Biomarkers Of Alcohol Consumption Research Articles

Selective detection of phosphatidylethanol homologues in blood as biomarkers for alcohol consumption by LC‐ESI‐MS/MS

A new validated method for the quantitation of the abnormal phospholipid phosphatidylethanol (PEth)--a biomarker for ethanol uptake--has been developed by LC-ESI-MS/MS following miniaturised organic solvent extraction and reversed phase chromatography with phosphatidylbutanol (PBut) as internal standard. PEth homologues with two fatty acid substituents-PEth 18:1/18:1, PEth 16:0/16:0-were determined in post-mortem blood collected from heavy drinkers at autopsy and also in whole blood samples from a volunteer after a single 60 g-dose of ethanol. Furthermore, PEth 18:1/16:0 or its isobaric isomer PEth-16:0/18:1 was detected. In comparison to previous high-performance liquid chromatography (HPLC) methods with evaporative light scattering detection (ELSD), the LC-MS/MS-method is more sensitive--with a limit of detection below 20 ng/ml--and more selective for single PEth homologues, while ELSD has been used for detection of the sum of PEth homologues with approximately 10 times less sensitivity. LC-MS/MS enables monitoring of PEth homologues as biomarkers for harmful and prolonged alcohol consumption as with HPLC/ELSD earlier, where PEth is measurable in blood only after more than 50 g ethanol daily intake for more than 2 weeks. Because of its higher sensitivity, there is a potential to detect single heavy drinking by LC-MS/MS, when PEth is formed in very low concentrations. This opens a new field of application of PEth to uncover single or multiple heavy drinking at a lower frequency and with a larger window of detection in blood than before by HPLC/ELSD or by use of other direct markers, e.g. ethyl glucuronide or ethyl sulfate.

Novel Objective Biomarkers of Alcohol Use: Potential Diagnostic and Treatment Management Tools in Dual Diagnosis Care

Alcohol use disorders are highly prevalent conditions that generate a large fraction of the total public health burden. These disorders are concentrated in mentally ill populations, in which reliability of self-reporting of alcohol consumption may be especially compromised. The application of objective biomarkers for alcohol use may therefore play an important role in these patients. This article provides a description and comparative overview of traditional versus novel biomarkers of alcohol consumption. Greater professional familiarity with and use of novel biomarkers as diagnostic and treatment management tools may enhance clinical standards and research on alcohol use in patients with a dual diagnosis.

Association Between the Stin2 VNTR Polymorphism of the Serotonin Transporter Gene and Treatment Outcome in Alcohol-Dependent Patients

The aim of this study was to investigate the potential association between functional polymorphisms of dopaminergic [dopamine receptor D2 (DRD2), dopamine receptor D3 (DRD3) and dopamine transporter (SLC6A3)] and serotonergic [serotonin 2A receptor (HTR2A) and serotonin transporter (SLC6A4)] genes and treatment outcome in alcohol-dependent patients. A total of 90 Spanish Caucasian alcohol-dependent outpatients (ICD-10 criteria) were enrolled in the study. The association between genotypes and drinking outcomes was measured over 6 months of treatment. Biomarkers of alcohol consumption, as well as alcohol consumption and its consequences, craving, disability and quality of life, were assessed. Based on those measures, we created a composite secondary measure to globally assess treatment outcome in alcoholism. No association was found between DRD2, DRD3, SLC6A3 or HTR2A gene variants and treatment outcome. However, SLC6A4 STin2 12/12 carriers showed poor 6-month time point treatment outcome [32.8% in the good outcome group versus 64.0% in the poor outcome group, chi(2) (df) = 7.20 (1), corrected P = 0.042, OR (95% CI) = 0.27 (0.10-0.72)]. Nevertheless, independent analysis of each treatment group reveals that the excess of 12/12 carriers in the poor outcome group was only found in the naltrexone-treated group [24.1% versus 64.7% chi(2) (df) = 7.41 (1), corrected P = 0.042, OR (95% CI) = 0.17 (0.05-0.64)]. In the whole sample, the L-10 repeats haplotype (5-HTTLPR-STin2 VNTR) is associated with good outcome (LRT = 3.88, df = 1, P = 0.049). Our findings suggest that functional polymorphism of the SLC6A4 gene may have an influence on treatment outcome in alcohol-dependent patients.

