consistent with chlorzoxazone as a specific probe of CYP2E1 that may be useful to alcohol researchers.
Quantitative Trait Locus Analyses of Sleep‐Times Induced by Sedative‐Hypnotics in LSXSS Recombinant Inbred Strains of Mice
Authors: Christensen Steven C. , Johnson Thomas E. , Markel Paul D. , Clark Vanessa J. , Fulker David W. , Corley Robin P. , Collins Allan C. , Wehner Jeanne M.
Published: April 2006

The long‐sleep (LS) and short‐sleep (SS) selected lines of mice show highly significant differences in sleep‐time for many sedative‐hypnotic drugs, and the quantitative genetic nature of these differences has been well‐established. Using an interval‐mapping approach, quantitative trait locus (QTL) analyses of LSXSS recombinant inbred (RI) strains have been applied to sleep‐time responses for various classes of sedative‐hypnotic drugs: alcohols (ethanol, n‐propanol, and n‐butanol), the atypical anesthetic chloral hydrate, barbiturates (pentobarbital and secobarbital), and benzodiazepines (chlordiazepoxide and flurazepam). Several provisional QTLs were mapped to similar locations within and between drug classes, suggesting that some common loci are involved in sleep‐times elicited by these drugs. Consistent with correlations of strain mean sleep‐times between drugs tested in the LSXSS recombinant inbred strains, the number of provisional QTLs mapping to the locations of highest significance for ethanol decreases when the lipid solubility of a particular drug becomes less similar to that of ethanol. Provisional QTLs mapped for the benzodiazepines, however, revealed considerable overlap with those mapped for ethanol, although these drugs represented the most lipid‐soluble category of sedative‐hypnotics tested. Provisional QTLs for pentobarbital and secobarbital differed from most of those mapped for the alcohols, which supports the hypothesis that alcohols and barbiturates exert their effects mainly through different biological mechanisms in the LS and SS lines. Blood ethanol concentrations at regaining the righting reflex also mapped to several provisional QTLs corresponding to ethanol‐induced sleep‐times that support the contention that sleep‐time is a reasonable index of the observed differences in central nervous system sensitivities to ethanol between LS and SS mice.
Ethanol‐Induced Alterations in the Posttranslational Processing, But Not Secretion of Luteinizing Hormone‐Releasing Hormone In Vitro
Authors: Uddin S. , Wilson T. , Emanuele M. A. , Williams D. , Kelley M. R. , Emanuele N.
Published: April 2006

The effects of ethanol (EtOH) on the male hypothalamic pituitary reproductive axis are multiple and varied. Although direct gonadal toxicity has been reported, hypothalamic‐pituitary perturbations have also been noted. The difficulty of sampling the hypothalamus has made direct investigation of EtOH‐induced alterations on luteinizing hormone‐releasing hormone (LHRH) fraught with interpretation problems. To circumvent this, we have conducted a series of experiments exploring the effect of 200 mg% EtOH in vitro on GT1‐7 cells, a newly developed LHRH secreting neural cell line. Cell lines were treated with EtOH‐containing or EtOH‐free media for 2,6,24, or 48 hr. EtOH caused no significant change in LHRH secretion at any time point, although there was a trend to increased secretion after 2 hr EtOH exposure when compared with control. Significantly increased total (i.e., cellular plus secreted) pro‐LHRH coupled with significantly reduced cellular LHRH after 6 hr only of EtOH exposure suggested that EtOH caused a transient decrease in processing from bioinactive pro‐LHRH to bioactive LHRH. However, even at this time point, LHRH secretion from these EtOH‐exposed cells was no different than from control cells. Steady‐state LHRH mRNA levels were not changed by EtOH at any time point. These findings are concordant with previous in vitro data using hypothalamic tissue that has similarly demonstrated no effect of EtOH on LHRH secretion. Taken together with the in vivo demonstration that EtOH reduces hypothalamic‐pituitary portal blood levels of LHRH, these data indicate that EtOH exerts its effect either at an extrahypothalamic locus and/or on non‐LHRH‐producing cells within the hypothalamus.
Effects of Alcohols on Murine Preimplantation Development: Relationship to Relative Membrane Disordering Potency
Authors: Kowalczyk Carole L. , Stachecki James J. , Schultz Jeffery F. , Leach Richard E. , Armant D. Randall
Published: April 2006

During in vitro culture of murine preimplantation embryos, we have observed that exposure to 0.1% ethanol induces an immediate increase in intracellular calcium levels and subsequently accelerates embryogenesis. If the observed effects of ethanol on developing embryos is mediated by its membrane disordering potency, we hypothesized that the relative membrane disordering potencies of related alcohols would correspondingly effect embryonic intracellular calcium levels and developmental rates. Two‐cell embryos were exposed to 0.1% ethanol or 0.05 to 1.0% (w/v) n‐butanol, n‐propanol, isopropanol, 1,2‐propanediol, glycerol, or methanol for 24 hr at 37°C, and development to the blastocyst stage was monitored after 5 days. n‐Butanol, n‐propanol, isopropanol, and methanol treatment caused a dose‐dependent inhibition (p < 0.01) of development to the blastocyst stage, whereas 1,2‐propanediol or glycerol neither accelerated nor inhibited development. In a second experiment, 8‐cell morulae were treated with 1,2‐propanediol or glycerol, and cavitation rates were examined. There was no significant difference from control embryos in the onset of cavitation or the blastocoel expansion rate of 1,2‐propanediol‐ or glycerol‐exposed embryos, whereas exposure to 0.1 % ethanol accelerated cavitation (p > 0.05). In a third experiment, morulae were exposed to 0.1% or 1.0% of each alcohol and were monitored for changes in intracellular calcium levels using the fluorescent indicator, fluo‐3‐acetoxymethyl ester. There was an immediate increase in intracellular calcium levels when morulae were treated with 1.0% ethanol or n‐butanol, but only ethanol induced an increase (p < 0.05) in the level of intracellular calcium at 0.1 %. These data suggest that ethanol is unique in its ability to accelerate embryogenesis and that the membrane disordering potency of ethanol does not directly underlie its effects on intracellular calcium release and the acceleration of preimplantation development.
Ethanol Inhibits Human Osteoblastic Cell Proliferation
Authors: Klein Robert F. , Fausti Kristina A. , Carlos Amy S.
Published: April 2006

