Marker Of Chronic Alcohol Abuse Research Articles

A Systematic Review of Solid-Phase Microextraction Applications in the Forensic Context.

Since the introduction in 1990, the solid-phase microextraction (SPME) technology has brought significant progress in many fields of forensic sciences due to the versatility of this fast and solventless alternative to conventional extraction techniques. A systematic review about SPME applications in forensic context from January 1995 to June 2018 was carried out according to systematic review guidelines. The majority of the reviewed articles (40/133) aimed to identify drugs (cannabinoids, cocaine, opiates, amphetamines, simultaneous detection of different drugs of abuse, prescribed drugs); 29 of the 133 articles focused on the investigation of fatalities; 28 of the 133 papers used headspace SPME technique for the identification of markers of chronic alcohol abuse. Sixteen papers involved this technique for the isolation of volatile organic compounds for the human odor profile and 20 concerned forensic applications regarding living people. Solid-phase microextraction was preferably employed in the headspace mode and many kinds of fibers were employed, although polydimethylsiloxane was the most adaptable to many forensic realities. Gas chromatography/mass spectrometry was more frequently used, probably for the well-established coupling with SPME. Most of the papers validated their method to harmonize the scientific approaches of procedures development. Good outcomes are reported on biological material collected from living people as well as on cadaveric samples. The results obtained by most of the studies about alcohol biomarkers on scalp hair have been adopted by the "Society of Hair Testing" to demonstrate abstinence over a pre-defined time period and to assess chronic excessive alcohol consumption.

Carbohydrate-deficient transferrin: utility of HPLC in handling atypical samples uninterpretable by capillary electrophoresis

Carbohydrate-deficient transferrin (CDT) is a marker of chronic alcohol abuse. Uninterpretable (atypical) CDT patterns have been detected by both capillary electrophoresis (CE) and HPLC. The aim of this study was to evaluate the performance of HPLC as a second-line test for the interpretation of most frequent atypical CDT profiles detected by CE. CDT was analyzed by CE (Capillarys 2, Sebia) on 9120 consecutive samples in a routine laboratory setting during a 2-year period. A commercial method (ClinRep CDT kit, Recipe) was employed to retest 123 (1.4%) samples with atypical CDT patterns on a Prominence LC-20AT HPLC (Shimadzu). CE-uninterpretable samples were categorized as having low transferrin (Tf) concentration (LT; n = 42, 0.5%), di-trisialotransferrin bridging (D-TB; n = 63, 0.7%) or atypical peak profile (APP; n = 18, 0.2%). CDT was detectable by HPLC in 58 of 123 (47%) samples including 21of 42 (50%) with LT, 27 of 63 (43%) with D-TB and 10 of 18 (56%) with APP. Second-line HPLC testing reduced uninterpretable samples by 47%, with similar rates of improvement regardless of the type of CDT pattern. The usefulness of HPLC as a second-line test for CDT should be evaluated according to cost-benefit considerations in the context of each laboratory.

WOMEN COINFECTED WITH HIV AND VHC. PART 1. PSYCHOSOCIAL CHARACTERISTIC AND READINESS TO ANTIVIRAL THERAPY

The objective of the present study was the analysis of medical and social status of women co-infected with HIV and VHC with the following assessment of their readiness to antiviral therapy of chronic hepatitis C virus infection (CHC) taking into account the mental health and living conditions and sustaining the adherence to antiviral therapy thereafter. Materials and methods. To clarify the social and psychological status, 100 outpatient cards of women co-infected with HIV/HCV were analyzed. The in-depth study with the assessment of the psychosocial and detoxification status and the adherence to antiretroviral therapy (ART) was held in fifty patients. Results. Most of the women had a long-standing history of HIV infection and HCV. Of these, 78% underwent ART and 84,6% had a high adherence to treatment. The main social problems of the women were connected with their work (34,2%) and family circumstances (26,3%). An increase in the concentration of carbohydrate-deficient transferrin (CDT), the marker of chronic alcohol abuse, was detected in three of 50 patients, and other three of the 50 had this parameter in the zone of unstable values. 72% of the patients showed mental disorders, while more than half of them (60%) had actively used psychoactive substances, and organic symptomatic mental disorder effects were observed in 20% of the patients. Most of the women were in remission on the use of psychoactive substances. Evaluation of the depression and anxiety levels has showed scattered results but in the meantime 54% of the women expressed complaints of asthenic nature, anxious mood was marked in 38% of the patients, 40% of them earlier had been suicidally inclined. Conclusion. Taking into account the social problems in one-third of patients that may negatively affect the treatment of HCV and HIV infection, it is essential to lend complimentary psychological support and assist with solving difficulties before the treatment assignment in order to increase the adherence to treatment. Due to mental health reasons, the patients need continuous observation and periodic condition correction. The majority of surveyed agree ART of CHC at this moment and in the short term but carefully relate to offered treatment regimens and its duration. Given the mental state of the patients, it can be assumed that virus C in women with HIV/HCV co-infection should be treated with direct-acting antiviral agents that do not exacerbate existing disorders. The next issue of the journal .HIV infection and immunosuppression disorders. (2018. Vol. 10, No. 4) will present the results of the study of clinical status features and the prospects for the therapy.

