bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Alcohol and Nicotine on Developing Olfactory Bulb: Loss of Mitral Cells and Alterations in Neurotransmitter Levels
Authors: Chen Wei‐Jung A. , Parnell Scott E. , West James R.
Published: May 2006

Previous research from our laboratory has shown that [ethanol (EtOH)] exposure during the brain growth spurt is detrimental to olfactory bulb development. This study extends those findings by examining the effects of EtOH, nicotine (NIC), and the combination of these drugs (EtOH/NIC) on olfactory bulb mitral cell numbers, as well as on various major neurotransmitter levels in neonatal rats. An artificial rearing paradigm was used in the present studies. These artificially reared pups were given 4 g/kg/day of EtOH and/or 6 mg/kg/ day of NIC on postnatal day (PD) 4 to PD 9, except in the case of the acute neurochemistry study, in which the pups received treatment on PD 9 only. An artificially reared gastrostomy control group (GC) and a suckle control group were included. The mean total numbers of mitral cells in the EtOH and NIC groups were significantly reduced from that of the GC, as well as the volume of the left main olfactory bulb. There was no difference among any of the groups in mitral cell density. As for neurochemistry data, there was no difference in neurotransmitter levels among any of the groups in the repeat exposure regimen. There were, however, changes after the acute exposure (exposure on PD 9 only). Both serotonin and GABA levels were significantly increased only after NIC exposure. However, norepinephrine levels were significantly decreased after acute exposure in all three drug treatment groups, compared with that of the control group. Except for the GC control group, dopamine levels were not detected consistently after acute exposure to EtOH, NIC, or EtOH/NIC. Collectively, these findings demonstrate that exposure to EtOH or NIC individually during the brain growth spurt results in developmental deficits in the olfactory bulb, suggesting that both EtOH and NIC are neuroteratogens. Furthermore, this study demonstrated the capability of NIC to antagonize (protect) EtOH‐induced mitral cell loss in the developing olfactory bulb.
Characterization of Adducts of Ethanol Metabolites with Cytochromec
Authors: Anni Helen , Israel Yedy
Published: May 2006

Cytochrome c (cyt c) is found in the mitochondria of all mammalian cells where hydrogen peroxide is produced as a byproduct of the electron transport chain. In the presence of peroxide cyt c generates a ferryl heme and radicals at Tyr residues (Barr et al., 1996). These radicals can be transferred to Trp residues within the protein or to Tyr‐ and Trp‐containing peptides (Deterding et al., 1998). We report that addition of ethanol to this system of cyt c plus peroxide results in replacement of the Tyr/Trp radicals by 1‐hydroxyethyl radicals (HER), and covalent binding of up to 10 mol of ethanol per mol of cyt c. In the absence of exogenously added peroxide, ethanol incorporation to cyt c is attained also with a reconstituted system of the ethanol‐inducible cytochrome PeE1 isozyme. Comparative studies with myoglobin and apomyoglobin suggest that the heme is necessary for ethanol adduction of the protein to occur. Structural analysis by mass spectrometry of the tryptic digestion fractions of adducted cyt c is consistent with several peptides bearing one‐to‐three acetaldehyde moieties on Lys residues, and three distinct Tyr/Trp‐containing peptides: P[28‐53], P[56‐73], P[73‐91] carrying one‐to‐two HER. The x‐ray crystallographic structure of cyt c shows that the Tyr/Trp residues in the adducted peptides are in close proximity to the heme. In conclusion, our data show that ethanol metabolites alkylate cyt c under oxidative stress and point to HER‐Tyr/Trp adducts as plausible markers of alcoholism.
Alcohol Rapidly Lowers Plasma Testosterone Levels in the Rat: Evidence that a Neural Brain‐Gonadal Pathway May Be Important for Decreased Testicular Responsiveness to Gonadotropin
Authors: Rivier Catherine
Published: May 2006

Alcohol is reported to suppress testosterone (T) secretion in the adult male rat. Decreases in the circulating levels of luteinizing hormone (LH) and/or the activity of testicular steroidogenic enzymes have been proposed as putative mechanisms underlying this inhibitory effect. We have recently provided functional evidence for a neural pathway between the brain and the male gonads that plays an important role in the ability of brain proinflammatory cytokines to blunt testicular responsiveness to human chorionic gonadotropin (hCG). The present work was designed to test the hypothesis that a similar pathway might be implicated in the inhibitory influence of alcohol on T secretion. Alcohol, administered intraperitoneally or intragastrically, significantly prevented the T response to the gonadotropin. This effect was significant within 15 min of drug treatment. In the intragastric model (the only one used for this type of experiment), the effect of alcohol was not altered by prior blockade of LH release, which suggests that it is independent of changes in the activity of the pituitary gonadotrophs. The lowest effective dose of alcohol, delivered intraperitoneally, was 2.0 g/kg. The intracerebroventricular injection of the α and β‐adrenergic receptor antagonists phentolamine and propranolol significantly reversed the inhibitory influence of alcohol when it was administered 15 min, but not 60 min, before hCG. Collectively, our results indicate that (1) alcohol induces a rapid and profound decrease in plasma T levels that is secondary to decreased testicular responsiveness to hCG; and (2) at least part of this acute inhibitory action of alcohol may depend on the activation of a neural, adrenergic‐dependent pathway between the brain and the testes.
Ethanol‐Induced Expression of c‐Fos Differentiates the FAST and SLOW Selected Lines of Mice
Authors: Demarest Kristin , Hitzemann Barbara , Phillips Tamara , Hitzemann And Robert
Published: May 2006