Measurement of direct ethanol metabolites suggests higher rate of alcohol use among pregnant women than found with the AUDIT—a pilot study in a population-based sample of Swedish women

The objective of the study was to investigate whether biomarkers of alcohol consumption would provide additional information to the use of a validated alcohol questionnaire in pregnant women. One hundred three pregnant women were included in the study. The women completed the Alcohol Use Disorders Identification Test (AUDIT) questionnaire, and a urine and hair sample was collected. The urine samples were used for determination of ethyl glucuronide (EtG) and ethyl sulfate and the hair samples for EtG and fatty acid ethyl esters (FAEE). Twenty-six women (25.2%) were identified as possible alcohol consumers by the combined use of AUDIT and direct ethanol metabolites. Seven subjects had EtG or FAEE levels in hair highly suspicious of heavy drinking, but only 1 of these were positive according to the AUDIT questionnaire The combined use of the AUDIT questionnaire and direct ethanol metabolites appear to identify more potential alcohol consumers among pregnant women than does the sole use of the AUDIT questionnaire.

Alcohol consumption, %CDT, GGT and blood pressure change during alcohol treatment

Blood pressure (BP) changes in alcohol-dependent individuals during a 12-week alcohol relapse prevention study were examined in light of drinking status and biomarkers of alcohol consumption [carbohydrate-deficient transferrin (%CDT) and gamma-glutamyl transpeptidase (GGT)]. Of 160 randomized alcoholic individuals, 120 who had hypertension and in whom daily drinking data was available, at 6 and 12 weeks of treatment were included. The impact of alcohol consumption on change in systolic BP (SBP) and diastolic BP (DBP) was examined. Further analysis determined the relationship between BP and alcohol-use biomarkers. A significant effect of complete abstinence on both SBP (-10 mmHg; P = 0.003) and DBP (-7 mmHg; P = 0.001) when compared to any drinking (SBP and DBP = -1 mmHg) was observed. At week 12, participants with a positive %CDT (> or =2.6) had 7 mmHg greater SBP (P = 0.01) and DBP (P < 0.001) than those with negative %CDT. Participants with positive GGT (> or =50 IU) had 10 mmHg greater SBP (P = 0.12) and 9 mmHg greater DBP (P = 0.03) than those with negative GGT. The percent change in SBP was correlated with percent change in %CDT (P = 0.003) but not GGT (P = ns). The percent change in DBP was correlated with both percent change in %CDT (P < 0.0001) and GGT (P = 0.03). Abstinence from alcohol significantly decreased the BP and a positive relationship between BP and both alcohol-use biomarkers was illustrated. Since %CDT is more specific than GGT for heavy alcohol consumption, clinicians may monitor the role of alcohol in hypertension using %CDT as a supplemental aid, providing an objective assessment of drinking to influence BP treatment decisions.

Co-Occurrence of IgA Antibodies Against Ethanol Metabolites and Tissue Transglutaminase in Alcohol Consumers: Correlation with Proinflammatory Cytokines and Markers of Fibrogenesis

IgA antibodies to tissue transglutaminase have been suggested to be specific indicators of celiac disease. However, no studies have addressed the relationships between such antibodies and alcohol abuse, which is also a common cause of IgA-isotype immune responses and tissue injury in the gastrointestinal tract and liver. Here, measurements of specific IgAs against tissue transglutaminase and proteins modified by acetaldehyde, the first metabolite of ethanol, showed significantly higher levels of both antibodies in alcoholic liver disease patients than in healthy controls or heavy drinkers without liver disease. These antibodies also significantly co-occurred in heavy drinkers without liver disease, moderate drinkers, and abstainers, and correlated with biomarkers of alcohol consumption, proinflammatory cytokines and markers of fibrogenesis. The data suggests a link between such immune responses, perturbations in cytokine profiles and fibrogenesis, which should be implicated in studies on the pathogenesis and diagnosis of ethanol-induced tissue injury and celiac disease.

