The rate and kinetic order of ethanol elimination

The rate and kinetic order of ethanol elimination was evaluated in human volunteers. Part I of the study involved dosing individuals with alcoholic beverages on two separate occasions. Breathalyzer tests were performed at 15-min intervals for a period of 5 h. Attention was focused on values obtained after peak blood ethanol levels had been reached. The second part of the study included having samples drawn from alcoholics at predetermined intervals during recovery from alcoholic intoxication. Blood ethanol concentration data was analyzed for kinetic order and a comparison of ethanol elimination rates of alcoholics and non-alcoholics was made. The predicative capability of estimating a BAC from both the zero and first order theories was also investigated.It was concluded that ethanol elimination is a zero order process. For subjects classified as non-drinkers (consume less than 6 ounces of ethanol/month), the mean ethanol elimination rate as determined in the study was 12 ± 4 mg/h. For subjects classified as social drinkers (consume more than 6 ounces but less than 30 ounces of ethanol/month), the mean ethanol elimination rate was 15 ± 4 mg%/h, and for alcoholics, the mean ethanol elimination rate was 30 ± 9 mg%/h. These results indicate that the rate of ethanol elimination increases with drinking experience.     

Vitreous humor carbohydrate-deficient transferrin concentrations in the postmortem diagnosis of alcoholism

Deaths from the effects of alcohol intoxication are encountered routinely in forensic practice. In an important number of cases difficulty may arise in interpreting the significance of results obtained in the autopsy. In clinical practice biochemical markers, particularly serum gamma-glutamyl-transpeptidase (GGT), alanine aminotransferase (ALT), aspartate transaminase (AST), carbohydrate-deficient transferrin (CDT), and erythrocyte mean corpuscular volume are used to diagnose heavy alcohol consumption. CDT is used as a reliable and specific marker. In postmortem diagnosis, because of the difficulty in interpreting blood alcohol levels and relatively non-specific pathological features, biochemical compounds have been studied for use as possible markers. The aim of this study was to evaluate the usefulness of the postmortem determination of CDT in vitreous humor as a confirmation of antemortem alcoholism. CDT levels were studied in 66 male cadavers with a mean age of 55.9 years (S.D. 17.0, range 22-87 years) with a mean postmortem interval of 17.9 h (S.D. 11.4, range 4-72 h). Cases were assigned to two diagnostic groups according to the antemortem diagnosis of alcoholism. Statistically significant differences were found for CDT and ALT concentrations between the two diagnostic groups. The highest vitreous humor levels of CDT and ALT were obtained in the group of cases with a previous diagnosis of alcoholism. Our results suggest that vitreous humor CDT levels are useful in cases where the postmortem diagnosis of alcoholism is hindered by the non-specificity of data.     

Comparison of Four Saliva Detection Methods to Identify Expectorated Blood Spatter

Blood spatter analysis is an important step for crime scene reconstruction. The presence of saliva in blood spatter could indicate expectorated blood which is difficult to distinguish from impact spatter. In this study, four saliva test methods (SALIgAE^®, Phadebas^® sheet, RSID^™-Saliva kit, and starch gel diffusion) were compared to identify the best method for detecting expectorated blood spatter. The RSID^™-Saliva kit showed the highest sensitivity even when saliva was mixed with blood, and was not inhibited by the presence of blood. The SALIgAE^® test provided easy and rapid results, but the yellow color of a positive reaction was overwhelmed by the red color of the blood. The starch gel diffusion method and the Phadebas^® sheet exhibited relatively low sensitivity and the assay took a long time. When using the RSID^™-Saliva kit for identifying saliva in blood, results should be read within 10 min.     

Forensic analysis of carbohydrate-deficient transferrin (CDT): implementation of a screening and confirmatory analysis concept is hampered by the lack of CDT isoform standards

The aim of the study was to test combinations of commercially available carbohydrate-deficient transferrin (CDT) assays for their usefulness as screening and confirmatory CDT analysis systems. A set of 292 serum samples from routine CDT analysis was analyzed by two assays based on anion-exchanger microcolumn CDT and non-CDT fractionation followed by a turbidimetric immunoassay (ChronAlcoI.D. and %CDT TIA) and a high-performance liquid chromatography with on-line sample preparation (ClinRep CDT on-line). The CDT analysis results were divided into four groups based on the test-specific borderlines of the compared methods: NN with negative CDT results by both tests, PN with positive screening but negative confirmation results, NP with negative screening and positive confirmation results, and PP with positive results by both tests. Regardless of the test combination and whether applying the lower or upper limits of the borderlines, approximately one-third of contradictory (positive screening and negative confirmation or vice versa corresponding to groups PN and NP) were obtained. This was not due to analytical outliers (only 6 of 292 serum samples). Indeed, parametric and non-parametric ANOVA analysis pointed to different calibrations and/or recoveries of the three CDT assays. Our data give again evidence for the urgent need of an international CDT isoform standard material. At this time, we cannot recommend a combination of the three tests for screening and confirmatory analysis in forensic CDT testing.     

