Ethyl glucuronide (EtG) is a non-volatile, water-soluble, direct metabolite of ethanol that can be detected in body fluids and hair. We investigated urine and serum samples from three patient groups: (1) 33 in-patients in acute alcohol withdrawal; (2) 30 detoxified in-patients (treated for at least 4 weeks) from a 'motivation station'; and (3) 43 neuro-rehabilitation patients (non-alcoholics; most of them suffering from stroke, traumatic brain injury, Parkinson's disease etc.) using gas chromatography/mass spectrometry (GC/MS) with deuterium-labelled EtG as the internal standard and additionally in the second group of patients using liquid chromatography (LC/MS-MS). We found no correlation between the concentration of EtG in urine at hospitalization and the blood-ethanol concentration (r = 0.17), the time frame of detection (r = 0.5) or the total amount of clomethiazole required for the treatment of withdrawal symptoms (r = 0.28). In four out of 30 in-patients from the 'motivation station'--where neither clinical impression nor routine laboratory findings gave indications of relapse--concentrations of EtG in urine ranged between 4.2 and 196.6 mg/l. EtG concentrations in urine of between 2.89 and 23.49 mg/l were found in seven out of 43 neuro-rehabilitation patients using GC/MS. The GC/MS and the LC/MS-MS results showed a correlation of 0.98 with Pearson's correlation test and 1.0 with Spearman's correlation test. We suggest that EtG is a marker of alcohol consumption that can be detected for an extended time period after the complete elimination of alcohol from the body. When used as a relapse marker with a specific time frame of detection intermediate between short- and long-term markers, EtG fills a clinically as well as forensically important gap. Its specificity and sensitivity exceed those of all other known ethanol markers.

The rat experiments indicated that the very high PEth concentrations found in the organs of the alcoholics were probably largely formed while the organs were frozen at -20 degrees C. Our data suggest that tissue material from bodies that were exposed to ethanol must be stored properly to obtain reliable results from subsequent analysis for PEth. Tissue should not be frozen at -20 degrees C but instead stored refrigerated until extraction, preferably within hours of autopsy, or frozen in liquid nitrogen and stored at -80 degrees C. Blood samples that contain ethanol can be stored refrigerated for up to 72 hr or frozen in liquid nitrogen and stored at -80 degrees C without affecting PEth levels.

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In German-speaking countries, blood ethanol concentrations (BECs) are usually calculated using Widmark's equation. The distribution factor r of this equation is a correction factor needed to obtain a reduced body mass and corresponds to the ratio of total body water and blood water content. To enhance the reliability of Widmark's model equation, the body weight, body height, blood water content and total body water of 256 women and 273 men were measured. The ratio of body water to blood water ranged from 0.44 to 0.80 in women and from 0.60 to 0.87 in men. For both sexes equations were developed by multiple regression analysis which allow the determination of the individual, more realistic distribution factors rFI (for females) and rMI (for males) even when only body height and body weight are known. Drinking experiments revealed a clearly higher congruence of calculated and measured blood ethanol concentrations when rFI or rMI were used instead of rigid distribution factors, i.e. 0.6 for women and 0.7 for men with or without the assumption of a 10% so-called resorption deficit. Additionally, Widmark's equation in combination with rFI or rMI allows a more accurate prediction of blood ethanol concentrations than the equations of Watson and Ulrich.

The significance of both Purkinje cell numbers and various neuronal changes for the diagnosis and timing of hypoxic-induced brain lesions was investigated in tissue samples from the cerebellar cortex of 52 individuals with a history of acute or prolonged cerebral hypoxia/ischemia before death. Furthermore, the area of the Purkinje cell somata (PC size) was measured using an automatic image processing and analysis system (LEICA QWin). Significantly reduced numbers of Purkinje cells (<6 cells/unit length of 1 mm) and a decreased portion (<50%) of intact Purkinje cells could be detected in individuals with a period of resuscitation of at least 2 h after acute circulatory arrest. Average cell numbers of less than 4 cells/unit were found in individuals who suffered from diffuse brain swelling and were ventilated for at least 3 days, as well as in individuals who died of brain death. Moreover, the Purkinje cells in these cases exhibited shrunken somata compared to the controls. Specimens that were stored at room temperature up to 30 h after removal at autopsy showed no significant autolytic changes of the Purkinje cells. After 46 h, however, reduced Purkinje cell numbers and shrunken cell bodies were found.