The habitual consumption of alcoholic beverages is clearly associated with low bone mass and an increased prevalence of skeletal fractures. Microscopic analysis of skeletal tissue from alcoholic patients reveals reduced osteoblast number and suppressed bone formation activity with a relative sparing of resorptive indices. The decreased number of osteoblasts observed in alcoholic subjects results from either impaired proliferation or accelerated senescence. Polyamines and ornithine decarboxylase (ODC), the rate‐limiting enzyme for polyamine synthesis, are essential for cell proliferation in a variety of cell types. To determine if the adverse effect of ethanol on osteoblast number involves modulation of polyamine biosynthesis, we examined the effect of ethanol on parameters of cell growth and ODC activity in a human osteoblast‐like osteosarcoma cell line (TE‐85). Ethanol markedly impaired DNA synthesis and cell proliferation in a dose‐dependent fashion, but alkaline phosphatase activity (a marker of differentiated osteoblast function) remained intact, and accelerated apoptosis was not evident. Thus, the reduced osteoblastic cell number was a result of a direct effect on proliferative processes rather than a nonspecific toxic effect of ethanol to accelerate cell death. Induction of ODC activity was impaired in ethanol‐exposed cell cultures in a dose‐dependent fashion that paralleled the antiproliferative effects. Finally, supplemental polyamine administration substantially improved DNA synthesis in ethanol‐exposed UMR 106‐01 cell cultures. These data confirm a direct inhibitory effect of ethanol on osteoblast proliferation without overt cellular toxicity that may, in part, explain the reduced bone mass observed in those who consume excessive amounts of alcohol.
Zonal Differences in Ethanol‐Induced Impairments in Fluid‐Phase Endocytosis in Rat Hepatocytes
Authors: Camacho Kenneth B. , Tuma Dean J. , Casey Carol A.
Published: April 2006

We have previously shown that ethanol administration impairs the processes of fluid‐phase endocytosis (FPE) and receptor‐mediated endocytosis (RME) in isolated rat hepatocytes after as early as 1 week of ethanol administration. The defects in RME were most prominent in the perivenule (PV) region of the liver lobule, the area wherein alcoholic liver disease has been shown to start and predominate. We undertook the present study to see if changes in FPE were likewise more apparent in the PV versus the periportal (PP) region of the liver. For these studies, we fed male Sprague‐Dawley rats with an ethanol‐supplemented liquid diet or an isocaloric control diet for 1 or 5 weeks. PV and PP hepatocytes were isolated using a digitonin‐collagenase perfusion method. Internalization and efflux of the marker dye, Lucifer Yellow, was then examined in the cell populations. After as early as 1 week of feeding, cells from the PV region in ethanol‐fed animals showed dramatic impairments in the net internalization of dye, compared with PV cells from the pair‐fed controls, and these changes persisted throughout the 5‐week feeding period. In contrast, internalization of Lucifer Yellow in cells from the PP region of the liver were not different between control and ethanol animals. Because net internalization represents the balance between uptake into the cells versus efflux from the cells, we examined these components individually. Early uptake of the dye into the cell was not altered by ethanol treatment. The decreased net internalization seemed to be caused by enhanced efflux of the dye, which was significantly increased in PV cells, compared with the same cell type in control animals. Cells from the PP region of the ethanol‐fed animals did not exhibit altered efflux after either 1 or 5 weeks of feeding. These results indicate that ethanol‐induced impairments in FPE are more dramatic in the PV region of the liver, and these impairments seem to result from an ethanol‐induced enhancement of efflux.
S‐Methyl‐N,N‐Diethylthiocarbamate Sulfoxide and S‐Methyl‐N,N‐Diethylthiocarbamate Sulfone, Two Candidates for the Active Metabolite of Disulfiram
Authors: Mays Dennis C. , Nelson Albert N. , Lam‐Holt Jennifer , Fauq Abdul H. , Lipsky James J.
Published: April 2006