First objective association between elevated carbohydrate-deficient transferrin concentrations and alcohol-related traffic accidents.

Carbohydrate-deficient transferrin (CDT) is a well-recognized highly specific marker of chronic alcohol abuse. The association of CDT with alcohol-related traffic accidents was evaluated to objectively validate the use of this marker for certifying the physical fitness for driving license regranting after its confiscation for drunk driving. The study was carried out on 468 injured drivers (InjDr), who underwent mandatory blood alcohol concentration (BAC) and drug analysis in biological fluids. The InjDr group was divided into 2 subgroups on the basis of BAC legal limit adopted in Italy (BAC ≤ 0.5 g/l: InjDr1 ; BAC >0.5 g/l: InjDr2 ). The control group (CntDr) included 236 subjects holding safety-sensitive job positions and undergoing mandatory toxicological analyses. The determination of BAC in blood and CDT in serum were performed using validated analytical methods based on head-space gas chromatography and high-performance liquid chromatography, respectively. The evaluation of CDT distribution in the 3 groups (CntDr, InjDr1 , InjDr2 ) showed that CDT distribution in the InjDr1 group was similar to that observed in the CntDr group (p = 0.159) and different from that observed in the InjDr2 group (p < 0.001). Partitioning the CDT data of each group into "CDT positives" and "CDT negatives" on the basis of the cut off (1.90%), it was possible to calculate the odds of the 3 groups and then the odds ratios. The odds ratio of InjDr1 versus CntDr was 4.56 (p = 0.158), whereas the odds ratio of InjDr2 versus CntDr was 132 (p < 0.001). Furthermore, a dose-response effect was found only when comparing InjDr2 with CntDr. The data of the present study strongly support the use of the CDT test to evaluate the risk of a subject to be involved in a road accident while driving under the influence of alcohol.

Microheterogeneity of Serum β‐Hexosaminidase in Chronic Alcohol Abusers in a Driver's License Regranting Program

Carbohydrate-deficient transferrin (CDT) is one of the best indicators for chronic alcohol abuse and detection of relapse. In this study, we explore the microheterogeneity of β-hexosaminidase (β-HEX) in chronic alcohol abusers in the framework of a driver's license regranting program. Studies have shown that increased serum activity of β-HEX B (isoforms P, I, and B) may be a sensitive marker for chronic alcohol abuse. Here, we describe methodology, limitations, and correlation of β-HEX isoforms with CDT. CDT was assayed at the central laboratory of the Ghent University Hospital by capillary zone electrophoresis, measured on the Capillarys 2™ system and was expressed as a percentage of total serum transferrin (%CDT). Serum of chronic alcohol abusers was compared to nonheavy drinkers using agarose gel isoelectric focusing (IEF). Total β-HEX activity was assayed fluorimetrically following preparative IEF in 81 subjects. β-HEX isoforms were investigated and compared between nonheavy drinkers and heavy drinkers. Agarose gel IEF shows additional cathodal bands in serum of chronic alcohol abusers. Mean total β-HEX activity between pH 6.8 and 7.7, designated as HEX-7, showed the highest correlation with %CDT (r = 0.70, p < 0.0001, n = 68). In a selected subgroup, where CDT could not be quantified (n = 13) because of an atypical electropherogram, HEX-7 was in concordance with either estimated %CDT value or liver enzyme activities. In this proof-of-concept study, we introduce a novel approach to quantify β-HEX isoforms using preparative IEF and fluorimetry. A highly significant correlation of HEX-7 and %CDT has been found. Because of exclusion of the P isoform, HEX-7 could be a useful supplementary marker for detecting chronic alcohol abuse.