The effect of ethanol on the number of Fos‐like immunoreactive (Fos‐li) neurons was previously studied in the C57BL/6J (B6) and DBA/2J (D2) inbred mouse strains (Hitzemann and Hitzemann, 1997). Data obtained suggested that the locomotor activation response to ethanol found in the D2 but not the B6 strain was associated with an increase in the number of Fos‐li neurons (a putative measure of synaptic activity) in the central nucleus of the amygdala (CeA), but not in other brain regions, including the basal ganglia. Supporting results were obtained in B6D2 F2 intercross animals (Demarest et al., 1998) those animals exhibiting a marked locomotor activation response to ethanol also showed a significant increase in the number of Fos‐li neurons in the CeA. The current study extends this line of investigation to the FAST and SLOW selected lines of mice (Shen et al., 1995). Twenty‐eight SLOW and FAST mice (taken evenly from both replicate lines) were randomly assigned to receive either saline or ethanol (1.5 g/kg). One hour later, the animals were sacrificed, and the number of Fos‐li neurons were determined using standard immunocytochemical techniques. Both the FAST and SLOW lines showed a marked increase (>3000%) in the number of Fos‐li neurons in the lateral aspect of the CeA; however, in the capsular division, only the FAST line showed an increase (>500%). In several brain regions, the basal (saline) response was markedly higher in the SLOW line; these regions included the subthalamic nucleus, the entopeduncular nucleus, the substantia nigra compacta, and the ventral tegmental area. Furthermore, it was found that ethanol decreased the number of Fos‐li neurons in the ventral tegmental area of the SLOW but not FAST mice. These data suggest a substantial involvement of the basal ganglia in the segregation of the FAST and SLOW lines.
Polyenylphosphatidylcholine Opposes the Increase of Cytochrome P‐4502E1 by Ethanol and Corrects Its Iron‐Induced Decrease
Authors: Aleynik Maria K. , Leo Maria Anna , Aleynik Semyon I. , Lieber Charles S.
Published: May 2006

Dietary iron overload damages membrane phospholipids and decreases microsomal cytochromes P‐450. We wondered whether this might also pertain to cytochrome P‐4502El (2E1) and whether polyenyiphosphatidylcholine (PPC), a 94‐96% pure mixture of linoleate‐rich polyunsaturated phosphatidylcholines that protects against alcohol‐induced liver injury, also affects 2E1, either in the presence or absence of iron. Accordingly, rats were fed for 8 weeks our standard liquid diet containing ethanol (36% of energy) or isocaloric carbohydrates, with either PPC (3 g/1000 Cal) or equivalent amounts of linoleate (as safflower oil). 2E1 was assessed by Westem blots and by two of its characteristic enzyme activities: the microsomal ethanol oxidizing system (MEOS), evaluated by the conversion of ethanol to acetaldehyde (determined by head space GC), and p‐nitrophenolhydroxylase (PNP) activity, measured by HPLC with W detection of 4nitrocatechol. With ethanol (36% of energy) replacing carbohydrates, 2E1 content increased 10‐fold, with a corresponding increase in PNP and MEOS activities, but when carbonyl iron (5 g/1000 Cal) was added, the induction was significantly reduced. This iron‐induced decrease was corrected by PPC. PPC is rich in linoleate, but when the latter was given as triglycerides (safflower oil), there was no effect, whereas hepatic nonheme iron content was the same in both these groups. It also was found that in the absence of iron, the ethanol‐mediated induction of 2E1 and its corresponding enzyme activities were significantly less with PPC (p < 0.001) than with safflower oil. In addition, in alcohol‐fed animals, PPC decreased the oxidative stress (as determined by F2‐isoprostanes), which reflects yet another hepatoprotective effect of PPC.
Localized Proton Magnetic Resonance Spectroscopy of the Cerebellum in Detoxifying Alcoholics
Authors: Seitz Dietmar , Widmann Uwe , Seeger Uwe , Nägele Thomas , Klose Uwe , Mann Karl , Grodd Wolfgang
Published: May 2006

An increased daily alcohol consumption results in neurological symptoms and morphological central nervous system changes, e.g. shrinkage of the frontal lobes and the cerebellar vermis. Brain shrink‐age can be due to neuronal loss, gliosis, or alterations of (cell) membrane constitutes/myelin. Neuronal, glial, and metabolic change can be measured in vivo with proton magnetic resonance spectroscopy. A total of 11 alcoholics and 10 age‐matched volunteers were examined by magnetic resonance imaging and localized magnetic resonance spectroscopy at an echo time of 135 and 5 msec. Pea integral values were calculated for N‐acetylaspartate (NAA), choline (Cho), myo‐inositol (ml), glutamate/glutamine (Glx), and normalize to phosphocreatine/creatine (Cr). Patients had a significant shrink‐age of the cerebellar vermis. NAA/Cr and Cho/Cr ratios were r duced in both sequences, but the NAA/Cr reduction was only significant in long echo time, although the Cho/Cr reduction WAS significant in short echo time. The ml/Cr and Glx/Cr ratios did not show any significant difference between volunteers and patients. The decrease of NAA/Cr in alcohol dependent patients is consistent with neuronal loss. The Cho/Cr decrease and an unchanged ml/C may reflect cell membrane modification or myelin alterations alcohol‐dependent patients. These changes lead to brain shrinkage although hydration effects and gliosis are less likely.
Drinking, Menstrual Cycle, and Female Sexuality: A Diary Study
Authors: Lindman Ralf E. , Koskelainen Betina M. , Peter Eriksson C. J.
Published: May 2006

Sexual feelings in connection with alcohol intake were investigated in 97 Finnish women aged 18 to 35 years. The subjects were drawn from a population sample and included both oral contraceptive users and nonusers. Structured diaries were used to monitor sexual feelings and alcohol intake during one menstrual cycle for each subject. Alcohol drinking was found to be significantly associated with elevated sexual interest and arousal in the post‐ and intermenstrual phases. We suggest that these phase‐dependent effects could be due to known alcohol‐mediated testosterone elevations.
Alcohol Blocks TNFα but Not Other Cytokine‐Mediated Neuroprotection to NMDA
Authors: Gahring Lorise C. , Carlson Noel G. , Wieggel Whitney A. , Howard Jill , Rogers Scott W.
Published: May 2006

Background: We have investigated the effects of ethanol/alcohol (ETOH) on the pro‐inflammatory CNS cytokine network that mediates neuroprotection to an excitotoxic challenge with the glutamate receptor agonist N‐methyl‐D‐aspartic acid (NMDA).