The mechanism of action of disulfiram involves inhibition of hepatic aldehyde dehydrogenase (ALDH). Although disulfiram inhibits ALDH in vitro, it is believed that the drug is too short‐lived in vivo to inhibit the enzyme directly. The ultimate inhibitor is thought to be a metabolite of disulfiram. In this study, we examined the effects of S‐methyl‐N,N‐diethylthiocarbamate (MeDTC) sulfoxide and S‐methyl‐N,N‐diethylthiocarbamate sulfone (confirmed and proposed metabolites of disulfiram, respectively) on rat liver mitochondrial low Km ALDH. MeDTC sulfoxide and MeDTC sulfone, in 10‐min incubations with detergent‐solubilized mitochondria, inhibited ALDH activity with an IC50 (mean ± SD) of 0.93 ± 0.04 and 0.53 ± 0.11 μM, respectively, compared with 7.4 ± 1.0 μM for the parent drug disulfiram. Inhibition by MeDTC sulfone and MeDTC sulfoxide, both at 0.6 μM, was time‐dependent, following apparent pseudo‐first‐order kinetics with a t1/2 of inactivation of 3.5 and 8.8 min, respectively. Dilution of ALDH inhibited by either sulfoxide or sulfone did not restore activity, an indication of irreversible inhibition. Addition of glutathione (50 to 1000 μM) to ALDH before the inhibitors did not alter the inhibition by MeDTC sulfoxide. In contrast, the inhibition by MeDTC sulfone was decreased > 10‐fold (IC50= 6.3 μM) by 50 μM of glutathione and almost completely abolished by 500 μM of glutathione. The cofactor NAD, in a concentration‐dependent manner, protected ALDH from inhibition by MeDTC sulfoxide and MeDTC sulfone. In incubations with intact mitochondria, the potency of the two compounds was reversed (IC50 of 9.2 ± 3.6 and 0.95 ± 0.30 μM for the MeDTC sulfone and sulfoxide, respectively). Our results suggest that MeDTC sulfone is highly reactive with normal cellular constituents (e.g., glutathione), which may protect ALDH from inhibition, unless this inhibitor is formed very near the target enzyme. In contrast, MeDTC sulfoxide is a better candidate for the ultimate active metabolite of disulfiram, because it is more likely to be sufficiently stable to diffuse from a distant site of formation, such as the endoplasmic reticulum, penetrate the mitochondria, and react with ALDH located in the mitochondrial matrix.
Quantitative Autoradiography Using Selective Radioligands for Central and Peripheral‐Type Benzodiazepine Receptors in Experimental Wernicke's Encephalopathy: Implications for Positron Emission Tomography Imaging
Authors: Leong Dorothy K. , Oliva Luis , Butterworth Roger F.
Published: April 2006

Wernicke's encephalopathy (WE) is difficult to diagnose during life, with up to 80% of cases being missed by routine neurological evaluation in both alcoholics and AIDS patients. Therefore, there is a need for noninvasive diagnostic procedures. Using the pyrithiaminetreated rat, an animal model of WE, we have studied, using radioligands for central (neuronal) and “peripheral‐type” (glial) benzodiazepine receptors, the regional distribution of changes in the densities of these receptors in relation to the degree of reactive gliosis accompanying neuronal loss. Histological studies revealed neuronal loss in selective regions, including the thalamus, inferior colliculus, inferior olivary nucleus, and mammillary body. Autoradiographic studies demonstrated increases in densities of [3H]PK11195 binding sites that closely paralleled the topographic distribution of neuronal cell loss and reactive gliosis. In contrast, [3H]Ro 15‐178 showed poor spatial correlation, with the neuronal loss seen in pyrithiamine‐induced thiamine deficiency. The positron emission tomography ligand [11C]PK11195 may be useful for the assessment of thiamine deficiency‐induced brain damage in human alcoholics.
Glutamate Receptors in the Frontal Cortex of Alcoholics
Authors: Freund Gerhard , Anderson Kevin J.
Published: April 2006

This study tests the hypothesis that glutamate receptors are altered in the brains of alcoholics as a result of chronic alcohol neurotoxicity. Release of the neurotransmitter glutamate after seizures or brain ischemia may damage postsynaptic neurons by increasing calcium flux through N‐methyl‐D‐aspartate (NMDA) receptor‐gated ion channels. Alcohol has two opposite effects on glutamate receptor ion channel complexes, depending upon the duration of exposure. Acute exposure to alcohol inhibits ion flow through these receptor‐ channel complexes, whereas chronic exposure up‐regulates the number of these receptors and thereby increases ion flow. Acute withdrawal from alcohol results in hyperexcitability and seizures in the presence of up‐regulated channels, thereby making postsynaptic neurons vulnerable to excitotoxic damage. We selected 13 histologically normal brains from alcoholics and 13 brains from controls from our brain bank that were matched for age, postmortem interval, and storage time. Maximal binding and affinities of glutamate receptor subtypes were determined by quantitative autoradiography in the superior frontal cortex, Brodmann area 8. The most alcohol‐sensitive subtype, NMDA receptor‐channel complexes, were modestly but consistently increased in alcoholics. This included agonist sites (NMDA‐sensitive [3H]glutamate), and antagonist site [3H]CGP‐39653), and a [3H]MK‐801 binding site in the channel interior, although the increase of the latter did not reach statistical significance. Age, autopsy delay, time in storage, liver diseases, thiamine deficiency, CNS medications, and various diseases causing acute and chronic hypoxia did not significantly affect receptor density or affinity. In contrast, the other two glutamate channel subtypes, AMPA and kainate receptors, were not significantly different in alcoholics compared with controls. In conclusion, chronic alcoholism moderately increases the density of the NMDA subtype of glutamate receptors in the frontal cortex. This up‐regulation may represent a stage of alcohol‐induced chronic neurotoxicity.
Eating Pathology among Women with Alcoholism and/ or Anxiety Disorders
Authors: Sinha Rajita , Robinson Jane , Merikangas Kathleen , Wilson G. Terence , Rodin J. , O'Malley Stephanie
Published: April 2006

