bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ethanol Effects on Nitric Oxide Production in Cerebral Pial Cultures
Authors: Shih Chin‐Lung , Chi Shu‐Ing , Chiu Ted H. , Sun Grace Y. , Lin Teng‐Nan 
Published: April 2006

Background: Although alcohol abusers are known to have higher incidences of hemorrhagic cerebrovascular diseases, it is not known whether these changes are associated with ethanol (EtOH) action on nitric oxide (NO) production in the cerebrovascular cells. The purpose of this study was to examine the effects of EtOH treatment on basal and cytokine‐induced NO production in cortical pial cultures.Methods: Cell cultures for this study included murine primary pial vascular cells, primary glial cells and cortical neurons. These cells were exposed to cytokines or EtOH for 24 to 48 hr. The culture media were used for measurement of nitrite, as an indication for NO release, and lactate dehydrogenase (LDH), as an index of cell membrane integrity. In addition, immunocytochemical determinations were carried out to identify cell types and to assess inducible nitric oxide synthase (iNOS).Results: Exposure of primary pial vascular cultures to cytokines that consisted of interleukin‐1β (IL‐1β; 250 pg/mL) and interferon‐γ (IFNγ; 2 ng/mL) or to EtOH (50 to 100 mM) for 24 to 48 hr significantly elevated NO production. NO production could be attenuated by N‐nitro‐L‐arginine (N‐arg), a nonspecific NOS inhibitor, or aminoguanidine (AG), an iNOS inhibitor. Increased iNOS immunoreactivity was observed in cytokines‐ or EtOH‐treated pial cells. When pial cells were cocultured with cortical neurons, prolonged EtOH exposure led to a large increase in NO production as well as LDH release. However, this increase was not observed in pial culture alone or in mixed cortical culture. Nevertheless, inhibition of NO production with N‐arg or AG did not alter the EtOH‐induced LDH release in the pial cells cocultured with cortical neurons.Conclusion: These results show that EtOH exposure led to increased production of NO in primary pial cell culture. In mixed culture that contained cortical neurons and pial cells, EtOH induced increase in NO as well as LDH release, which is an indication of loss of cell membrane integrity. However, EtOH‐mediated LDH release in mixed cortical pial cultures was not a consequence of the increase in NO production by these cells. Studies that use mixed cortical‐pial cultures may provide a unique in vitro system for examining the interactions among glial cells, neurons, and cerebrovascular cells.
Identification of a 251‐bp Fragment of the PAI‐1 Gene Promoter That Mediates the Ethanol‐Induced Suppression of PAI‐1 Expression
Authors: Grenett Hernan E. , Wolkowicz Paul E. , Benza Raymond L. , Tresnak Jennifer K. , Wheeler Crystal G. , Booyse Francois M. 
Published: April 2006

Background: Moderate alcohol consumption reduces the risk for coronary heart disease. This cardioprotection may be due to ethanol enhancement of fibrinolysis. Fibrinolysis involves the interaction of plasminogen activators (PAs) and the plasminogen activator inhibitor type‐1 (PAI‐1). Factor(s) that decrease endothelial cell (EC) PAI‐1 expression increase fibrinolysis and may decrease the risk for cardiovascular disease.Methods: Five promoter deletion fragments were generated from a 1.1‐kb PAI‐1 promoter fragment and ligated to a luciferase reporter gene. Cultured human umbilical vein endothelial cells (HUVECs) were transiently transfected with these PAI‐1 deletion constructs. A 251‐base pair (bp) fragment of the PAI‐1 promoter, positions −800 to −549, was cloned upstream of a heterologous promoter/enhancer. ECs luciferase activity was measured in the absence/presence of 20 mM ethanol. Electrophoresis mobility shift assays were performed with nuclear extracts from untreated and ethanol‐treated ECs using this 251‐bp fragment.Results: Deletion analysis showed a region between position −800 and −549 mediated ethanol repression of luciferase activity. This 251‐bp promoter fragment also repressed the activity of a heterologous promoter/enhancer in the presence of ethanol. Using the labeled 251‐bp fragment, nuclear extracts from ethanol‐treated ECs contained two inducible bands and one enhanced band. Non‐ethanol treated nuclear extracts also contained a band that was not observed in ethanol‐treated samples. Competition using 100‐fold molar excess of unlabeled probe abolished these four bands.Conclusions: Repression of PAI‐I gene transcription in cultured HUVECs exposed to ethanol may involve the interaction of several transcription factors with binding sites localized between positions −800 and −549 of the PAI‐1 gene promoter.
NMDA Receptor Subunit mRNA and Protein Expression in Ethanol‐Withdrawal Seizure‐Prone and ‐Resistant Mice
Authors: Mason John N. , Eshleman Amy J. , Belknap John K. , Crabbe John C. , Loftis Jennifer M. , Macey Tara A. , Janowsky Aaron 
Published: April 2006