Methods: Cultured murine cortical neurons were incubated with either TNFα, IL‐1α, IL‐1β, or IL‐6 in the presence or absence of 20 mM ETOH, maintained in an alcohol equilibrated humidified chamber, and the effects of these cytokines on neuronal survival after a chronic 20 hr exposure to NMDA was quantified.

Results: Neuroprotection induced by TNFα, but not IL‐1α, IL‐1β, or IL‐6, was inhibited by a concentration of alcohol (20 mM) that alone did not neuroprotect. Alcohol also affected the paracrine/autocrine induction of cytokine transcripts in neuronal cell cultures, which included enhancing the ability of TNFα to stimulate IL‐6 transcripts. This result supports distinct cytokine‐modulated neuroprotective pathways of which only TNFα is sensitive to low alcohol concentrations. We have shown previously that nicotine, acting through an α‐bungarotoxin sensitive receptor, is also neuroprotective, but it too specifically abolishes TNFα‐mediated neuroprotection. However, alcohol did not affect nicotine‐induced neuroprotection.

Conclusions: We suggest that the effects of low concentrations of alcohol on neuronal cytokine networks proceed through antagonism of neuroprotective pathway(s) unique to TNFα.
Breakpoint Determination and Ethanol Self‐Administration Using an Across‐Session Progressive Ratio Procedure in the Rat
Authors: Czachowski Cristine L. , Samson Herman H.
Published: May 2006

Background: Progressive ratio schedules are used to determine the “breakpoint” or limit to the amount of “work” that a subject is willing to perform to obtain a reinforcer. Reinforcing efficacy is inferred from the breakpoint values, which are typically measured in a single session by increasing the number of responses required for successive reinforcer presentations. This procedure is not feasible, however, when assessing the reinforcing efficacy of a substance that can change as a function of its physiological actions during self‐administration, as in the case of ethanol.

Methods: The present study made use of a procedure that increased the response requirement across single daily sessions rather than within a session. Completion of the response requirement in each daily session resulted in the presentation of a drinking tube that allowed for self‐administration of ethanol for a 20‐min period. This procedure made possible the assessment of ethanol‐directed appetitive (number of lever presses) and consummatory (number of licks and intake volume) behaviors. Reliable responding for 10% ethanol was initiated using sucrose‐substitution on a fiwed ratio (FR) 4 schedule in male Long Evans rats. Then four successive breakpoint determinations were made which were separated by a return to the FR4 schedule to re‐establish baseline responding.

Results: The results indicated that there was an increase in breakpoint values from the first to the second determination, which was then stable over the following three determinations. Individual rats reached breakpoints as high as 240 lever presses to receive access to 10% ethanol and maintained ethanol intake over sessions in the 1.0 g/kg range. Ethanol intake (g/kg), however, was stable across all four determinations (mean 0.86 ± 0.06 to 1.01 ± 0.10). Moreover, ethanol intake was not related to the preceding appetitive responding, as no differences between intake on the session before a breakpoint (high FR) and the following baseline period (FR4) were observed.

Conclusions: This model provides an assessment of the distinct mechanisms that mediate ethanol‐seeking versus ethanol consumption in subjects that drink measurable amounts of ethanol, with the appetitive behaviors not altered by the pharmacological effects of ethanol.
Alcohol and Fatal Injury: The Use of Routinely Collected Fatality Data in Community Prevention Evaluation
Authors: Treno Andrew J.
Published: May 2006

Background: This study analyzed patterns of alcohol involvement among coroners’cases (which typically include blood alcohol content information) to develop a method for weighting death‐certificate cases (which typically do not include blood alcohol content information) for likelihood of alcohol involvement for purposes of alcohol‐involved injury intervention evaluation.

Methods: The coroners’ data analyzed here were collected from four California communities and correspond to all injury coroner cases between 1987 and 1996 in those communities. The death‐certificate data were provided by the State of California, Department of Health Services, and correspond to all injury deaths occurring in that state between 1980 and 1996. Each injury fatality in the death‐certificate data was assigned a probability of alcohol involvement based upon the coroners’ data. These were then summed to provide an estimate of, or “surrogate measure” for, the total number of alcohol‐involved fatal injuries per 10,000 California residents aggregated across the state of California by month for the period from January 1980 to 1996. As a test of this estimate, we examined its responsiveness to an intervention designed to reduce alcohol‐involved injuries which was implemented in California in 1990 using a time series analysis technique (ARIMA) that corrects for serial autocorrelation typically found in time ordered data.

Results: This analysis found an effect during the postintervention period (p= 0.046). An alternative model testing for intervention effects on all injury fatalities did not find an effect.

Conclusions: This surrogate measure seems responsive to intervention effects and may provide a useful tool for interventions designed to reduce alcohol‐involved injuries.
Reduced Gene Expression for Dopamine Biosynthesis and Transport in Midbrain Neurons of Adult Male Rats Exposed Prenatally to Ethanol
Authors: Szot Patricia , White Sylvia S. , Veith Richard C. , Rasmussen Dennis D.
Published: May 2006

Background: Prenatal ethanol exposure affects brain dopaminergic neuronal systems, and many of these alterations are permanent.