The effect of chronic alcohol intoxication and smoking on the activity of oral peroxidase

Peroxidase is the most important antioxidant enzyme in saliva. Through peroxidation of thiocyanate in the presence of H2O2, peroxidase catalyses the formation of bacteriocidic compounds such as hypothiocyanate. The purpose of this study was to evaluate the effect of chronic alcohol intoxication and smoking on the activity of oral peroxidase (OPO). A total of 37 volunteers participated in the study. This cohort consisted of 17 male alcohol-dependent smoking patients after chronic alcohol intoxication (AS group, alcohol + smoking) (mean age: 42 years; range: 26–55) (100–700 g/day of alcohol; 10–20 cigarettes/day) and 20 control male social drinkers (CNS group, control non-smokers) with no history of alcohol abuse or smoking (mean age: 42 years; range: 30–53). Salivary peroxidase activity was measured by the colorimetric method. The differences between groups were evaluated using the Mann–Whitney U test. There was significantly higher activity of OPO (p = 0.00001) and significantly lower salivary flow (SF) (p = 0.007) in alcohol-dependent smokers after chronic alcohol intoxication compared to the control group. OPO activity significantly correlated with the number of days of alcohol intoxication, but not with smoking. Gingival index (GI) was significantly higher in smoking alcohol-dependent persons than in the control group, and correlated with OPO activity. The sensitivity of the OPO test was 70% in smoking alcoholics, while specificity was 95%. The increased activity of OPO suggests chronic oxidative stress is more likely due to ethanol action than to smoking. Smoking alcohol-dependent persons have a worse periodontal status than controls. OPO activity as a marker of chronic alcohol abuse may help in the diagnosis of alcoholism.

The effect of chronic alcohol intoxication and smoking on the activity of oral peroxidase

Peroxidase is the most important antioxidant enzyme in saliva. Through peroxidation of thiocyanate in the presence of H₂O₂, peroxidase catalyses the formation of bacteriocidic compounds such as hypothiocyanate.The purpose of this study was to evaluate the effect of chronic alcohol intoxication and smoking on the activity of oral peroxidase (OPO). A total of 37 volunteers participated in the study. This cohort consisted of 17 male alcohol-dependent smoking patients after chronic alcohol intoxication (AS group, alcohol + smoking) (mean age: 42 years; range: 26-55) (100-700 g/day of alcohol; 10-20 cigarettes/day) and 20 control male social drinkers(CNS group, control non-smokers) with no history of alcohol abuse or smoking (mean age: 42 years; range:30-53). Salivary peroxidase activity was measured by the colorimetric method. The differences between groups were evaluated using the Mann-Whitney U test. There was significantly higher activity of OPO (p = 0.00001)and significantly lower salivary flow (SF) (p = 0.007) in alcohol-dependent smokers after chronic alcohol intoxication compared to the control group. OPO activity significantly correlated with the number of days of alcohol intoxication, but not with smoking. Gingival index (GI) was significantly higher in smoking alcohol-dependent persons than in the control group, and correlated with OPO activity. The sensitivity of the OPO test was 70% in smoking alcoholics, while specificity was 95%. The increased activity of OPO suggests chronic oxidative stress is more likely due to ethanol action than to smoking. Smoking alcohol-dependent persons have a worse periodontal status than controls. OPO activity as a marker of chronic alcohol abuse may help in the diagnosis of alcoholism.