Urine/blood ratios of ethanol in deaths attributed to acute alcohol poisoning and chronic alcoholism

The concentrations of ethanol were determined in femoral venous blood (BAC) and urine (UAC) and the UAC/BAC ratios were evaluated for a large case series of forensic autopsies in which the primary cause of death was either acute alcohol poisoning (N=628) or chronic alcoholism (N=647). In alcohol poisoning deaths both UAC and BAC were higher by about 2g/l compared with chronic alcoholism deaths. In acute alcohol poisoning deaths the minimum BAC was 0.74 g/l and the distribution of UAC/BAC ratios agreed well with the shape of a Gaussian curve with mean+/-standard deviation (S.D.) and median (2.5th and 97.5th centiles) of 1.18+/-0.182 and 1.18 (0.87 and 1.53), respectively. In alcoholism deaths, when the BAC was above 0.74 g/l (N=457) the mean+/-S.D. and median (2.5th and 97.5th centiles) UAC/BAC ratios were 1.30+/-0.29 and 1.26 (0.87 and 2.1), respectively. When the BAC was below 0.74 g/l (N=190), the mean and median UAC/BAC ratios were considerably higher, being 2.24 and 1.58, respectively. BAC and UAC were highly correlated in acute alcohol poisoning deaths (r=0.84, residual S.D.=0.47 g/l) and in chronic alcoholism deaths (r=0.95, residual S.D.=0.41 g/l). For both causes of death (N=1275), the correlation between BAC and UAC was r=0.95 and the residual S.D. was 0.46 g/l. The lower UAC/BAC ratio observed in acute alcohol poisoning deaths (mean and median 1.18:1) suggests that these individuals died before absorption and distribution of ethanol in all body fluids were complete. The higher UAC/BAC ratio in chronic alcoholism (median 1.30:1) is closer to the value expected for complete absorption and distribution of ethanol in all body fluids.     

Tolerance at high blood alcohol concentrations: a study of 110 cases and review of the literature

One hundred ten consecutive alcoholics who voluntarily entered a detoxification center were studied to determine their ability to perform certain designated functions (a) while under the influence of alcohol at admission and (b) four days later, after they had undergone detoxification. Their blood alcohol concentrations (BACs) within an hour of admission were determined by using the DuPont Automated Clinical Analyzer (ACA), following standard procedures. The results of the tests performed by these subjects were then studied to ascertain the level of psychomotor impairment in alcoholics at elevated BACs. The results were broken down by age, sex, race, and BAC. The findings indicate that alcoholics develop an increased tolerance to alcohol at BACs, which are extremely high, including levels generally considered to be potentially fatal.     

Variability in the Blood/Breath Alcohol Ratio and Implications for Evidentiary Purposes

The breath analyzer is an indispensable tool for identifying alcohol levels among drivers. While numerous studies have shown high correlations between blood and breath alcohol concentrations, most are limited by the study design. This study seeks to assess this relationship by minimizing potential measurement bias, document time from alcohol consumption to testing, and adjusting for potential confounders. A blinded study was performed using conditions closely resembling those in the field. The Draeger 7110 MKIII IL breath analyzer was used to assess breath alcohol concentrations (BrAC). Participants were 61 healthy volunteers aged 21–37 years with body mass index ≤30 and no history of alcoholism. A total of 242 valid blood/breath tests were performed in four test sets. The study results showed a high correlation coefficient between BrAC and blood alcohol concentration (BAC) levels (r = 0.983) with high sensitivity (97%) and specificity (93%). This strong association between the breath analyzer and BAC persisted even after adjustment for various stages of alcohol absorption. These results illustrate the high diagnostic sensitivity of the breath analyzer in field‐tested conditions.     

The course of alcoholism. Long-term prognosis in different types

The disease concept of alcoholism was first introduced into the medical literature by Magnus Huss in 1984. Since that time many authors have attempted to define therapy-related sub-groups of chronic alcoholics but have been unsuccessful until now because alcoholism represents a complex development and becomes apparent in various clinical pictures. As the method of cross-sectional investigations does not seem to be able to produce reliable results, we performed a prospective long-term study of 444 alcoholics. The methodological claims in literature were taken into consideration like selection criteria, programmes and goals of therapy, follow-up rate and time, etc. Careful observation of the pertinent parts of the pathogenetic pathway leading to chronic alcoholism enabled the authors to establish 4 sub-groups of alcoholics relevant for treatment.     