Background: Ethanol withdrawal seizure‐prone (WSP) and ‐resistant (WSR) mice have been genetically selected for differences in handling‐induced convulsion severity during withdrawal from chronic ethanol administration. Importantly, drug‐naïve mice from these selected lines also differ in handling‐induced convulsion severity. Different N‐methyl‐D‐aspartate (NMDA) receptor subunit and splice variant associations confer varying sensitivities to ethanol, and may play a role in the different behavioral responses of the WSP and WSR mice.Methods: In situ hybridization of riboprobes was used to characterize NMDA receptor subunit and splice variant mRNA expression in cortex and hippocampus from WSP and WSR mice. In addition, immunoblotting and immunohistochemistry were used to examine the expression of specific NMDA receptor subunits and splice variants in hippocampus and cortex from the selected mouse lines.Results: In situ hybridization of riboprobes indicated that, in brain sections from both WSP and WSR mice, there was a differential regional distribution of mRNA for the mouse NR1, NR2A, NR2B, and NR2C NMDA receptor subunits. However, there were no differences between the selected lines in the hybridization of riboprobes to hippocampal subfields or cortical layers. In addition, hybridization of the probe for a 63‐base N1‐terminal cassette of ethanol‐sensitive NR1 splice variants labeled both cortex and hippocampus. The level of hybridization did not differ across subfields of the hippocampus. Results from Western blot and immunohistochemical experiments also indicated that there were no differences between selected lines in NMDA receptor subunit protein expression. However, there was a correlation between mRNA and protein expression in hippocampus and cortex for each NMDA receptor subunit that was examined.Conclusions: The data suggest that at the level of both mRNA and protein, NMDA receptor subunit and splice variant expression can be uncoupled from convulsion severity in mice that have been selectively bred for symptoms of ethanol withdrawal.
Familial Transmission of Alcoholism Among Nonalcoholics and Mild, Severe, and Dyssocial Subtypes of Alcoholism
Authors: Johnson Eric O. , Pickens Roy W. 
Published: April 2006

Background: In the field of alcohol studies, there are many typologies attempting to reduce the heterogeneity of expression of this complex disorder to better understand its natural history and etiology. However, few typologies have included empirical assessment of the degree of familial liability. To the extent there is variability in genetic vulnerability to alcoholism, inclusion of measures of this variability in proposed typologies is important to their validity and utility. We test whether the mild, severe, and dyssocial typology distinguished cases of alcohol dependence with high familial liability from those with low familial liability to alcoholism.Methods: Data came from the National Longitudinal Alcohol Epidemiologic Survey–1992, a household probability sample representative of those 18 years of age and older in the contiguous US Response rate was 92%. Only whites were included here because the typology under study has been successfully applied to this race/ethnic group only. The total number of respondents were 32,447 and included 13,825 men and 18,622 women. Identification of a biological relative as alcoholic was based on the proband's report. All analyses were weighted to adjust for sampling under a multistage stratified design.Results: Familial density of alcoholism (number of alcoholics/number of adult family members) substantially differed by proband alcohol dependence status for both men and women (male probands—nonalcoholics 7%, mild 13%, severe 25%, dyssocial 19%; female probands—nonalcoholics 8%, mild 18%, severe 33%, dyssocial 24%;p < .001). Cross‐fostering analysis of the probands with adoptive/stepparents indicated little difference between nonalcoholic and mild alcoholic probands and suggests greater influence of biological parents for severe subtype probands compared to other probands.Conclusions: These results suggest construct validity for the alcoholism typology as distinguishing subtypes with differing degrees of familial liability to alcoholism. The typology may be useful when employing an extreme comparison strategy in genetic studies of alcohol dependence.
Effects of Parathyroid Hormone on Bone Formation in a Rat Model for Chronic Alcohol Abuse
Authors: Turner Russell T. , Evans Glenda L. , Zhang Minzhi , Sibonga Jean D. 
Published: April 2006