A reduction in alcohol consumption is associated with reduced plasma F 2-isoprostanes and urinary 20-HETE excretion in men

There is considerable evidence that chronic moderate-to-high alcohol consumption increases blood pressure. The mechanisms by which this occurs are not clear. Alcohol consumption can induce oxidative stress and cytochrome P450 (CYP450) isoforms that are associated with oxidative stress and may influence vascular tone. To study the role of such mechanisms we examined whether reducing alcohol intake in moderate-to-heavy drinkers (40–110 g/day) resulted in changes in urinary excretion of 20-HETE, a CYP450 metabolite of arachidonic acid, and plasma and urinary F 2-isoprostanes as markers of lipid peroxidation. After a 4-week run-in period during which healthy men maintained their usual drinking pattern they were randomized to a two-way crossover intervention study. In each of the 4-week treatment periods subjects either substituted their usual alcohol intake with a 0.9% alcohol beer or maintained their usual alcohol intake. Plasma and urinary F 2-isoprostanes and urinary 20-HETE were measured by gas chromatography mass spectrometry, and serum γ-glutamyl transpeptidase (γ-GT) was measured as a biomarker of alcohol consumption, at the end of each study period. Sixteen healthy men age 51.0 ± 2.7 years and with a BMI of 26.4 ± 0.61 kg/m 2 completed the study. The reductions in alcohol intake (72.4 ± 5.0 vs 7.9 ± 1.6 g/day, p < 0.001) and serum γ-GT (geometric mean 24.4 U/L (95% CI 19.7, 30.2) vs 18.6 U/L (95% CI 15.5, 22.2, p < 0.01) were accompanied by a significant fall in blood pressure as well as urinary 20-HETE excretion (158 ± 23 vs 109 ± 19 pmol/mmol creatinine, p < 0.001) and plasma F 2-isoprostanes (3438 ± 158 vs 2929 ± 145 pmol/L, p = 0.01). A substantial reduction in alcohol consumption in healthy men lowered plasma F 2-isoprostanes and urinary 20-HETE. Increased oxidative stress and 20-HETE production may be linked, at least in part, to the pathogenesis of alcohol-related hypertension.

Biomarkers of alcohol consumption in patients classified according to the degree of liver disease severity

Objective. In the search for optimal biomarkers of excessive drinking, only a few studies have been conducted to compare the relationships between ethanol consumption, liver status, and various laboratory markers of ethanol‐induced diseases. Material and methods. Concentrations of carbohydrate‐deficient transferrin (%CDT and CDTect methods), serum sialic acid (SA), γ‐glutamyl transferase (γ‐GT), aspartate aminotransferase (ASAT), mean corpuscular volume (MCV), and a marker of fibrogenesis (PIIINP) were studied in 102 alcoholics with (n = 59) or without (n = 43) alcoholic liver disease. Controls were 34 healthy volunteers who were either social drinkers or abstainers. Results. Although concentrations of all markers were significantly higher in the alcoholic patients than in the healthy controls, their diagnostic characteristics showed a considerable degree of variation. The %CDT, SA, and MCV showed the strongest correlations with the amount of recent alcohol intake. The presence of liver pathology notably influenced the results of CDTect, GT, ASAT, and PIIINP. In ROC analyses, the highest rates of diagnostic accuracy for detecting hazardous drinking were reached with GT (0.94), CDT (0.86), and SA (0.85), followed by MCV (0.79) and ASAT (0.77). Upon abstinence, the estimated times for normalization varied between 10 days (CDTect) and 25 days (GT). Conclusions. Our data suggest distinct differences in the clinical characteristics of biological markers of ethanol consumption. While the overall accuracy of CDT and GT appear to be highest in the detection of problem drinking, serum SA and PIIINP measurements are of further value when the effects of liver pathology and ethanol drinking need to be differentiated.

Behavioral patterns of contemporary biomarkers of alcohol consumption during relapse episodes after discharge from treatment: Pamela Bean, Tracy Debruin, Rogers Memoral Hosp, Madison, WI; James Harasymiw, Alcohol Detection Svcs, Brookfield, WI; James Mundt, Healthcare Technology Systems, Madison, WI

Behavioral patterns of contemporary biomarkers of alcohol consumption during relapse episodes after discharge from treatment: Pamela Bean, Tracy Debruin, Rogers Memoral Hosp, Madison, WI; James Harasymiw, Alcohol Detection Svcs, Brookfield, WI; James Mundt, Healthcare Technology Systems, Madison, WI

A COMPARISON OF TWO INTENSITIES OF PSYCHOSOCIAL INTERVENTION FOR ALCOHOL DEPENDENT PATIENTS TREATED WITH ACAMPROSATE.