Sexual abuse and anti-wrinkle cream: evidence from octocrylene

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 910pg/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. The hair lotion was transmitted to our laboratory. The ethanol concentration in this lotion was determined with 35g/L. The EtG immunoassay gave a positive result indicating EtG, which could be confirmed by GC-MS/MS-NCI. In a follow-up experiment the lotion was applied to the hair of a volunteer over a period of six weeks. After this treatment, EtG could be measured in the hair at a concentration of 72pg/mg suggesting chronic and excessive alcohol consumption. Overnight incubation of EtG free hair in the lotion yielded an EtG concentration of 140pg/mg. In the present case, the positive EtG hair findings could be interpreted as the result of an EtG containing hair care product. To our knowledge, the existence of such a product has not yet been reported, and it is exceptionally unusual to find EtG in cosmetics. Therefore, external sources for hair contamination should always be taken into account when unusual cosmetic treatment is mentioned. In those cases, it is recommended to analyze the hair product for a possible contamination with EtG. The analysis of body hair can help to reveal problems occurring from cosmetic treatment of head hair. As a consequence, the assessment of drinking behavior should be based on more than one diagnostic parameter.     

Validation of benzodiazepine β-glucuronide primary reference materials for hydrolysis and quality assurance controls

The major metabolite of hydroxylated benzodiazepines is the β-glucuronide metabolite, thus a hydrolysis step is included in benzodiazepine analyses in urine. We evaluated the new Alltech (R,S) oxazepam and lorazepam β-glucuronide primary reference materials as hydrolysis controls and for use in proficiency testing. Using HPLC analysis and GC/MS analysis following acid and β-glucuronidase hydrolysis, we found that the lorazepam β-glucuronide material was >95% pure, but the oxazepam β-glucuronide material was only 54.0% pure. Recovery after hydrolysis with β-glucuronidase at 60 °C for two hours and 22 °C for 24 h was compared. There was no difference in the recovery of either drug incubated under both conditions. Using HPLC analysis to separate the R and S isomers of the glucuronides, we found that the S isomer hydrolyzes faster than the R isomer, but at optimal conditions hydrolysis of both isomers is complete. Alltech's glucuronide materials can be valuable as hydrolysis controls in method development and routine benzodiazepine analyses, but each laboratory must validate the purity of the materials and determine the acceptable references ranges for in-house controls made from these materials. The reference materials were also evaluated for possible use as quality control material for urine drug screening. The material had to be added at 130% of the reported cutoff concentration to produce a positive result on the Triage^™ Panel for Drugs of Abuse. For the Triage^™ Plus TCA Panel for Drugs of Abuse lorazepam β-glucuronide was added at 154% of the cutoff concentration, but addition of the oxazepam β-glucuronide at >200% of the reported cutoff concentration did not produce a positive result. Discrepancies may be due to preferential binding of antibodies to the S isomer, because the Alltech glucuronide materials do not contain the isomers in the same ratio as that excreted in human urine.     

Comparison of Catalytic and Immunological Amylase Tests for Identifying of Saliva from Degraded Samples

The stability of salivary α‐amylase is a critical factor in both catalytic and immunological method‐based forensic saliva identification. This study aimed to assess the sensitivity of catalytic and immunological tests on degraded saliva samples. Degraded saliva stains were prepared by microbial decomposition using humid soil. Salivary α‐amylase activity was catalytically detected both qualitatively and quantitatively using the Phadebas^® amylase test. As immunological methods, we conducted qualitative and quantitative tests using the RSID?‐saliva test and ELISA, respectively. Salivary α‐amylase activity of degraded samples (incubated at 37°C for 12 h) was significantly lower than that of controls in the quantitative tests. All the degraded samples obtained by the humid soil produced negative results in the Phadebas^® tests, but showed positive results in the RSID?‐saliva test and ELISA. These results suggest that immunological tests are effective for testing degraded saliva samples that have lost their enzymatic activity.     