Background: Alcoholism is a risk factor for osteoporosis and it is not clear whether the detrimental effects of alcohol on bone are reversible. Parathyroid hormone (PTH) is a potent stimulator of bone matrix synthesis and is being investigated as a therapeutic agent to reverse bone loss. The present investigation was designed to determine the effects of PTH on bone formation in a rat model for chronic alcohol abuse.Methods and Results: Alcohol was administered in the diet of female rats (35% caloric intake) for 2 weeks. Human (1–34) PTH (80 μg/kg/day) was administered subcutaneously during the second week of the study. Alcohol resulted in a transient reduction in steady‐state mRNA levels for the bone matrix proteins type 1 collagen, osteocalcin, and osteonectin compared with rats that were fed an alcohol‐free (control) diet. As expected, alcohol decreased and PTH increased histologic indices of bone formation. Additionally, two‐way ANOVA demonstrated that alcohol antagonized PTH‐induced bone formation. Despite antagonism, bone formation and mRNA levels for bone matrix proteins in alcohol‐fed rats treated with PTH greatly exceeded the values in rats fed the control diet.Conclusions: The results of this study contribute to a growing body of evidence that alcohol‐induced bone loss is primarily due to reduced bone formation. We conclude that alcohol does not prevent the stimulatory effects of PTH on bone formation. This is evidence that the effects of alcohol on the skeleton are reversible. Additionally, the positive effects on bone formation in rats that consumed high concentrations of alcohol suggested that PTH may be useful as an intervention to treat alcohol‐induced osteoporosis.
Effects of Age and Parental History of Alcoholism on EEG Findings in Mission Indian Children and Adolescents
Authors: Ehlers Cindy L. , Wall Tamara L. , Garcia‐Andrade Consuelo , Phillips Evelyn 
Published: April 2006

Background: Many, but not all, Native American tribes have some of the highest rates of alcohol abuse and dependence. Yet, risk factors for the development of problem drinking in these high risk groups remain largely unknown. In primarily Euroamerican populations, electrophysiological variables have been associated with risk for alcoholism. The EEG has a specific developmental time course that has been described in a diverse set of ethnic groups, but it has not been described in Native American youth. In addition, the relationship between EEG development and risk for alcoholism in Indian youth has not been previously studied.Methods: Clinical ratings and spectral characteristics of the resting EEG were investigated in 140 Native American Mission Indian children and adolescents between the ages of 7 and 13 years. The specific aims of the study were to (1) investigate the relationship of age and gender with EEG spectral variables to determine if this population conforms to similar trends from previously published data in other ethnic groups and (2) to determine whether children with a parental history of alcoholism differ from those without alcoholic parents on EEG spectral parameters.Results: No excess of abnormal EEG activity was found in this sample of Native American youth. Age, but not gender, was found to have a significant effect on EEG spectral characteristics with younger children (7–11 years old), having significantly more power in slow activity (0.5–7.5 Hz) and in alpha power (8–12 Hz) as well as slower alpha frequencies than older children (12–13 years old). Consistent with other studies of Native American youth, 66% of the children and adolescents participating in this study had at least one parent who had a lifetime diagnosis of alcohol dependence. However, an ANCOVA that covaried for age and gender revealed no significant differences in power or frequency characteristics of the EEG on the basis of parental history of alcoholism.Conclusions: These studies suggest that this sample of Mission Indian children, despite high levels of parental alcohol dependence and low socioeconomic status, show normal EEG development. As yet, no relationship has been found between any specific EEG phenotype and parental history of alcoholism in this population, however, further EEG maturation may be necessary before any relationships can be fully delineated.
Effects of Alcohol Deprivation on Alcohol Consumption Using a Sipper‐Tube Procedure
Authors: Samson Herman H. , Chappell Ann 
Published: April 2006

