Alcohol

In Memoriam: Robert D. Myers: Founding Editor, Alcohol: An International Biomedical Journal

Brian A. McMillen — 2011-10-24

It is with sadness that I note the passing of Robert D. Myers, the founding Editor-in-Chief of this journal, on July 16, 2011. Bob was always ebullient, and his charm and wit will be missed. He was born on October 25, 1931 in Philadelphia. He received his BS degree from Ursinus College (Pennsylvania) in 1953 and the MS and PhD degrees from Purdue University (Indiana) in 1954 and 1956. It was at Ursinus College that he met Marjorie Fretz, married her in 1953, and they remained inseparable for 57 years. Rather than a recitation of facts about his life that included more than 500 scholarly publications, including the first reports that opiate antagonists would reduce alcohol consumption in rat and monkey drinking models (), plus numerous awards and honors, I will give the reader some anecdotes that trace Bob’s journey.

An in vitro model for studying the effects of continuous ethanol exposure on N-methyl-d-aspartate receptor function

2011-09-19

Abstract: Long-term ethanol exposure has deleterious effects on both glial and neuronal function. We assessed alterations in both astrocytic and neuronal viability, and alterations in N-methyl-d-aspartate receptor (NMDAR) function, in cocultures of rat cerebellar granule cells (CGCs) and astrocytes after continuous ethanol exposure (CEE). Treatment of cells with 100 mM EtOH once every 24 h for 4 days resulted in a mean ethanol concentration of 57.3 ± 2.1 mM. Comparisons between control and post–ethanol-treated cells were made 4 days after the last ethanol treatment. CEE did not alter glial cell viability, as indicated by the absence of either changes in astrocytic morphology, actin depolymerization, or disruption of astrocytic intracellular mitochondrial distribution at any day postethanol treatment. The CGCs were healthy and viable after CEE, as indicated by phase-contrast microscopy and the trypan-blue exclusion method. Whole-cell patch-clamp experiments indicated that NMDA-induced currents (INMDA) were altered by CEE treatment. Similar to previous results obtained during the withdrawal phase from chronic ethanol exposure, INMDA from CEE-treated cells were significantly larger than INMDA from NMDARs in control CGCs, but returned to control values by the fourth day post-CEE. However, after the last ethanol dosing and during a time when ethanol concentrations remained high, INMDA were significantly smaller than control values. Identical results were observed in CGCs expressing the NR2A or NR2B subunit. In summary, both neurons and astrocytes remained healthy following exposure to CEE with no signs of neurotoxicity at the cellular level, and modulation of NMDAR function is consistent with findings from prior experiments. Thus, we conclude that the CEE paradigm in glial–neuronal cocultures readily lends itself to long-term in vitro studies of ethanol effects that include glial–neuronal interactions and the ability to study ethanol withdrawal-induced neurotoxicity.

Effects of naltrexone and LY255582 on ethanol maintenance, seeking, and relapse responding by alcohol-preferring (P) rats

2011-10-03

Abstract: Research indicates opioid antagonists can reduce alcohol drinking in rodents. However, tests examining the effects of opioid antagonists on ethanol seeking and relapse behavior have been limited. The present study examined the effects of two opioid antagonists on ethanol maintenance, seeking, and relapse responding by alcohol-preferring (P) rats. Adult P rats were self-trained in two-lever operant chambers to self-administer 15% (vol/vol) ethanol on a fixed-ratio 5 (FR5) versus water on a FR1 concurrent schedule of reinforcement in daily 1-h sessions. After 10 weeks, rats underwent extinction training, followed by 2 weeks in their home cages. Rats were then returned to the operant chambers without ethanol or water to measure responses on the ethanol and water levers for four sessions. After a subsequent 2 weeks in the home cage, without access to ethanol, rats were returned to the operant chambers with ethanol and water available. Effects of antagonists on maintenance responding were tested after several weeks of daily 1-h sessions. Naltrexone (NAL; 1–10mg/kg, subcutaneously [s.c.]; n=8/dose), LY255582 (LY; 0.03–1mg/kg, s.c.; n=8/dose), or vehicle were injected 30min before the first session (in the absence of ethanol), following 2 weeks in their home cages, and for four consecutive sessions of ethanol self-administration under maintenance and relapse conditions. Both NAL and LY reduced responses on the ethanol lever without any fluids present, and ethanol self-administration under relapse and on-going drinking conditions, with LY being more potent than NAL. Both NAL and LY were less effective in reducing responding in the absence of ethanol than in reducing ethanol self-administration. Overall, the results indicate that the opioid system is involved in mediating ethanol seeking, and ethanol self-administration under relapse and on-going alcohol drinking, but that different neurocircuits may underlie these behaviors.