Serum and plasma brain-derived neurotrophic factor (BDNF) in abstinent alcoholics and social drinkers

Although the effects of alcohol on brain-derived neurotrophic factor (BDNF) have been extensively studied in rodents, BDNF levels have rarely been measured in abstinent, alcohol-dependent (AD) individuals. Interpretation of reported group comparisons of serum BDNF levels is difficult due to limited information regarding analytical variance, biological variability, and the relative contribution of platelet and plasma pools to serum BDNF. Analytical variance (intra- and inter-assay coefficients of variation) of the enzyme-linked immunosorbent assay (ELISA) was characterized. Within- and between-subject variability, and group differences in serum and plasma BDNF, was assessed on three separate days in 16, 4-week abstinent AD individuals (7M/9F) and 16 social drinkers (SDs; 8M/8F). Significantly higher mean (±sd) serum BDNF levels were observed for the AD group compared to the SD (p = 0.003). No significant difference in mean baseline plasma BDNF levels was observed between AD and SD groups. The low analytical variance, high day-to-day within-individual stability and the high degree of individuality demonstrates the potential clinical utility of measuring serum BDNF levels. The low correlations that we observed between plasma and serum levels are congruent with their representing separate pools of BDNF. The observation of higher basal serum BDNF in the AD group without a concomitant elevation in plasma BDNF levels indicates that the elevated serum BDNF in AD patients is not due to greater BDNF exposure. Further research is warranted to fully elucidate mechanisms underlying this alteration and determine the utility of serum BDNF as a predictor or surrogate marker of chronic alcohol abuse.

A novel C2 transferrin variant interfering with the analysis of carbohydrate-deficient transferrin

BackgroundCarbohydrate-deficient transferrin (CDT) is used as a marker for chronic alcohol abuse. The presence of genetic transferrin variants might affect an individual's iron status and can interfere with CDT analysis. We report on the identification of a patient carrying a novel transferrin variant. We describe the performance of the various CDT methods in its detection and the associated iron status. MethodsDNA of the coding region of transferrin was sequenced and CDT levels were analysed using four different methods: high pressure liquid chromatography (HPLC), capillary zone electrophoresis (CZE), immunochemistry and iso-electric focussing (IEF). ResultsA novel transferrin variant, T139M, was found as a heterozygous genotype in the index patient and all of his four living direct family members (c.416C>p.Thr139Met). CDT analysis of the variant by HPLC and CZE was compromised as a result of the coelution of the different isoforms. CDT levels could be quantified by immunochemistry. Similar results were obtained using IEF analysis. The presence of the C2 transferrin variant did not affect iron status in any of the investigated patients. ConclusionsTransferrin T139M, present as a heterozygous genotype, interferes with CDT analysis by HPLC and CZE but not by immunochemistry. Physiologically, it appears to be functionally normal.

Improved capillary electrophoresis determination of carbohydrate-deficient transferrin including on-line immunosubtraction

The instrumental analysis of carbohydrate-deficient transferrin (CDT), a recognized marker of chronic alcohol abuse, is most commonly carried out by high-performance liquid chromatography (HPLC) or capillary zone electrophoresis (CZE). Between these two techniques, CZE shows higher efficiency and productivity, but is often reported to be inferior to HPLC in terms of selectivity, because of a less specific ultraviolet detection wavelength than HPLC. On these grounds, the present work was aimed at the development of an improved CZE method for CDT determination, including an on-line immunosubtraction step specifically aimed at enhancing the analytical specificity of CZE determination. The analytical conditions were as follows: uncoated fused silica capillary, 30 microm x 60 cm (L = 50 cm to detector); running buffer, 100 mmol/L borate and 6 mmol/L DAB (1,4-diaminobutane), pH 8.3; voltage, 30 kV; temperature, 25 degrees C; detection, 200 nm. Under the described CZE conditions, a baseline separation between all the CDT related peaks was achieved with good analytical performances in terms of both precision and accuracy. In order to achieve unequivocal recognition of the CDT peaks, an in-capillary immunosubtraction step was included by loading a plug of anti-human transferrin antibody solution after the sample plug. This analytical approach was applied successfully to recognize CDT peaks in the presence of potential interferences.

