Article

Detecting alcohol abuse: Traditional blood alcohol markers compared to ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEEs) measurement in hair

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  • Lipidomix GmbH
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Abstract

Alcohol abuse is a common problem in society; however, the technical capabilities of evaluating individual alcohol consumption using objective biomarkers are rather limited at present. In recent years research has focused on alcohol markers using hair analysis but data on performance and reliable cut-off values are still lacking. In this study 169 candidates were tested to compare traditional biomarkers, such as carbohydrate-deficient-transferrin (CDT), gamma glutamyl transferase (GGT), aspartate amino transferase, alanine amino transferase and the mean corpuscular volume of the erythrocytes, with alcohol markers detectable in hair such as ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEEs). This study revealed that EtG, GGT and CDT showed the best results, demonstrating areas under the curve calculated from receiver operating characteristics of 0.941, 0.943 and 0.899 respectively. The lowest false-negative and false-positive rates were obtained by using a combined interpretation system for hair EtG and FAEEs. All markers demonstrated only low to moderate correlations. Optimum cut-off values for differentiation between social and chronic excessive drinking calculated for hair EtG and FAEEs were 28 pg/mg and 0.675 ng/mg, respectively. The critical values published in the "Consensus on Alcohol Markers 2012" by the Society of Hair Testing were confirmed.

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... Total energy ex- penditure (kcal/24 h) considered resting metabolic rate and was mea- sured by the Actical® activity monitor (Mini Mitter Co., Inc., Bend). Serum γ-glutamyl transferase (γ -GT) was measured as a marker for alcohol consumption [23], while serum cotinine values were taken as a measure of smoking status [24]. ...
... Normal distribution of data was tested, and logarithmic transformations were done for physical activity, cotinine, γ-GT, CRP, estradiol, renin, aldos- terone, eGFR and PHQ-9 data. A priori covariates included age, body surface area, physical activity, γ-GT [23] and cotinine [24]. ...
... Both ethnic groups showed co-morbidity for chronic depression plus DBP-HT, but depression was related to RAAS desensitization only in the Blacks. The higher alcohol consumption in Blacks should be considered, as levels at baseline and follow-up exceeded the value of chronic al- cohol abuse according to Hastedt et al. (> 41 U/L; sensitivity 83%, specificity 92%) [23]. This trend may explain the higher eGFR levels suggesting hyper-filtration due to endothelial dysfunction, which was not evident in Whites. ...
... Epidemiological studies consistently report worldwide increases in the percentage of alcohol users, in the amount consumed per capita and in the extent of alcohol-related mortality and disability, along with a decrease in the age of first use [1]. In 2015, the World Health Organization stated that almost 6% of worldwide deaths are linked to alcohol abuse, with mortality related to hazardous drinking increasing significantly (up to 25%) among young people (age group [20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37][38][39] [1,2]. ...
... A noteworthy limitation of PEth when compared to hEtG, however, is its shorter window of detection, as it is completely eliminated from circulating blood in 2-3 weeks for long-term alcohol abusers and less for social drinkers, with diagnostic sensitivity decreasing over time [2,3,23]. Measurement of FAEEs in hair (hFAEEs), on the other hand, shows comparable detection window and sensitivity for chronic alcohol abuse as hEtG, but it has poor diagnostic specificity as it is highly susceptible to false positives secondary to routine use of ethanol-containing hair products [3,10,22,24]. hFAEE analysis is therefore currently indicated only as a complementary test to hEtG, as it is less susceptible than the latter to false negatives caused by chemical hair treatments [8]. Finally, EtS and EtP are not routinely employed in toxicological analysis because of the paucity of studies about them, their short window of detection (in blood and urine), poor correlation with amount of alcohol consumed, and low specificity (EtS) and sensitivity (EtP). ...
... These tests are routinely available and relatively inexpensive, however they are not specific to alcohol consumption and can be influenced by several factors, including gender, age, comorbidities (especially non-alcoholic liver disease), medications and smoking [2,3,6,22,29]. As compared to traditional markers, hEtG displays higher diagnostic sensitivity and specificity (respectively 0.85 and 0.97, when using a cut-off of 30 pg/mg in the proximal 3-cm hair segment to detect alcohol consumption >60 g/day [18]), longer window of detection (3-6 months compared to few weeks for indirect markers) and better correlation with amount of alcohol consumed [3,10,22,24,29]; combining it with any other biomarker does not improve the diagnostic performance of hEtG alone [3,4], except in selected circumstances (e.g. suspected false negatives due to hair treatments [30,31] and suspected false positives due to EtG-contaminated hair products [32,33]), in which case concomitant hair FAEE analysis can confirm the result or prompt further investigations [8]. ...
Article
Ethyl glucuronide (EtG) is a minor, non-oxidative ethanol metabolite that can be detected in several matrices (e.g. blood, urine, hair, meconium) for variable periods of time. Quantification of EtG in hair (hEtG) has established itself, over recent years, as one of the most reliable biomarkers of long-term alcohol consumption habits, with the Society of Hair Testing (SoHT) offering cut-off values for assessment of both abstinence and heavy drinking (>60 g/day). Despite its high diagnostic performance, however, issues concerning inter- and intra-laboratory variability as well as data interpretation are still being investigated and represent the ultimate barrier to widespread acceptance of hEtG in the forensic context. The aim of this review is to summarize currently available analytical methods of hEtG testing, provide a framework to understand current hEtG cut-offs and their possible upcoming changes (in particular, a lower abstinence cut-off has been proposed for the 2019 revision of the SoHT consensus), and offer a schematic but exhaustive overview of the pitfalls in result reproducibility and interpretation that may limit applications of hEtG testing in the forensic context. Ultimately, the purpose of the authors is not to undermine the reliability of hEtG as an alcohol use marker, but rather to enhance it by promoting familiarization with all aspects related to it, from ethanol pharmacokinetics and EtG incorporation into hair, to sample preparation and analytical methods, to specific cases warranting close attention and additional tests for correct interpretation of hEtG results.
... An important goal of forensic and clinical toxicology is to identify appropriate biological markers of ethanol consumption to evaluate harmful drinking [1][2][3][4][5] or to ascertain alcohol abstinence [6][7][8]. With these aims, the detection of ethylglucuronide (EtG) in hair represents nowadays the approach with the highest diagnostic performances [9,10]. ...
... The Receiver Operation Characteristics (ROC) curve [11] is an efficient statistical method, used to evaluate the discrimination power of a certain biomarker and select a suitable cutoff with optimized sensitivity and specificity. This technique was consistently applied in hair EtG studies to compare the performance of this molecule with that of other excessive alcohol consumption biomarkers [4,9,[12][13][14][15][16]. In all these studies, the diagnosis of chronic alcohol abuse by means of hair EtG determination was based on a cut-off ranging from 25 and 30 pg/mg, which resulted highly accurate (AUC > 0.9). ...
... This technique was consistently applied in hair EtG studies to compare the performance of this molecule with that of other excessive alcohol consumption biomarkers [4,9,[12][13][14][15][16]. In all these studies, the diagnosis of chronic alcohol abuse by means of hair EtG determination was based on a cut-off ranging from 25 and 30 pg/mg, which resulted highly accurate (AUC > 0.9). In five of the aforementioned papers [4,9,[13][14][15], the sample treatment The quantification of ethylglucuronide (EtG) in hair is nowadays recognized as the approach with the highest diagnostic performance to evaluate harmful drinking. A widely accepted cut-off of 30 pg/mg has been selected after several accurate compared studies. ...
Article
The quantification of ethylglucuronide (EtG) in hair is nowadays recognized as the approach with the highest diagnostic performance to evaluate harmful drinking. A widely accepted cut-off of 30pg/mg has been selected after several accurate compared studies. While most of the studies that were used to establish the appropriate cut-off value prescribed to cut hair into small segments before their extraction, hair milling has subsequently been identified as the most efficient pretreatment procedure and was therefore recommended in the last Consensus document issued by the Society of Hair Testing. In this study, we initially compared the results obtained with the two sample preparations, namely cutting and milling, both being applied to the same specimens (n=781). Among these, 205 samples produced measurable EtG values with both methods, with differences ranging from -41.7% up to +415% (the mean increase in EtG concentration, switching from cutting to milling, was +62.1% and the median was +42.3%). Among the aforementioned 205 samples, 29 specimens (3.7% of the total 781 samples) produced significantly different outcome, being classified as negative (i.e., below 30pg/mg) if the cutting procedure is used, but largely positive (above 40pg/mg) when milling is used. Subsequently, the positivity rates obtained on a large population dataset (>27,000 samples) with the two procedures, were retrospectively compared using variable cut-offs values. The percentage of head hair samples with EtG concentration exceeding 30pg/mg upon application of the milling procedure shows a 45% increase (from 10.9% to 15.8%) with respect to cutting procedure, whereas the fraction of hair samples with EtG exceeding 40pg/mg (10.5%) overlaps the percentage of positive samples obtained after cutting pretreatment and applying a cut-off of 30pg/mg. On the basis of these results, it would be worth considering the application of cut-off values linked with the pretreatment procedure, taking into account the results of forthcoming inter-laboratory calibrations.
... Indeed, these compounds have proven to allow effectively for the estimation of alcohol exposure surpassing some limitations, such as the lack of specificity presented by indirect biomarkers, such as enzyme analysis and mean corpuscular volume (Hastedt et al., 2013;Morini et al., 2011;Pragst et al., 2010). ...
... determination of several drugs of abuse, therein including alcohol biomarkers, as mostly these compounds present higher stability in this sample when compared with others as is the case of blood (Auwärter et al., 2001;Barroso, Gallardo, Vieira, López-Rivadulla, & Queiroz, 2011). Hair also presents other advantages, for example, the drinking behaviour for months before sample collection can be evaluated through segmental hair analysis of 0-3 or 0-6 cm proximal segments (Hastedt et al., 2012, Hastedt et al., 2013Kintz, 2015;Pragst, 2015;Suesse, Blueml, & Pragst, 2015). Additionally, the sample collection procedure is non-invasive, and the chances of adulteration are very low (Barroso et al., 2011;Pragst & Balikova, 2006). ...
... GC coupled to MS or GC-MS/MS methods are also used, but require previous derivatization (Crunelle et al., 2014;Oppolzer et al., 2016;Pragst et al., 2010). FAEEs are hydrophobic compounds, and their extraction from hair is usually performed using non-polar solvents; sample clean-up is mostly performed using headspace solid-phase micro-extraction (HS-SPME) and analysis by GC-MS (Auwärter et al., 2001;Cabarcos et al., 2009;Hastedt et al., 2012Hastedt et al., , 2013Pragst, Auwaerter, Sporkert, & Spiegel, 2001;Pragst et al., 2010;Oppolzer et al., 2016;Suesse et al., 2012), while a few papers describe solid phase extraction (SPE) procedures (Bertol, Bravo, Vaiano, Mari, & Favretto, 2014;Caprara, Klein, & Koren, 2005;Politi, Mari, Furlanetto, Del Bravo, & Bertol, 2011). Combined use of both biomarkers mentioned above has shown to increase the accuracy of alcohol consumption interpretation (Hastedt et al., 2012;Pragst et al., 2010;Suesse et al., 2012). ...
Article
Hair testing for alcohol biomarkers is an important tool for monitoring alcohol consumption. We propose two methods for assessing alcohol exposure through combined analysis of ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEEs) species (ethyl myristate, palmitate, stearate and oleate) in hair (30 mg). EtG was analysed by liquid chromatography-tandem mass spectrometry, while FAEEs were analysed by gas chromatography-tandem mass spectrometry using electron impact ionization. Both methods were validated according to internationally accepted guidelines. Linearity was proven between 3-500 pg/mg for EtG and 30-5000 pg/mg for FAEEs, and the limits of quantification were 3 pg/mg for EtG and 30 pg/mg for each of the four FAEEs. Precision and accuracy were considered adequate, processed EtG samples were found to be stable for up to 96 h left in the injector and processed FAEEs samples for up to 24 h. Matrix effects were not significant. Both methods were applied to the analysis of 15 authentic samples, using the cut-off values proposed by the Society of Hair Testing for interpretation. The results agreed well with the self-reported alcohol consumption in most cases, and demonstrated the suitability of the methods to be applied in routine analysis of alcohol biomarkers, allowing monitoring consumption using low sample amounts.
... Health care expenditures, business and criminal justice costs associated to alcohol-related problems amount to hundreds of billions of dollars yearly, and even a greater economic burden is sustained when alcohol addictive behaviors remain untreated. Over the last decade, numerous scientific studies focused on improving the diagnosis of chronic excessive alcohol consumption to efficiently identify individuals in need of recovery programs, health care, therapeutic monitoring, etc. [1][2][3]. ...
... No changes were observed in the correct classification rates nor in the information loss (Table 1).To stretch the system even further, a naïve multivariate model with only the FAEEs and EtG variables was built (LR 2 , Fig. 3a). In general, FAEEs and EtG proved to provide the best discrimination between non-chronic and chronic alcohol abusers, while the contribution of indirect biomarkers is negligible, as several other independent studies concluded [1,3,8,9,13,45]. ...
... Moderate (M) support to H 2 10 À1 LR < 1 Limited (L) support to H 2 LR = 1 Inconclusive support to both hypotheses 1 < LR 10 1 Limited (L) support to H 1 10 1 < LR 10 2 Moderate (M) support to H 1 10 2 < LR 10 3 Moderately strong (MS) support to H 1 10 3 < LR 10 4 Strong (S) support to H 1 LR > 10 4 Very strong (VS) support to H 1 of the experimental data. Furthermore, LR approaches take into account the information about the rarity of the physicochemical data, allowing the identification of anomalous values that might have been influenced by external factors (e.g. ...
Article
The detection of direct ethanol metabolites, such as ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEEs), in scalp hair is considered the optimal strategy to effectively recognize chronic alcohol misuses by means of specific cut-offs suggested by the Society of Hair Testing. However, several factors (e.g. hair treatments) may alter the correlation between alcohol intake and biomarkers concentrations, possibly introducing bias in the interpretative process and conclusions.
... A 3 cm segment of hair corresponds roughly to a 3-month history of drinking pattern [143,144]. Results of recent studies correlating hair EtG with alcohol consumption patterns are shown in Table 3 [130,[145][146][147][148][149][150][151][152][153][154][155][156][157][158]. ...
... FAEEs in hair samples show 59.3% sensitivity and 91.0% specificity for heavy drinking at a cut-off level of 0.675 ng/mg [157]. FAEEs were assessed in a large sample of 1057 autopsy cases (168 social drinkers, 502 alcohol abusers and 387 unknown). ...