Effects of ethanol administration on corticosterone levels in adolescent and adult rats

2011-09-15

Abstract: Adolescent humans and rodents have been shown to consume more alcohol than their adult counterparts. Given that corticosterone (CORT) has been shown to be related to the intake of several drugs of abuse, this study assessed the ontogenetic effects of low–moderate doses of ethanol on CORT increases and recovery. Despite no significant differences in baseline (home cage) CORT levels, CORT responses to ethanol were greater in females than in males and in adult females than in adolescent females; males, however, showed less marked age differences in CORT levels after ethanol consumption. Adolescent blood ethanol concentrations (BECs) were lower than those of adults, although these BEC differences appear insufficient to account for the ontogenetic differences in CORT levels. Collectively, these findings suggest that it is unlikely that age differences in CORT elevations provide a major contribution to the ontogenetic differences in alcohol intake seen between adolescents and adults.

Expression pattern, ethanol-metabolizing activities, and cellular localization of alcohol and aldehyde dehydrogenases in human large bowel: association of the functional polymorphisms of ADH and ALDH genes with hemorrhoids and colorectal cancer

2011-09-23

Abstract: Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) are principal enzymes responsible for metabolism of ethanol. Functional polymorphisms of ADH1B, ADH1C, and ALDH2 genes occur among racial populations. The goal of this study was to systematically determine the functional expressions and cellular localization of ADHs and ALDHs in human rectal mucosa, the lesions of adenocarcinoma and hemorrhoid, and the genetic association of allelic variations of ADH and ALDH with large bowel disorders. Twenty-one surgical specimens of rectal adenocarcinoma and the adjacent normal mucosa, including 16 paired tissues of rectal tumor, normal mucosae of rectum and sigmoid colon from the same individuals, and 18 surgical mixed hemorrhoid specimens and leukocyte DNA samples from 103 colorectal cancer patients, 67 hemorrhoid patients, and 545 control subjects recruited in previous study, were investigated. The isozyme/allozyme expression patterns of ADH and ALDH were identified by isoelectric focusing and the activities were assayed spectrophotometrically. The protein contents of ADH/ALDH isozymes were determined by immunoblotting using the corresponding purified class-specific antibodies; the cellular activity and protein localizations were detected by immunohistochemistry and histochemistry, respectively. Genotypes of ADH1B, ADH1C, and ALDH2 were determined by polymerase chain reaction-restriction fragment length polymorphisms. At 33mM ethanol, pH 7.5, the activity of ADH1C*1/1 phenotypes exhibited 87% higher than that of the ADH1C*1/*2 phenotypes in normal rectal mucosa. The activity of ALDH2-active phenotypes of rectal mucosa was 33% greater than ALDH2-inactive phenotypes at 200μM acetaldehyde. The protein contents in normal rectal mucosa were in the following order: ADH1>ALDH2>ADH3≈ALDH1A1, whereas those of ADH2, ADH4, and ALDH3A1 were fairly low. Both activity and content of ADH1 were significantly decreased in rectal tumors, whereas the ALDH activity remained unchanged. The ADH activity was also significantly reduced in hemorrhoids. ADH4 and ALDH3A1 were uniquely expressed in the squamous epithelium of anus at anorectal junctions. The allele frequencies of ADH1C*1 and ALDH2*2 were significantly higher in colorectal cancer and that of ALDH2*2 also significantly greater in hemorrhoids. In conclusion, ADH and ALDH isozymes are differentially expressed in mucosal cells of rectum and anus. The results suggest that acetaldehyde, an immediate metabolite of ethanol, may play an etiological role in pathogenesis of large bowel diseases.