Methods: The primary objective of this study was to determine the effects of prenatal ethanol exposure on adult mRNA expression for two key regulatory proteins in the mesolimbic and nigrostriatal dopaminergic cell groups which mediate behavioral responses to alcohol and other drugs of abuse: tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the substantia nigra pars compacta (SNpc) and ventral tegmental area (VTA). To also address the effects on noradrenergic regulation, we quantitated mRNA expression for TH and norepinephrine transporter (NET) in the noradrenergic loci of the locus cocruleus (LC).

Results: Daily dietary ethanol consumption by female Sprague‐Dawley rats for 3 weeks before, and continuing throughout, pregnancy decreased both DAT (∼ 68%, p < 0.002) and TH (∼ 45%, p < 0.002) mRNA expression in the VTA of adult male offspring. This prenatal exposure also suppressed DAT mRNA expression in the SNpc (∼ 81%; p < 0.03), although TH mRNA expression in this region was not significantly altered. Prenatal ethanol exposure did not alter significantly either TH or NET mRNA expression in the LC of adult male offspring, which suggests that this brain catecholaminergic response may be limited to DA neurons.

Conclusion: These results demonstrated that prenatal maternal ethanol consumption suppresses mRNA expression for important regulatory proteins in the mesolimbic and nigrostriatal dopaminergic systems of adult male rat offspring. These persistent prenatal ethanol‐induced changes in mRNA expression may thus contribute to the persistent effects of fetal ethanol exposure on the diverse behavioral and/or metabolic responses meditated by the mesolimbic and nigrostriatal dopaminergic systems in the adult.
A Pilot Study of a New Chicken Model of Alcohol‐Induced Cardiomyopathy
Authors: Morris Nancy , Kim Catherine S. , Doye Angelia A. , Hajjar Roger J. , Laste Nancy , Gwathmey Judith K.
Published: May 2006

Background: Excessive alcohol consumption is recognized as a common cause of left ventricular (LV) dysfunction. It is currently thought that 36% of all cases of dilated cardiomyopathy are due to excessive alcohol intake. Suitable animal models are needed to study the pathogenic mechanisms of ethanol‐induced LV dysfunction. We have therefore created a new model of ethanol‐induced LV dysfunction in the chicken.

Methods: For 12 weeks, adult chickens were given, twice a day, by gavage, 73% of their total calculated daily water intake containing a 20% ethanol concentration. Twenty percent ethanol also was placed in the water and provided ad libitum. Control chickens received the same volume of water by gavage twice a day without ethanol. Water without ethanol was given ad libitum to control birds.

Results: Our study shows that after a relatively short duration of ethanol ingestion, chickens developed LV dilatation and LV dysfunction. The serum concentrations of ethanol attained in this new model were similar to those reported in humans. Furthermore, unlike other currently available animal models of ethanol‐induced cardiac disease, this model demonstrates myocyte hypertrophy, interstitial fibrosis, and myocytolysis, similar to observations in human ethanol‐induced cardiac dysfunction.

Conclusions: We conclude that this new avian model should provide a useful tool for investigating the mechanism(s) and pathophysiology of ethanol‐induced dilated cardiomyopathy and heart failure.
Ethanol Decreases Glial Derived Neurotrophic Factor (GDNF) Protein Release but Not mRNA Expression and Increases GDNF‐Stimulated Shc Phosphorylation in the Developing Cerebellum
Authors: McAlhany Robert E. , Miranda Rajesh C. , Finnell Richard H. , West James R.
Published: May 2006

Background: Ethanol exposure during development leads to substantial neuronal loss in multiple regions of the brain. Although differentiating Purkinje cells of the cerebellum are particularly vulnerable to ethanol exposure, the mechanisms underlying ethanol‐induced Purkinje cell loss have not been well defined. Our previous research indicated that exogenous Glial‐Derived Neurotrophic Factor (GDNF) attenuated ethanol‐induced Purkinje cell loss in cerebellar explant cultures, which suggests that ethanol, in turn, may decrease endogenous trophic factor‐mediated survival mechanisms.

Methods: The present experiments used an explant culture model of the developing rat cerebellum to test the hypothesis that ethanol decreases endogenous trophic support by limiting the availability of trophic factors, such as GDNF, or by altering the activation of key adapter proteins such as Shc (Src homology domain carboxy‐terminal) that couple GDNF binding to multiple intracellular signaling pathways. GDNF mRNA and protein levels were measured by reverse northern blot analysis and sandwich enzyme‐linked immunosorbent assay respectively, whereas Shc phosphorylation was measured by immunoprecipitation/western immunoblot analysis.

Results: The developing cerebellum expresses both GDNF mRNA and protein in vitro. Ethanol exposure (68, 103, or 137 mM) had no effect on cerebellar levels of GDNF mRNA. However, ethanol (68 and 137 mM) decreased levels of GDNF protein released into culture medium. In addition, ethanol itself had no effect on Shc phosphorylation. However, in the presence of the highest dose of ethanol (137 mM) GDNF did stimulate Shc phosphorylation.

Conclusions: Together, these results suggest that ethanol decreases GDNF‐mediated trophic support of Purkinje cells in the developing cerebellum. However, GDNF in turn activates intracellular signaling pathways throughout the developing cerebellum as part of its Purkinje cell‐selective neuroprotective response to ethanol exposure.
Effect of Repeated Ethanol Withdrawal on Glutamate Microdialysate in the Hippocampus
Authors: Dahchour Abdelkader , Witte Philippe De
Published: May 2006

Background: Our previous studies, which identified that ethanol withdrawal is associated with increases in glutamate microdialysate in the nucleus accumbens and reaches a maximum at 12 hr, have now been extended in order to assess whether repeated cycles of chronic ethanol intoxication followed by 12 hr withdrawal periods on three occasions alters glutamate release in the hippocampus of male rats.