... ng/mg) among social drinkers and 1.346 ng/mg (range 0.010-83.7 ng/mg) among alcohol abusers. Based on these findings, the optimal cut-off value for differentiating social drinkers from alcohol abusers was calculated at 1.08 ng/mg [157]. ...
Article
Full-text available
The quantitative, measurable detection of drinking is important for the successful treatment of alcohol misuse in transplantation of patients with alcohol disorders, people living with human immunodeficiency virus that need to adhere to medication, and special occupational hazard offenders, many of whom continually deny drinking. Their initial misconduct usually leads to medical problems associated with drinking, impulsive social behavior, and drunk driving. The accurate identification of alcohol consumption via biochemical tests contributes significantly to the monitoring of drinking behavior. A systematic review of the current methods used to measure biomarkers of alcohol consumption was conducted using PubMed and Google Scholar databases (2010-2015). The names of the tests have been identified. The methods and publications that correlate between the social instruments and the biochemical tests were further investigated. There is a clear need for assays standardization to ensure the use of these biochemical tests as routine biomarkers. Alcohol ingestion can be measured using a breath test. Because alcohol is rapidly eliminated from the circulation, the time for detection by this analysis is in the range of hours. Alcohol consumption can alternatively be detected by direct measurement of ethanol concentration in blood or urine. Several markers have been proposed to extend the interval and sensitivities of detection, including ethyl glucuronide and ethyl sulfate in urine, phosphatidylethanol in blood, and ethyl glucuronide and fatty acid ethyl esters in hair, among others. Moreover, there is a need to correlate the indirect biomarker carbohydrate deficient transferrin, which reflects longer lasting consumption of higher amounts of alcohol, with serum γ-glutamyl transpeptidase, another long term indirect biomarker that is routinely used and standardized in laboratory medicine.
... Specific gamma glutamyl transferase (g-GT) cut-points associated with 24-h hypertension have been developed accordingly (Oosthuizen et al., 2016). To further explore the detrimental effect of excessive alcohol consumption between ethnicities e regardless of the presence of fatty liver disease and oxidative stress (Hastedt et al., 2013;Tsai, Ford, Li, & Zhao, 2012) e these ethnic-specific g-GT cutpoints may be utilized to determine the degree of alcohol consumption and associated risk markers pertaining to cardiac perfusion as well as electrical and structural alterations. ...
... The Actical ® accelerometer (Montr eal, Qu ebec) determined total energy expenditure (TEE) calculated in calories per 24 h; the resting metabolic rate was also taken into account. The degree of alcohol consumption was determined using g-GT as a biochemical marker (Hastedt et al., 2013;Oosthuizen et al., 2016). The following cut-points were used: <19.5 U/L was indicative of low alcohol consumption, and !19.5 U/L was indicative of alcohol abuse in Caucasian participants; while <55.7 U/L and !55.7 U/L indicated low and excessive alcohol consumption, respectively, in Africans (Oosthuizen et al., 2016). ...
... Metabolic perturbation in a smaller cohort of hypertensive Black men was associated with alcohol abuse and support our findings where alcohol abusers revealed a vulnerable cardio-metabolic profile in high g-GT vs. low g-GT in both African and Caucasian groups (van Deventer et al., 2015). According to Hastedt et al. (2013) and Tsai et al. (2012), g-GT accurately indicates the degree of alcohol consumption, regardless of the presence of fatty liver disease and oxidative stress. ...
Article
Alcohol contributes greatly to vascular and structural modifications. Due to differences in the metabolism and tolerance of alcohol between ethnic groups, the manner of these modifications may differ. We investigated the association between alcohol consumption – measured via ethnic specific gamma glutamyl transferase (γ-GT) cut-points – and markers of cardiac perfusion, electrical activity and pre-clinical structural alterations. A South African target population study was performed in a bi-ethnic gender cohort (N=405). Alcohol consumption was determined according to previously defined ethnic-specific γ-GT cut-points, where γ-GT ≥ 19.5U/L and γ-GT ≥55U/L indicated excessive alcohol consumption in Caucasians and Africans respectively. Ambulatory 24h BP and-electrocardiograms (ECG), 12-lead ECG-left ventricular hypertrophy (LVH), ischemic events, N-terminal pro-brain natriuretic peptide (NT-proBNP) and QTc prolongation were assessed. Fasting blood samples were obtained. A poorer cardio-metabolic profile, mean 24h hypertensive and ECG-LVH values were evident in high γ-GT groups of both ethnicities, when compared to their low counterparts. The African high γ-GT group reported a higher intake of alcohol high and presented significant increases in NT-proBNP (p<0.001), QTc prolongation (p=0.008) and ischemic events (p=0.013). Regression analyses revealed associations between ECG-LVH and NT-proBNP, QTc prolongation, ischemic events and SBP, in the African high γ-GT group exclusively. High alcohol consumer’s presented delayed electrical conduction in the heart accompanied by ECG-LVH, ischemic events, and increased vaso-responsiveness predominantly in Africans. Ultimately, increased left ventricular distension on a pre-clinical level may elevate the risk for future cardiovascular events in this population.
... [6] However, supportive evidence for γ-GT as behavioral adjustment to psychosocial stress in Africans has been demonstrated. [7] It was also found that γ-GT correlated well with other markers for alcohol abuse. [7] Another important factor in the development of cardiovascular disease (CVD) is the effect of daily stress and how each individual reacts to and copes with such stress. ...
... [7] It was also found that γ-GT correlated well with other markers for alcohol abuse. [7] Another important factor in the development of cardiovascular disease (CVD) is the effect of daily stress and how each individual reacts to and copes with such stress. [8] Different coping styles exist, each exhibiting a different psychological stress response. ...
... The data was then converted into 1-second intervals for analysis. γ-GT levels were measured as marker for alcohol abuse, [7,20] and for each participant's smoking habits, cotinine was measured. ...
Chapter
Full-text available
Objective: Vulnerability for hypertension in Africans increases when they are exposed to psychosocial stress or urbanization. Inevitable cultural and lifestyle changes may impact on cardiomyopathy risk markers. Therefore, we aimed to assess the relationship between blood pressure, silent ischemia, glycated hemoglobin (HbA1c), QTc dispersion and ECG Cornell Product left ventricular hypertrophy (LVH) in a cohort of African men. Methods: We included 100 Caucasian and 82 African men, aged 25- 60 years, from the Sympathetic activity and Ambulatory Blood Pressure in Africans (SABPA) Study. HbA1c as well as ambulatory blood pressure (BP) and -silent ischemia values were evaluated, compared and correlated. QTc dispersion and Cornell product data were obtained via 12-lead ECG. Results: African men revealed (p = 0.05) pre-diabetic status with a mean HbA1c (> 6%). In addition, they revealed increased ambulatory BP, silent ischemia, QTc and LVH values compared to the Caucasian men. In forward stepwise regression analyses [Adjusted R2 for model, 0.33], ambulatory systolic BP (0.53, 95% CI 0.12 to 0.94) and silent ischemia (0.26, 95% CI 0.06 to 0.46) were associated with LVH in African men. Conclusion: Systolic BP and silent ischemia were associated with LVH in a cohort of African men. Increased cardiovascular disease risk markers may reveal progressing cardiomyopathy.
... [6] However, supportive evidence for γ-GT as behavioral adjustment to psychosocial stress in Africans has been demonstrated. [7] It was also found that γ-GT correlated well with other markers for alcohol abuse. [7] Another important factor in the development of cardiovascular disease (CVD) is the effect of daily stress and how each individual reacts to and copes with such stress. ...
... [7] It was also found that γ-GT correlated well with other markers for alcohol abuse. [7] Another important factor in the development of cardiovascular disease (CVD) is the effect of daily stress and how each individual reacts to and copes with such stress. [8] Different coping styles exist, each exhibiting a different psychological stress response. ...
... The data was then converted into 1-second intervals for analysis. γ-GT levels were measured as marker for alcohol abuse, [7,20] and for each participant's smoking habits, cotinine was measured. ...
Chapter
Full-text available
Introduction: Alcohol abuse is among the Westernized world's most common lifestyle factors contributing to sensitization of vascular structure and pathology. Gamma glutamyl transferase (γ-GT), as a marker of alcohol abuse, may indicate increased sensitization and vascular disease risk when using chronic defense coping (DefS). DefS was contradicted as a promoter of health, as it was related with dissociation between self-reported behavioral ("in-control") albeit physiological ("loss-of-control") responses. Therefore, we aimed to assess if DefS may mask autonomic dysfunction and induce alcohol-related subclinical atherosclerosis. Methods: A black African male teacher cohort with similar socioeconomic standing and without atrial fibrillation (N=101) from the North-West Province of South Africa, was recruited and investigated. Fasting blood values, 24h ambulatory blood pressure (ABPM) and -ECG, Coping Strategy Indicator (CSI) scores and ultrasound left carotid intima media far wall scanning values, were obtained. The standard deviation between successful R-R intervals (SDNN) was utilized as a marker of time-domain 24h heart rate variability (HRV). Multivariate regression analyses were computed. Results: A hypertensive state in the men was evident at a much lower ROC γ-GT cut point than the suggested MERCK cut-point of 78 U/L [55.7 U/L, sensitivity/specificity 60%/86% (AUC 0.78; 95% CI: 0.67; 0.88]. Hereafter, males were stratified into low (<55.7 U/l) and high (≥ 55.7 U/l) γ-GT groups. Overall, the high γ-GT group, revealed a poorer lifestyle profile, mean higher 24h ABPM, depressed 24h HRV and more 24h silent ischemic events (14 vs. 2) compared to their low γ-GT counterparts (P ≤ 0.01). HRV responses were inversely associated with γ- GT when utilizing DefS (r = -0.28; P = 0.015). Additionally, ABPM predicted 24h silent ischemia (OR 1.11; P ≤ 0.01) whereas 24h silent ischemia predicted a trend for structural vascular disease in the DefS African men (OR 1.06; p ≤ 0.06). Conclusion: DefS revealed behavioral "in-control" responses directly opposing or masking physiological "loss-of-control" responses, underpinning a dissociative response. Alcohol, which is a central nervous system depressant, enhanced vasoconstriction in the vasculature. Indeed, alcohol abuse was related to depressed heart rate variability, when utilizing DefS, sensitizing the vasculature, induced silent ischemia and ultimately structural vascular remodeling. The detrimental effects of DefS acting in tandem with autonomic dysfunction augment alcohol-related ischemia and may increase the risk for early sub-clinical atherosclerosis.
... Objective screening for alcohol use can provide an opportunity to intervene for improving patient health and quality of life. However, in the absence of specific and sensitive biomarkers for alcohol abuse, physicians often fail to recognize such patterns 1,7,8 . Questionnaires sometimes provide unreliable information or may not be applicable to all patients. ...
... Compared to other short term and long term biomarkers, it is a more specific and sensitive marker of alcohol abuse 10,11 , which can be applied for both clinical and forensic analysis 14,15 . Many analytical methods have been developed and validated for detection and quantization of EtG in the biological matrices, such as nuclear magnetic resonance (NMR) 16 , capillary zone electrophoresis (CZE) 7 , liquid chromatography-mass spectrometry (LC-MS) 14 and gas chromatography-mass spectrometry (GC-MS) 8,15 . But because of the complex nature of urine and serum, these methods are frequently hindered by the matrix compounds co-eluting with EtG in urine and serum. ...
... For extraction of biological samples liquid as well as solid phase extraction techniques was applied. SPE has advantage over liquidliquid extraction (8) as it is humble, rapid accurate with high degree of recovery (≥85%) clear extracts were obtained. This simple, sensitive & specific method can be functional for extraction and quantification of many samples at a time. ...
... Gamma-glutamyl transferase (γ-GT) was measured as a marker for alcohol consumption (19) and cotinine levels as indicator of smoking habits (20). Anthropometric measures were conducted in triplicate by level II anthropometrists (as accredited by the International Society for the Advancement of Kinanthropometry) using calibrated instruments and standardized methods. ...
... The use of γ-GT as a marker of alcohol consumption has been questioned as levels may also be increased in people for non-alcohol-related liver conditions, such as fatty liver disease (10). However, the use of γ-GT has recently been compared to direct and exclusive alcohol markers and it was declared suitable as an indicator of alcohol abuse (19). Whether genetic variations influence the effect of alcohol on the cardiovascular system between ethnicities is still unclear. ...
Article
The behavioral defense coping response (DefS) as a measure of coping with emotional stress may increase alcohol intake (gamma glutamyl transferase (γGT)), the risk for coronary artery disease (CAD) and insulin sensitivity (homeostasis model assessment, HOMA). We assessed associations between coping and cardiometabolic risk markers in a bi-ethnic cohort (N = 390) from South Africa. Ambulatory blood pressure (BP) and ECG, fasting blood and coping scores were obtained. Africans, and mostly when utilizing DefS, showed higher 24h BP, a low-grade inflammatory state, central obesity, increased HOMA [4.07 (3.66, 4.47)] and more ST events compared to their Caucasian counterparts. ROC γ-GT analyses predicting 24-h ambulatory hypertension showed a higher γ-GT cut-point in Africans (55.4 U/l) than in Caucasians (19.5 U/l). Odds ratios (ORs) of γ-GT cut-points predicting 24-h ambulatory hypertension was evident in DefS African men [OR: 7.37 (95% CI: 6.71-8.05), p = 0.003] and in DefS Caucasians, albeit at a lower γ-GT cut-point (19.5 U/l). Higher γ-GT cut-points in DefS Africans or Caucasians were not associated with HOMA > 3. DefS accompanied by alcohol abuse in taxing emotional situations, if no social support is forthcoming, underscores a profile of reduced coronary perfusion. It may enhance vasoconstriction of the coronary arteries, with compensatory increases in BP, and induce a risk for future coronary artery disease.
... Alcohol abuse was independently associated with BP, transient 24 h ischemic events, autonomic dysfunction and structural vascular disease in Ghana (Agyemang 2006), Uganda (Bosu 2010;Kotwani et al. 2013) and South Africa (Malan et al. 2013Oosthuizen et al. in press). Gamma glutamyl transferase, a marker of alcohol consumption (Hastedt et al. 2013 (Zatu et al. 2016). ...
... The impact of alcohol abuse in Blacks may thus be detrimental to subcortical and vascular health in Blacks. Alcohol is a central nervous system depressant, which can induce higher metabolic demands and vascular depression (Hastedt et al. 2013;Kobayashi et al. 1997;Van Deventer and Lindeque 2015). Therefore, statistical adjustment for alcohol abuse in data analyses fails to provide a clear answer when considering the effect of ethanol on the brain, vasculature and metabolism. ...