Malondialdehyde–acetaldehyde-adducted protein inhalation causes lung injury

2011-09-28

Abstract: In addition to cigarette smoking, alcohol exposure is also associated with increased lung infections and decreased mucociliary clearance. However, little research has been conducted on the combination effects of alcohol and cigarette smoke on lungs. Previously, we have demonstrated in a mouse model that the combination of cigarette smoke and alcohol exposure results in the formation of a very stable hybrid malondialdehyde–acetaldehyde (MAA)-adducted protein in the lung. In in vitro studies, MAA-adducted protein stimulates bronchial epithelial cell interleukin-8 (IL-8) via the activation of protein kinase C epsilon (PKCɛ). We hypothesized that direct MAA-adducted protein exposure in the lungs would mimic such a combination of smoke and alcohol exposure leading to airway inflammation. To test this hypothesis, C57BL/6J female mice were intranasally instilled with either saline, 30μL of 50μg/mL bovine serum albumin (BSA)–MAA, or unadducted BSA for up to 3 weeks. Likewise, human lung surfactant proteins A and D (SPA and SPD) were purified from human pulmonary proteinosis lung lavage fluid and successfully MAA-adducted in vitro. Similar to BSA–MAA, SPD–MAA was instilled into mouse lungs. Lungs were necropsied and assayed for histopathology, PKCɛ activation, and lung lavage chemokines. In control mice instilled with saline, normal lungs had few inflammatory cells. No significant effects were observed in unadducted BSA- or SPD-instilled mice. However, when mice were instilled with BSA–MAA or SPD–MAA for 3 weeks, a significant peribronchiolar localization of inflammatory cells was observed. Both BSA–MAA and SPD–MAA stimulated increased lung lavage neutrophils and caused a significant elevation in the chemokine, keratinocyte chemokine, which is a functional homologue to human IL-8. Likewise, MAA-adducted protein stimulated the activation of airway and lung slice PKCɛ. These data support that the MAA-adducted protein induces a proinflammatory response in the lungs and that the lung surfactant protein is a biologically relevant target for malondialdehyde and acetaldehyde adduction. These data further implicate MAA-adduct formation as a potential mechanism for smoke- and alcohol-induced lung injury.

Alcohol drinking, mean corpuscular volume of erythrocytes, and alcohol metabolic genotypes in drunk drivers

2011-09-15

Abstract: Regular and irregular abuse of alcohol are global health priorities associated with diseases at multiple sites, including cancer. Mechanisms of diseases induced by alcohol are closely related to its metabolism. Among conventional markers of alcohol abuse, the mean corpuscular volume (MCV) of erythrocytes is prognostic of alcohol-related cancer and its predictivity increases when combined with functional polymorphisms of alcohol dehydrogenase (ADH1B [rs1229984] and ADH1C [rs698]) and the mitochondrial aldehyde dehydrogenase (ALDH2 [rs671]). Whether these genetic variants can influence abuse in alcohol drinking and MCV has never been examined in drunk-driving traffic offenders. We examined 149 drunk drivers, diagnosed as alcohol abusers according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth edition Text Revision (DSM-IV-TR) and enrolled in a probation program, and 257 social drinkers (controls), all Caucasian males. Alcohol intake was assessed according to self-reported drink-units/d and MCV unadjusted and adjusted for age, smoking, and body mass index. Multivariable models were used to compute MCV adjusted means. Genotype analyses were performed by PCR on DNA from blood. The adjusted MCV mean was higher in drunk-driving abusers than in controls (92 vs. 91fL; P<.0001) and increased with the number of drink-units/d in both abusers and controls (P-trend=.0316 and .0089) already at intermediate quantities (0–1 vs. 2–4 drink-units/d: P=.054 and .024). Carriers of the common ADH1B*1/*1 (rs1229984) genotype were more likely to be drunk-driving abusers (P=.008), reported higher drink-units/d (P=.0126), and had larger MCV (P=.035). The rs698 ADH1C and rs671 ALDH2 polymorphisms were not associated with MCV. ADH1B*1/*1 polymorphism is significantly associated with being a drunk-driving abuser, higher alcohol drinking, and MCV enlargement. This suggests that drunk drivers with augmented MCV modulated by the alcohol metabolic ADH1B*1/*1 genotype may be at higher risk of driving incapability and of alcohol-related cancer.

