Background. Alcohol is a carcinogen for the liver. Chronic alcohol consumption increases Cytochrome P450 2E1 (CYP2E1) in liver which is associated with a generation of highly carcinogenic DNA lesions. The purpose of the present study was to investigate whether the CYP2E1 inhibitor chlormethiazole (CMZ) has an effect on hepatic carcinogenesis induced by diethylnitrosamine (DEN). Methods. Rats received Lieber–DeCarli ethanol containing control diets with and without CMZ and a single intraperitoneal injection of a low dose of DEN (20 mg per kg body weight). The formation of hepatic preneoplastic foci was assessed by immunostaining of placental glutathione-S-transferase (p-GST). In addition, hepatic proliferation was assessed by immunohistochemistry for proliferating cellular nuclear antigen, Ki-67 and cyclin D1. In addition, hepatic expression of TNF-α and activation of NF-κB and CYP2E1 were assessed by either real-time PCR or western blotting analysis. Results. Ethanol feeding resulted in a significant induction of hepatic CYP2E1 increased nuclear accumulation of NF-κB protein and TNF-α expression which was associated with increased hepatocyte proliferation markers and cyclin D1 protein expression as well as increased p-GST-positive altered hepatic foci in rat liver tissues. Chlormethiazole treatment significantly inhibited ethanol-induced CYP2E1 expression, TNF-α mRNA expression, NF-κB activation, hepatocyte proliferation and altered hepatic foci formation after a 1-month period. Conclusion. These data indicate that the inhibition of ethanol-induced CYP2e1 can block hepatic preneoplastic lesion by decreasing TNF-α expression and NF-κB activation in DEN-treated rats.

BACKGROUND Apolipoprotein J (ApoJ) is a component of plasma high-density lipoproteins. Previous studies have shown progressive recovery of ApoJ sialic acid content with increased duration of alcohol abstinence. Therefore, the sialic acid index of plasma apolipoprotein J (SIJ) seems to be a promising alcohol biomarker. Phosphatidylethanol (PEth) is a direct ethanol metabolite and has recently attracted attention as a biomarker of prolonged intake of higher amounts of alcohol. The aim of the pilot study was to explore sensitivity, specificity, and normalization of SIJ and PEth in comparison with traditional and emerging biomarkers. METHODS Five male alcohol-dependent patients (International Classification of Diseases 10, F 10.25) were included (median: 40 years old; Alcohol Use Disorders Identification Test value, 30; alcohol consumption in the previous 7 days, 1,680 g). SIJ, PEth, urinary ethyl glucuronide (UEtG), urinary ethyl sulfate (UEtS), and gamma glutamyl-transpeptidase (GGT) were determined at days 1, 3, 7, 10, 14, 21, and 28. RESULTS At study entry, SIJ, PEth, UEtG, and UEtS were positive in all subjects, whereas GGT and mean corpuscular volume were positive in 3 of 5 (60%) of the subjects. Individual SIJ levels increased between day 1 and 28 between 13.7 and 44.3%, respectively. For SIJ and PEth, the ANOVA (p < 0.005) showed a significant trend with the average subject's SIJ and PEth changing 1.22 and 1.02, respectively, per week. CONCLUSIONS Our preliminary data suggest that SIJ and PEth might hold potential as markers of heavy ethanol intake.

ABSTRACTPhosphatidylethanol (PEth) is a direct ethanol metabolite, and has recently attracted attention as biomarker of ethanol intake. The aims of the current study are: (1) to characterize the normalization time of PEth in larger samples than previously conducted; (2) to elucidate potential gender differences; and (3) to report the correlation of PEth with other biomarkers and self‐reported alcohol consumption. Fifty‐seven alcohol‐dependent patients (ICD 10 F 10.25; 9 females, 48 males) entering medical detoxification at three study sites were enrolled. The study sample was comprised of 48 males and 9 females, with mean age 43.5. Mean gamma glutamyl transpeptidase (GGT) was 209.61 U/l, average mean corpuscular volume (MCV) was 97.35 fl, mean carbohydrate deficient transferrin (%CDT) was 8.68, and mean total ethanol intake in the last 7 days was 1653 g. PEth was measured in heparinized whole blood with a high‐pressure liquid chromatography method, while GGT, MCV and %CDT were measured using routine methods. PEth levels at day 1 of detoxification ranged between 0.63 and 26.95 µmol/l (6.22 mean, 4.70 median, SD 4.97). There were no false negatives at day 1. Sensitivities for the other biomarkers were 40.4% for MCV, 73.1% for GGT and 69.2% for %CDT, respectively. No gender differences were found for PEth levels at any time point. Our data suggest that PEth is (1) a suitable intermediate term marker of ethanol intake in both sexes; and (2) sensitivity is extraordinary high in alcohol dependent patients. The results add further evidence to the data that suggest that PEth has potential as a candidate for a sensitive and specific biomarker, which reflects longer‐lasting intake of higher amounts of alcohol and seemingly has the above mentioned certain advantages over traditional biomarkers.