Chapter
Globally most interventions focus on improving lifestyle habits and treatment regimens to combat hypertension as a non-communicable disease (NCD). However, despite these interventions and improved medical treatments, blood pressure (BP) values are still on the rise and poorly controlled in sub-Saharan Africa (SSA). Other factors contributing to hypertension prevalence, such as chronic emotional stress, might provide some insight for future health policy approaches. Currently, Hypertension Society guidelines do no mention emotional stress as a probable cause for hypertension. Recently the 2014 World Global Health reports, suggested that African governments should consider using World Health Organization hypertension data as a proxy indicator for social well-being. However, the possibility that a stressful life and taxing environmental factors might disturb central neural control of BP regulation has largely been ignored in SSA. Linking emotional stress to vascular dysregulation is therefore one way to investigate increased cardiometabolic challenges, neurotransmitter depletion and disturbed hemodynamics. Disruption of stress response pathways and subsequent changes in lifestyle habits as ways of coping with a stressful life, and as probable cause for hypertension prevalence in SSA, may be included in future preventive measures. We will provide an overview on emotional stress and central neural control of BP and will include also implications thereof for clinical practice in SSA cohorts.
... The trustworthy assessment of harmful drinking represents a major commitment for forensic and clinical toxicologists and requires both a careful evaluation of suggestive medical evidence and the execution of laboratory analysis aimed at the detection of excessive alcohol consumption biomarkers. [1][2][3][4][5][6][7][8][9][10][11] The latter include indirect biomarkers and direct alcohol metabolites, with a wide range of reliability, depending on the proportion of correct classification of the investigated individuals among harmful drinkers, social drinkers, and teetotallers. 3,[12][13][14][15][16] In recent years, the determination of ethyl glucuronide (EtG) in the keratinous matrices has gained increasing appreciation, since it achieves the highest combination of sensitivity and specificity in the discrimination among alcohol consumers with different drinking habits. ...
... [1][2][3][4][5][6][7][8][9][10][11] The latter include indirect biomarkers and direct alcohol metabolites, with a wide range of reliability, depending on the proportion of correct classification of the investigated individuals among harmful drinkers, social drinkers, and teetotallers. 3,[12][13][14][15][16] In recent years, the determination of ethyl glucuronide (EtG) in the keratinous matrices has gained increasing appreciation, since it achieves the highest combination of sensitivity and specificity in the discrimination among alcohol consumers with different drinking habits. 2,[16][17][18] Thus, the determination of EtG in hair is nowadays widely accepted for testing and monitoring of chronic excessive alcohol intake and used in different areas of forensic and clinical toxicology, including workplace testing, firearms and driving license re-granting, and post-mortem investigation. ...
Article
The assessment of chronic excessive alcohol consumption by ethyl glucuronide (EtG) determination in hair is generally based on a cut‐off value of 30 pg/mg recognized by regulatory authorities and scientific societies that guide the decision process. The ongoing debate about the risks connected with the straightforward application of this cut‐off refers to the factors that may influence the detected EtG concentration. The present contribution to this debate evaluates the seasonal variation of the averaged EtG values along a seven‐years period. Over 65,000 data points have been statistically analysed to provide a mathematical model that interprets the data, gives insight into several influencing factors, and forecasts progressive data‐points of the time series. This model shows that there is an annual pattern in the data exhibiting lower EtG concentrations during warm seasons and higher values in cold seasons. The estimated EtG cycles are characterized by the seasonal variation of ±2.78 pg/mg above and below the overall mean (with 5.56 pg/mg absolute difference overall). This seasonal factor associated with EtG quantification might result in a potential source of bias, at least in the regional/climatic conditions observed in the samples’ collection area. Moreover, the EtG time series reveals that the change in the sample pre‐treatment procedure has an effect on the modelled pattern as an abrupt increment (+38%) in the mean value of the EtG concentration. This change corresponds to the time when the former protocol of cutting hair into small segments before their extraction was substituted by their pulverization with a ball mill.
... The current toxicology state of the art identifies a person who falls into the category of excessive alcohol consumer through the analysis of direct biomarkers in hair, i.e. ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEEs) [1][2][3][4][5][6][7][8][9]. Although traditional indirect biomarkers (i.e., not formed by alcohol metabolic processes), including aspartate transferase (AST), alanine transferase (ALT), gammaglutamyl transferase (GGT), mean corpuscular volume of the erythrocytes (MCV) and carbohydratedeficient-transferrin (CDT) on blood/serum are still utilized to evaluate chronic excessive alcohol intake [5,[10][11][12], they rather reveal the damaging effects of alcohol on target organs, but exhibit unsatisfactory sensitivity and specificity [11,13]. In fact, indirect effects largely depend on the interindividual variability, resulting in a high rate of false positive and false negative outcomes. ...
... Consequently, the correlation observed between alcohol consumption and biomarkers' concentrations is not exact and partly depends also on their hydrophilic (EtG) or lipophilic (FAEEs) nature. Other sources of bias are the use of alcohol-based hair care products [34] and lipophilic hair waxes [13], respectively leading to FAEEs increase or decrease. ...
Article
The chronic intake of an excessive amount of alcohol is currently ascertained by determining the concentration of direct alcohol metabolites in the hair samples of the alleged abusers, including ethyl glucuronide (EtG) and, less frequently, fatty acid ethyl esters (FAEEs). Indirect blood biomarkers of alcohol abuse are still determined to support hair EtG results and diagnose a consequent liver impairment. In the present study, the supporting role of hair FAEEs is compared with indirect blood biomarkers with respect to the contexts in which hair EtG interpretation is uncertain. Receiver Operating Characteristics (ROC) curves and multivariate Principal Component Analysis (PCA) demonstrated much stronger correlation of EtG results with FAEEs than with any single indirect biomarker or their combinations. Partial Least Squares Discriminant Analysis (PLS-DA) models based on hair EtG and FAEEs were developed to maximize the biomarkers information content on a multivariate background. The final PLS-DA model yielded 100% correct classification on a training/evaluation dataset of 155 subjects, including both chronic alcohol abusers and social drinkers. Then, the PLS-DA model was validated on an external dataset of 81 individual providing optimal discrimination ability between chronic alcohol abusers and social drinkers, in terms of specificity and sensitivity. The PLS-DA scores obtained for each subject, with respect to the PLS-DA model threshold that separates the probabilistic distributions for the two classes, furnished a likelihood ratio value, which in turn conveys the strength of the experimental data support to the classification decision, within a Bayesian logic. Typical boundary real cases from daily work are discussed, too.
... Indirect ethanol exposure biomarkers such as serum activities of enzymes aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyltransferase (GGT), and carbohydratedeficient-transferrin (CDT) were used to evaluate alcohol consumption behavior [4][5][6][7]. However, the measurements of these non-specific markers are strongly affected by age and the presence of several pathologies, which limits their use to estimate the abusive use of ethanol [6][7][8]. ...
... Indirect ethanol exposure biomarkers such as serum activities of enzymes aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyltransferase (GGT), and carbohydratedeficient-transferrin (CDT) were used to evaluate alcohol consumption behavior [4][5][6][7]. However, the measurements of these non-specific markers are strongly affected by age and the presence of several pathologies, which limits their use to estimate the abusive use of ethanol [6][7][8]. The measurement of more specific biomarkers of ethanol consumption in hair is a more specific and reliable alternative to evaluate excessive exposure [9,10]. ...
Article
The presence of Ethyl glucuronide (EtG) in hair provides a strong indication of ethanol consumption and its investigation is of interest in both clinical and forensic contexts because of the wide window of detection. However, due to the possibility of false negative results in cases of small ethanol intake or excessive hair washing, the combined measurement of ethyl palmitate (EtP) with EtG could be useful. In this study, a sensitive UHPLC-MS/MS procedure for the measurement of EtG in hair was developed and validated, using optimized sample preparation and chromatographic separation. Milled hair was extracted with water for 24 h at room temperature, followed by clean-up of the extract by ion-exchange solid phase extraction (SPE). Extraction was highly efficient, with yield of 96.93-101.06%. Chromatographic separation was performed with a Fluoro-Phenyl stationary phase. The assay was linear from 4 to 500pgmg-1, with accuracy in the range of 100.30-106.16%. Matrix effects (-0.87 to 5.89%) were adequately compensated by the use of deuterated EtG as internal standard. EtG was measured in hair samples of 46 volunteers, and results were compared with hair concentrations of ethyl palmitate (EtP) and the score in the AUDITC questionnaire. EtG hair concentrations were significantly correlated to the AUDIT-C classification (rs=0.365, p<0.05), but not to EtP hair levels. The diagnostic performance of EtG hair concentrations to identify excessive or moderate ethanol use was similar to the capability of AUDIT-C to identify severe and high health risk (Kappa, p=0.013). The developed assay is suitable for clinical use, providing a useful tool to evaluate chronic ethanol consumption.
... Objective screening for alcohol use can provide an opportunity to intervene for improving patient health and quality of life. However, in the absence of specific and sensitive biomarkers for alcohol abuse, physicians often fail to recognize such patterns 1,7,8 . Questionnaires sometimes provide unreliable information or may not be applicable to all patients. ...
... Compared to other short term and long term biomarkers, it is a more specific and sensitive marker of alcohol abuse 10,11 , which can be applied for both clinical and forensic analysis 14,15 . Many analytical methods have been developed and validated for detection and quantization of EtG in the biological matrices, such as nuclear magnetic resonance (NMR) 16 , capillary zone electrophoresis (CZE) 7 , liquid chromatography-mass spectrometry (LC-MS) 14 and gas chromatography-mass spectrometry (GC-MS) 8,15 . But because of the complex nature of urine and serum, these methods are frequently hindered by the matrix compounds co-eluting with EtG in urine and serum. ...
Article
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Alcohol misuse has now become a serious public health problem and early intervention is important in minimizing the harm. Biochemical markers of recent and high levels of alcohol consumption can play an important role in providing feedback regarding the health consequences of alcohol misuse. Existing markers are not sensitive to recent consumption and in detecting early relapse. Ethyl glucuronide (EtG), a phase-II metabolite of ethanol is a promising marker of recent alcohol use and can be detected in body fluids. In this study an analytical technique for quantitation of EtG in body fluids using solid-phase extraction (SPE) and gas chromatography (GC) with mass spectrometric detection (MS) was developed and validated. De-proteinization of serum and urine samples was done with perchloric acid and hydrochloric acid, respectively. Serum samples were passed through phospholipids removal cartridges for further clean up. EtG was isolated using amino propyl solid phase extraction columns. Chromatographic separation was achieved by gas chromatography with mass spectrometry. Limit of detection and limit of quantitation were 50 and 150 ng/ml for urine and 80 and 210 ng/ml for serum, respectively. Signal to noise ratio was 3:1, mean absolute recovery was 80-85 per cent. Significant correlation was obtained between breath alcohol and serum EtG levels (r=0.853) and urine EtG and time since last abuse (r = -0.903) in clinical samples. In the absence of other standardized techniques to quantitate EtG in biological samples, this gc - ms method was found to have high throughput and was sensitive and specific.
... The aim of this scoping review is to present a critical overview of the current knowledge and research on blood biomarkers of alcohol use. Although different types of biospecimen, such as the urine, saliva, and hair, have been examined to detect alcohol use [8,9], the blood is by far the most commonly used sample for laboratory measurements in general practice and primary care settings [10]. In addition, specific to alcohol, biomarkers are generally detectable in the blood earlier and offer a wider range of detection, compared to other biospecimens [11]. ...
... We included those records that reported development, characteristics, and/or use of blood biomarker for alcohol use. Studies that focused on non-blood (e.g., hair and urine) biomarkers for alcohol use were excluded; for an overview of such non-blood biomarkers, see: [8,9]. Studies geared toward the use and/or effects of alcohol in specific conditions, such as comorbid health problems (e.g., HIV, heart disease, cancer, obesity), mental health problems (e.g., bipolar disorder, depression, psychosis), or other alcohol-related diseases (e.g., fetal alcohol syndrome, alcohol-associated liver disease) were also excluded. ...
Article
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Purpose of Review Understanding whether a person has consumed alcohol or not, as well as quantitative assessment of alcohol use, are often based on self-reported measures, which may be subject to recall bias, among other challenges. Although not without limitations, blood biomarkers may complement self-reported assessments to provide a more accurate determination of the presence and quantity of alcohol use. The aim of this review is to provide a critical overview of the current knowledge and research on biomarkers of alcohol use, with a particular focus on blood tests. Recent Findings This scoping review summarizes the published work on blood tests currently used in clinical practice, including phosphatidyl ethanol (PEth), fatty acid ethyl ester (FAEE), carbohydrate-deficient transferrin (CDT), total serum sialic acid (TSA), mean corpuscular volume (MCV), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma glutamyl transpeptidase (GGT), and cholesteryl ester transfer protein (CETP). Emerging blood biomarkers with a potential use to assess alcohol drinking are also briefly reviewed, including N-Acetyl-β-Hexosaminidase (Beta-Hex), macrophage migration inhibitory factor (MIF), and d-dopachrome tautomerase (DDT). We discuss the aforementioned biomarkers in the context of their clinical implications, characteristics, strengths, and limitations. Summary The available blood biomarkers considerably vary in the time period in which they detect alcohol use and the amount of alcohol they are sensitive to. While currently available biomarkers provide useful information, especially in combination with self-reported measures, future work is needed to identify more sensitive and specific blood biomarkers for different levels and patterns of alcohol use. Integration of such biomarkers into clinical practice and research will increase the accuracy and richness of the data and may guide more effective and targeted strategies for prevention, diagnosis, and treatment of excessive alcohol use.
... Sodium fluoride glucose, serum levels of insulin, triglycerides, total cholesterol:highdensity lipoprotein, cotinine (nicotine metabolite as indicator of smoking habits [36], gamma-glutamyl-transferase/GGT (as marker of alcohol consumption [37] and ultrasensitive C-reactive protein (CRP) were analyzed with an enzyme rated method (Unicel DXC 800-Beckman and Coulter, 4300 N. Harbor Blvd., Fullerton, CA 92835 U.S.A), homogeneous immunoassay (Modular ROCHE Automized systems, Basel, Switzerland) and a particle-enhanced turbidimetric assay (Cobas Integra 400 plus, Roche, Basel, Switzerland), respectively. A dysregulated hypothalamic-pituitary-adrenal cortex axis (HPA axis) contributes to cognitive decline and dysfunctional growth hormones [38], e.g., serum total insulin-like growth factor 1 (IGF-1) and insulin-like growth factor binding protein 3 (IGFBP-3). ...