Methods: In this study, the microdialysis technique has been used with the HPLC and electrochemical detection.

Results: During the first cycle of ethanol withdrawal, glutamate content increased significantly 8 hr after withdrawal (198.4%± 89.14%) by comparison with control rats. During the second period of ethanol withdrawal, 1 week after the initial withdrawal episode, glutamate microdialysate content increased significantly 10 hr after withdrawal, but to a much lower degree than in the first episode (179.08 ± 25.68%), by comparison with control rats. During the third cycle of ethanol withdrawal, the concentration of glutamate in the hippocampus microdialysate did not significantly change at either of these time points except at 12 hr when glutamate was significantly decreased by comparison with control rats (52.09 ± 14.38%). Apart from arginine, which was significantly decreased both at the cessation of alcoholization and during the 12 hr of the three withdrawal episodes, none of the other neurotransmitters assayed, aspartate, taurine, alanine, or GABA, showed any significant alteration.

Conclusion

: These results clearly indicate that elevated glutamate release during the first withdrawal episode is not paralleled in subsequent withdrawal episodes.
Alcohol Inhibition of Cell Adhesion in BMP‐Treated NG108‐15 Cells
Authors: Wilkemeyer Michael F. , Pajerski Mandy , Charness Michael E.
Published: May 2006

Background

The L1 cell adhesion molecule is expressed as alternatively spliced neuronal and nonneu‐ronal isoforms. We have reported that in transfected fibroblasts, ethanol variably inhibits cell‐cell adhesion mediated by the nonneuronal isoform of human L1. In contrast, ethanol consistently inhibits morphoge‐netic changes and cell‐cell adhesion in NG108‐15 cells treated with OP‐1 (BMP‐7), a powerful inducer of L1 and N‐CAM gene expression.

Methods

All studies were performed by using NG108‐15 cells cultured in serum‐free medium. Cell morphology was assessed by a quantitative assay of cell clustering. Cell adhesion was measured by a short‐term re‐aggregation assay, and isoforms of L1 were characterized by RT‐PCR and sequencing.

Results

We show that ethanol inhibits the morphogenetic effects of BMP‐2, BMP‐4, BMP‐5, and BMP‐6, each of which increases the expression of L1 and N‐CAM. Pretreatment of NG108‐15 cells with 25‐100 mM ethanol did not induce tolerance to ethanol's inhibition of OP‐1 morphogenesis or cell‐cell adhesion. Ethanol or anti‐L1 Fab fragments partially inhibited cell‐cell adhesion in OP‐1‐treated NG108‐15 cells. The combination of ethanol and Fab fragments did not inhibit cell‐cell adhesion more than Fab fragments alone. As in L1‐transfected fibroblasts, a series of n‐alcohols displayed a cutoff between butanol and pentanol for inhibition of cell‐cell adhesion in OP‐1‐treated NG108‐15 cells. RT‐PCR and direct sequencing revealed that the neuronal isoform was the sole or predominant L1 isoform in OP‐1‐treated NG108‐15 cells.

Conclusions

These data suggest that ethanol inhibits cell‐cell adhesion in OP‐1‐treated NG108‐15 cells by interacting directly or indirectly with the neuronal isoform of L1.
MK‐801‐Induced Locomotor Activity in Long‐Sleep × Short‐Sleep Recombinant Inbred Mouse Strains: Correlational Analysis With Low‐Dose Ethanol and Provisional Quantitative Trait Loci
Authors: Zahniser Nancy R. , Negri Cori A. , Hanania Taleen , Gehle Vaughn M.
Published: May 2006

Background

Law doses of theN‐methyl‐D‐aspartate receptor (NMDAR) antagonist MK‐801 (dizocilpine) or ethanol increase locomotor activity to a lesser extent in long‐sleep (LS), than in short‐sleep (SS), mice. LS mice also have fewer brain (3H]MK‐801 binding sites than SS mice. In this study, LSXSS recombinant inbred (RI) mice were used to investigate whether different NMDAR densities contribute to differential MK‐801 activation and whether common genes are involved in initial sensitivity to MK‐801‐ and ethanol‐induced activation.

Methods

Locomotor activity was measured for 90 min after saline or MK‐801 injection. Quantitative autoradiographic analysis of [3H]MK‐801 binding was used to measure densities of NMDARs in seven brain regions. The ethanol (1–2 gkg) activation scores from Erwin and colleagues (1997) were used for correlational analysis, as was their method for quantitative trait loci (QTL) analysis.

Results

Both saline and MK‐801 (0.3 mgkg, given intraperitoneally) induced a continuum of locomotor responses across the LSXSS RI strains. There was a 4‐fold range of MK‐801 difference scores (MK‐801 score‐saline baseline), with the RI 9 and RI 4 strains representing low and high responders, respectively. Dose‐response experiments with these two strains confirmed that 0.3 mgkg MK‐801 produced significant activation, similar to previous results with LS and SS mice. However, unlike previous LS/SS results, lower densities of NMDARs were not observed in the RI 9 than in the RI 4 mouse brains. No significant genetic correlations were observed between MK‐801‐induced and ethanol‐induced responses in the LSXSS RI mice. Two provisional MK‐801 activation QTLs were identified (p< 0.01) on chromosomes 11 and 19, neither in common with those mapped for ethanol activation.