Phosphatidylethanol (PEth) is a direct ethanol metabolite, and has recently attracted attention as biomarker of ethanol intake. The aims of the current study are: (1) to characterize the normalization time of PEth in larger samples than previously conducted; (2) to elucidate potential gender differences; and (3) to report the correlation of PEth with other biomarkers and self-reported alcohol consumption. Fifty-seven alcohol-dependent patients (ICD 10 F 10.25; 9 females, 48 males) entering medical detoxification at three study sites were enrolled. The study sample was comprised of 48 males and 9 females, with mean age 43.5. Mean gamma glutamyl transpeptidase (GGT) was 209.61 U/l, average mean corpuscular volume (MCV) was 97.35 fl, mean carbohydrate deficient transferrin (%CDT) was 8.68, and mean total ethanol intake in the last 7 days was 1653 g. PEth was measured in heparinized whole blood with a high-pressure liquid chromatography method, while GGT, MCV and %CDT were measured using routine methods. PEth levels at day 1 of detoxification ranged between 0.63 and 26.95 µmol/l (6.22 mean, 4.70 median, SD 4.97). There were no false negatives at day 1. Sensitivities for the other biomarkers were 40.4% for MCV, 73.1% for GGT and 69.2% for %CDT, respectively. No gender differences were found for PEth levels at any time point. Our data suggest that PEth is (1) a suitable intermediate term marker of ethanol intake in both sexes; and (2) sensitivity is extraordinary high in alcohol dependent patients. The results add further evidence to the data that suggest that PEth has potential as a candidate for a sensitive and specific biomarker, which reflects longer-lasting intake of higher amounts of alcohol and seemingly has the above mentioned certain advantages over traditional biomarkers.

OBJECTIVE:The aim of this study was to investigate the congruence of biomarkers, questionnaires, and interviews as instruments to assess adolescent alcohol consumption.METHOD:The methodology used was a cross-sectional study with a randomized sample. Four different methods were used to estimate high adolescent alcohol consumption. The concordance of the results was investigated. Surveys were performed, and biological specimens were collected at all schools in the county of Västmanland, Sweden, in 2001. Eighty-one boys and 119 girls from a population of 16- and 19-year-old adolescents were randomly selected from quartiles of volunteers representing various degrees of psychosocial risk behaviors. Using a questionnaire (for a 1-hour session) and in-depth interviews, subjects were assessed regarding their alcohol-use habits. Blood and hair samples were analyzed for phosphatidylethanol (PEth) and fatty acid ethyl esters (FAEEs), respectively.RESULTS:High alcohol consumption was underreported in the questionnaire compared with the interviews. PEth and FAEE analyses weakly confirmed the self-reports, and the results of the two biochemical tests did not overlap. The PEth blood test was the most specific but the least sensitive, whereas the FAEE hair test revealed low specificity and an overrepresentation of positive results in girls.CONCLUSIONS:The expected higher self-report of high alcohol consumption by interview rather than by questionnaire was confirmed partly because of the influence of a bogus pipeline procedure. The absence of overlap between PEth and FAEE results and their poor agreement with self-reports suggested that biomarkers are unsuitable as screening tools for alcohol consumption in adolescents.

Phosphatidylethanol (PEth) represents a group of glycerophospholipid homologues where ethanol by phospholipase D has been bound at the position that normally contains an amino-alcohol. Since the formation of PEth is specifically dependent on ethanol, the diagnostic specificity of PEth as an alcohol biomarker is theoretically 100%. The half-life of PEth in blood is approximately 4 days. The amount of alcohol consumed correlates to blood concentration of PEth and PEth has been shown to be a more sensitive indicator of alcohol consumption than traditional alcohol markers, such as CDT (carbohydrate-deficient transferrin), GGT (γ-glutamyl transferase), and MCV (mean corpuscular volume) or a combination of these. Almost all clinical data so far available are based on a high performance liquid chromatography (HPLC) method with limited analytical sensitivity. With the advent of methods with considerably higher analytical sensitivity (e.g. mass spectrometric methods), clinical sensitivity will increase correspondingly. The possibility of determining very low concentrations of PEth by new sensitive analytical techniques may, however, have both ethical and legal consequences that have to be considered.