Carbohydrate-Deficient Transferrin - A Contemporary Biomarker in Comparison with Traditional Laboratory Markers of Chronic Alcohol Abuse

Carbohydrate-Deficient Transferrin - A Contemporary Biomarker in Comparison with Traditional Laboratory Markers of Chronic Alcohol AbuseTimely identification of excessive alcohol use and its potential complications is a prerequisite for successful treatment. Several routine tests have been used in laboratories that may help in diagnosing alcoholism, such as determination of MCV, AST, ALT, GGT, but it has been shown that they lack specificity and sensitivity. Contemporary bio-markers are increasingly being used today that may due to their unique characteristics help in discovering the onset of chronic alcohol abuse, as well as in abstinence and relapse monitoring. The term carbohydrate-deficient transferrin (CDT) stands for a small group of human transferrin isoforms (asialo, monosialo, and disialotran sferrin) with a lower degree of glycosylation in comparison to the dominant transferrin isoform (tetrasialotransferrin). Persons consuming large quantities of alcohol (≥50-80 g daily) over a period of at least two weeks have increased concentrations of transferrin isoforms lacking one (disialotransferrin) or both (asialotransferrin) carbohydrate chains. In this paper the traditional markers of chronic alcohol abuse (GGT, AST, ALT, and MCV) were determined, as well as the new biomarker CDT, after which diagnostic evaluation was performed and their usability and clinical value in routine laboratory practice were estimated. These markers were also determined in heavy alcoholics on admission into hospital and after two weeks of therapy, with the aim of estimating their diagnostic value for abstinence and relapse monitoring.

11-Nor-Δ 9-tetrahydrocannabinol-9-carboxylic acid ethyl ester (THC-COOEt): Unsuccessful search for a marker of combined cannabis and alcohol consumption

11-Nor-Δ 9-tetrahydrocannabinol-9-carboxylic acid ethyl ester (THC-COOEt) can be presumed to be a mixed metabolite formed during combined consumption of cannabinoids and alcohol. In order to examine this hypothesis, THC-COOEt and its deuterated analogue D 3-THC-COOEt were synthesized as reference substance and internal standard from the corresponding carboxylic acids and diazoethane and methods were developed for the sensitive detection of THC-COOEt in plasma and hair based on gas chromatography–electron impact mass spectrometry after silylation with N-methyl- N-tert-butyldimethylsilyl-trifluoroacetamide and gas chromatography–negative chemical ionization mass spectrometry (GC-NCI-MS) as well as tandem mass spectrometry (GC-NCI-MS-MS) after derivatization with pentafluoropropionyl anhydride. The methods were applied for THC-COOEt determination to plasma samples from 22 drunk driving cases which contained both ethanol (0.30–2.16 mg/g) and THC-COOH (15–252 ng/mL) as well as to 12 hair samples from drug fatalities which were both positive for THC (0.09–2.04 ng/mg) and fatty acid ethyl esters as markers of chronic alcohol abuse (0.70–6.3 ng/mg). In none of these samples THC-COOEt could be found with limits of detection of 0.3 ng/mL in plasma and 2 pg/mg in hair in 11 samples using GC-NCI-MS and 0.2 pg/mg in one sample using GC-NCI-MS. Therefore, the use of this compound as a marker for combined cannabis and alcohol consumption could not be achieved.

Carbohydrate-deficient transferrin (CDT): A reliable indicator of the risk of driving under the influence of alcohol when determined by capillary electrophoresis

Carbohydrate-deficient transferrin (CDT) is a marker of chronic alcohol abuse, which has recently been introduced to evaluate the physical fitness for obtaining a driving license. The aim of the present study was to evaluate the prevalence of elevated CDT levels in subjects stopped while driving under the influence of alcohol by using a validated method based on capillary electrophoresis. The study was carried out on a group of 40 drunken drivers (group A) and on a control group ( n = 51) of subjects chosen from the general population (group B). CDT was directly determined by capillary electrophoresis in free solution and UV detection at 200 nm. CDT results from both groups were classified as “negative” or “positive” on the basis of the cut-off set at 2.00% (CDT index). The subjects classified as “positive” in group A were 24 (60%), whereas in group B were 2. The subjects classified as “negative” in group A were 16 (40%), whereas in group B was 49 (96.1%). The comparison of the observed percentages, evaluated with the χ 2-test, was highly significant ( p < 0.001). The present study confirms the high prevalence of chronic alcohol abusers among drunken drivers and the usefulness of CDT as a predictor of the risk of drunk driving.