Conclusions

Different densities of brain NMDARs are unlikely to account for the differential activation of LSXSS RI mice by MK‐801. Additionally, in the RI mice either separate sets of genes regulate low dose MK‐801‐ and ethanol‐induced locomotor responses or the overlapping subset of genes controlling these two behaviors is small (≤ 10%).
Genetic Association Between Chronic Ethanol Withdrawal Severity and Acoustic Startle Parameters in WSP and WSR Mice
Authors: Ponomarev Igor , Crabbe John C.
Published: May 2006

Background

: The present study examined the genetic association between chronic ethanol withdrawal severity and acoustic startle response (ASR) in replicated lines of mice selected for high (Withdrawal Seizure‐Prone; WSP) and low (Withdrawal Seizure‐Rent; WSR) susccptihility to handling‐induced convulsions after withdrawal from chronic exposure to ethanol. Any differences on a nonselected (correlated) trait between the opposite‐selected lines is strong evidence for pleiotropic effects of the genes fixed by selection.

Methods

: Naïve WSP and WSR micc of both replicates were placed in startle chambers and exposed to a series of white noise stimuli of different intensities. In Experiment 1, two parameters [the maximal acoustic startle response (Rmax), and the sound intensity ncccssary to produce 50% of the maximal startle response (dB50)] were obtained from a least‐squares nonlinear regression by fitting data for each subject to a sigmoidal function that best described the relationship betwcen sound intensity and mean ASR. Response hahituation of WSP and WSR mice to a repeated acoustic stimulus of high intensity was examined in Experiment 2.

Results

: When ASR amplitude was plotted versus sound intensity, the sigmoid intensity‐response curves of both WSP replicates were shifted to the right rclative to the responses of WSR mice, which suggested decreased startle sensitivity in the WSP animals. Statistical analysis showed that naivc WSP mice were less sensitive (higher dB50) to acoustic stimulation than Scizure‐Resistant animals whereas Rmax was similar for both lines. The selected lines also diffcred in their responses to repeated acoustic stimulation, with WSP mice demonstrating grcater habituation.

Conclusion

: Results of the present study suggest some common genetic mechanisms underlying behavioral responsiveness to acoustic stimulation and severity of ethanol withdrawal.
Executive Functioning in Children With Heavy Prenatal Alcohol Exposure
Authors: Mattson Sarah N. , Goodman Amy M. , Caine Chip , Delis Dean C. , Riley Edward P.
Published: May 2006

Background

: Children with heavy prenatal alcohol exposure have well documented deficits in overall cognitive ability. Recently, attention has turned to the executive function (EF) domain in this population. Until recently, comprehensive measures of EF have not been available within one test battery. This study used a battery of tests to assess four domains of EF in alcohol‐exposed children.

Methods

: The Delis‐Kaplan Executive Function Scale was used to evaluate EF in 18 children with heavy prenatal alcohol exposure, with and without a diagnosis of fetal alcohol syndrome (FAS), and 10 nonexposed controls. Children ranged in age from 8 to 15 years. Measures from four domains of executive functioning were analyzed: planning ability, cognitive flexibility, selective inhibition, and concept formation and reasoning. Tasks consisted of primary EF measures as well as measures of secondary component skills.

Results

: Alcohol‐exposed children were deficient on EF measures compared with nonexposed controls. Furthermore, in most cases, children with and without the FAS diagnosis did not differ from one another. These deficits were not entirely explainable by concomitant deficits on component skills. Specific impairments were identified within the domains of planning and response inhibition, with additional deficits in abstract thinking and flexibility.

Conclusions

: Deficits in executive functioning were observed in alcohol‐exposed children with or without the diagnosis of FAS and in the absence of mental retardation. Performance on these EF tasks provides insight into the cognitive processes driving overall performance and has implications for adaptive and daily functions. These results arc consistent with anecdotal and empirical reports of deficits in behavioral control and with neuroanatomical evidence of volumetric reductions in structures within the frontal‐subcortical system in children with heavy prenatal alcohol exposure.
Altered GABAA‐Benzodiazepine Receptor Number and Pharmacology in the Adult Guinea Pig Cerebral Cortex After Chronic Prenatal Ethanol Exposure
Authors: Bailey Craig D.C. , Brien James F. , Reynolds James N.
Published: May 2006

Background

: The effects of chronic prenatal ethanol exposure on GABAA‐benzodiazepine receptor number and binding pharmacology were examined in the cerebral cortex of the postnatal guinea pig.

Methods

: [3H]Flunitrazepam binding to GABAA‐benzodiazepine receptors was measured in a cerebral cortical cell membrane preparation obtained at postnatal days 11 (preweaning), 21 (postweaning), and 61 (adulthood). Zolpidem, a GABAA‐benzodiazepine type 1 receptor‐selective ligand, was used in a [3H]flunitrazepam compctition study. 3α‐Hydroxy‐5α‐pregnan‐20‐one (allopregnanolone) potentiation of [3H]muscimol and [3H]flunitrazepam binding, and GABA potentiation of [3H]flunitrazepam binding were measured in these same animals.

Results

: At postnatal day 61, but not at the younger ages studied, the following was observed: (1) [3H]Flunitrazepam binding exhibited an increased receptor number (Bmax) and decreased affinity (increased KD) in the ethanol‐treated offspring compared with isocaloric‐sucrose (with pair‐feeding) and water‐treated controls; and (2) the relative proportion of GABAA‐benzodiazepine receptors that had high‐affinity binding sites for zolpidem was decreased in the ethanol‐treated offspring by 31% and 38% compared with the isocaloric‐sucrose/pair‐fed and water‐treated controls, respectively. Chronic prenatal ethanol exposure did not alter the efficiency of coupling between GABA, benzodiazepine, and neurosteroid binding sites at any postnatal ages studied.