... The lower IGF-1 and IGFBP-3 and higher HOMA-IR levels in the Stressed group might further support inactivation or downregulation of retinal glia insulin receptors and subsequent endothelial dysfunction or glaucoma risk [50]. Decreases in IGF-1 concentration, a neurotrophic peptide in the central nervous system, in the Stressed group might therefore be regarded as a potential biomarker for early diagnosis or even prognosis of impaired insulin signaling [37]. IGF-1 and insulin together with cytokines activate common signaling pathways necessary for the reprogramming of Müller glia cells and retinal redevelopment upon injury [51]. ...
Article
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Citation: Malan, L.; Hamer, M.; von Känel, R.; van Wyk, R.D.; Sumner, A.E.; Nilsson, P.M.; Lambert, G.W.; Steyn, H.S.; Badenhorst, C.J.; Malan, N.T. A Stress Syndrome Prototype Reflects Type 3 Diabetes and Ischemic Stroke Risk: The SABPA Study. Biology 2021, 10, 162. https://doi.org/10.3390/biology10020162
... For instance, the black eumelanin in Orientals tends to have stronger binding affinity to specific drug compared to the reddish-brown pheomelanin in Westerners, and thus resulting in false positive tests [22]. In this regard, the evaluation of cut-off value for the drug misuse among multiracial populations is a relatively less well-studied area [31][32][33][34][35][36]. ...
... Several studies [31][32][33] have determined the cut-off value for biomarkers of hair among European alcohol misusers based on different sample sizes varied from several hundreds to thousands. The cut-off values of fatty acid ethyl ester and ethyl glucuronide in hair samples were obtained at different levels of concentration, which has became a valuable resource for distinguishing chronic alcohol misuser from the general population with reference to the comparable cut-off value recommended by SoHT. ...
Article
Methamphetamine (MAMP) is one of the most commonly abused illicit drugs in Asian countries, which belongs to the amphetamine-type stimulant class of substances. To detect the chronic drug misuse, human hairs have often been used as analytical specimens due to their long detection windows and easy accessibility. However, there is no investigation regarding the cut-off value of MAMP detection used in black-hair Chinese populations. Based on the analytical data obtained from 563 MAMP users, the cut-off value was found to be 0.97 ng/mg for the simultaneous detection of MAMP and amphetamine (AMP) ≥0.004 ng/mg (LOD). Through the established HPLC-MS/MS analytical method, the limits of detection and quantification of MAMP were 0.004 and 0.01 ng/mg, respectively. The cut-off value was optimized by AMP detection rate and receiver operating characteristic analysis, and the results were consistent with the previously reported MAMP/AMP ratio.
... The discrimination power and the SFAEE cut-offs for excessive drinking were repeatedly evaluated by receiver operation characteristics analysis (ROC analysis) [55][56][57] and recently reviewed [4]. ...
Article
The consensus on alcohol markers in hair was revised for the fourth time by an expert group of the Society of Hair Testing based on current state of research. This revision was adopted by the members of the Society during the business meeting in Brisbane on August 29th 2016. For both markers, ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEEs), two cut-off values for discrimination between teetotalers or occasional low amount consumption and moderate alcohol drinking (low cut-off), and between non-excessive (abstinence up to moderate alcohol intake) and chronic excessive drinking (high cut-off value) were critically examined. For the current revision, the cut-off values for EtG (7 pg/mg and 30 pg/mg, respectively) remained unchanged despite different findings or discussions published in the meantime. This was mainly due to the lack of broader data collections from new studies with great numbers of volunteers following thorough study concepts. In contrast, an essential change of the consensus was accepted for the FAEEs, where the concentration of ethyl palmitate (E16:0) can be used autonomously for interpretation instead of the concentration sum (ƩFAEE) of the four esters ethyl myristate, ethyl palmitate, ethyl oleate and ethyl stearate, as previously applied. After evaluation of the data from seven laboratories, the E16:0 cut-off for abstinence assessment was defined at 0.12 ng/mg for the 0–3 cm segment and at 0.15 ng/mg for the 0–6 cm segment. The cut-off for chronic excessive drinking was fixed at 0.35 ng/mg for the 0–3 cm segment and at 0.45 ng/mg for the 0–6 cm segment. The use of E16:0 with these cut-offs in place of ƩFAEE for alcohol intake assessment produces only a minor loss in discrimination power, leads to no essential difference in the interpretation concerning chronic excessive alcohol consumption and is suitable to confirm EtG results in abstinence assessment if ethanol containing hair sprays or lotions are excluded.
... Serum cotinine and gammaglutamyl transferase (GGT) were respectively used as biochemical markers for smoking status and alcohol consumption. 38,39 Cotinine was measured by means of a modular Roche automised (Switzerland) apparatus using a homogeneous immunoassay. Cotinine, a metabolite of nicotine, defined habitual smokers when levels were ≥ 14.99 ng/ml. ...
Article
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Background: In a South African bi-ethnic cohort, defensive (DefS)/social support/avoidance coping strategies have been shown to influence cardiac troponin T (cTnT) levels through different stress signalling pathways. Personality traits (extra-version, neuroticism, conscientiousness, openness to experience , agreeableness) partially control stress coping responses and may affect prospective cardiac responses. Hence in this cohort, we aimed to examine relationships between personality traits and coping strategies, and to assess associations between cTnT changes over time, personality traits and coping strategies. Methods: A cohort of African and Caucasian male and female teachers (n = 359) participating in both phases of the Sympathetic activity and Ambulatory Blood Pressure in Africans (SABPA) study, was prospectively followed for three years. Personality traits (Basic Traits Inventory) and coping (Coping Strategy Indicator) scores were determined. Fasting serum samples for cTnT determination were collected. Established hypertension-related cTnT cutoff points of 4.2 pg/ml (Africans) and 5.6 pg/ml (Caucasians) were applied. Results: Higher neuroticism and lower conscientiousness scores were found in the Africans than in the Caucasians (p < 0.05). Both traits correlated with all three coping strategies in Caucasians, but only with DefS and avoidance coping in Africans. Over a period of three years, cTnT levels decreased in both races. Compared to Africans, Caucasians showed a greater recovery from the ethnic-specific cTnT cut-off point over time. In the Africans with high DefS scores, cTnT level changes were inversely associated with conscientiousness (adjusted R2 = 0.14; β = –0.26). In Caucasians scoring high in avoidance coping, conscientiousness (odds ratio 0.84) and neuroticism (odds ratio 0.90) showed a lower likelihood of predicting the cTnT cut-off point. Conclusion: In both races, conscientiousness may contribute to healthier stress coping responses and protect against cardiac ischaemia and risk of hypertension.
... Cotinine was measured to determine participants' smoking status, with participants regarded as smokers when cotinine values exceeded 14.99 ng/ml [23]. Gamma-glutamyltransferase (γ-GT) was measured as a marker for alcohol use [24]. Anthropometrists, with level II accreditation from the International Society for the Advancement of Kinanthropometry, determined body mass and height in triplicate to the nearest 0.1 kg and 0.1 cm, respectively, whilst subjects wore minimal clothing and no shoes (Precision Health Scale, A & D Company, Tokyo, Japan; Invicta Stadiometer IP 1465, Invicta, London, UK). ...
... 47,48 Recent investigations pointed out that homocysteine levels on admission might be a useful screening method for the risk of seizures in AWS, particular in combination with CDT. 41,43,[49][50][51] Several days after alcohol abstinence, homocysteine plasma levels decrease to normal. 50 The TWEAK is an acronym of the first letter of the key words in the questions of this screening tool: Tolerance, Worried, Eye-opener, Amnesia, K (cutdown) and represents a modification of the CAGE. ...
Article
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The alcohol withdrawal syndrome is a well-known condition occurring after intentional or unintentional abrupt cessation of heavy/constant drinking in patients suffering from alcohol use disorders (AUDs). AUDs are common in neurological departments with patients admitted for coma, epileptic seizures, dementia, polyneuropathy, and gait disturbances. Nonetheless, diagnosis and treatment are often delayed until dramatic symptoms occur. The purpose of this review is to increase the awareness of the early clinical manifestations of AWS and the appropriate identification and management of this important condition in a neurological setting.
... This is a limitation for hair analysis that may be overcome in the future performing new studies and analyzing new or more markers. A further limitation for hair analysis is the risk of false positivity, mainly due to external contamination or hair treatments (21,(43)(44)(45)(46). Various measures can be adopted to limit this drawback, such as effective decontamination procedures and detection of metabolites (47). ...
Article
Background: Clinical reliability of self-reported data for alcohol, caffeine, and nicotine consumptions is lacking, particularly in adolescents. Objectives: To compare a self-report questionnaire and hair analysis to assess the reliability and effectiveness of the self-report. Methods: A cross-sectional study on 14-15-year-old Italian students (n = 874, 38% males, 62% females) was performed comparing self-reported data to hair analysis. The latter quantified hair concentrations of caffeine, nicotine, cotinine, ethyl glucuronide (EtG), and fatty acid ethyl esters (FAEEs) using mass spectrometry. Results: Concordance between self-report and hair testing ranged from good to poor across substances and levels of use: poor for heavy alcohol intake (EtG: k = 0.36, 20 positive cases by hair analysis, false negative by self-report, 2.3% of total sample; FAEE k = 0.31, 25 positive cases, 2.9% of total sample); fair to poor for active smokers (k = 0.40, 125 positive cases, 14.3% of total sample); and moderate for caffeine (k = 0.57, 56 positive cases, 6.4% of total sample). Conclusions: Epidemiological studies on alcohol, caffeine, and nicotine consumption in adolescents may benefit from the inclusion of toxicological analysis on hair samples to overcome the under-reporting phenomenon of questionnaires and detect more cases of problematic substance use.
... Numerous studies about the quantification of FAEEs in hair have been published [7,9,221,233,226,[234][235][236][237][238][239][240][241][242][243][244][245]. Because of their lipophilic character, most authors explain the incorporation of FAEEs into the hair mainly through sebum [19]. ...
Thesis
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In Belgium, the analysis of indirect biomarkers such as carbohydrate deficient transferrin (CDT%), gamma-glutamyltransferase (GGT), aspartate aminotransferase/alanine aminotransferase (AST/ALT) and mean corpuscular volume (MCV), is currently used to monitor the alcohol consumption in cases of fitness to drive assessment. To estimate how the use of direct ethanol markers (e.g. ethylglucuronide (EtG), ethylsulfate (EtS) and phosphatidylethanol species (PEths)) could improve the current process, three quantitative methods (EtG in hair; EtG and EtS in urine; and PEth 16:0/18:1, PEth 18:1/18:1 and PEth 16:0/16:0 in blood, venous (V) and capillary (C) dried blood spots (DBSs)) were developed, validated and tested. Fifty volunteers, for whom fitness to drive had to be assessed and for whom a blood analysis for indirect biomarkers was requested, were included in the study. The sampling and analysis of hair, urine and C-DBS were added to the process currently used. Hair EtG and C-DBS PEths are more sensitive to detect alcohol abuse than the currently used indirect biomarkers and allow to disprove an abstinence period. EtG and EtS in urine form a relevant parameter to detect recent alcohol intake (even one single alcohol consumption) during the days (up to 5 days) prior to the sampling and can thus be used to disprove strict abstinence. The three analyses tested here provide different levels of information and can be used separately or combined. The combined use of the three strategies allows better inference about the evolution of the alcohol consumption prior to the sampling. Moreover, the exclusive use of non- or minimally invasive sampling (hair, urine and C-DBS) allows this to be performed directly during the fitness to drive assessment by regular staff members. In conclusion, the three approaches that were evaluated in this work offer the potential to improve the Belgian driver’s licence regranting process.
... Second, further prospective studies are needed to evaluate the efficacy of the NC cutoff points for predicting atherosclerosis and cardiovascular diseases based on this cross-sectional study. Thirdly, it is noted that liver enzyme levels were closely correlated with metabolic disorders [32], however, the subjects with hepatic and renal dysfunction were excluded from our study, which eliminating the influence of those illnesses to metabolic disorders to some extent. ...
Article
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Neck circumference is a new anthropometric index for estimating obesity. We aimed to determine the relationship between neck circumference and body fat content and distribution as well as the efficacy of neck circumference for identifying visceral adiposity and metabolic disorders. A total of 1943 subjects (783 men, 1160 women) with a mean age of 58 ± 7 years were enrolled in this cross-sectional study. Metabolic syndrome was defined according to the standard in the 2013 China Guideline. Analyses were conducted to determine optimal neck circumference cutoff points for visceral adiposity quantified by magnetic resonance imaging, and to compare the performance of neck circumference with that of waist circumference in identifying abdominal obesity and metabolic disorders. Visceral fat content was independently correlated with neck circumference. Receiver operating characteristic curves showed that the area under the curve for the ability of neck circumference to determine visceral adiposity was 0.781 for men and 0.777 for women. Moreover, in men a neck circumference value of 38.5 cm had a sensitivity of 56.1 % and specificity of 83.5 %, and in women, a neck circumference value of 34.5 cm had a sensitivity of 58.1 % and specificity of 82.5 %. These values were the optimal cutoffs for identifying visceral obesity. There were no statistically significant differences between the proportions of metabolic syndrome and its components identified by an increased neck circumference and waist circumference. Neck circumference has the same power as waist circumference for identifying metabolic disorders in a Chinese population.
... The use of EtG, EtS, and ethanol as reliable and valid markers for alcohol has been established though numerous studies [57,65,67]. However, the establishment of the most valid cutoff points and the establishment of specific detection windows for each of these markers is still being debated. ...
... Clinical characteristic profile of a South African bi-ethnic sex cohort at baseline.Abbreviations: CVD, history of cardiovascular disease; Active smoking (Cotinine > 50 ng/ml)[31]; Alcohol abuse (gamma-glutamyl transferase ! 41 U/l)[32]; PHQ-9 ! 10, Patient Health Questionnaire[26]; Central obesity, waist !94 cm men and !80 cm women[30]; Pre-diabetes, HbA1C ! 5$7[30]; Low grade inflammation, CRP ! ...