The diagnostic power of direct carbohydrate-deficient transferrin immunoassay in alcoholics. Absolute or relative values?

Lech Chrostek, Bogdan Cylwik, Ewa Gruszewska, Jolanta Tobolczyk — 2011-09-26

Abstract: The objective of this study was to compare the diagnostic power of direct carbohydrate-deficient transferrin (CDT) immunoassay in alcohol abuse expressed in relative units with the diagnostic power of the results expressed in absolute units. Serum CDT was determined in 127 alcoholics using N Latex CDT direct immunonephelometric assay (Siemens Healthcare Diagnostics, Marburg, Germany). The diagnostic sensitivity, specificity, negative and positive predictive value, and also the positive and negative likelihood ratios do not differ between results expressed in relative or absolute units independently of cutoff chosen. Finally, the area under the receiver operating characteristic (ROC) curves for N Latex CDT test expressed in absolute units does not differ from the area for results expressed in relative units. We conclude that the diagnostic usefulness of N Latex immunonephelometric assay using the relative or absolute values is the same.

Blood alcohol concentration at 0.06 and 0.10% causes a complex multifaceted deterioration of body movement control

Fredrik Modig, Per-Anders Fransson, Måns Magnusson, Mitesh Patel — 2011-08-05

Abstract: Alcohol-related falls are recognized as a major contributor to the occurrence of traumatic brain injury. The control of upright standing balance is complex and composes of contributions from several partly independent mechanisms such as appropriate information from multiple sensory systems and correct feedback and feed forward movement control. Analysis of multisegmented body movement offers a rarely used option for detecting the fine motor problems associated with alcohol intoxication. The study aims were to investigate whether (1) alcohol intoxication at 0.06 and 0.10% blood alcohol concentration (BAC) affected the body movements under unperturbed and perturbed standing; and (2) alcohol affected the ability for sensorimotor adaptation. Body movements were recorded in 25 participants (13 women and 12 men, mean age 25.1 years) at five locations (ankle, knee, hip, shoulder, and head) during quiet standing and during balance perturbations from pseudorandom pulses of calf muscle vibration over 200s with eyes closed or open. Tests were performed at 0.00, 0.06, and 0.10% BAC. The study revealed several significant findings: (1) an alcohol dose-specific effect; (2) a direction-specific stability decrease from alcohol intoxication; (3) a movement pattern change related to the level of alcohol intoxication during unperturbed standing and perturbed standing; (4) a sensorimotor adaptation deterioration with increased alcohol intoxication; and (5) that vision provided a weaker contribution to postural control during alcohol intoxication. Hence, alcohol intoxication at 0.06 and 0.10% BAC causes a complex multifaceted deterioration of human postural control.