Conclusions

: These results suggest that, in the cerebral cortex of the adult guinea pig, chronic prenatal exposure to ethanol results in increased GABAA‐benzodiazepine receptor number, decreased affinity for flunitrazepam, and decreased relative proportion of the GABAA‐benzodiazepine type 1 receptor.
Alcohol Metabolism and Cardiovascular Response in an Alcoholic Patient Homozygous for theALDH2*2Variant Gene Allele
Authors: Chen Yi‐Chyan , Lu Ru‐Band , Peng Giia‐Sheun , Wang Ming‐Fang , Wang Hsu‐Kun , Ko Huei‐Chen , Chang Yue‐Cune , Lu Jang‐Jih , Li Ting‐Kai , Yin Shih‐Jiun
Published: May 2006

Background: Alcohol metabolism is one of the biological determinants that can influence drinking behavior. Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) are the principal enzymes involved in ethanol metabolism. Allelic variation of theADHandALDHgenes can significantly affect vulnerability for the development of alcoholism. Homozygosity of the variantALDH2*2allele previously was believed to fully protect East Asian populations against the development of alcoholism.

Methods: Eighty Han Chinese alcoholics who met DSM‐III‐R criteria for alcohol dependence and 144 nonalcohol‐dependent subjects were recruited and their data combined with data from 340 alcohol‐dependent and 545 nonalcohol‐dependent subjects described in an earlier report (Chen et al., 1999) to assess risk for alcoholism by logistic regression analysis. Genotypes ofADH2, ADH3, andALDH2were determined by polymerase chain reaction and restriction fragment length polymorphism. TheALDH2genotype was confirmed by direct nucleotide sequencing. Blood ethanol concentration was determined by headspace gas chromatography and acetaldehyde concentration by high‐performance liquid chromatography with fluorescence detection of the derivatized product. Cardiovascular hemodynamic parameters were measured by two‐dimensional Doppler echocardiography and sphygmomanometry. Extracranial arterial blood flow was measured by Doppler ultrasonography.

Results: An alcohol‐dependent patient was identified to beALDH2*2/*2, ADH2*2/*2, andADH3*1/*2.Following challenge with a moderate oral dose of ethanol (0.5 g/kg of body weight), the patient exhibited peak concentrations for ethanol (55.7 mg/dl) and acetaldehyde (125 μM). During 130 min postingestion, the patient generally displayed similar or even less intense cardiovascular hemodynamic alterations when compared to a previously published study of nonalcoholic individuals withALDH2*2/*2who had received a lower dose of ethanol (0.2 g/kg). Logistic regression analysis of the combinatorial genotypes ofADH2andALDH2in 420 alcohol‐dependent and 689 nonalcohol‐dependent subjects indicated that risk for alcoholism was 100‐fold lower for theADH2*2/*2—ALDH2*2/*2individuals than theADH2*1/*1—ALDH2*1/*1individuals.

Conclusions: The gene status ofALDH2*2/*2alone can tremendously but not completely (as thought previously) protect against development of alcohol dependence. Individuals carrying the combinatorial genotype ofADH2*2/*2—ALDH2*2/*2are at the least risk for the disease in East Asians. Physiological tolerance or innate insensitivity to the accumulation of blood acetaldehyde following alcohol ingestion may be crucial for the development of alcoholism in individuals homozygous forALDH2*2.
CYP2E1 Expression in Human Lymphocytes From Various Ethnic Populations
Authors: Raucy Judy L. , Schultz Eric D. , Kearins Margaret C. , Arora Sanjeev , Johnston David E. , Omdahl John L. , Eckmann Lars , Carpenter Susan P.
Published: May 2006

Background: Monitoring CYP2E1 levels in alcoholic individuals holds inherent appeal because such determinations might indicate individuals at increased risk for alcoholic liver disease. We previously demonstrated that lymphocyte CYP2E1 expression reflects in vivo activity of the hepatic enzyme.

Methods: To further validate this approach, the current investigation compared lymphocyte CYP2E1 content and chlorzoxazone pharmacokinetics in 51 alcoholic and nonalcoholic White, Navajo, and Mexican American subjects. After an oral dose of chlorzoxazone, blood samples were collected and lymphocytes isolated.

Results: Alcoholics exhibited a 2‐fold clevation in lymphocyte CYP2E1 messenger ribonucleic acid (mRNA) and protein compared to nonalcoholics. Chlorzoxazone clearance rates were 1.9‐fold higher and area under the concentration curve (AUC) values 1.8‐fold lower in alcoholic individuals compared to nonalcoholics. Furthermore, chlorzoxazone clearance rates correlated (r= 0.55, p < 0.01, n= 38) with lymphocyte CYP2E1 mRNA content, and transcript levels further correlated (r= 0.52, p < 0.001, n= 38) with CYP2E1 protein content. To compare phenotype with genotype, restriction fragment length polymorphism analyses on deoxyribonuclcic acid samples were performed to identify polymorphisms in the CYP2E1 gene. No subjects were homozygous for rare alleles c2 or C. Nonetheless, 27% of the Navajos and 15% of the Mexican Americans were heterozygous for the c2 allele. Two White subjects appeared heterozygous (c1/c2) when RsaI was used to characterize CYP2E1 genotype but homozygous (c1/c1) at the PstI locus. Fifteen percent of Mexican American subjects, 20% of Navajo subjects, and 6% of White subjects were heterozygous for the C allele. Neither CD nor c1/c2 genotypes were associated with alcoholism.

