Determination of ethyl glucuronide in human hair by SPE and LC-MS/MS

A method for the sensitive and selective determination of ethyl glucuronide (EtG) in hair has been developed using solid-phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Washed and cut hair segments were extracted by ultrasonication (3h, 50 degrees C) and the extracts were cleaned-up with aminopropyl SPE columns. LC-MS/MS analysis was performed using a polar-endcapped phenyl-hexyl-RP-phase with negative mode electrospray ionisation (ESI) using a triple quadrupole mass spectrometer (Sciex API 365) with a turboionspray source and post-column addition of acetonitrile for enhanced sensitivity. The MS/MS transitions monitored were m/z 221 -->75 for EtG and 226 -->75 for D(5)-EtG as an internal standard. The method was selective and sensitive, with a detection limit of 51 pg/mg hair at a signal-to-noise ratio of 3:1. The mean recovery was 96%, with an intra- and inter-day precision of less than 11.7% at a concentration of 200 pg/mg. The linearity was assessed in the range of 25-2000 pg/mg hair, with a correlation coefficient of 0.997. The method was successfully applied to 97 human hair samples which were taken at autopsies from persons with known alcoholism or were obtained from alcoholics who were hospitalized for ethanol withdrawal, from social drinkers and from children having not consumed any alcohol. Although, approximately two-third of the alcoholics showed EtG concentrations in hair of higher than 51 pg/mg (up to >4000 pg/mg), in one-third the EtG concentration was below the detection limit. However, only in one of five hair samples of "social drinkers", the EtG concentration was above the detection limit (51 pg/mg). No EtG has been detected in the hair of children. These investigations demonstrate that heavy alcohol consumption may be but not necessarily has to be detectable by EtG analysis in hair.     

Time-dependent immunohistochemical detection of proinflammatory cytokines (IL-1β, IL-6, TNF-α) in human skin wounds

The proinflammatory cytokines interleukin-1beta (IL-1β), interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-α) hold important functions in the early and late courses of inflammation, trauma and wound healing. In the present study, human skin wounds due to sharp force (n=105) were collected during surgery and autopsy. The wound age mainly varied from several minutes to 5 h, some specimens aged up to 6 weeks. Control specimens from uninjured skin were available in each case. After preparation of cryostat sections, immunohistochemistry was performed according to the APAAP technique, using monoclonal and polyclonal antibodies. The results were evaluated semiquantitatively. All markers were weakly expressed in normal human skin constitutively. However, the staining pattern changed significantly in vital wounds concerning epidermal layers, subepidermal cells, vessels and sweat glands. IL-1β and IL-6 showed enhanced expression after 15 and 20 min at the earliest (increase of epidermal reactivity). After 30–60 and 60–90 min, respectively, marked expression was observed with these markers. Similar alterations were detectable with TNF-α after 15 and 60–90 min. The reactivity of all three markers persisted over several hours, then decreased to basal levels again and sometimes reappeared after days and in granulation tissue. Leukocytes reacting with IL-1β and IL-6 appeared after approximately 2 h. Conclusion: proinflammatory cytokines can serve as a useful tool for the estimation of vitality and wound age, in particular in the early post-traumatic interval prior to leukocyte reaction. Autolysis did not play a role in the samples investigated (postmortem interval up to 8 days). Problems could sometimes rise from constitutive expression. Therefore, it is recommended to examine control samples from the same individual and to compare the reactivity with wound specimens.     

Brain amino acid abnormalities in liver disease--a postmortem study

In a postmortem exploratory study, we examined whether specific amino acid abnormalities associated with liver diseases in vivo may also be detected in human brain samples obtained at clinical autopsies. The branched-chain amino acids (BCAA: valine, leucine, isoleucine) were decreased in the group of patients with liver diseases compared with the control group, whereas the aromatic amino acids (AAA: phenylalanine, tyrosine) were increased. However, the ranges overlapped significantly and were not statistically different. The molar ratio BCAA/AAA was determined to be 1.92 in the collection of patients with liver diseases compared with 2.27 in the control group. In patients with liver disease, ornithine concentrations in the brain appeared significantly decreased whereas glutamine was significantly increased. No significant difference was found in the brain concentrations of proline. Amino acid analysis may contribute to the understanding of pathophysiological mechanisms of liver disease, which are discussed, and may supplement the postmortem diagnosis.     

What differentiates anal rapists from vaginal rapists?

During the daily assessment routine of the Federal Documentation Centre for Sexual Offenders, important differences between rapists who commit their offense through anal penetration and other rapists became apparent. The hypothesis following these findings assumed that these and offenders would show a more sexualized behaviour than other rapists. The following differences between these two groups could be shown: violence and hypersexual desire. And rapists show a significantly higher degree in both areas, whereas, on the other side, no difference concerning psychopathic behaviour of the offenders could be proved. The preliminary conclusions that could be drawn according to these results express that anal rapists are more sexualized and, furthermore, they have a more obsessive sex life.     

Mixed fatal poisoning caused by taking L-methadone and chloral hydrate

A 42-year-old female drug user who was enrolled in a methadone maintenance program was found dead in her apartment. Cause of death was an intoxication with chloral hydrate and L-methadone. Trichloroethanol (TCE), the primary metabolite of chloral hydrate, was quantified by solid phase microextraction (SPME) and GC/MS in heartblood (27 micrograms/ml) and urine (338 micrograms/ml). D- and L-methadone were differentiated by chiral HPLC, which showed that only L-methadone had been taken. The quantitation of L-methadone and its metabolite EDDP was carried out by GC/MS from heartblood (1300 ng/ml and 86 ng/ml, respectively), urine (5239 ng/ml and 4960 ng/ml, respectively) and gastric contents (159 ng/ml and 122 ng/ml, respectively). The concentrations of both--trichloroethanol and methadone--were in toxic ranges.     

An analysis of toxic deaths, 1982 to 1985, Pima County, Arizona

Toxic deaths in Pima County, Arizona, were studied over a four-year period. The deaths were analyzed according to cause and manner of death, toxic substance, and demographic data. The age group 40 to 49 years had the highest rate of suicide from toxic substances. The accident death rate was highest for ages 20 to 29 years. Carbon monoxide was most often found to be the cause of deaths in this study. The most prevalent drugs were narcotics followed by antidepressants, cocaine, and barbiturates. Comparisons are made with similar studies.