Aims: To compare two levels of psychosocial intervention in combination with acamprosate medication for the treatment of alcohol dependence. Methods: Patients (n = 70) were prescribed acamprosate and randomized to Minimal Psychosocial Intervention (MPI) or Extended Psychosocial Intervention (EPI). MPI patients met a psychiatrist for 20-30 min sessions on four occasions during a 6 month period. EPI patients were offered 10-15 sessions with a psychiatric nurse in addition to the visits to the psychiatrist. EPI patients were trained to use behavioural and cognitive coping skills to deal with high-risk situations in line with a manual developed for relapse prevention. Patients were assessed four times during the 24-week study by self-report and laboratory tests. Results: Patients on average reported a decline in days with heavy drinking and in cumulative number of drinking days. No significant differences between patients in MPI and EPI were found with respect to heavy drinking, cumulative number of drinking days, number of days to first drink, or biomarkers of alcohol consumption. Higher age and lower level of education were significant predictors of treatment success. Conclusions: Adding more intensive individual treatments appears to add no extra improvement beyond that obtained by prescribing acamprosate and offering an infrequent consultation with a physician.

Association of a GNAS1 Gene Variant with Hypertension and Diabetes Mellitus

Previous studies have shown that the T allele of the GNAS1 T393C polymorphism is associated with poor responsiveness to beta-blockade and that the T393C polymorphism interacts with cigarette smoking and alcohol consumption in the pathogenesis of hypertension. Thus, the T393C polymorphism is likely to interact with beta-adrenoceptor (beta-AR) stimulation in the pathogenesis of hypertension. Although this interaction might be caused by a direct effect of Gs proteins on the cardiovascular system, it could also result from an indirect effect of Gs proteins mediated by glucose metabolism. Moreover, association studies are often irreproducible. We therefore examined the possible interaction between the T393C polymorphism and gamma-glutamyl transpeptidase (GGT), which is an established biomarker of alcohol consumption, in the association with glucose metabolism as well as with hypertension in a Japanese population. Genotyping for GNAS1 was performed by using the polymerase chain reaction-restriction fragment length polymorphism method in all 821 samples. The present study showed a significant interaction between the T393C polymorphism and GGT in the association with hypertension (p =0.033). This interaction was even more significant after adjustment for all confounding factors (p =0.0025). In contrast, analysis of the possible interaction of the T393C polymorphism with GGT in the association with diabetes mellitus or fasting plasma glucose failed to show a significant result. These results did not support the hypothesis that the interaction between the T393C polymorphism and GGT in the association with hypertension could be caused by an indirect effect of Gs proteins mediated by glucose metabolism.

Oxidative stress in human hypertension: association with antihypertensive treatment, gender, nutrition, and lifestyle

There is growing evidence that oxidative stress contributes to the pathogenesis of hypertension. Our aim was to measure markers of oxidative stress in hypertensive subjects, and assess the potential confounding influences of antihypertensive therapy, other cardiovascular risk factors, gender, lifestyle, and nutrition. Markers of oxidative stress, including plasma and 24 h urinary F 2-isoprostanes, were measured in 70 untreated (men = 43, women = 27) and 85 treated (men = 43, women = 42) hypertensive subjects and 40 normotensive controls (men = 20, women = 20). Overall, F 2-isoprostanes were not elevated in hypertensive subjects compared with controls. However, urinary and plasma F 2-isoprostanes were significantly lower in treated compared with untreated hypertensive men, but not women. In hypertensive men, the number of antihypertensive drugs taken was inversely associated with both urinary and plasma F 2-isoprostanes ( p < .05). Self-reported alcohol intake and biomarkers of alcohol consumption (γ-glutamyl transpeptidase and high-density lipoprotein cholesterol) were positively associated with plasma but not urinary, F 2-isoprostanes in men. Several nutrients were independently associated with plasma and urinary F 2-isoprostanes in women. The results do not support the hypothesis that treated or untreated hypertensive subjects are under increased oxidative stress compared with normotensive controls.