Two hundred one non‐treatment seeking women with alcoholism, anxiety disorders, alcoholism and anxiety disorders, or neither alcoholism nor anxiety disorders were interviewed to assess core psychopathology associated with eating disorders using the Eating Disorders Examination and DSM‐IIIR psychiatric diagnoses using the Schedule of Affective Disorders and Schizophrenia‐Lifetime version. Alcoholic women had significantly higher mean scores on each of the Eating Disorders Examination subscales of Restraint, Overeating, Eating Concern, Shape Concern, and Weight Concern compared with nonalcoholic women. Women with anxiety disorders had significantly elevated scores on subscales of Overeating, Eating Concern, and Weight Concern compared with women without anxiety disorders. Women with both alcoholism and anxiety disorders had higher rates of bulimia nervosa and/or eating disorder NOS compared with women with either disorder alone. Implications of these findings are discussed in the context of the co‐morbid association between alcoholism, eating disorders, and anxiety disorders.
Effects of Chronic Alcohol Consumption on Enzyme Activities and Active Methionine Absorption in the Small Intestine of Pregnant Rats
Authors: Polache A. , Martin‐Algarra R. V. , Guerri C.
Published: April 2006

The present study evaluates the effect of chronic alcohol intake on the intestinal transport of methionine during pregnancy. For this purpose, we have used an in vitro technique that allows measurement of the unidirectional influx of the amino acids across the brush‐border membrane of the rat mid‐jejunum, and the basolateral membrane enzyme Na+,K+‐ATPase was also evaluated in the duodenum and jejunum. For chronic alcohol treatment, the rats were fed a liquid diet containing ethanol (36% of calories) or an isocaloric diet (pair‐fed control) for 5 weeks before and during pregnancy. Animals were killed at 21 days of gestation. Results from the kinetic analysis revealed that chronic ethanol treatment reduces the maximum transport (Jm) of methionine uptake when compared with controls. Further experiments performed in the presence and absence of sodium have shown that ethanol selectively inhibited Na+‐dependent methionine transport. At the same time, this treatment significantly reduced the levels of Na+,K+‐ATPase in ethanol‐fed rats compared with the controls. Alterations in methionine intestinal transport in pregnant alcohol‐fed rats may contribute to the ethanol‐induced fetal growth abnormalities.
Effects of Buthionine Sulfoximine on the Outcome of the In Utero Administration of Alcohol on Fetal Development
Authors: Reyes E. , Ott S.
Published: April 2006

The adverse effects of the maternal consumption of alcohol on the fetus have been recognized for centuries. Fetal alcohol syndrome is characterized by pre‐ and postnatal growth retardation, mental retardation, behavioral deficits, and facial deformities. Despite numerous animal studies, the biochemical mechanism(s) by which alcohol produces teratogenic effects on the developing fetus are not well understood. Several studies have shown that administration of alcohol to adult rats produces a decrease in hepatic levels of glutathione (GSH). In utero administration of alcohol has also been shown to produce a decrease in GSH levels, as well as prenatal growth retardation and intrauterine death. In an effort to determine if GSH may have a vital role in protecting the fetus against the teratogenic effects of alcohol, buthionine (SR)‐sulfoximine (BSO) was used to deplete GSH levels in the mother and fetus. Timed pregnant Sprague‐Dawley rats were placed on a liquid BioServ diet containing either 0%, 11 YO, =YO, 29%, 31 Yo, 33%, or 35% ethanol‐derived calories, with or without BSO (888 mglkgl24 hr), starting on day 1 of pregnancy. Another set of mothers were fed lab chow and water as a control group for the liquid diet. The mothers were maintained on the diet until gestation day 21 when they were anesthetized with sodium pentobarbital and the pups delivered by cesarean section. The offspring were counted, weighed, killed, and the brain and liver weighed. The effects of BSO on the alcohol dose‐response curves (body weights, brain weights, and litter number) were then determined to ascertain if a depletion in GSH potentiated the effects of alcohol. In utero administration of BSO, aside from the depletion of GSH in the liver and brain in the developing fetus, produced a shift to the left in the alcohol dose‐response curve.
Ethanol and Mitotic Inhibitors Promote Differentiation of Trophoblastic Cells
Authors: Karl Peter I. , Harvey Barrington , Fisher Stanley E.
Published: April 2006

Chronic ethanol abuse during pregnancy can cause fetal injury. A contributing factor in this fetal injury may be the effect of ethanol on the placenta. Ethanol treatment increases human chorionic gonadotropin (hCG) production by cultured human placental trophoblasts. In this study, we show that ethanol treatment reduces total DNA and total protein while stimulating hCG production in term trophoblasts. Ethanol treatment inhibits growth in rapidly proliferating trophoblastic cells from a first trimester placenta and JEG‐3 choriocarcinoma cells. In both cell types, the normal increases in total DNA were inhibited in an ethanol dose‐dependent manner. Normal increases in total protein were inhibited as well. In contrast, hCG production, an indicator of differentiation, was stimulated by ethanol treatment. Treatment of JEG‐3 cells with antimitogenic agents, methotrexate (MTX) or cytosine arabinoside (Ara‐C), inhibited cell growth as indicated by decreased total DNA and total protein accumulation. Similar to that with ethanol treatment, inhibition of cell proliferation was accompanied by increases in hCG production. Taken together, these data suggest that one mechanism by which ethanol increases hCG production in human placental trophoblasts may involve alterations in cellular growth and/or differentiation; such alterations may also occur in other proliferating cells in the growing fetus.
Intra‐amygdala Muscimol Decreases Operant Ethanol Self‐administration in Dependent Rats
Authors: Roberts Amanda J. , Cole Maury , Koob George F.
Published: April 2006