Current role of capillary electrophoretic/electrokinetic techniques in forensic toxicology

The current application of capillary electrophoresis in forensic toxicology has been critically reviewed with special focus on the areas where this technique has shown real advantages over chromatographic methods. For example, capillary electrophoresis has been most successfully applied to the chiral analysis of some drugs of forensic interest, including amphetamines and their congeners. Another typical application field of capillary electrophoresis is represented by protein analysis. Recently, special interest has been paid to carbohydrate deficient transferrin (CDT), the most important biological marker of chronic alcohol abuse. Other specific applications of capillary electrophoresis of potential forensic toxicological concern are also discussed. The review includes 62 references.

Relationship between serum sialic acid and sialylated glycoproteins in alcoholics

Total sialic acid (TSA) has been suggested as a marker for chronic alcohol abuse. It seems that the elevation of TSA during excessive alcohol consumption reflects the changes in sialylated glycoproteins in the sera. On the other hand, chronic ethanol consumption increases the desialylation rate of many serum glycoproteins. The aim of this study was to evaluate the relationship between the total and free form of sialic acid levels (FSA), and the concentration of sialylated glycoproteins in alcoholics. We determined the serum concentration of many glycoproteins (alpha1-antitrypsin, alpha1-acid glycoprotein, haptoglobin, ceruloplasmin, transferrin, complement C3 protein, fibrinogen and immunoglobulin G) in a sample of 100 alcoholics and 30 healthy controls. Total sialic acid was determined by the enzymatic method and its free form by using a modification of the thiobarbituric acid method. Among alcoholics, we found increased concentrations of alpha1-antitrypsin and alpha1-acid glycoprotein but decreased levels of transferrin. The concentrations of TSA and FSA were significantly higher in alcoholics than in healthy controls. The tested glycoproteins, except for transferrin and immunoglobulin G, positively correlated with TSA and FSA. The correlations with TSA were higher than that with FSA. Chronic alcohol abuse alters the concentrations of some sialylated glycoproteins in the sera. The alpha1-antitrypsin, alpha1-acid glycoprotein, and transferrin are the only affected glycoproteins. The serum level of total and free form of sialic acid in the sera of alcoholics depends on the concentration of the most sialylated glycoproteins.

Forensic analysis of carbohydrate-deficient transferrin (CDT) by HPLC—Statistics and extreme CDT values

Background Carbohydrate-deficient transferrin (CDT) is the most specific serum marker of chronic alcohol abuse so far. There is little knowledge about extreme CDT values of >20% and the more >30% CDT. Methods Serum CDT/transferrin ratios from 19,236 serum samples sent to our laboratory for routine CDT analysis were determined by HPLC. About 75% of these serum samples were from traffic or employment medicine investigations. A CDT value frequency histogram was computed and extreme CDT values were clinically validated. Results Fourteen thousand four hundred and sixty-one CDT results were normal (≤1.7%) and 4775 increased (1.8–36.9% CDT). Most frequent normal and increased results were 0.9% CDT ( n = 1964) and 1.8% CDT ( n = 356). CA. 70% of the pathological results were between 1.8% and 5.0% CDT, ca. 88% between 1.8% and 10.0% CDT, and 98% between 1.8% and 20.0%. CDT values >20.0% appeared in 79 cases and results >30.0% in two cases (33.8% and 36.9%). In each case of CDT values >20%, chronic alcohol abuse was the underlying cause as confirmed by anamnestic exploration. Conclusions CDT/transferrin ratios are usually <20%. Higher values can appear in rare cases. CDT results >30% can be due to alcohol abuse but should be considered as remarkable single observations. Visualization of the transferrin isoform patterns by HPLC allows the detection of pathological transferrin isoform patterns and of genetic transferrin variants. This is essential for a reliable interpretation of (extreme) CDT values. CDT analysis by immunoassays without physico-chemical confirmatory analysis is no longer acceptable.

Post-mortem stability and redistribution of carbohydrate-deficient transferrin (CDT)

Post-mortem diagnosis of chronic alcohol abuse is a challenge for forensic experts due to the lack of pathognomonic morphological findings and often also inadequate background information. Objective methods demonstrating chronic excessive alcohol consumption would therefore be a useful tool for forensic pathologists. In clinical practice, several markers of chronic alcohol abuse have recently been introduced, among which carbohydrate-deficient transferrin (CDT) is the most accepted, but the use of these markers in autopsy has not yet been established. We examined post-mortem stability and possible post-mortem redistribution of CDT and compared two analytical methods, capillary zone electrophoresis and high-performance liquid chromatography. According to our results, CDT remains stable for an appreciable time after death. The results further indicate that CDT is not subject to major post-mortem redistribution.