AimTo identify alcohol biomarker and psychometric measures that relate to drivers' blood alcohol concentration (BAC) patterns from ignition interlock devices (IIDs).Design, setting, participants, measurementsIn Alberta, Canada, 534 drivers, convicted of driving under the influence of alcohol (DUI), installed IIDs and agreed to participate in a research study. IID BAC tests are an established proxy for predicting future DUI convictions. Three risk groups were defined by rates of failed BAC tests. Program entry and follow-up blood samples (n = 302, 171) were used to measure phosphatidyl ethanol (PETH), carbohydrate deficient transferrin (%CDT), gamma glutamyltransferase (GGT) and other biomarkers. Program entry urine (n = 130) was analyzed for ethyl glucuronide (ETG) and ethyl sulphate (ETS). Entry hair samples were tested for fatty acid ethyl esters (FAEE) (n = 92) and ETG (n = 146). Psychometric measures included the DSM-4 Diagnostic Interview Schedule Alcohol Module, Alcohol Use Disorders Identification Test (AUDIT), the time-line follow-back (TLFB), the Drinker Inventory of Consequences (DRINC) and the Temptation and Restraint Inventory (TRI).FindingsExcept for FAEE, all alcohol biomarkers were related significantly to the interlock BAC test profiles; higher marker levels predicted higher rates of interlock BAC test failures. PETH, the strongest with an overall analysis of variance F ratio of 35.5, had significant correlations with all nine of the other alcohol biomarkers and with 16 of 19 psychometric variables. Urine ETG and ETS were correlated strongly with the IID BAC tests.ConclusionsThe findings suggest that several alcohol biomarkers and assessments could play an important role in the prediction and control of driver alcohol risk when re-licensing.

A 37-year-old female subject had been convicted of driving under the influence of alcohol, and 19 months later, claimed abstinence after supervised disulfiram treatment. Our aim was to elucidate the value of direct ethanol metabolites as measures of abstinence. Ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEE) in hair, phosphatidylethanol in whole blood and EtG and ethyl sulphate in urine were measured. The results were compared with self-report of alcohol consumption and traditional blood biomarkers for chronically elevated alcohol consumption as carbohydrate deficient transferrin (CDT), gamma glutamyl transpeptidase, mean corpuscular erythrocyte volume, aspartate aminotransferase and alanine aminotransferase. EtG was found in distal parts of hair only, whereas the proximal parts were negative. Furthermore, FAEE concentrations were found in the typical distribution over the hair length and showed values typical for either moderate social drinking or abstinence. CDT was above cut-off in 9 out of 16 analyses with a decreasing tendency and the lowest values in the last 2 months before the end of sampling. The data suggest that in addition to traditional markers, a combination of direct ethanol metabolites can be useful in the expert assessment of judging driving ability. A careful individual interpretation of the results for the different markers, however, is an absolute necessity.

BACKGROUND:A major part of medical pathology in internal medicine is associated with chronic alcoholism. The aim of the current study was to investigate whether screening for Alcohol Use Disorders (AUD) can be improved through determination of direct ethanol metabolites compared to traditional biological state markers, the Alcohol Use Disorders Identification Test (AUDIT) and additional self-reports beyond the detection time period of a positive blood alcohol concentration (BAC). METHODS:A total of 74 blood alcohol negative male patients who presented at the emergency room with either thoracic or gastrointestinal complaints were included. Phosphatidylethanol (PEth) was determined in whole blood, and ethyl glucuronide (EtG) in serum and urine samples. Traditional biological state markers [carbohydrate deficient transferrin (%CDT), gamma glutamyl transpeptidase (GGT), mean corpuscular volume (MCV)] were determined. The AUDIT was obtained and furthermore, all patients completed an additional self-report of alcohol consumption. Patients were divided into two (2) groups: AUDIT scores < 8 and AUDIT scores >or= 8. RESULTS:After assessment of the AUDIT, patients were allocated to one of the following groups: patients with AUDIT scores < 8 (n = 52) and with AUDIT scores >or= 8 (n = 22). Twenty-five percent of the patients with AUDIT scores below the cut-off (n = 13/52) were tested positive for both PEth and UEtG. Of the patients who declared to be sober during the past 12 months, 38.5% were tested positive for PEth and UEtG. PEth discriminated similarly as %CDT for AUDIT scores >or= 8 (AUC: 0.672; 95%CI 0.524 to 0.821). Self-reports of alcohol consumption were unreliable. CONCLUSION:Determination of direct ethanol metabolites such as PEth and UEtG provides additional evidence in screening for AUD in an ER setting. Determination of PEth might be considered complementary with or alternatively to %CDT.