Ethanol causes endoplasmic reticulum stress and impairment of insulin secretion in pancreatic β-cells

Khanh Hoa Nguyen, Jong Han Lee, B.L. Grégoire Nyomba — 2011-08-16

Abstract: Chronic ethanol consumption increases the risk of type 2 diabetes mellitus, and ethanol has been reported to cause insulin resistance and, inconsistently, to reduce insulin secretion. The mechanism(s) underlying the reduction of insulin secretion by ethanol is not known. We used β-cell lines and isolated murine islets to determine the effect of ethanol on insulin content and secretion at low- and high-glucose concentrations, in the presence of KCl, diazoxide, tolbutamide, and regulators of cyclic AMP and protein kinase C (PKC). We also determined the gene expression of insulin; pancreas duodenum homeobox 1; and endoplasmic reticulum (ER) stress markers, such as Chop, ERp57, glucose-regulated protein 78/binding immunoglobulin protein, and inositol 1,4,5-triphosphate receptors. Ethanol reduced insulin secretion by interfering with muscarinic signaling and PKC activation but not the K-ATP channels. In addition, ethanol reduced insulin content and caused ER stress. The deleterious effects of ethanol on β-cells were prevented by 4-methyl pyrazole, an inhibitor of alcohol dehydrogenase, suggesting that ethanol metabolism is required for these effects.

Proceedings of the 2009 annual meeting of the Fetal Alcohol Spectrum Disorders Study Group

Feng C. Zhou, Cynthia J.M. Kane, Susan M. Smith — 2011-05-30

Abstract: The annual meeting of the Fetal Alcohol Spectrum Disorders Study Group (FASDSG) was held on June 20, 2009 in San Diego, CA, as a satellite of the Research Society on Alcoholism Meeting. The FASDSG membership includes clinical, basic, and social scientists who meet to discuss recent advances and issues in Fetal Alcohol Spectrum Disorders research. The main theme of the meeting was “Epigenetics and Development.” Two keynote speakers, Dr. Randy Jirtle and Dr. Michael Skinner, addressed the role of epigenetics and environmental inputs, including alcohol, during critical stages of development and their potential critical and long-lasting effects. Members of the FASDSG provided new findings through brief “FASt” data reports, and national agency representatives provided updates on activities and funding priorities. Scientific presentations were made by recipients of the Student Research Merit Award and Rosett Award.

Proceedings of the 2010 annual meeting of the Fetal Alcohol Spectrum Disorders Study Group

Cynthia J.M. Kane, Susan M. Smith, Rajesh C. Miranda, Julie Kable — 2011-09-02

Abstract: The annual meeting of the Fetal Alcohol Spectrum Disorders Study Group (FASDSG) was held on June 26, 2010 in San Antonio, TX, as a satellite of the Research Society on Alcoholism meeting. The FASDSG membership includes clinical, basic, and social scientists who meet to discuss recent advances and issues in Fetal Alcohol Spectrum Disorder (FASD) research. The central theme of the meeting was “Glia and Neurons: Teamwork in Pathology and Therapy.” Alcohol disruption of neuron development and alcohol-induced neurodegeneration is central to the pathology and clinical expression of FASD. The active role of glia as perpetrator, victim, or bystander in neurotoxicology and neurodegenerative processes has emerged at the forefront of adult central nervous system (CNS) disorders and therapy. Glia– and neuron–glial interactions hold the potential to elucidate causes and offer treatment of FASD as well. Growing evidence indicates that neurons and glia are direct targets of alcohol, but may also be vulnerable to molecules produced in peripheral systems as a result of alcohol exposure. Diagnostics and therapies can take advantage of these processes and biomarkers, and these may be applicable to CNS pathology in FASD. Two keynote speakers, Howard E. Gendelman, M.D., and Ernest M. Graham, M.D, addressed the role of glia and neuroinflammation in brain development and neurodegeneration. The invited speakers and FASDSG members discussed new paradigms in CNS development and discuss new strategies for understanding and treating neurodegenerative disease. Members of the FASDSG provided updates on new findings through presentation of breaking research in the FASt data sessions. Representatives of national agencies provided updates on programs, activities, and funding priorities. The Henry Rosett Award was presented to R. Louise Floyd, R.N., D.S.N., for her career contributions to the field of fetal alcohol research. The Student and Postdoctoral Fellow Research Merit Award was presented to Shonagh O’Leary-Moore, Ph.D., for her contributions to the field as a young investigator.