High prevalence of increased trisialotransferrin concentrations in patients with anorexia nervosa: Implications for determination of carbohydrate-deficient transferrin

Background Carbohydrate-deficient transferrin (asialo- + monosialo- + disialotransferrin, CDT) is currently the most specific laboratory marker of chronic alcohol abuse. We tested whether previous findings of false-positive CDT results for anorexia nervosa patients have been due to invalid CDT analysis methods or anorexia nervosa by itself. Methods Serum CDT from 49 anorexia nervosa patients, 14 bulimia nervosa patients and 22 healthy controls (all adolescent, female and age-matched) was determined in a retrospective study by HPLC (Clin-Rep®-CDT-in-serum-online, cut-off ≥ 1.8%, Recipe), by capillary electrophoresis (Capillarys-CDT, cut-off ≥ 1.3%, Sebia) and (due to limited surplus serum volume for a subset of 18 anorexia nervosa patients with increased trisialotransferrin detected by HPLC) by immunoassay based on anion-exchange CDT and non-CDT fractionation (%CDT-TIA, cut-off ≥ 2.6% CDT, Bio-Rad). Results HPLC and capillary electrophoresis: No false-positive CDT results were obtained. Asialo- and monosialotransferrin were not detected and disialotransferrin (CDT) was in each case clearly below the test-specific cut-offs. Trisialotransferrin (a non-CDT isoform) was increased (cut-off ≥ 5.0% for HPLC) in 33 anorexia patients, 2 bulimia patients and 2 controls. %CDT-TIA: 8 false-positive CDT results of ≥ 2.6% out of the 18 samples tested (CDT-range/mean/median 2.6–4.6/3.2/2.8%). Conclusions Anorexia nervosa does not cause by itself increased CDT results. False-positive CDT values from the past are most likely due to an incomplete separation of trisialotransferrin from CDT and thus overdetermination of CDT. Immunological CDT testing without confirmatory analysis by HPLC or CE is no longer acceptable.

Alanine Aminopeptidase Activity in Urine: A New Marker of Chronic Alcohol Abuse?

Alanine Aminopeptidase Activity in Urine: A New Marker of Chronic Alcohol Abuse?

Combining markers of nephrotoxicity and hepatotoxicity for improved monitoring and detection of chronic alcohol abuse

Most of the commonly used markers of chronic alcohol abuse reflect alcohol hepatotoxicity; however, such abuse is deleterious to the kidneys as well. Combined use of serum markers of liver origin and urinary markers of kidney origin may be of diagnostic advantage. The study was performed in 73 male alcoholics undergoing detoxification and 36 male alcoholics who had maintained abstinence for > or =6 weeks. Factor analysis, discriminant analysis and receiver operating characteristic (ROC) analysis were used to assess the discriminative power of two urinary markers of alcohol nephrotoxicity, namely beta-N-acetylhexosaminidase (Hex, EC 3.2.1.52) and alanine aminopeptidase (EC 3.4.11.2), and of three serum markers of alcohol hepatotoxicity, namely aspartate aminotransferase (EC 2.6.1.1), alanine aminotransferase (EC 2.6.1.2) and gamma-glutamyltransferase (GGT, EC 2.3.2.2), and of their quantitative combinations. The discriminative power of the urinary markers matched that of the serum markers. Hex and GGT appeared to be the best for discriminating the study groups. Their combination given by the equation G&H=0.62 x ln(GGT)+0.72 x ln(Hex) showed excellent discriminative ability (ROC area under the curve 0.92) that was significantly higher than that of any single marker in this report, indicating superior diagnostic accuracy of the compound marker. Kidney-derived urinary markers, particularly Hex, can complement or replace, if necessary, serum markers of chronic alcohol abuse that relate to alcohol hepatotoxicity. The compound marker proposed seems a promising tool for the monitoring and perhaps detection of chronic alcohol abuse and warrants further studies.