Article
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Background: Depression has been associated with impaired nitric oxide (NO)-mediated vasodilation and vascular dysregulation (VD). Whether depression and NO levels will disturb retinal hemodynamics is not clear. Objectives and methods: Associations between the retinal vasculature, diastolic ocular perfusion pressure (DOPP) as measure of hypoperfusion, NO metabolites (NOx) and depression symptoms were assessed. Chronic VD risk markers [depression symptoms (Patient Health Questionnaire/PHQ-9 ≥ 10) and 24h pulse pressure] were determined in a bi-ethnic cohort (n=313; 48.6 ± 9 years; 53.9% men). At 3 year follow-up, retinal vessel calibre and retinopathy signs were quantified from digital images. Salivary NOx was obtained pre- and post-flicker light-induced provocation (FLIP). DOPP was defined as diastolic blood pressure minus intraocular pressure. Results: Chronic VD risk was evident in Blacks opposed to acute risk in Whites (P<0.05). At follow-up, retinopathy (Blacks 60.4%/Whites 39.6%), lower pre-FLIP (μM) and higher post-FLIP NOx (changes from baseline, %), arteriolar narrowing and wider venular calibre values were evident in Blacks compared to Whites, independent of confounders. A wider venular calibre, an index of stroke risk, was associated with chronic depression symptoms [cut point 248 MU: Area under the curve 0·61 (95% CI: 0·51, 0·72); 71% sensitivity; 55% specificity] as well as with hypoperfusion in the Blacks. In this group, arteriolar narrowing was associated with hypoperfusion; and attenuated arteriolar dilation with increased post-FLIP NOx responses. Conclusions: Chronic depression symptoms may alter NO regulation and facilitate VD. NO-mediated vasoconstriction presumably impeded perfusion, retinal hemodynamics and -remodelling; potentiating stroke risk in Blacks.
... Following detection in keratinous specimens, EtG presents the potential to reflect and monitor patterns of ethanol consumption. This is inclusive of periods of abuse or abstinence (Pragst and Yegles, 2008;Hastedt et al., 2013), which can be highly significant. ...
Article
Aims: This work aimed to assess the performance of hair and fingernail ethyl glucuronide (EtG) measurement for use as a biomarker of alcohol consumption in persons with known drinking history across a range of drinking behaviours. Methods: EtG concentrations were assessed from the hair and fingernails of 50 study participants. Alcohol consumption of the previous 90 days was assessed by participant interview using the alcohol timeline follow-back method. EtG concentration was determined using LC-MS-MS using a method which was validated and accredited to ISO/IEC 17025 standards. Results: There was significant correlation between alcohol consumption and EtG concentrations found in hair and fingernail samples across the study group (n = 50). From participants testing positive for EtG (male n = 14, female n = 13) no significant difference was found between male and female EtG levels in either hair or fingernails. Across all participants there was no significant difference in hair or fingernail EtG concentration between male (n = 23) and females (n = 27). Conclusions: Our results support the use of EtG to indicate alcohol consumption over the previous 90 days, or ~3 months as is the normal practice in hair analysis. The results confirm that fingernails can be a useful alternative matrix where hair samples are not available.
... Numerous studies about the quantification of FAEEs in hair have been published [37,171,179,[186][187][188][189][190][191][192][193][194][195][196][197][198][199]. Because of their lipophilic character, most authors explain the incorporation of FAEEs into the hair mainly through sebum [2]. ...
Article
Monitoring of alcohol consumption by living persons takes place in various contexts, amongst which workplace drug testing, driving under the influence of alcohol, driving licence regranting programs, alcohol withdrawal treatment, diagnosis of acute intoxication or fetal alcohol ingestion. The matrices that are mostly used today include blood, breath and urine. The aim of this review is to present alternative sampling strategies that allow monitoring of the alcohol consumption in living subjects. Ethanol itself, indirect (carbohydrate deficient transferrin, CDT%) as well as direct biomarkers (ethyl glucuronide, EtG; ethyl sulphate, EtS; fatty acid ethyl esters, FAEEs and phosphatidylethanol species, PEths) of ethanol consumption will be considered. This review covers dried blood spots (CDT%, EtG/EtS, PEths), dried urine spots (EtG/EtS), sweat and skin surface lipids (ethanol, EtG, FAEEs), oral fluid (ethanol, EtG), exhaled breath (PEths), hair (EtG, FAEEs), nail (EtG), meconium (EtG/EtS, FAEEs), umbilical cord and placenta (EtG/EtS and PEth 16:0/18:1). Main results, issues and considerations specific to each matrix are reported. Details about sample preparation and analytical methods are not within the scope of this review.
... This procedure is based on headspace solid-phase microextraction (HS-SPME) in which the hair extract is placed in headspace vials, and phosphate buffer and sodium chloride (NaCl) are added. A 65 µm polydimethylsiloxane/divinylbenzene (PDMS/DVB) fibre is used in a HS-SPME robotic system, being the entire procedure automatic [59,64,[67][68][69][70][76][77][78][79][80]. ...
Chapter
Fatty acid ethyl esters (FAEEs) are formed through the reaction of ethanol with endogenous free fatty acids, triglycerides, lipoproteins or phospholipids, catalysed by cytosolic and microsomal FAEE synthases as well as by unspecific enzymes. Since the 1960s, these compounds have been known as biotransformation products of ethanol, and in the 1990s they were proposed as promising biomarkers for monitoring ethanol exposure. Indeed, they present a linear structure which contains the intact ethyl group of ethanol, which translates into biomarkers with high specificity and sensitivity towards alcohol consumption since it excludes other reasons than alcohol consumption for their formation. The FAEEs include over 20 species, while not all the species are used as alcohol biomarkers, several species have been successfully applied as alcohol biomarkers in different biological specimens, including blood, serum, plasma, hair, meconium, and tissues such as skin surface lipids, sebum, liver and adipose tissue. The FAEEs species used as alcohol biomarkers vary in accordance with the analysed biological specimen, depending on their correlation with alcohol consumption. In this paper, we intend to review the mechanisms of formation and structure of FAEEs to their use as alcohol biomarkers. Focusing on the developed analysis methods, fields of application and published articles, including a critical overview on the methodological challenges and considerations for their analysis; the present limitations and advantages relatively to other alcohol biomarkers; challenges and directions in the research of FAEEs as alcohol biomarkers. Keywords: alcohol biomarkers, fatty acid ethyl esters, biological specimens, toxicological analysis
... Currently, two cutoffs are accepted and used in forensic toxicology: 7 pg/mg assesses the occasional and moderate ethanol consumption, while 30 pg/mg indicates a chronic excessive alcohol consumption (ethanol daily intake -EDI: > 60 g/day) [1]. During the last decade, several studies proved the higher specificity of hEtG whenever compared to other ethanol biomarkers, such as carbohydrate-deficient transferrin (CDT) in serum [3][4][5][6][7], and fatty acid ethyl esters (FAEEs) in hair [8][9][10]. To date, only phosphatidylethanol (PEth) in whole blood proved a better performance in the diagnosis of chronic excessive alcohol consumption [11]. ...
Article
Background: Ethyl glucuronide in hair (hEtG) is a commonly used biomarker in the diagnosis of chronic excessive alcohol consumption. Though the high diagnostic specificity, some cosmetic treatments may influence the hEtG concentration, leading to false positive results. Aim: The aim of this study was to evaluate three cases of alleged false positive hair samples due to the exposure to perfumes. Methods: Three subjects were monitored for up to 3 years; at least one sample collection was performed each year; all three men declared to have exposed hair to alcohol-based perfumes during all the period, except during the last three months before last sample collection. A volunteer treated the right side of his scalp every day during a month period with alcohol-based perfumes. Collection of hair was done at both sides before the first sample treatment and at the end of the month. Hair were processed following the method routinely used in the laboratory for the determination of hEtG (double washing with methanol/dichloromethane, pulverization, overnight incubation in water, and LC-MS/MS analysis, LLOQ: 3pg/mg). Results: EtG levels in hair collected from the three men during the declared exposition period ranged from < 3.0 pg/mg to 1130 pg/mg. hEtG concentrations, at the end of the three-month period without any use of perfumes, were < 3.0 pg/mg for the two men declaring an abstinence from alcoholic beverages and 26.3 pg/mg for the subject declaring an average drinking habit of about 35 g/day ethyl alcohol. Non-treated hair of the volunteer provided negative results, while hEtG levels in treated hair were 14.6 and 24.7 pg/mg Conclusions: prolonged exposition of hair to alcohol-based perfumes may increase hEtG levels, resulting in false positive results.
... g-Glutamyltransferase (g-GT) was measured as a marker for alcohol abuse. [24] Cotinine was measured to determine participants' smoking status, with subjects regarded as smokers if cotinine values exceeded 14.99 ng/ml. [25] Anthropometric measurements were performed in triplicate by level II registered biokineticists using standardized methods (Precision Health Scale; A&D Company, Tokyo, Japan; and Invicta Stadiometer IP 1465; Invicta, London, UK). ...
Article
Objectives: A dissociation between behavioural (in-control) and physiological parameters (indicating loss-of-control) is associated with cardiovascular risk in defensive coping (DefS) Africans. We evaluated relationships between DefS, sub-clinical atherosclerosis, low-grade inflammation and hypercoagulation in a bi-ethnic sex cohort. Methods: Black (Africans) and white Africans (Caucasians) (n = 375; aged 44.6 ± 9.7 years) were included. Ambulatory BP, vascular structure (left carotid cross-sectional wall area (L-CSWA) and plaque counts), and markers of coagulation and inflammation were quantified. Ethnicity/coping style interaction was revealed only in DefS participants. Results: A hypertensive state, less plaque, low-grade inflammation, and hypercoagulation were more prevalent in DefS Africans (27-84%) than DefS Caucasians (18-41%). Regression analyses demonstrated associations between L-CSWA and 24 hour systolic BP (R(2) = 0.38; β = 0.78; p < 0.05) in DefS African men but not in DefS African women or Caucasians. No associations between L-CSWA and coagulation markers were evident. Conclusion: Novel findings revealed hypercoagulation, low-grade inflammation and hyperkinetic BP (physiological loss-of-control responses) in DefS African men. Coupled to a self-reported in-control DefS behavioural profile, this reflects dissociation between behaviour and physiology. It may explain changes in vascular structure, increasing cerebrovascular disease risk in a state of hyper-vigilant coping.
... The Mosteller formula [14] was used to determine body surface area (m 2 ) from body mass and height measurements. Gamma glutamyl transferase (GGT) was used to determine alcohol consumption [15]. Serum cotinine (a nicotine metabolite) values were used to determine smoking status [16]. ...
Article
Background: Defensive coping (DefS) was associated with cardiovascular disease (CVD) susceptibility in Blacks. Whether coping strategies will associate with sub-clinical left ventricular hypertrophy (electrocardiographic-left ventricular hypertrophy [ECG-LVH] or Cornell product), cardiomyocyte injury and blood pressure (BP), is unclear. Therefore, we assessed relationships between ECG-LVH, cardiac troponin T (cTnT) and 24-hour BP in bi-ethnic groups when habitually utilising a certain coping style, and these groups when having a stress-related cTnT cut-point of 4.2ng/L. Methods: A target population study included a Black (n=190) and White (n=204) teachers' gender cohort (20-65years) from South Africa. The Coping Strategy Indicator determined DefS, social support and avoidance coping scores. Fasting blood samples, 10-lead ECG, 24-hour BP and -ECG data were obtained. Results: Interaction effects showed no gender, social support and avoidance coping differences. Stratification of groups was done for ethnicity and DefS. Blacks sought more social support, used less avoidance coping and presented with higher CVD susceptibility. Hypertension prevalence and ECG-LVH levels in DefS Blacks (63%) were higher compared to DefS Whites (40%). Multivariate regression analyses showed positive associations between Cornell product, cTnT and BP [p≤0.05] in DefS Blacks only. Their 24-hour systolic blood pressure (SBP) was associated with time-domain depressed heart-rate-variability and prolonged ST-segment-depression especially when applying an established stress-related cTnT ≥ 4.2ng/L cut-point. Conclusions: Defensive coping facilitated autonomic hyperactivity, myocardial injury and subsequent compensatory BP elevations as possible homeostatic reflexes to alleviate myocardial perfusion deficits. The resulting pressure overload increased sub-clinical wall remodelling and ischaemic heart disease risk in Blacks utilising habitual defensiveness. We therefore recommend regular ECG and high sensitivity cTnT screening in asymptomatic patients with emotional stress susceptibility. Longitudinal evidence is needed to confirm causality and progression of cardiomyopathy risk.
... In addition to traditional biomarkers, direct alcohol markers, such as phosphatidylethanol in the blood, and fatty acid ethyl esters or ethylglucuronide in hair have drawn attention as they are highly specific for chronic alcohol use. 40,41 Additional studies using biomarkers other than GGT are still needed. Second, caution should be taken to interpret the contribution of age and gender in comparison of combined the CAGE and GGT with AUDIT. ...
Article
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Purpose: The CAGE is a convenient test for alcohol-related disorder due to its brevity, but it is not as effective as the alcohol use disorders identification test (AUDIT). Gamma-glutamyl transferase (GGT) is an objective blood biochemical marker of excessive alcohol intake; however, it has low sensitivity. This study tested the performance of the combined use of CAGE and GGT to screen problem drinking (PD), alcohol use disorder (AUD), and alcohol dependence (AD). Methods: A total of 394 subjects composed of 91 normal controls and 303 subjects with PD were enrolled in this study. Of the PD subjects, 147 were diagnosed with AUD (77 alcohol abuse and 70 AD). A series of multiple logistic regression models for PD, AUD, and AD discrimination were used to obtain new combined CAGE and GGT scores after adjusting for age and gender (CAGE+GGT). A receiver operating characteristic curve analysis was conducted to determine how well the CAGE+GGT score discriminated between individuals with PD, AUD, and AD. Results: The discrimination accuracy of the AUDIT for PD was significantly better than that of the CAGE or the CAGE+GGT (z=6.927, p<0.0001; z=5.301, p<0.0001, respectively). The CAGE and the CAGE+GGT were better than the AUDIT at discriminating AUD (z=2.535, p=0.0112; z=2.894, p=0.0038, respectively). The discrimination accuracy of the AUDIT for AD was significantly better than that of the CAGE and GGT (z=3.233, p=0.0012; z=6.529, p<0.0001, respectively), but the CAGE+GGT was comparable with the AUDIT (z=1.652, p=0.0985). Conclusion: Our findings support the combined use of the CAGE questionnaire and serum GGT level as a sensitive and useful tool for AD screening.
... Gamma-glutamyl transferase (γ-GT) levels were measured as a marker of alcohol abuse and cotinine levels reflected smoking status. 20,21 All anthropometric measures were conducted in triplicate by ISAK (International Society for the Advancement of Kinanthropometry) level II accredited anthropometrists using calibrated instruments and standardised methodology to obtain waist circumference, with inter-and intra-observer variability less than 10%. Each participant's individual coping style ( Fig. 2) was identified with the coping strategy indicator (CSI). ...