Verbal Aggression Among Male Alcoholic Patients and Their Wives in the Year Before and Two Years After Alcoholism Treatment

Our earlier reports on 88 male alcoholics and their wives showed that domestic violence decreased significantly in the first and second year following a behavioral marital therapy (BMT) alcoholism treatment program. The present study examined verbal aggression in this same sample. In the year before BMT, verbal aggression was significantly greater—being five to seven times more prevalent for clinically elevated aggression and substantially more frequent—for the alcoholic husbands and their wives than for a demographically matched, nonalcoholic comparison sample. In the two years after BMT, both alcoholic men and their wives showed significant and substantial reductions in verbal aggression as compared with the year before BMT. Despite these significant reductions from the year before BMT, verbal aggression in the two years after BMT remained significantly elevated relative to demographically similar nonalcoholic controls. As predicted, relapsed alcoholics and their wives showed more verbal aggression in the 2 years after BMT than both couples with a remitted alcoholic husband and demographically similar nonalcoholic controls, whereas remitted alcoholics and their wives had similar levels of verbal aggression to the nonalcoholic controls. Further, frequency of drinking was positively correlated with verbal aggression in the 2 years after BMT; verbal aggression was greater when the alcoholic husband drank more frequently.     

Drugs and chronic alcohol abuse in drivers

Blood specimens from 210 drivers (179 male and 31 female) apprehended in Luxembourg from autumn 2001 to spring 2002 and requested for the determination of their blood alcohol concentration (BAC) were tested for medicinal drugs, illicit drugs, and chronic alcohol abuse (by quantification of the carbohydrate-deficient transferrin: CDT). These additional analyses were performed anonymously and with permission of state prosecutor. The 22.8% had consumed medicinal drugs, with benzodiazepines and antidepressants (10.9 and 7.6%, respectively) as main psychoactive classes. Cannabis was the most detected illicit drug (9.5%) but only one in three had THC detectable in their blood. Association of two or more psychoactive substances (poly-drug use) was observed in 27.6% of drivers (90.6% of drug consumers). On the basis of CDT values, 29.5% of drivers investigated were assumed to be chronic alcohol abusers. Statistical analysis revealed that chronic alcohol abuse and medicinal psychoactive drugs were associated with significantly higher BAC. Medicinal psychoactive drugs were clearly associated with poly-drug use, and were furthermore detected at supra-therapeutic levels in 34.9%.     

Direct PCR Improves the Recovery of DNA from Various Substrates

This study reports on the comparison of a standard extraction process with the direct PCR approach of processing low-level DNA swabs typical in forensic investigations. Varying concentrations of control DNA were deposited onto three commonly encountered substrates, brass, plastic, and glass, left to dry, and swabbed using premoistened DNA-free nylon FLOQswabs^™. Swabs (n = 90) were either processed using the DNA IQ^™ kit or, for direct PCR, swab fibers (~2 mm²) were added directly to the PCR with no prior extraction. A significant increase in the height of the alleles (p < 0.005) was observed when using the direct PCR approach over the extraction methodology when controlling for surface type and mass of DNA deposited. The findings indicate the potential use of direct PCR for increasing the PCR product obtained from low-template DNA samples in addition to minimizing contamination and saving resources.     

Role of alcoholization in pathogenesis of morphological changes in substance abusers

Morphological manifestations of combined intoxication with ethanol and narcotics are described. Special attention is paid to changes in the liver which were studied both qualitatively and quantitatively. Specific features in the pathological processes concomitant with combined intoxication are described; high incidence and expression of morphological markers of alcoholism in subjects abusing both ethanol and narcotics in comparison with "pure" alcoholics were determined. These features can be used in forensic medical and pathological (autopsy) diagnosis of chronic narcotic intoxication and its combination with alcoholism.     

Statistical evaluation of standardized field sobriety tests

Standardized Field Sobriety Tests (SFSTs) are used as qualitative indicators of impairment by alcohol in individuals suspected of DUI. Stuster and Burns authored a report on this testing and presented the SFSTs as being 91% accurate in predicting Blood Alcohol Concentration (BAC) as lying at or above 0.08%. Their conclusions regarding accuracy are heavily weighted by the large number of subjects with very high BAC levels. This present study re-analyzes the original data with a more complete statistical evaluation. Our evaluation indicates that the accuracy of the SFSTs depends on the BAC level and is much poorer than that indicated by Stuster and Burns. While the SFSTs may be usable for evaluating suspects for BAC, the means of evaluation must be significantly modified to represent the large degree of variability of BAC in relation to SFST test scores. The tests are likely to be mainly useful in identifying subjects with a BAC substantially greater than 0.08%. Given the moderate to high correlation of the tests with BAC, there is potential for improved application of the test after further development, including a more diverse sample of BAC levels, adjustment of the scoring system and a statistically-based method for using the SFST to predict a BAC greater than 0.08%.     