Conclusions: Human lymphocyte CYP2E1 mRNA levels may be useful predictors of alcohol‐mediated alterations in hepatic CYP2E1 activity. Moreover, ethnicity does not appear to play a major role in the levels of expression of lymphocyte CYP2E1.
Role of Nitric Oxide‐Dependent Pathways in Ethanol‐Induced Anxiolytic Effects in Rats
Authors: Ferreira Vania M.M. , Valenzuela C. Fernando , Morato Gina S.
Published: May 2006

Background: Nitric oxide (NO) is a neuromodulator and an intercellular and retrograde messenger that mediates several functions in the central nervous system. The effects of ethanol (EtOH) on neuronal NO‐dependent pathways have been the focus of recent research. Most studies have concentrated on the actions of chronic EtOH exposure. In this study, we examined the role of NO‐dependent pathways in the acute actions of EtOH.

Methods: We used the elevated plus‐maze test to study the role of NO‐dependent pathways in the behavior of rats treated with acutc EtOH. We tested the effects of 7‐nitroindazole, a reversible competitive inhibitor of nitric oxide synthasc. We also studicd the effects of the cyclic guanylate monophosphate (cGMP) analog, 8‐Bromoguanosine cyclic 3′, 5′‐monophosphate sodium salt, and the NO donors S‐nitroso‐N‐acetylpenicillamine or sodium nitroprusside.

Results: When injected by either intraperitoneal (6 mg/kg) or intrahippocampal (20 nmol) routes, 7‐nitroindazole increased the percentage of open arm entries and time spent in open arms for rats injected with EtOH (1.0 g/kg, ip). This dose of EtOH did not produce an anxiolytic effect when administered alone. Additional experiments were performed with a dose of 1.2 g/kg of EtOH (ip), which produced an anxiolytic effcct. Intrahippocampal administration of the cGMP analog, 8‐Bromoguanosine cyclic 3′3′‐monophosphate sodium salt (40 nmol), or the NO donors S‐nitroso‐N‐acetylpenicillamine (20 or 40 nmol) or sodium nitroprusside (20 or 40 nmol) blocked the anxiolytic effect of this dose of EtOH.

Conclusions: These results indicate that inhibition of NO‐dependent pathways enhances, whereas stimulation of these pathways decreases, the efficacy of EtOH to produce anxiolytic effects in rats. We postulate that NO‐dependent increases in guanylate cyclase activity and cGMP levels oppose the anxiolytic effects produced by acute EtOH administration.
The Role of Somatic Disorders and Physical Injury in the Development and Course of Alcohol Withdrawal Delirium
Authors: Wojnar Marcin , Bizoń Zdzisław , Wasilewski Dariusz
Published: May 2006

In a retrospective study, we evaluated the role of somatic disease and physical injury in the development and course of alcohol withdrawal delirium. Medical records of 1179 patients treated for alcohol withdrawal in Nowowiejski Hospital in Warsaw from 1973 to 1987 were reviewed using a structured questionnaire. Development, symptoms’ severity, and the course of alcohol withdrawal delirium were assessed in possible relation to the somatic state of patients and other variables of alcohol dependence. Development of the first episode of delirium tremens (DT) was associated with the incidence of somatic disease or injury in 19% of cases. Somatic disorders directly preceded the second episode of DT in 73% and the third in 57% of cases. A positive correlation was found between the greater severity and/or longer duration of DT symptoms, and occurrence of pneumonia, coronary heart disease, alcohol liver disease, and anemia, as well as daily amount of alcohol consumed during the last drinking bout. There was no relationship of severity of DT with the duration of alcohol abuse. Early development and severe course of alcohol withdrawal delirium correlated with the late beginning of excessive drinking (over the age of 40) and concomitant abuse of benzodiazepines or barbiturates. We concluded that somatic disorders or physical injury might trigger delirium during alcohol withdrawal, and have essential influence on the symptoms’ severity and duration of DT. A more severe course of DT is also correlated with the quantity of alcohol consumed and concomitant abuse of sedatives.
The Oklahoma Postmenopausal Women's Health Study: Recruitment and Characteristics of American Indian, Asian, Black, Hispanic, and Caucasian Women
Authors: Gavaler Judith S. , Bonharn‐Leyba Marilyn , Castro Cesar A. , Harman Susan E.
Published: May 2006

Since 1994, the National Institutes of Health has required the inclusion of women and minorities in all of its sponsored clinical research. This study describes a workable recruitment strategy that embraces the National Institutes of Health requirement. We describe the recruitment pattern of the Oklahoma Postmenopausal Women's Study conducted in the general community of Oklahoma City and in surrounding areas that are both urban and rural. For the period 1994 through 1997, 491 postmenopausal women from all racial/ethnic groups in the community have participated in this study. Over 4 years of recruitment, the percentage of minority women in the study population has risen annually from 31% in 1994 to 81% in 1997. The overall percentage of minority women in the study population is currently 63.3%: American Indian, 21.8%; Asian, 3.7%; Black, 14.9%; Hispanic, 9.4%; White/American Indian Blend, 13.6%; and White, 36.7%. The recruitment approach described may be implemented in a variety of research settings. Specific recruitment approaches are described, as well as the distribution of sociodemographic and health behaviors across and within ethnic/racial groups.
Innovations in Adolescent Substance Abuse Intervention
Authors: Wagner Eric F. , Brown Sandra A. , Monti Peter M. , Myers Mark G. , Waldron Holly B.
Published: May 2006

Adolescent alcohol and other drug abuse is an important public health concern, and the past two decades has seen a dramatic increase in the demand for interventions to address substance use problems among teenagers. This demand has led to the development of multiple primary, secondary, and tertiary substance abuse prevention programs, some of which have little theoretical basis and most of which currently operate in the absence of data supporting their effectiveness. Very recently, there has been increased emphasis on the goal of developing and testing theoretically based and empirically supported intervention approaches for adolescent substance abuse. We describe five ongoing research programs devoted to meeting this goal. The background and rationale for each research program are discussed, and preliminary efficacy data concerning the specific interventions are presented.