Background: When rats with prior experience drinking ethanol solutions are deprived of ethanol for various time periods, a transitory increase in alcohol consumption is observed when ethanol solutions are again made available. This has been termed the alcohol deprivation effect (ADE). The ADE has been observed in limited‐access operant procedures in which small dippers of ethanol are presented following completion of a lever press requirement. However, it has not been determined if the effect occurs in an operant model of ethanol self‐administration that separates the ethanol‐seeking responding from the consummatory drinking (the sipper procedure).Methods: Rats were initiated to drink ethanol from a sipper tube using the sucrose‐substitution procedure. Once initiated, the rats had to make 30 lever presses to gain access to a sipper‐tube containing the ethanol solution for 20 min. The effects of 2–3 days, 7 days, and 14 days of ethanol deprivation were examined on ethanol consumption and extinction responding (seeking response).Results: There were no effects of deprivation on intake at any deprivation period examined. Contrary to expectations, the alcohol‐seeking response as measured by extinction responding was decreased after 7 and 14 days of deprivation.Conclusions: The data from this study and others using the limited access operant procedures suggest that the ADE may be more related to consummatory control (i.e., loss of control) and not to ethanol‐seeking behaviors (i.e., craving).
The Combined Dexamethasone‐Suppression/CRH‐Stimulation Test in Alcoholics During and After Acute Withdrawal
Authors: Hundt W. , Zimmermann U. , Pöttig M. , Spring K. , Holsboer F. 
Published: April 2006

Background: Chronic alcoholism is often accompanied by disturbances of the hypothalamic‐pituitary‐adrenal (HPA) system. Patients with alcoholism frequently show nonsuppression in the dexamethasone (Dex) suppression test and also a blunted increase of adrenocorticotropin (ACTH) after injection of corticotropin‐releasing hormone (h CRH). However, the underlying mechanisms have not been fully elucidated. The combined Dex/CRH test (pretreatment with 1.5 mg dexamethasone at 2300 hr, injection of 100 μg h CRH at 1500 hr the next day) has been established as a more sensitive tool to investigate HPA system regulation in depressed patients.Methods: We studied the effect of the combined Dex/CRH test in 19 alcoholic inpatients (9 male, 10 female) during and after withdrawal along with 19 healthy controls.Results: Compared to normal controls, patients showed a severely dysregulated HPA system during withdrawal, with significantly elevated cortisol and ACTH response to h CRH after pretreatment with dexamethasone. After completed withdrawal, cortisol levels after injection of h CRH were almost normalized while ACTH values were partially lower in patients, compared to controls.Conclusions: We conclude that the HPA system is severely disturbed during alcohol withdrawal, possibly reflecting an exaggerated release of hypothalamic corticotropin and vasopressin.
Chronic Ethanol Consumption Alters the Glutathione/Glutathione Peroxidase‐1 System and Protein Oxidation Status in Rat Liver
Authors: Bailey Shannon M. , Patel Vinood B. , Young Tracey A. , Asayama Kohtaro , Cunningham Carol C. 
Published: April 2006

Background: Alcohol‐induced liver damage is associated with oxidative stress, which might be linked to disturbances in liver antioxidant defense mechanisms. The effect of chronic ethanol consumption on the mitochondrial and cytosolic glutathione/glutathione peroxidase‐1 (GSHPx‐1) system and oxidative modification of proteins was therefore studied in the rat.Methods: Male Sprague‐Dawley® rats were fed liquid diets that provided 36% total calories as ethanol for at least 31 days. Pair‐fed controls received isocaloric diets with ethanol calories substituted with maltose‐dextrins. Mitochondrial and cytosolic fractions were prepared from livers and assayed for GSHPx‐1 and glutathione reductase activities and total and oxidized concentrations of glutathione. Catalase activity was measured in the postmitochondrial supernatant. Levels of GSHPx‐1, lactate dehydrogenase, and the beta subunit of the F1 portion of the ATP synthase protein were determined by western blot analysis. Concentrations of mitochondrial and cytosolic protein carbonyls were measured to assess ethanol‐induced oxidation of proteins.Results: Chronic ethanol consumption significantly decreased cytosolic and mitochondrial GSHPx‐1 activities by 40% and 30%, respectively. Levels of GSHPx‐1 protein in cytosol were unaffected by ethanol feeding, whereas there was a small decrease in GSHPx‐1 protein levels in mitochondria isolated from ethanol‐fed rats. Glutathione reductase activities were increased in both intracellular compartments and catalase activity was increased as a consequence of ethanol exposure. Cytosolic total glutathione was mildly decreased, whereas ethanol feeding increased mitochondrial levels of total glutathione. Chronic ethanol feeding significantly increased both cytosolic and mitochondrial concentrations of protein carbonyls by 30% and 60%, respectively.Conclusions: This study demonstrates that chronic ethanol‐induced alterations in the glutathione/GSHPx‐1 antioxidant system might promote oxidative modification of liver proteins, namely those of the mitochondrion, which could contribute to the adverse effects of ethanol on the liver.
Alcohol Exposure During the Brain Growth Spurt Promotes Hippocampal Seizures, Rapid Kindling, and Spreading Depression
Authors: Bonthius Daniel J. , Pantazis Nicholas J. , Karacay Bahri , Bonthius Nancy E. , Taggard Derek A. , Lothman Eric W. 
Published: April 2006