Background Levels of Salivary-α-amylase Plus Foreign DNA in Cases of Oral Intercourse: a Female Perspective

Saliva plus DNA from a suspect is commonly encountered in sexual assault cases on bodily swabs. However, without background knowledge, the weight of this evidence is unknown. It may indicate the presence of saliva resulting from cunnilingus, or it may represent indirect transfer. In this study, females who refrained from cunnilingus donated 43 items of underwear and 19 vaginal swabs. The samples were subjected to Phadebas^®, RSID^™-Saliva and mRNA profiling and were subsequently DNA-profiled to determine the prevalence of background saliva in the female population. The results report that 15.8% of females who refrained from cunnilingus were positive for saliva and a further 10.5% also had DNA from unknown source(s). These findings of the rate of indirect transfer were evaluated with the Bayesian approach, and it was found that the evidence of saliva plus a high foreign DNA source adds moderately strong support to the allegation of cunnilingus.     

The association of alcohol-induced blackouts and grayouts to blood alcohol concentrations

The primary aim of this study was to investigate the association between measured blood alcohol concentration (BAC) and the presence and degree of amnesia (no amnesia, grayout, or blackout) in actively drinking subjects. A secondary aim was to determine potential factors other than BAC that contribute to the alcohol-induced memory loss. An interview questionnaire was administered to subjects regarding a recent alcohol associated arrest with a documented BAC greater than 0.08 g/dL for either public intoxication, driving under the influence, or under age drinking was administered. Demographic variables collected included drinking history, family history of alcoholism, presence of previous alcohol-related memory loss during a drinking episode, and drinking behavior during the episode. Memory of the drinking episode was evaluated to determine if either an alcohol-induced grayout (partial anterograde amnesia) or blackout (complete anterograde amnesia) occurred. Differences in (1) mean total number of drinks ingested before arrest, (2) gulping of drinks, and (3) BAC at arrest were found for those having blackouts compared with no amnesia; while differences in drinking more than planned were found between the no amnesia and grayout groups. A strong linear relationship between BAC and predicted probability of memory loss, particularly for blackouts was obvious. This finding clinically concludes that subjects with BAC of 310 g/dL or greater have a 0.50 or greater probability of having an alcoholic blackout.     

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

Carbohydrate-deficient transferrin (CDT) is a marker of chronic alcohol abuse, which has recently been introduced to evaluate the physical fitness for obtaining a driving license. The aim of the present study was to evaluate the prevalence of elevated CDT levels in subjects stopped while driving under the influence of alcohol by using a validated method based on capillary electrophoresis. The study was carried out on a group of 40 drunken drivers (group A) and on a control group (n = 51) of subjects chosen from the general population (group B). CDT was directly determined by capillary electrophoresis in free solution and UV detection at 200 nm. CDT results from both groups were classified as “negative” or “positive” on the basis of the cut-off set at 2.00% (CDT index). The subjects classified as “positive” in group A were 24 (60%), whereas in group B were 2. The subjects classified as “negative” in group A were 16 (40%), whereas in group B was 49 (96.1%). The comparison of the observed percentages, evaluated with the χ²-test, was highly significant (p < 0.001). The present study confirms the high prevalence of chronic alcohol abusers among drunken drivers and the usefulness of CDT as a predictor of the risk of drunk driving.     

A "back door" approach to analysis of ethanol-associated risks and behavior

Ethanol has been detected in the majority of medical examiner cases involving nonnatural death and in a substantial number of natural deaths. The higher the serum alcohol concentration (SAC), the greater were the odds that death was due to external (traumatic) causes. We identified 149 medical examiner cases that were positive for ethanol and negative for 11 major drug of abuse groups, and studied those cases to further clarify ethanol-related risk factors and behaviors. Based on terms of frequency of occurrence, the data identify SACs of 201 mg% to 300 mg% as a particularly high risk factor for sudden, unexpected, traumatic death. Traumatic deaths associated with SACs below 100 mg% were infrequent, and attendant opinions concerning the contributory role of ethanol must be made with circumspection. SACs below 150 mg% could not be used to predict whether or not the decedents had been engaged in active or sedentary behavior prior to death because both situations occurred with equal frequency. At SACs between 151 mg% and 350 mg%, active situations were observed three times as often as sedentary situations. When SACs were 350 mg%, sedentary situations were observed twice as often as active situations. Suicide victims and driver fatalities rarely showed SACs above 350 mg%; therefore finding such SACs in apparent suicidal or driver fatality situations should prompt a thorough investigation to rule out other possibilities.(ABSTRACT TRUNCATED AT 250 WORDS)