Dependence is an important factor motivating continued alcohol use in human alcoholics. Development of a model of ethanol (EtOH) consumption in dependent animals would advance the understanding of reinforcement after chronic EtOH exposure and allow for the investigation of the neuropharmacological mechanisms mediating reinforcement in dependent versus nondependent animals. In the present study, rats were trained to lever press for 10% EtOH, surgically implanted with bilateral guide cannulae in the amygdala, and either made dependent on EtOH by exposure for 2 weeks to EtOH or exposed to air in identical vapor chambers. Upon removal, the rats were placed in operant boxes and allowed to respond on levers for 10% EtOH or water during a 12‐hr period. Rats were removed briefly at ∼6.5 hr for intra‐amygdala injections of saline or the GABAA, receptor agonist muscimol. After the test period, rats were returned to the vapor Chambers for 4 days before retest. EtOH‐dependent animals responded more for EtOH across the 12‐hr test period than did air control nondependent rats; this difference became more pronounced with repeated test sessions. Intra‐amygdala muscimol significantly decreased responding for EtOH in EtOH‐dependent rats, but had no effect in nondependent controls. These data suggest that the reinforcing effects of EtOH and neurotransmitter pathways mediating reward are altered after the development of dependence, and they support the use of this paradigm for further investigations into the neuropharmacology of EtOH dependence.
Differential Effects of Ethanol in Adolescent and Adult Rats
Authors: Little Patrick J. , Kuhn Cynthia M. , Wilson Wilkie A. , Swartzwelder H. Scott
Published: April 2006

Alcohol use in children and adolescents is widespread. However, very little is known about the effects of alcohol exposure during this period of postnatal development. The goal of the present study was to compare the relative sensitivity to the sedative effects of alcohol in periadolescent and adult rats. After treatment with either 4 or 5 glkg ethanol, both 20‐ and 30‐day‐old rats regained their righting reflex significantly earlier than 80‐day old rats. In 30‐day‐old rats, serum ethanol concentrations (SECs) were significantly greater at the time of the recovety of the righting reflex than 80‐day‐old rats. Developmental differences in the effects of ethanol on locomotor activity were also observed. In 60‐day‐old rats, 2.5 glkg ethanol generally decreased locomotor activity. Ethanol did not significantly alter locomotor activity in 20‐ and 30‐day‐old rats. Finally, there were sig nificant developmental differences in the pharmacokinetics of ethanol with a significant delay in the time to peak SECs in 80‐day‐old rats relative to 20‐ and 30‐day‐old rats. These findings indicate that periadolescent rats are less sensitive to the sedative effects of ethanol as they recovered their righting reflex earlier and at significantly higher SECs than adult rats.
Cultured Postnatal Rat Medial Septal Neurons Respond to Acute Ethanol Treatment and Nerve Growth Factor By Changing Intracellular Calcium Levels
Authors: Webb B. , Suarez S.S. , Heaton M.B. , Walker D.W.
Published: April 2006

Ethanol neurotoxicity results in the loss of neurons during the development of the nervous system. Nerve growth factor (NGF) can ameliorate the neurotoxic effects of ethanol (EtOH) in rat medial septal (MS) neurons. These experiments study the effects of EtOH and NGF on neuronal calcium (Ca2+) homeostasis in cultured postnatal day of birth (PO) rat MS neurons. Previously, we observed that EtOH and NGF modulate intracellular Ca2+ levels ([Ca2+]i) in unstimulated and high potassium stimulated (30 mM KCI) cultured rat embryonic day 21 (E21) MS neurons (Webb et al., Brain Res 701:61‐74, 1995). The purpose of the present study was to explore whether the effects of EtOH and NGF on Ca2+ homeostasis were altered by developmental stage. The hypotheses tested were the following: treatment with EtOH affects Ca2+ homeostasis in postnatal day of birth (PO) rat MS neurons by causing transient and persistent changes in [Ca2+]i; NGF modulates Ca2+ homeostasis in MS neurons by regulating [Ca2+]i; the action of NGF changes the response of MS neurons to EtOH, thus altering Ca2+ homeostasis; and that EtOH and or NGF effects on Ca2+ homeostasis are developmentally regulated. Our results indicated that behaviorally relevant levels of EtOH caused a rapid transient increase in basal [Ca2+]i, whereas there was no effect of NGF on basal [Ca2+]i. Ethanol and NGF interacted, resulting in the lowering of [Ca2+]i. During stimulation with high K+, EtOH inhibited the change in [Ca2+]i. NGF partially ameliorated this effect of higher levels of EtOH, allowing [Ca2+]i to increase. NGF and the lowest level of EtOH potentiated the high K+ stimulated increase in [Ca2+]i. Ethanol and NGF effects on [Ca2+]i were different in the PO neurons compared with our previously published observations in E21 neurons. Therefore, these data suggest that EtOH neurotoxicity and NGF protection involve mechanisms that regulate neuronal Ca2+ homeostasis, and the magnitude of these effects depend on developmental stage.
Medical Consequences of Alcohol Consumption—United States, 1992
Authors: Chou S. Patricia , Grant Bridget F. , Dawson Deborah A.
Published: April 2006