Article
Full-text available
Objective: In past studies, a lack of social support has been associated with cardiovascular disease (CVD) risk, particularly in black Africans. However, whether or not coping strategies have beneficial effects on blood pressure (BP) and emotional well-being is not clear. We therefore assessed the relationship between BP levels, depression and coping strategies. Methods: A prospective bi-ethnic cohort followed 359 black and white South African school teachers (aged 20-65 years) over a three-year period. Data on ambulatory 24-hour blood pressure, depression, coping strategies (defensiveness, social support, avoidance) and culture-specific coping scores (cognitive/emotional debriefing, spiritual-, collectivistic and ritualcentred) were obtained. Results: Over three years, chronic depression (38 vs 19%) and hypertension (68 vs 35%) were apparent in blacks ( d-values > 0.3) as opposed to whites. In both groups, depression was accompanied by more avoidance (loss-of-control) coping. Consistent spiritual and increasing collectivistic coping were apparent in whites. Over time, increasing defensiveness (OR 1.08, p ≤ 0.05) and ritual coping (OR 1.27, p ≤ 0.01; d-values > 0.5), predicted chronic depression in blacks. The change in their symptoms of depression predicted 24-hour hypertension (OR 1.11, p = 0.04). No similar associations existed in whites. Conclusions: Blacks showed increasing defensiveness and ritual- and spiritual-centred coping in an attempt to combat chronic depression, which may be costly, as reflected by their chronic hypertensive status. Whites showed consistent spiritual- centred coping while utilising avoidance or loss-of-control coping, with a trend of seeking less social support or isolation as a coping mechanism. During counselling of depressed patients with hypertension, the beneficial effects of social support and spiritual coping may be of great importance.
... Fatty acid ethyl esters (FAEE), which do not cross the placental barrier, accumulate in meconium as a result of ethanol metabolism by the fetus (Burd & Hofer, 2008). FAEE in meconium have demonstrated substantial sensitivity and specificity (Bearer et al., 2003) for heavy PAE, and are widely used in prevalence studies (Hastedt et al., 2013;Himes et al., 2015;Pichini et al., 2012). Bearer and co-authors reported that a positive test for meconium FAEE correctly identifies 72% of pregnant women who consume ≥1 drink per week during the third trimester (Bearer et al., 1999). ...
Article
Background Although fetal alcohol spectrum disorders represent a significant public health problem, Native Americans are underrepresented in population and targeted screening programs. Prior reports suggest that Native American tribal communities may have a higher prevalence of alcohol use during pregnancy; however, systematic examination using ethanol biomarkers is lacking. Methods This study utilized data collected through the Navajo Birth Cohort Study (NBCS)—a birth cohort study of a Southwestern tribal community. Prevalence of prenatal alcohol exposure (PAE) was assessed by a battery of meconium biomarkers among 333 NBCS participants. Meconium samples were analyzed for nine individual fatty acid ethyl ester (FAEE) species, ethyl glucuronide (EtG), and ethyl sulfate (EtS) by LC–MS/MS. Results Participants were recruited from five hospitals at the Navajo Nation located in Arizona (Chinle, Tséhootsooí, Tuba City) and New Mexico (Gallup, Shiprock). All participants identified as Native American; most reported a personal income of <$20,000 per year (71.3%), and ≤high school education (55.3%). The most prevalent biomarker was EtS (7.8%) followed by ethyl oleate (6.9%); 5.4% of the sample were positive for at least two biomarkers. Conclusions Results of this study on the prevalence of PAE in the Navajo Nation, obtained for the first time with an objective comprehensive panel of meconium biomarkers, indicate that the rates in the NBCS may be comparable to the general U.S. population and are in accord with recent U.S. national survey estimates. Our findings emphasize that drinking behaviors among Native American communities in the United States can vary, and generalization across all Native American populations is not warranted.
... The African group showed a higher mean body mass index, and higher levels of mean systolic, diastolic and hypertension prevalence values when compared to the Caucasian group. The levels of Gamma-glutamyl transferase (GGT), which is a liver enzyme indicating hepatic dysfunction, and that could be associated with alcohol use (Hastedt, Büchner, Rothe, Gapert, Herre, Krumbiegel, Tsokos, Kienast, Heinz & Hartwig, 2013), were also elevated in the African group. Conversely, the Caucasian group showed higher levels of physical activity than the African group. ...
Article
The aim of this study was to validate the Teacher Stress Inventory (TSI) for use in a South African context. The process of scale validation also sheds significant light on this culturally diverse group of participants’ levels of psychological well-being and physical health, and its association with the level of stress that teachers reported. Using a cross-sectional survey design, Caucasian (n = 209) and African (n = 200) educators’ subsiding in the North-West Province of South Africa, completed the TSI, together with a number of self-report and physiological measures of stress and well-being. In contrast to the five factors of the TSI identified in US samples, statistical analysis yielded a two-factor model (i.e. General circumstance-related stress and Learner-related stress) with satisfactory reliability indices. Significant correlation with measures of psychological and physiological health also reflected positively on the criterion-related validity of the scale. The TSI proved to be a useful, brief self-report questionnaire for the assessment of teacher stress in this cohort of South African teachers. © 2018, South African Journal Of Education. All rights reserved.
... It has been reported that the lowest false negative and false positive rates were obtained using the results of hair ethyl glucuronide and fatty acid ethyl esters together. But even here the markers showed relatively poor correlation [94]. It is stated that an ethyl glucuronide concentration of !30 pg/mg hair in the 0-3 up to 0-6 cm proximal scalp hair segment strongly suggests chronic excessive alcohol consumption. ...
Article
Introduction: Head hair analysis for drugs and drug metabolites has been used widely with the aim of detecting exposure in the weeks or months prior to sample collection. However, inappropriate interpretation of results has likely led to serious miscarriages of justice, especially in child custody cases. Objective: The aim of this review is to assess critically what can, and perhaps more importantly, what cannot be claimed as regards the interpretation of hair test results in a given set of circumstances in order to inform future testing. Methods: We searched the PubMed database for papers published 2010–2016 using the terms “hair” and “drug” and “decontamination”, the terms “hair” and “drug” and “contamination”, the terms “hair” and “drug-facilitated crime”, the terms “hair” and “ethyl glucuronide”, and the terms “hair”, “drug testing” and “analysis”. Study of the reference lists of the 46 relevant papers identified 25 further relevant citations, giving a total of 71 citations. Hair samples: Drugs, drug metabolites and/or decomposition products may arise not only from deliberate drug administration, but also via deposition from a contaminated atmosphere if drug(s) have been smoked or otherwise vaporized in a confined area, transfer from contaminated surfaces via food/fingers, etc., and transfer from sweat and other secretions after a single large exposure, which could include anesthesia. Excretion in sweat of endogenous analytes such as γ-hydroxybutyric acid is a potential confounder if its use is to be investigated. Cosmetic procedures such as bleaching or heat treatment of hair may remove analytes prior to sample collection. Hair color and texture, the area of the head the sample is taken from, the growth rate of individual hairs, and how the sample has been stored, may also affect the interpretation of results. Toxicological analysis: Immunoassay results alone do not provide reliable evidence on which to base judicial decisions. Gas or liquid chromatography with mass spectrometric detection (GC- or LC-MS), if used with due caution, can give accurate analyte identification and high sensitivity, but many problems remain. Firstly, it is not possible to prepare assay calibrators or quality control material except by soaking “blank” hair in solutions of appropriate analytes, drying, and then subjecting the dried material to an analysis. The fact that solvents can be used to add analytes to hair points to the fact that analytes can arrive not only on, but also in hair from exogenous sources. A range of solvent-washing procedures have been advocated to “decontaminate” hair by removing adsorbed analytes, but these carry the risk of transporting adsorbed analytes into the medulla of the hair therefore confounding the whole procedure. This is especially true if segmental analysis is being undertaken in order to provide a “time course” of drug exposure. Proposed clinical applications of hair analysis: There have been a number of reports where drugs seemingly administered during the perpetration of a crime have been detected in head hair. However, detailed evaluation of these reports is difficult without full understanding of the possible effects of any “decontamination” procedures used and of other variables such as hair color or cosmetic hair treatment. Similarly, in child custody cases and where the aim is to demonstrate abstinence from drug or alcohol use, the issues of possible exogenous sources of analyte, and of the large variations in analyte concentrations reported in known users, continue to confound the interpretation of results in individual cases. Conclusions: Interpretation of results of head hair analysis must take into account all the available circumstantial and other evidence especially as regards the methodology employed and the possibility of surface contamination of the hair prior to collection.
Chapter
With some delay, alcohol biomarkers have taken their position in hair analysis appropriate to the widespread use and abuse of alcohol. In this chapter, the progress of knowledge about biochemical and physiological basics, of methods for analytical determination, of interpretation, and of practical application are reviewed for ethyl glucuronide (EtG), fatty acid ethyl esters (FAEEs), and cocaethylene. Practical application in larger scale was reported for driving ability examination, workplace testing, prenatal alcohol exposure, child protection cases, liver transplantation, and postmortem alcohol exposure. Despite enormous progress further efforts are necessary to harmonize the analytical methods and to increase sensitivity and accuracy for EtG and FAEE, to improve quality control, to optimize the cutoffs and to increase reliability of interpretation. As generally in hair analysis, the toxicologist should be aware of the limitations of these markers and should avoid formal and noncritical application.
Article
Ineffective stress-coping in Africans is associated with cardiac ischemia during acute mental stress. Ischemic conditions may be worsened by stress-induced release of glial-derived S100‑calcium-binding-protein β (S100B), which is pro-apoptotic for cardiomyocytes Whether estradiol as coping regulator and cardio-protective factor will protect against pro-apoptotic effects, remains unclear. Therefore, we aimed to investigate stress-induced associations between cardiac troponin T/cTnT (cardiac ischemic marker), S100B and estradiol in a bi-ethnic cohort of defensive copers of both sexes. The target population study included African and Caucasian teachers of both sexes (n = 344; aged 20–65 years). The Stroop-color-word-conflict-test was administrated for 1 min to induce acute mental stress in the participants. A chronic stress risk phenotype score was obtained. The Coping Strategy Indicator determined habitual defensive/avoidance/seeking social support coping scores. Fasting blood samples were obtained prior to and 10 min post-Stroop-stress to assess cTnT, S100B and estradiol levels. An interaction between ethnicity, sex and defensive coping (p˂0.05) was found for acute stress-induced percentage changes in estradiol. In defensive coping African men, the Stroop-color-word-conflict-test elicited decreases in S100B and increases in estradiol. Again, in this group, S100B decreases were related to unchanged cTnT, a chronic stress risk phenotype and acute estradiol increases (p < 0.05). No associations among main markers were apparent in the African women or the Caucasian defensive copers of both sexes. In the defensive coping African men, the markers studied may play a relevant role in the brain-cardiovascular system interaction during stress exposure. Further research is needed to elaborate on potential mechanisms and to establish clinical relevance.
Article
Forensic investigation involves gathering the information necessary to understand the criminal events as well as linking objects or individuals to an item, location or other individual(s) for investigative purposes. For years techniques such as presumptive chemical tests, DNA profiling or fingermark analysis have been of great value to this process. However, these techniques have their limitations, whether it is a lack of confidence in the results obtained due to cross-reactivity, subjectivity and low sensitivity; or because they are dependent on holding reference samples in a pre-existing database. There is currently a need to devise new ways to gather as much information as possible from a single trace, particularly from biological traces commonly encountered in forensic casework. This review outlines the most recent advancements in the forensic analysis of biological fluids, fingermarks and hair. Special emphasis is placed on analytical methods that can expand the information obtained from the trace beyond what is achieved in the usual practices. Special attention is paid to those methods that accurately determine the nature of the sample, as well as how long it has been at the crime scene, along with individualising information regarding the donor source of the trace.
Article
The use of hair as a matrix for the evaluation of chronic ethanol drinking behavior presents the advantage of a longer window of detection and higher specificity when compared to classical biochemical markers. The most recent recommendations the Society of Hair Testing (SOHT) indicate that ethyl palmitate (EtP) hair levels can be used to estimate the ethanol drinking behavior, alternatively to the combined measurement of four main fatty acid ethyl esters. In this study, solid-phase microextraction (SPME) conditions for the extraction of EtP from hair were optimized using response surface analysis, after a Box-Behnken experiment. Analyses were performed by GC-MS. The optimized HS-SPME conditions, using a PDMS-DVB (65 μm) fiber, were pre-adsorption time of 6 min, extraction time of 60 min and incubation temperature of 94°C. The linear range was 0.05 to 3 ng mg-1, with accuracy within 95.15-109.91%. Between-assay and within-assay precision were 8.58-12.53 and 6.12-6.82%, respectively. The extraction yield was 61.3-71.9%. The assay was applied to hair specimens obtained from 46 volunteers, all presenting EtP levels within the linear range of the assay. Using a statistically designed experiment, a sensitive SPME-GC-MS assay for the measurement of EtP in hair was developed and validated, requiring only 20 mg of hair.
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Sympathetic activation may trigger acute coronary syndromes. We examined the relation between circulating neurotrophic factors and hemostatic risk factors of atherothrombotic cardiovascular disease at baseline and in response to acute mental stress to establish a brain-heart link. In 409 black and white South Africans, brain-derived neurotrophic factor (BDNF) and fibrinolytic measures were assessed at baseline. Glial cell-derived neurotrophic factor (GDNF), S100 calcium-binding protein (S100B), von Willebrand factor (VWF), fibrinogen and D-dimer were assessed at baseline and 10 min after the Stroop test. Neurotrophins were regressed on hemostatic measures adjusting for demographics, comorbidities, cardiometabolic factors and health behaviors. Higher baseline BDNF was associated with greater stress-induced increase in fibrinogen (p = 0.003) and lower D-dimer increase (p = 0.016). Higher baseline S100B was significantly associated with higher baseline VWF (p = 0.031) and lower fibrinogen increase (p = 0.048). Lower baseline GDNF was associated with higher baseline VWF (p = 0.035) but lower VWF increase (p = 0.001). Greater GDNF (p = 0.006) and S100B (p = 0.042) increases were associated with lower VWF increase. All associations showed small-to-moderate effect sizes. Neurotrophins and fibrinolytic factors showed no significant associations. The findings support the existence of a peripheral neurothrophin-hemostasis interaction of small-to-moderate clinical relevance. The implications for atherothrombotic cardiovascular disease need further exploration.