ABSTRACT Phosphatidylethanol (PEth), a direct ethanol metabolite, is detectable in blood for more than 2 weeks after sustained ethanol intake. Our aim was to assess the usefulness of PEth [comparing sensitivity, specificity and the area under the curve (AUC)] as compared with carbohydrate‐deficient transferrin (CDT), gamma‐glutamyl transpeptidase (GGT) and mean corpuscular volume (MCV), calculating the results from sober patients against those from alcohol‐dependent patients during withdrawal. Fifty‐six alcohol‐dependent patients (ICD‐10 F 10.25) in detoxification, age 43 years, GGT 81 U/l, MCV 96.4 fl, %CDT 4.2, 1400 g ethanol intake in the last 7 days (median), were included in the study. Over the time of 1 year, 52 samples from 35 sober forensic psychiatric addicted in‐patients [age 34 years, GGT 16 U/l, MCV 91 fl, CDT 0.5 (median)] in a closed ward were drawn and used for comparison . PEth was measured in heparinized whole blood with a high‐performance liquid chromatography method. GGT, MCV and %CDT were measured using routine methods. A receiver operating characteristic curve analysis was carried out, with ‘current drinking status’ (sober/drinking) as the state variable and PEth, MCV, GGT and CDT as test variables. The resulting AUC was 0.974 ( P < 0.0001, confidence interval 0.932–1.016) for PEth. At a cut‐off of 0.36 µmol/l, the sensitivity was 94.5% and specificity 100%. The AUC for CDT, GGT and MCV were 0.931, 0.894 and 0.883, respectively. A significant Spearman’s rank correlation was found between PEth and GGT ( r = 0.739), CDT ( r = 0.643), MVC ( r = 0.639) and grams of ethanol consumed in the last 7 days ( r = 0.802). Our data suggest that PEth has potential to be a sensitive and specific biomarker, having been found in previous studies to indicate longer lasting intake of higher amounts of alcohol.

The risk of impaired driving can be estimated through the use of several technologies but few programs utilize the full spectrum of predictive technologies and often underutilize the ones they do employ. The record of alcohol ignition interlock breath alcohol (BAC) tests are good predictors of future DUI risk, but still not widely used. These are a good, but imperfect predictor, since drivers other than the DUI offender of record can use the interlock car. Biological markers of alcohol use can more directly target the impairment risk of specific individuals. Use of biomarkers to monitor progress among DUI offenders is growing in Europe but less so in North America. A sample of 537 interlock using DUI offenders from Alberta, Canada have been providing self-report assessments as well as biological specimens at two time points, the beginning of an interlock installation, and typically 6 months later, to determine: a) the predictive relationship between the biomarkers and the record of interlock BAC tests, b) the cross-relationship among the biomarkers, c) the relationship between the biomarkers and self-report assessments, and d) the extent to which the markers, self-report, and interlock BAC tests can correctly predict future (post-interlock) recidivism. Biomarkers under study include conventional alcohol markers such as GGT, ALT, AST, MCV, and percent carbohydrate deficient transferrin (%CDT), but also markers that directly reflect ethanol consumption such phosphatidyl ethanol (PEth), fatty acid ethyl esters (FAEE), and ethyl glucuronide (EtG). Self-report assessments include AUDIT, Drinker Inventory of Consequences (Drinc), the DSM4 Alcohol Dependence assessment from the DIS-C, and Timeline Followback (TLFB) and Temptation Restraint Inventory (TRI). Interlock BAC test data are represented as a proportion of all tests taken that are elevated above .02 g/dL over the full six months as well as for each 2 month interval since installation. Preliminary results show many markers correlate significantly and all in the right direction with interlock BAC tests and self-report indicators. TLFB self-report variables (such as % drinking days, drinks per week) show significant correlations (Spearman and Pearson) with markers and BAC tests. Direct markers, especially PEth, EtG and FAEE, but also GGT, are significantly related to the proportion of elevated interlock BAC tests. Sample size is variable across measures, but continues to grow; post-interlock subsequent DUI events will also grow, but currently with just 9 repeat DUI events there are too few for predictive profiling. Biomarkers are an important monitoring tool and should be used as a supplement to conventional estimates of driver risk. These should be more widely utilized especially in conjunction with alcohol harm reduction or medical interventions.