There is plenty of evidence in the alcohol literature that chronic excessive use of alcohol poses a threat to every organ system in the body. At the same time, there is a growing consensus that drinking in moderation protects against cardiovascular disease. This study was based on the most recent national household survey of the United States general population on drinking practices, alcohol use disorders, and their associated disabilities. The prevalences of major alcohol‐related diseases were examined across different categories of drinking status. Excess morbidity caused by heavy intake of alcohol was also studied. Results were generally in agreement with the popular belief that light or moderate drinking is beneficial relative to abstention, particularly that moderate alcohol consumption confers a beneficial cardiovascular effect. Our findings also pointed toward the injurious effect of heavy alcohol use. However, results on benefits of drinking must be interpreted with caution.
No Association of PROP Taster Status and Paternal History of Alcohol Dependence
Authors: Kranzler Henry R. , Moore Pamela J. , Hesselbrock Victor M.
Published: April 2006

Taste sensitivity to bitter substances, including ethanol, may play a moderating role both in the initiation of drinking and in the intensity and frequency of drinking once it is initiated. A study (Pelchat and Danowski, Physiol Behav 51:1261‐1266, 1992) showed an association between the capacity to taste PROP (6‐n‐propylthiouraciI), a bitter tasting compound, and a family history of alcoholism. The implication of that finding is that family‐history‐negative individuals may be protected from early initiation of drinking or heavy consumption of alcohol once drinking is initiated. The present study sought to replicate those findings by using direct interview methods (versus history methods) to obtain alcohol use and alcohol problem information from the parents of “at risk” individuals and by examining a larger number of subjects. A bimodal distribution of the ability to taste PROP was found, similar to that observed in the general population. No association was found between the ability to taste PROP and a paternal history of alcoholism, regardless of the taste threshold employed or after controlling for several potentially confounding variables. A lack of association with alcoholism risk was also observed for subjects' self‐reports of reasons for drinking or not drinking alcohol based upon taste preference: high‐risk and low‐risk for alcoholism groups were not distinguished by either a stated preference for the taste of alcohol as a reason for drinking or a stated dislike for the taste of alcohol as a reason for limiting drinking. Thus, neither taste sensitivity for PROP nor the palatability of ethanol appear to influence the choice to drink among adolescent and young adult individuals at high risk for developing alcohol‐related problems.
Voices of the Afflicted
Authors: Erickson Carlton K.
Published: April 2006

Over the past 10 years, I have been privileged to conduct educational forums for audiences containing many recovering alcoholics or otherwise chemically dependent persons. In these forums about the addictive diseases and their treatment and research possibilities, significant interaction with the audience members occurs. During these interactions, certain anecdotal phenomena seem to predominate. The repetitive nature of these reports suggests the need for systematic investigation. As with editorial comments in major medical journals, observed phenomena and unanswered questions from those afflicted can be valuable in the generation of testable hypotheses. Perhaps the ideas presented herein will be useful in the development of future research on alcohol abuse and alcohol dependence.
Use of Serotonin Selective Pharmacotherapy in the Treatment of Alcohol Dependence
Authors: Pettinati Helen M.
Published: April 2006

This study presents a review of existing human studies that examine the role of serotonin [5‐hydroxytryptamine (5‐HT)] selective agents in the treatment of alcohol dependence. Findings of abnormally low 5‐HT levels in many alcohol‐dependent patients led investigators to explore the use of 5‐HT selective pharmacotherapy in the treatment of alcoholism. Several studies reported that the administration of 5‐HT selective pharmacological agents to alcohol‐dependent patients effectively reduced alcohol intake, although results have been relatively modest. The most recent study by Kranzler et al. (Am. J. Psychiatry 152:391–497, 1995) found no added benefit over cognitive‐behavioral treatment. However, the original relationship between the presence of alcohol dependence and low 5‐HT levels in the brains of patients with alcoholism was predicated on subtypes of alcohol dependence. Research has suggested that 5‐HT plays an important role in some forms of alcoholism that may be genetically or developmentally mediated. Thus, additional studies are needed to determine if treatment with 5‐HT selective agents is a viable therapeutic option for alcoholism, specifically in types of alcohol‐dependent patients who are most likely to benefit from treatment with 5‐HT selective pharmacotherapy.
N‐Methyl‐d‐Aspartate‐Stimulated Increases in Intracellular Calcium Exhibit Brain Regional Differences in Sensitivity to Inhibition by Ethanol
Authors: Randoll Lynne A. , Wilson Walter R. , Weaver Melanie S. , Spuhler‐Phillips Karen , Leslie Steven W.
Published: April 2006