Article
Introdução/ enquadramento/ objetivos: A generalidade dos profissionais a exercer em Saúde Ocupacional já vivenciou situações em que algum funcionário consumia álcool em quantidades superiores às consideradas adequadas para exercer as suas tarefas profissionais. Por vezes é muito relevante perceber se o indivíduo realmente parou de consumir e se já se encontra apto. Além de nem todos os gabinetes médicos terem instrumentos homologados para testar o etanol exalado ou urinário, tal apenas dá informação relativa a consumos nas últimas horas. Pode haver assim a necessidade de saber se nos últimos dias, semanas ou meses, houve consumo. Ao longo dos últimos anos foram surgindo diversos doseamentos em vários substratos (urina, sangue, cabelo, unha, ar exalado) que têm a capacidade de, com uma margem razoável de segurança, perceber qual tem sido o consumo etílico até os últimos seis meses, sensivelmente, em alguns casos. Pretendeu-se com esta revisão fazer um resumo do que mais recente e pertinente se publicou sobre o tema. Metodologia: Trata-se de uma Revisão Bibliográfica Integrativa, iniciada através de uma pesquisa realizada em junho de 2019 nas bases de dados “CINALH plus with fullt ext, Medline with full text, Database of Abstracts of Reviews of Effects, Cochrane Central Register of ControlledTrials, Cochrane Database of Systematic Reviews, Cochrane Methodology Register, Nursing and Allied Health Collection: comprehensive, MedicLatina e RCAAP”. Conteúdo: Entre os marcadores clássicos ou indiretos (que quantificam algumas lesões associadas ao álcool) destacam-se o ALT, AST, GGT, plaquetas e volume corpuscular médio, bem como o CDT. Os principais biomarcadores diretos são o Etanol, PEth, FAEEs, EtG e EtS, entre outros muito recentes e ainda em estudo (nota: todas estas siglas são referidas por extenso, na primeira vez que forem referidas no texto). Discussão e Conclusões: Ainda que não existam consensos absolutos, parece claro que os marcadores diretos dão informação mais fidedigna sobre o consumo agudo e sobretudo crónico, além de que são menos modulados pela patologia hepática, razoavelmente frequente entre indivíduos com dependência. Contudo, os marcadores indiretos ou clássicos são mais facilmente doseados em praticamente qualquer laboratório e a um custo muito mais acessível. Para além destas questões técnico/ científicas, a Equipa de Saúde Ocupacional terá de criar previamente uma infraestrutura, acordada com o empregador e um laboratório, que permita a requisição destes doseamentos com sigilo, ou seja, pagando o empregador o valor combinado (fixo ou em função do que for realmente doseado), sem saber o que foi testado ou a quem, mesmo que a dependência seja do conhecimento de todos na empresa, como por vezes acontece.
Article
The use of alcohol, despite it being a psychoactive substance, is widely accepted. This acceptance and the ease of access may lead to alcohol abuse and dependence. Misuse of alcohol is one of the leading causes of preventable illness, injury or death, has social consequences, and can be linked to various types of criminal activities. Upon monitoring alcohol consumption, the question is not necessarily solely whether or not someone is or was abstinent, but may also imply what kind of drinking behavior or patterns are displayed. To reveal the answer, a portfolio of different biomarkers for alcohol (ab)use has been established. A distinction is made between indirect and direct markers. Indirect biomarkers are proteins, enzymes, or cells of which the level or activity undergoes typical changes as a result of (excessive) alcohol intake [e.g., carbohydrate‐deficient transferrin (CDT)], while direct biomarkers are ethanol itself and molecules which are directly related to the metabolism of alcohol [e.g., ethyl glucuronide (EtG), ethyl sulphate (EtS), fatty acid ethyl esters (FAEE), and phosphatidylethanol (PEth)]. These direct biomarkers have the potential to differentiate between drinking behaviors because they allow a higher sensitivity and specificity, in particular for the detection of abstinence or any drinking, and because they can be measured in different matrices, with different detection windows. These assets open the possibilities for testing strategies based on a combination of different markers that can be used both in retrospective and prospective assessments of alcohol intake. This article is categorized under: Toxicology > Alcohol Identification of alcohol consumption behavior or patterns can be achieved using a portfolio of different (in)direct biomarkers with a detection window of up to 6 months.
Article
Early detection of alcohol misuse in OLT recipients is essential to offer patients support and prevent organ damage. Here, ethyl glucuronide, a metabolite of ethanol found in hair (hEtG) was evaluated for detection of alcohol consumption. In 104 transplant recipients, 31 with underlying alcoholic liver disease (ALD) and 73 with non-ALD, hEtG was determined in addition to the alcohol markers urine EtG, blood ethanol, methanol and carbohydrate deficient transferrin. Results were compared with patients' self-reports in a questionnaire and physicians' assessments. By physicians' assessments, 22% of patients were suspected of consuming alcohol regularly, although only 6% of patients acknowledged consumption of a moderate or high amount of alcohol. By testing all markers except for hEtG, alcohol consumption was detected in 7% of patients. When hEtG testing was added to the assessment, consumption was detected in 17% of patients. Hair-EtG determination alone revealed chronic alcohol consumption of >10g/day in 15% of patients. ALD patients had a positive hEtG result significantly more often than non-ALD patients did (32% vs 8%; P=0.003). Also the concentration of hEtG was higher in ALD patients (P=0.049) and revealed alcohol abuse with consumption of >60g ethanol/day in 23% of ALD and 3% of non-ALD patients. Patients' self-reports and physicians' assessments had a low sensitivity of 27% and 67%, respectively for detecting regular alcohol intake as indicated by hEtG. Hair-EtG determination improved the detection of liver transplant patients who used alcohol, revealing regular alcohol consumption in 32% of ALD and 8% of non-ALD patients.
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The applicability of fatty acid ethyl esters (FAEEs) and ethyl glucuronide (EtG) in hair in a workplace alcohol testing program was investigated. A total of 78 hair samples from employees in jobs with a high endangering potential were tested for EtG and FAEEs. In most cases excessive drinking was suspected. For 59 of these cases additional data of the traditional alcohol markers aspartate aminotransferase, alanine aminotransferase and gamma-glutamyltransferase and of the mean corpuscular volume of the erythrocytes (58 cases) were available. By application of the cut-offs of the Consensus of the Society of Hair Testing and of a gradual system for combined interpretation of FAEEs and EtG in hair no indications of alcohol abuse were obtained in 50 cases (64%), slight indications were seen in 13 cases (17%) and clear indications in 11 cases (14%). In four cases, the results were inconclusive with strongly conflicting results of both markers, the reason for which could not be cleared. The traditional markers confirmed the hair results only partly and displayed altogether a lower portion of positive results. EtG and FAEEs in hair, especially when interpreted in combination, are suitable for application in workplace alcohol testing programs. Nevertheless, the results obtained by hair analysis for alcohol markers can only be one part of a proper assessment aiming at the question whether an employee is addicted to alcohol or not.
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Ethyl glucuronide (EtG) in hair has emerged as a useful biomarker for detecting alcohol abuse and monitoring abstinence. However, there is a need to establish a reliable cutoff value for the detection of chronic and excessive alcohol consumption. One hundred and twenty-five subjects were classified as teetotalers, low-risk drinkers, at-risk drinkers, or heavy drinkers. The gold standard for subjects' classifications was based on a prospective daily alcohol self-monitoring log. Subjects were followed for a 3-month period. The EtG diagnostic performance was evaluated and compared with carbohydrate-deficient transferring (CDT) and the activities of aspartate aminotransferase, alanine aminotransferase, and γ-glutamyl-transferase (γGT). A cutoff of >9 pg/mg EtG in hair, suggesting an alcohol consumption of >20/30 g (at-risk drinkers), and a cutoff of >25 pg/mg, suggesting a consumption of >60 g (heavy drinkers), were determined by receiver operating characteristic analysis. The EtG diagnostic performance was significantly better (P < 0.05) than any of the traditional biomarkers alone. EtG, as a single biomarker, yielded a stronger or similar diagnostic performance in detecting at-risk or heavy drinkers, respectively, than the best combination of traditional biomarkers (CDT and γGT). The combination of EtG with traditional biomarkers did not improve the diagnostic performance of EtG alone. EtG demonstrated a strong potential to identify heavy alcohol consumption, whereas the traditional biomarkers failed to do so. EtG was not significantly influenced by gender, body mass index, or age. Hair EtG definitively provides an accurate and reliable diagnostic test for detecting chronic and excessive alcohol consumption. The proposed cutoff values can serve as reference for future cutoff recommendations for clinical and forensic use.
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Previous studies have indicated that the use of high-ethanol-content (>65%) hair-care products may elevate fatty acid ethyl ester (FAEE) concentrations in hair. In this case series, nine individuals were identified by FAEE analysis to be chronic alcohol abusers in the context of child-welfare substance abuse monitoring. Based on patient claims of moderate or no alcohol consumption, the presence of ethanol in the patients' hair-care regimens was investigated. Samples were additionally tested for the presence of ethyl glucuronide (EtG). From a total of nine patients, 12 hair samples were submitted for analysis. Patient histories were obtained as well as Material Safety Data Sheets (MSDS) listing hair-care product ethanol content. Hair samples were pre-washed to remove external contamination and analyzed for FAEE and EtG by GC-MS. According to the Society of Hair Testing consensus guidelines, FAEE levels exceeding 0.50 ng/mg and/or EtG levels exceeding 30 pg/mg indicate chronic excessive alcohol consumption. Upon initial analysis, the nine samples exhibited positive FAEE findings ranging from 0.496 to 4.984 ng/mg. MSDS review revealed the presence of ethanol from 10% to 95% by volume in at least one hair-care product used by each individual. Results of the EtG analysis ranged from 1.9 to 23.5 pg/mg. These findings indicate that regular use of products with ethanol content as low as 10% can impact FAEE results. EtG analysis should be used to confirm FAEE findings and appears to be unaffected by hair-care products, likely due to alternative mechanisms of incorporation.
Article
Ethyl glucuronide (EtG) in hair is a proposed biomarker for alcohol consumption. This study compares hair EtG concentrations with self-reported alcohol consumption data, in individuals with a range of alcohol use. Hair was collected from 100 participants with a range of alcohol use. Participants completed an Alcohol Use Disorders Identification Test C questionnaire to record alcohol consumption. Participants were categorized into one of the four groups: tee-totallers (consuming 0 units a week), lower-risk drinkers (1-21 units a week), increasing-risk drinkers' consuming (22-50 units a week) and high-risk drinkers (over 50 units a week). Hair from the proximal 3 cm was analysed for EtG using gas chromatography-mass spectrometry/mass spectrometry. EtG was detected in 29 out of 100 hair samples. Based on the Society of Hair Testing (SOHT) threshold of 30 pg/mg EtG, the hair test identified alcohol consumption in 57.9% of high-risk drinkers, 45.5% of increasing-risk drinkers and only 9.8% of lower-risk drinkers. EtG sensitivity was highest for high-risk drinkers (consuming more than 50 units a week), identified to be 0.52 using a 30 pg/mg threshold and 0.58 using a 45 pg/mg threshold. A positive result is highly likely to indicate any drinking (positive predictive value, 1.00). A negative result does not provide good evidence for abstinence (negative predictive value, 0.23). EtG has been identified to be a low sensitivity marker that cannot be used quantitatively to determine alcohol exposure. EtG can be used qualitatively to indicate alcohol consumption with a positive result providing strong evidence for an individual drinking within the past 3 months.
Article
Ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEE), non-volatile, direct metabolites of ethanol have been shown to be suitable markers for the evaluation of social and chronic excessive alcohol consumption. Previous investigations have shown that the regular use of hair-care products with high alcohol content lead to an increase of FAEE concentration and consequently gave false-positive results for the determination of FAEE in hair. In this study we investigated the influence of a long-term hair treatment with EtOH containing lotion, on the EtG concentrations in hair. In this study 7 volunteer subjects (classified as either rare, social or heavy drinkers) treated the right side of their scalp every day during a one or two month period with a commercial hair tonic (Seborin), which contains 44.0% ethanol (vol%). Collection of hair specimens from both sides of the scalp was done one day before hair treatment, one week and one month after treatment (for 5 subjects also after two months of treatment). A hair segment of 3 centimeters (cm) was cut and then washed with water and acetone, and then pulverized. EtG was quantified by GC/MS after pulverization and 2h of ultrasonication in water, extraction by solid phase extraction using Oasis MAX columns and derivatization with HFBA. Measurements were done in negative chemical ionization mode using EtG-D5 as internal standard. Comparison of EtG concentration in the treated and in the non-treated hair specimens did not show any increase at the different dates of collection for the 7 subjects. In conclusion, these results show that there is no indication for an increase of EtG after use of ethanol containing hair cosmetics.
Article
This article presents results from 1872 hair samples, which were analyzed for fatty acid ethyl esters (FAEEs) and ethyl glucuronide (EtG). The results were evaluated in the context of self-reported drinking behavior, the use of hair cosmetics, the gender of the sample donors and hair sample length. For comparison, CDT and GGT in serum were available in 477 and 454 cases, respectively. A number of alcohol abstainers or low moderate drinkers and excessive drinkers were selected for assessment of cut-offs for FAEEs in the proximal 6 cm hair segments and for EtG in the proximal 3 cm hair segments. Cut-off values were assessed by ROC analysis. It was found that the cut-offs of 1.0 ng/mg FAEE and 30 pg/mg EtG presently used for excessive drinking lead to a low portion of false positives (4% and 3% respectively) but to a higher portion of false negatives (23% and 25% respectively). Comparison of the mean and medium concentrations in samples without any reported hair cosmetics (N = 1079) and in samples with reported use of hair spray (N = 79) showed an increase by the factor of about two for FAEE but no significant difference for EtG. Mean values of EtG were decreased by 80% in bleached samples (N = 164) and by 63% in dyed samples (N = 96). There was no significant effect of bleaching and dyeing on FAEE. Hair gel and hair wax, oil or grease showed no significant effect on both FAEE and EtG.
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In a case of a driving ability assessment, hair analysis for ethyl glucuronide (EtG) was requested by the authorities. The person concerned denied alcohol consumption and did not present any clinical sign of alcoholism. However, EtG was found in concentrations of up to 910 pg/mg in hair from different sampling dates suggesting an excessive drinking behavior. The person declared to use a hair lotion on a regularly base. To evaluate a possible effect of the hair lotion, prospective blood and urine controls as well as hair sampling of scalp and pubic hair were performed. The traditional clinical biomarkers of ethanol consumption, CDT and GGT, were inconspicuous in three blood samples taken. EtG was not detected in all collected urine samples.