Background: Epilepsy is a prominent sign of brain dysfunction and a cause of substantial disability in some children with fetal alcohol syndrome. The hippocampal formation is vulnerable to alcohol‐induced pathologic changes and is the source of seizure activity in a variety of epileptic conditions. This study tests the hypothesis that developmental alcohol exposure facilitates epileptic activity and promotes kindling within hippocampal circuitry.Methods: Rat pups received either a moderate dose (2.0 g/kg) or a high dose (3.75 g/kg) of alcohol via intragastric intubation over postnatal days 4 to 10. Intubated control and suckle control groups were also included. Upon reaching adulthood (postnatal days 85–100), the rats underwent electrophysiologic testing. A double‐barrel potassium‐sensitive microelectrode was placed into the right dentate gyrus stratum granulosum for the recording of extracellular field potential and extracellular potassium concentration. Stimuli were delivered via an electrode positioned in the CA3 subregion of the left hippocampus. To assess whether alcohol promotes hippocampal seizures and rapid kindling, the parameters of maximal dentate activation (MDA) were measured before, during, and after a series of stimulation‐induced seizures.Results: Developmental exposure to the high dose of alcohol permanently altered several parameters of MDA. Time to onset of MDA and stimulus threshold for afterdischarge production were both decreased, whereas the duration of the afterdischarge was increased. Although the moderate alcohol dose reduced time to onset of MDA, it did not affect any other MDA parameters. Over the course of the repeated induced seizures, spreading depression occurred more often and with fewer stimuli in the high‐dose alcohol group than in any other group. The series of repeated electrographic seizures induced rapid kindling in all of the treatment groups. However, the kindling effect was enhanced in a dose‐response manner by the previous alcohol exposures.Conclusions: These findings demonstrate that exposure to alcohol during brain development can permanently alter the physiology of the hippocampal formation, thus promoting epileptic activity, enhancing kindling, and facilitating spreading depression. The relative roles of alcohol intoxication and withdrawal in these abnormal physiologic responses remain unknown.
Adult‐Onset Alcohol Consumption Induces Osteopenia in Female Rats
Authors: Hogan Harry A. , Argueta Floryluz , Moe Louise , Nguyen Lyndon P. , Sampson H. Wayne 
Published: April 2006

Background: Alcohol is a known risk factor for osteopenia and fracture in humans, and its effects on the skeleton have been studied extensively in animal models. Almost all studies of rats, however, have begun rats on alcohol diets while the animals were young and still growing. The purpose of the current study was to examine the effects of alcohol consumption on rats that began drinking alcohol as adults, so that the confounding effects of growth might be minimized.Methods: Nine‐month‐old female Sprague‐Dawley rats were studied for two durations (8 and 14 weeks). The following diet groups were used for both durations:alcohol (n= 7), in which rats were fed a liquid diet containing ethanol (8.1% v/v; Lieber‐DeCarli method);pair‐fed (n= 7), in which rats were fed a caloric‐equivalent liquid diet matched to the alcohol‐fed animals; and pellet (n= 6), in which rats consumed standard rat chow and water. A cessation protocol was also used in which alcohol‐ and pair‐fed groups were fed liquid diets for 8 weeks and then given pellet chow and water for 6 weeks, with pair feeding maintained during the cessation period.Results: Only minor effects developed in the rats in the 8‐week group, but after 14 weeks, the cancellous bone of the proximal tibia was severely osteopenic in the alcohol‐fed animals. The bone volume and trabecular number were both significantly lower in the alcohol‐fed animals than in the pair‐fed and pellet‐fed control animals and also lower than in the alcohol‐fed animals in the 8‐week group. Mechanical properties of the cancellous bone in the distal femur also were significantly diminished in the 14‐week alcohol‐fed group. Composition and mechanical properties of the cortical bone in the femur diaphysis were largely unaffected, but the yield stress was significantly lower in the 14‐week alcohol‐fed group than in the 8‐week alcohol‐fed group. No significant effects were found in the cessation groups with regard to almost all parameters measured.Conclusions: Our study results demonstrate that chronic adult‐onset alcohol consumption leads to significantly diminished cancellous bone properties and that these effects depend on the duration of alcohol use.