This study compared N‐methyl‐d‐aspartate (NMDA)‐stimulated increases in intracellular calcium in fura‐2–loaded neurons dissociated from newborn rat brainstem (EC50 in μM; 6.4), cerebellum (9.5), forebrain (6.3), and hippocampus (10.6). Ethanol inhibition of the response to 25 μM NMDA differed among the regions. The NMDA response in hippocampus was inhibited by 20 mM ethanol; cortex and cerebellum responses were inhibited by 80 mM ethanol, and no inhibition was seen in the brainstem. Addition of glycine (15 μM) failed to attenuate ethanol inhibition of the NMDA response. These results demonstrate that ethanol inhibition of NMDA‐stimulated responses varies according to brain region. In contrast to previous findings from this laboratory using dissociated neurons from whole brain, the addition of glycine did not reverse the inhibitory effects of ethanol on NMDA‐stimulated responses.
Role of Protein Synthesis on Ethanol Regulation of Adenylyl Cyclase Activity in Wild‐Type S49 Murine Lymphoma Cells
Authors: Nakamura Jiro
Published: April 2006

Adenylyl cyclase activity was determined in membranes from wild‐type S49 murine lymphoma cells that had been exposed to ethanol for 4 hr. Mn‐, NaF‐, and forskolin‐stimulated adenylyl cyclase activities of cells—pretreated with cycloheximide, puromycin, or serum deprivation—were significantly decreased by treatment with 50 mM of ethanol. As demonstrated for Mn‐stimulated activity, the decrease was dose‐dependent on ethanol and was temporal; a normal activity recovered after 16–24 hr treatment, even in the presence of cycloheximide and ethanol. Studies with a cell‐free membrane system of S49 cells revealed a similar activity decrease after treatment of the membranes with ethanol. In contrast, cells treated with 50 mM of ethanol in a regular culture condition showed no decrease in adenylyl cyclase activity over 24 hr. These results indicate that ethanol regulation of adenylyl cyclase activity in S49 cells depends on reduced or impaired protein synthesis.
Diabetic Patients' Alcohol Use and Quality of Life: Relationships with Prescribed Treatment Compliance among Older Males
Authors: Cox W. Miles , Blount Joseph P. , Crowe Paul A. , Singh Sant P.
Published: April 2006

One hundred fifty‐four male patients attending an outpatient diabetes clinic were interviewed to determine the incidence of alcohol use among them and to identify mechanisms by which alcohol use adversely affects the control of diabetes. Specifically, we determined how diabetic patients' use of alcohol is related to compliance with their diabetes treatment regimen, and diabetic patients' expectations of benefits from drinking alcohol and actual alcohol use is related to their quality of life. Greater alcohol use was related to poorer prescribed insulin injection compliance (p < 0.01), and stronger expectations of immediate, positive consequences of drinking alcohol were related to several indices of lower quality of life (p < 0.005). These results imply interactive relationships among diabetics' alcohol use and expectancies, treatment compliance, and quality of life.
Moderate Alcohol Consumption and Disorders of Human Spermatogenesis
Authors: Pajarinen Jarkko , Karhunen Pekka J. , Savolainen Vesa , Lalu Kaisa , Penttilä Antti , Laippala Pekka
Published: April 2006

Environmental factors are suspected to be responsible in part for the deterioration in semen quality observed worldwide during the recent few decades. Alcohol might be one factor, considering the frequent changes in testicular function associated with heavy drinking. The dose‐dependent effects of alcohol on human spermatogenesis are, however, not well known. We analyzed spermatogenesis and testicular tissue morphology of 195 men, aged 35–69 years, with computer‐assisted microscopy in this autopsy study. The men were categorized into controls and four “consumption groups” according to the average daily alcohol consumption, which was determined on the basis of blind interviews with relatives and acquaintances. When the average daily alcohol consumption was 40 g or less, 59 (66%) of the 90 men showed normal spermatogenesis, whereas 31 (34%) had partial spermatogenic arrest (SA). Of the 31 men with average daily intake between 40 and 80 g, 17 (54%) showed normal spermatogenesis, 13 (42%) had partial or complete SA, and 1 man exhibited more severe testicular damage—“Sertoli cell only” (SCO) syndrome. Among men with daily intake between 80 and 160 g, only 13 of 35 men showed normal spermatogenesis (37%), 19 (54%) had partial or complete SA (odds ratio = 2.92), and 3 (9%) had the SCO syndrome (odds ratio = 16.85). The frequencies of spermatogenic disorders were similar in men drinking in excess of 160 g. Both SA and the SCO syndrome showed a clear dependence on daily dose; p < 0.0001 and p < 0.0004, respectively. We conclude that long‐term average daily consumption of <40 g of alcohol seems not to be associated with disorders of spermatogenesis. Consumption of moderate amounts of alcohol may affect semen quality more often than previously thought, whereas high alcohol consumption may even be associated with serious disorders of spermatogenesis.
Relationship between Alcohol Withdrawal Seizures and Temporal Lobe White Matter Volume Deficits
Authors: Sullivan Edith V. , Marsh Laura , Mathalon Daniel H. , Lim Kelvin O. , Pfefferbaum Adolf
Published: April 2006