Article
The study aimed to evaluate the diagnostic sensitivity (SE) and specificity (SP) of chronic alcohol misuse diagnosis by comparing traditional biomarkers with ethyl glucuronide (EtG), an ethanol direct metabolite, detected in the keratinic matrix. Seventy-six subjects tested for chronic alcohol abuse for different purposes were recruited. EtG was detected in hair, whereas the analyses of carbohydrate-deficient transferrin (CDT), alanine transaminase, aspartate transaminase, gamma-glutamyl transferase, mean corpuscular volume, and mean corpuscular hemoglobin were performed in serum samples. Of the 76 patients examined, 26 were judged by the medical doctors as subjects with alcohol abuse problems and, therefore, not eligible for driving license renewal or liver transplant. EtG in hair (SE = 0.68, SP = 1.00) showed the best diagnostic SE and SP compared with the other biomarkers investigated. Among the traditional biomarkers, only CDT proved to be suitable for forensic purposes because of the high diagnostic specificity (SP = 1.00) although it showed poor diagnostic SE (0.27). The percentage of positive samples decreased for all the biomarkers by excluding the subjects with hepatic diseases, except for EtG and CDT, suggesting that these 2 biomarkers could be less affected by false positive results, because of hepatic diseases. This study showed that when EtG in hair and CDT results are combined, diagnostic SE in chronic alcohol abuse diagnosis clearly improved, suggesting that complementary analysis of both these biomarkers provides the best diagnostic tool in suspected cases of chronic excessive alcohol consumption.
Article
An important goal of forensic and clinical toxicology is to identify biological markers of ethanol consumption that allow an objective diagnosis of chronic alcohol misuse. Blood and head hair samples were collected from 175 subjects—objectively classified as non-drinkers (N = 65), social drinkers (N = 51) and active heavy drinkers (N = 59)—and analyzed to determine eight traditional indirect biomarkers of ethanol consumption [aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyltransferase (γ-GT), alkaline phosphatase (ALP), mean corpuscular volume (MCV), carbohydrate-deficient transferrin (CDT), and cholesterol and triglycerides in blood] and one direct biomarker [ethyl glucuronide (EtG) in head hair]. The experimental values obtained from these determinations were submitted to statistical evaluations. In particular, Kruskal–Wallis, Mann–Whitney and ROC curve analyses, together with principal component analysis (PCA), allowed the diagnostic performances of the various biomarkers to be evaluated and compared consistently. From these evaluations, it was possible to deduce that EtG measured in head hair is the only biomarker that can conclusively discriminate active heavy drinkers from social and non-drinkers, using a cut-off value of 30 pg/mg. In contrast, a few indirect biomarkers such as ALP, cholesterol, and triglycerides showed extremely low diagnostic abilities and may convey misleading information. AST and ALT proved to be highly correlated and exhibited quite low sensitivity and specificity. Consequently, either of these parameters can be discarded without compromising the classification efficiency. Among the indirect biomarkers, γ-GT provided the highest diagnostic accuracy, while CDT and MCV yielded high specificity but low sensitivity. It was therefore concluded that EtG in head hair is the only biomarker capable of supporting a confirmatory diagnosis of chronic alcohol abuse in both forensic and clinical practice, while it was found that γ-GT, CDT, MCV, and AST—whether used alone or in combination—do not allow the conclusive classification of subjects according to ethanol consumption. However, a diagnostic strategy combining these four parameters could be formulated in order to create a multivariate model capable of screening suspected active heavy drinkers. Figure PCA of 175 patients clinically classified as non-drinkers (green dots, N = 65), social drinkers (yellow dots, N = 51) and active heavy drinkers (red dots, N = 59). Score and loading (six parameters) bi-plot of PC1 and PC2.
Article
Drug and alcohol abuse is a concern for many European companies, especially those having safety-critical jobs. It is not uncommon for European companies to establish a drug policy with little or no provision for drug testing. The European Union (EU) has launched a number of initiatives in its fight against drugs. There is, however, no specific EU legislation and no generally accepted guidelines. Since the outcome of workplace drug testing (WDT) can have serious consequences for the employee, it is of utmost importance that WDT be performed in a defined quality standard and in a legally secured way. In order to fulfil this, the European Workplace Drug Testing Society (EWDTS) has formulated WDT guidelines in order to ensure that the entire drug testing process is of high quality, accredited, and legally defensible, hence giving accurate and reliable information about employee drug use while respecting the privacy of the employee. The aim of this paper is to present the recently formulated guidelines for workplace drug testing in hair.
Article
Ethyl glucuronide (EtG) is a minor and specific metabolite of ethanol. It is incorporated into growing hair, allowing a retrospective detection of alcohol consumption. However, the suitability of quantitative EtG measurements in hair to determine the quantity of alcohol consumed has not clearly been demonstrated yet. The purpose of this study was to evaluate the influence of ethanol dose and hair pigmentation on the incorporation of EtG into rat hair. Ethanol and EtG kinetics in blood were investigated after a single administration of ethanol.
Article
In this study the combined use of fatty acid ethyl esters (FAEE) and ethyl glucuronide (EtG) for diagnoses of chronically excessive alcohol abuse is investigated at 174 hair samples from driving ability examination, workplace testing and child custody cases for family courts and evaluated with respect to the basics of interpretation. Using the cut-off values of 0.50 ng/mg for FAEE and 25 pg/mg for EtG, both markers were in agreement in 75% of the cases with 103 negative and 28 positive results and there were 30 cases with FAEE positive and EtG negative and 13 cases with FAEE negative and EtG positive. As the theoretical basis of interpretation, the pharmacokinetics of FAEE and EtG is reviewed for all steps between drinking of ethanol to incorporation in hair with particular attention to relationships between alcohol dose and concentrations in hair. It is shown that the concentrations of both markers are essentially determined by the area under the ethanol concentration in blood vs. time curve AUC(EtOH), despite large inter-individual variations. It is demonstrated by calculation of AUC(EtOH) on monthly basis for moderate, risky and heavy drinking that AUC(EtOH) increases very strongly in the range between 60 and 120 g ethanol per day. This specific feature which is caused by the zero-order elimination of ethanol is a favorable prerequisite for a high discrimination power of the hair testing for alcohol abuse. From the consideration of the different profiles of FAEE and EtG along the hair and in agreement with the literature survey, a standardized hair segment 0-3 cm is proposed with cut-off values of 0.5 ng/mg for FAEE and 30 pg/mg for EtG. This improves also the agreement between FAEE and EtG results in the cases of the present study. A scheme for combined interpretation of FAEE and EtG is proposed which uses the levels of abstinence and the double of the cut-off values as criteria in addition to the cut-off's. Considering the large variations in the relationship between ethanol dose and FAEE and EtG concentrations in hair, the combined use of both parameters strongly increases the accuracy of the diagnosis by mutual confirmation and identification of false positive or false negative results due to biological variations or analytical errors.
Article
Ethyl glucuronide in hair (HEtG) has recently gained great attention, because of its high sensitivity and specificity in the diagnosis of chronic alcohol abuse. Due to its high polarity hydrophilicity, a strong hair treatment followed by a shampooing may lead to removal/degradation of this molecule from hair matrix. To set up an in vitro study in order to evaluate the ability of bleaching of modifying HEtG test results. Thirty hair samples from teetotalers (n=5), social drinkers (n=4) and heavy drinkers (n=21), after an informed written consent, were collected and divided longitudinally into four aliquots. The first aliquot was kept untreated and was processed following the method routinely used in our lab for the determination of HEtG (double washing with methanol/dichloromethane, overnight incubation in water, and LC-MS/MS analysis, LLOQ: 3pg/mg). To the other three aliquots a commercially available bleaching solution was applied, according to the manufacturer's instructions. One out of the three aliquots was submitted to the analysis by following the same procedure used for the untreated sample. The other two were submitted to a purification step before LC-MS/MS analysis, by using two different SPE cartridges (aminopropyl and dimethyl butylamine). HEtG levels in the untreated samples from social drinkers and heavy drinkers ranged from 7.7 to 149.0pg/mg. All the samples from teetotalers tested negative. The treated samples processed without any SPE extraction and with aminopropyl cartridges showed a relevant ion suppression for both EtG and D(5)-EtG (IS) signals. Samples treated with the bleaching solution and extracted with dimethyl butylamine cartridge allowed to sensitively reduce ion suppression (less than 35%) and to verify that EtG, after a strong treatment like bleaching, completely disappears. This in vitro study showed that HEtG disappears from hair matrix after a strong hair treatment. It is not clear whether the mechanism involved is chemical degradation or physical removal from the damaged keratinic matrix. However, owing to the highly hydrophilic character of the compound, the second mechanism seems more likely to occur. Finally, bleaching solutions could lead to a heavy ion suppression of this metabolite that may be avoided by using an SPE purification before instrumental analysis.
Article
An apparent decrease in deaths attributed to sudden infant death syndrome (SIDS) has been noted in a number of diverse geographical areas during the past several years. At the same time the definition of SIDS has been in a state of flux and some observers have raised the possibility that the fall in SIDS deaths is due to diagnostic transfer rather than to a genuine decrease in numbers. The present study was undertaken to investigate this possibility. All sudden and unexpected deaths in infants under 1 year of age in South Australia during a 10 year period from 1984 to 1993 were reviewed. The number of deaths due to SIDS fell from 40 in 1984 to 17 in 1993, with a maximum of 52 cases per year in 1987. In contrast, the number of cases of sudden death not due to SIDS remained under 10 per year. The overall infant death rate also fell, while the total number of births per year remained relatively unchanged. The lack of major change in sudden infant death rates from other causes, combined with the fall in SIDS deaths, is not supportive of diagnostic transfer being a major determinant of the declining SIDS death rate. Therefore, other factors are likely to be responsible for the falling SIDS rate in this population.
Article
Carbohydrate-deficient transferrin (CDT) has been demonstrated to be a marker of prolonged heavy alcohol consumption. We compared this marker with gamma-glutamyltranspeptidase (GGT) and mean corpuscular volume (MCV) in alcohol and nonalcohol abusers with liver disease. Our results confirm that the sensitivity of CDT in alcoholics is high, although lower than that of GGT and MCV; however, the specificity of CDT was higher than that of the other two markers. This finding supports the notion that CDT is only partially influenced by the presence of liver damage, whereas increases of GGT and MCV are greatly affected by several factors, including liver damage and drugs. Moreover, we observed that the sensitivity and the specificity of CDT were greater than those of GGT and MCV in younger drinkers.
Article
Fatty acid ethyl esters (FAEEs) are products of nonoxidative ethanol metabolism. After incorporation in hair, they should be suitable long-term markers of alcohol abuse. Hair samples from 19 alcoholics in a treatment program, 10 fatalities with verified excessive alcohol consumption, 13 moderate social drinkers who consumed up to 20 g ethanol/day, and 5 strict teetotalers were analyzed in 1-12 segments for four FAEEs (ethyl myristate, ethyl palmitate, ethyl oleate, and ethyl stearate) by external degreasing with n-heptane, extraction with a dimethyl sulfoxide-n-heptane mixture, headspace solid-phase microextraction of the extracts, and gas chromatography-mass spectrometry with deuterated internal standards. The n-heptane washings were analyzed in the same way for FAEEs from the hair surface. The sum of the four ester concentrations in hair calculated for the proximal 0-6 cm segment was 2.5-13.5 ng/mg (mean, 6.8 ng/mg) for the fatalities, 0.92-11.6 ng/mg (mean, 4.0 ng/mg) for 17 of the alcoholics in treatment, 0.20-0.85 ng/mg (mean, 0.41 ng/mg) for the moderate social drinkers, and 0.06-0.37 ng/mg (mean, 0.16 ng/mg) for the teetotalers. In almost all cases the segmental concentrations increased from proximal to distal. There was no agreement between the self-reported drinking histories of the participants and the FAEE concentrations along the hair length. Ethyl oleate was the dominant ester in all samples. FAEEs are deposited in hair mainly from sebum. Despite large individual differences, FAEE hair concentrations can be used as markers for excessive alcohol consumption with relatively high accuracy.
Article
Fatty acid ethyl esters (FAEE) can be used as alcohol markers in hair. It was investigated in this study whether this diagnostic method is disturbed by hair care and hair cosmetics. Traces of ethyl myristate, ethyl palmitate, ethyl oleate and ethyl stearate were detected in all of 49 frequently applied hair care products by headspace solid phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS). The highest concentration was 0.003% in a hair wax. From experiments with separated hair samples of alcoholics as well as from the evaluation of the FAEE concentrations and the data about hair care of 75 volunteers (alcoholics, social drinkers and teetotalers) follows that usual shampooing, permanent wave, dyeing, bleaching or shading are of minor importance as compared to the drinking amount and other individual features. However, false positive results were found after daily treatment with a hair lotion containing 62.5% ethanol, with a deodorant and with a hair spray. As an explanation, it is assumed that FAEE are formed in the sebum glands also after regular topical application of products with a higher ethanol content.
Article
Alcohol and alcohol-related diseases have become a major cause of death in Western countries. The most sensitive and specific of the commonly used biomarkers of alcohol intake are carbohydrate-deficient transferrin (CDT), and the combination of gamma-glutamyltransferase (GGT) and CDT. Other widely used laboratory markers are GGT, mean corpuscular volume of erythrocytes and the ratio of aspartate aminotransferase to alanine aminotransferase. Blood ethanol levels reveal recent alcohol use. However, more specific and sensitive biomarkers to improve the detection of excessive alcohol use at an early stage are needed. New biomarkers, not yet used in routine clinical work, include phosphatidylethanol, fatty acid ethyl esters, ethyl glucuronide, sialic acid, and acetaldehyde adducts.
Article
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.
Germany e-mail: rene.gapert@ucd.ie; renegapert@daad-alumni
  • M Hastedt
  • M Bü
  • R Gapert
  • S Herre
  • F Krumbiegel
  • M Tsokos
M. Hastedt Á M. Bü Á R. Gapert (&) Á S. Herre Á F. Krumbiegel Á M. Tsokos Á S. Hartwig Institute of Legal Medicine and Forensic Sciences, Charité, Medical University Berlin, Berlin, Germany e-mail: rene.gapert@ucd.ie; renegapert@daad-alumni.de M. Rothe Lipidomix GmbH, Berlin, Germany R. Gapert Human Anatomy, UCD School of Medicine & Medical Science, University College Dublin, Dublin, Ireland T. Kienast Psychiatrische Universitä der Charité im St. Hedwig-Krankenhaus, Berlin, Germany A. Heinz Klinik fü Psychiatrie, Psychotherapie und Psychosomatik der Charité, Universitä Berlin, Berlin, Germany References
Urteilsbildung in der Medizinisch-Psychologischen Fahreignungsdiagnostik
  • W Schubert
  • Mattern R Beurteilungskriterien