AIMSPhosphatidylethanol (PEth) is an abnormal phospholipid formed only in the presence of ethanol by the enzyme phospholipase D. PEth in blood is a promising new marker for ethanol abuse. None of the biological markers used at the present time is sensitive and specific enough for the diagnosis of alcoholism.METHODSThe most frequently used alcohol markers [carbohydrate deficient transferrin (CDT), gamma-glutamyltransferase (GGT), and mean corpuscular volume (MCV)] were studied together with PEth in actively drinking alcohol-dependent patients (inpatients and outpatients), with regard to correlation to ethanol intake and diagnostic sensitivity of the markers. The relation between the markers was also studied.RESULTSPEth, CDT, and GGT correlated to ethanol intake, with the strongest correlation found for PEth. The diagnostic sensitivity for PEth was 99%, and for other markers it varied between 40 and 77%. Only when CDT was combined with GGT was a sensitivity of 94% reached. PEth correlated to CDT and GGT but not to MCV. CDT did not correlate to GGT or MCV.CONCLUSIONSBlood concentrations of PEth are highly correlated to ethanol intake, and the present results indicate that its diagnostic sensitivity is higher than that for previously established alcohol markers.

BACKGROUND:S100B, a protein abundant in astroglial cells within the central nervous system, has been shown to increase in cerebrospinal fluid and serum after various neurologic diseases. However, the cerebral specificity of S100B has been questioned. This study aims to show serum S100B levels after uncomplicated bone fractures in patients without current or prior neurologic diseases. METHODS:Blood for sampling was drawn from patients seeking care at the emergency department presenting with various uncomplicated orthopedic fractures no older than 24 hours and having no previous or suspected neurologic disorder or head injury. RESULTS:Fifty-five consecutive patients with acute fractures were included in the study. Serum S100B levels were raised above 0.15 microg/L in 16 of 55 (29%) patients (range, 0.02-0.51 microg/L; mean, 0.13 +/- 0.11 microg/L). CONCLUSION:S100B levels were raised in 29% of patients with acute fractures without apparent cerebral injury, which suggests an extracerebral source of S100B. This information should be taken into account when interpreting S100B levels when dealing with brain damage.

This article summarizes content proceedings of a symposium held at the 2004 Research Society on Alcoholism Scientific Annual Meeting in Vancouver, Canada. The chairs were Friedrich M. Wurst and Raye Litten. The presentations were (1) Introduction, by Raye Litten; (2) Direct Ethanol Metabolites--On the Threshold From Science to Routine Use, by Friedrich M. Wurst; (3) Sialic Acid Index of Plasma Apolipoprotein J (SIJ) as a Viable Marker for Chronic Alcohol Consumption, by Philippe Marmillot; (4) The Emergence of Ethyl Glucuronide (EtG) Testing as a Tool in Monitoring Healthcare Professionals, by Gregory E. Skipper; (5) Application of Biomarkers for Alcohol Use Disorders in Clinical Practice, by Tim Neumann; (6) Utility of Biomarkers in Assessing the Efficacy of Medications for Treating Alcoholism, by Marty Javors; and (7) Discussion, by Raye Litten.

This article summarizes content proceedings of a symposium held at the 2004 International Society for Biomedical Research on Alcoholism Congress in Mannheim, Germany. The chairs were Boris Tabakoff and Friedrich M. Wurst. The presentations were (1) Genetic associations with alcoholism and affective disorders, by Paula Hoffman; (2) Proteomic analysis of blood constituents in alcoholism, by Boris Tabakoff; (3) Contrasts between the responses of GGT and CDT to high alcohol intake, and a test of their combined use, by John Whitfield; (4) Direct ethanol metabolites such as ethyl glucuronide, fatty acid ethyl esters, phosphatidylethanol and ethyl sulfate: a new line of sensitive and specific biomarkers, by Friedrich Martin Wurst; and (5) Genetic studies of alcoholism subtypes in a Han Taiwanese population, by Ru-Band Lu.