A previous magnetic resonance imaging study from our laboratory reported significant temporal lobe volume deficits in cortical gray matter, white matter, and anterior hippocampus in chronic alcoholic men relative to controls. In the present study, we reexamined these data and asked whether withdrawal seizure history was predictive of either the hippocampal or the extrahippocampal volume deficits. A review of the medical charts indicated that 11 alcoholics had experienced one or more alcohol‐related seizures and 35 were seizure‐free; no patient had a seizure disorder unrelated to alcohol. The two alcoholic groups did not differ significantly in age, education, alcohol consumption variables, premorbid intelligence, Memory Quotient, Trail Making, or detection of hidden figures. Although each alcoholic group showed significant bilateral volume deficits of the anterior hippocampus and frontal‐parietal and temporal gray matter, relative to controls, the seizure group had significantly smaller temporal lobe white matter volumes than either the control or the seizure‐free groups; the latter two groups did not differ from each other. Both alcoholic groups, however, had white matter volume deficits in the frontal‐parietal region. Thus, the seizure group accounted for the white matter volume deficits in the temporal lobe previously reported in the full sample of alcoholics. It seems, then, that reduced white matter volume in the temporal lobes may be either a risk factor for or sequela of alcohol withdrawal seizures.
Erythropoietin and Cytokine Levels in the Anemia of Severe Alcoholic Liver Disease
Authors: Means Robert T. , Mendenhall Charles L. , Worden Bradley D. , Moritz Thomas E. , Chedid Anthony
Published: April 2006

Purpose: The anemia of chronic disease is mediated by the cytokines that modulate the immune response, such as tumor necrosis factor (TNF) and γ‐interferon (γ‐IFN), and is associated with a blunted serum erythropoietin (sEPO) response to anemia. Previous reports suggest that patients with liver disease (LD) also exhibit a blunted sEPO response to anemia, and that patients with alcoholic LD had altered cytokines, including elevated TNF levels. To investigate the pathogenesis of anemia in alcoholic LD, sEPO, TNF, and γ‐IFN levels were determined in patients who had participated in a Department of Veterans Affairs Cooperative study of alcoholic LD. Methods: sEPO, serum TNF‐α, and serum γ‐IFN levels were evaluated in 40 patients with severe biopsy‐proven alcoholic LD whose serum had been stored during the Department of Veterans Affairs Cooperative Study 275, and in 18 patients with iron deficiency (controls). Results: Mean hemoglobin (Hgb) was 11.2 ± 0.3 g/dl for LD patients versus 11.4 ± 0.4 g/dl for controls (p= 0.84). sEPO levels measured by ELISA were 29.6 ± 4.1 units/liter in LD patients versus 25.4 ± 5.4 units/liter in controls (p= 0.64). In both sets of patients, sEPO and Hgb were inversely related; the slopes of the two regression lines did not differ significantly (p= 0.92). TNF was detected in 3 of 40 LD patients and in 0 of 18 iron‐deficient patients. Detection of TNF did not correlate with sEPO or Hgb, but did correlate strongly with severe caloric malnutrition (marasmus) and mortality at 6 months (p= 0.049 and 0.04, respectively).γ‐IFN was not detected. Conclusions: These findings indicate that the sEPO response is preserved in patients with severe alcoholic LD, and suggest that anemia in LD arises from different mechanisms than does the anemia of chronic disease. TNF production in severe alcoholic LD is strongly correlated with caloric malnutrition and mortality.
Alcohol Use and T‐Lymphocyte Subsets among Injection Drug Users with HIV‐1 Infection: A Prospective Analysis
Authors: Crum Rosa M. , Galai Noya , Cohn Sylvia , Celentano David D. , Vlahov David
Published: April 2006

Objectives: Alcohol use is known to alter immune function and has immunosuppressive effects that may modify T‐lymphocyte subpopulations. However, there is no clear evidence regarding the relationship of alcohol use with the progression of immunodeficiency in human immunodeficiency virus‐type 1 (HIV‐1)‐seropositive individuals, particularly injection drug users (IDUs). Methods: Using prospective data from a cohort of IDUs in a study of the natural history of HIV infection, we examined the relationship of alcohol use and changes in T‐lymphocyte subsets. Among the 2921 IDUs followed semiannually in outpatient clinics, 188 were documented HIV‐1 seroconverters with known time of seroconversion. At each visit, all study participants were interviewed, underwent physical examinations, and had blood drawn for laboratory studies. Alcohol use was measured by reported frequency and quantity of alcoholic beverages. Longitudinal analyses included data for up to 5 years postseroconversion. To formally test the association of alcohol use with change in levels of CD4+ and CD8+ cells subsequent to HIV seroconversion, regression models incorporating autocorrelation structure were applied. Results: Alcohol use was not appreciably related to age, gender, marital status, income, education, or the duration of intravenous drug use. CD4% decreased for all IDUs within the first 5 years after seroconversion, with no significant differences between alcohol categories. CD8% increased for all IDUs, with no significant differences by alcohol category within the first 2 years after seroconversion. However, between 2 to 5 years postseroconversion, there was a statistically significant increase among the heaviest drinkers: CD8% increased 6.9%/year [95% confidence interval (CI): 4.7, 8.0] for the IDUs who reported <21 drinks/week, 2.4%/year (95% CI: 0.8, 4.0) for IDUs who drank 21 drinks/week or less, and 0.4% (95% CI: ‐2.1, 2.9) for abstainers. Similar results were obtained for CD4 and CD8 absolute counts. Conclusions: In this study population of IDUs, CD8% (but not CD4%) is associated with alcohol consumption early after HIV seroconversion. This is the first prospective study to date to assess the relationship of alcohol use with HIV progression from the time of HIV seroconversion among a cohort of IDUs. If confirmed in future investigations, the findings may have significant implications for prevention and early intervention programs aimed at inhibiting disease progression among HIV‐positive IDUs.