Estrogen increases endothelial carbon monoxide, heme oxygenase 2, and carbon monoxide-derived cGMP by a receptor-mediated system.

Carbon monoxide, a gaseous activator of soluble guanylyl cyclase formed by a subtype of the enzyme heme oxygenase designated heme oxygenase-2 in vascular endothelium, has been found to dilate blood vessels independently from nitric oxide. Because of the parallels between nitric oxide and carbon monoxide, we speculated that estrogen might affect carbon monoxide production in vascular endothelium. Endothelial cells of human origin (umbilical vein and uterine artery) were incubated for 4 or 24 h with 10(-12)-10(-6) M 17beta-estradiol. 17beta-Estradiol, at a concentration such as that attained during the ovulatory phase of the menstrual cycle (10(-10) M), administrated for 4 h led to a 2-fold increase in intracellular carbon monoxide production and heme oxygenase-2 protein levels (P < 0.05). A reporter assay, measuring the formation of cGMP as the direct product of carbon monoxide-induced activation of soluble guanylyl cyclase in endothelial cells, also revealed a 56% increase in cellular cGMP after treatment with 10(-10) M E2 17beta-estradiol (P < 0.05). By contrast, higher 17beta-estradiol concentrations had no significant respective effects due to nitric oxide synthase inhibition of carbon monoxide release. This 17beta-estradiol effect appeared to be ER dependent, as preincubation with tamoxifen (10(-6) M) blocked the stimulatory effect of 17beta-estradiol in each instance. Our preliminary data indicate a potential role for carbon monoxide as a biological messenger molecule in estrogen-mediated regulation of vascular tone.

Different fatal toxicity of neuroleptics identified by autopsy.

Autopsies and toxicological analyses at the Institute of Forensic Medicine revealed 85 fatal intoxications with neuroleptics in Vienna from 1991 to 1997. A total of 17 cases were linked to a single neuroleptic (NL) alone, while 68 deaths were attributed to a combination of NLs with other drugs. The most frequently detected agent was prothipendyl (n=41). During the study period the number of defined daily doses of high-potency NLs prescribed increased significantly (P< or =0.001) due to increased prescribing of new atypical antipsychotics. The quantity of intermediate- and low-potency NLs dispensed remained stable. The most frequently prescribed NL was haloperidol. The relative toxicities of different NLs were calculated by dividing the number of deaths caused by this NL into the number of defined daily doses prescribed in the observation period (f-value). Single-substance intoxications and multiple-substance intoxications were distinguished. The highest f-values were associated with low-potency NLs, especially with prothipendyl, chlorprothixene and levomepromazine. Low f-values were found for the group of high-potency NLs, including flupentixol, fluphenazine, haloperidol and pimozide, as well as olanzapine. Compared to the f-values for all NLs prescribed, f-values for low-potency NLs were shown to be significantly higher concerning single-substance intoxications (P< or = 0.05) and multiple-substance intoxications (P < or = 0.001), while f-values for high-potency NLs were significantly lower (P< or = 0.05 and P< or = 0.001). We are not aware of the psychiatric diagnoses in our post-mortem sample. However, the present results indicate that careless use of low-potent NLs should be avoided in patients with a potential risk of accidental or suicidal overdose.

Suicide by antidepressant intoxication identified at autopsy in Vienna from 1991-1997: the favourable consequences of the increasing use of SSRIs.

In the area of Vienna, any person dying under questionable circumstances is examined at the Institute of Forensic Medicine, where the cause of death is determined by means of autopsy and chemical analysis. Our study on fatal intoxications was performed in the period between 1991 and 1997, when selective serotonin reuptake inhibitors (SSRIs) were establishing themselves on the market, reaching the top of prescription statistics. Tricyclic antidepressants (TCAs) were involved in 30 single- and 127 multiple-substance intoxications, with amitriptyline and doxepin being the most frequently used drugs. SSRIs were involved in five multiple-substance intoxications. The f-value, which refers to the number of deaths per million defined daily doses prescribed, was found to be significantly (P

Quality of heroin and heroin-related deaths from 1987 to 1995 in Vienna, Austria.

AIMS: To investigate the quality of heroin seized in Vienna between 1987 and 1995 and to examine whether there was a relationship between the quality of heroin and the rate of drug-related deaths. DESIGN: Reports of heroin seizure analysis and post-mortem reports of heroin-related deaths in Vienna from 1987 to 1995 were analysed. RESULTS: There were 386 seizures of heroin comprising a total weight of 25 640.12 g heroin. All the seizures were in the base form. All seizures also contained a diluent, mainly lactose. Additionally, in 95 seizures caffeine, in four seizures paracetamol and in three seizures metaqualon were detected. Of a total of 764 drug-related deaths 506 cases were classified as heroin-related deaths. In 336 cases other central nervous system-depressant drugs and/or alcohol could be detected in the blood in addition to morphine (polydrug heroin-related deaths). There was evidence of a trend towards greater polydrug involvement during the study period. The age of victims of polydrug heroin-related deaths remained unchanged over time while the age of victims of pure heroin-related deaths decreased significantly. The morphine concentration in the medulla oblongata in heroin-related deaths decreased significantly. Finally, there was no statistically significant relationship between the rate of heroin-related deaths per year and the diacetylmorphine concentration of heroin seizures in that year. CONCLUSION: The results did not substantiate the widely held belief that increases in heroin-related deaths could be explained by an increase in the quality of heroin.

Drug-related emergencies and drug-related deaths in Vienna, 1995-1997.

The demographics of drug-related emergencies and drug-related deaths that occurred in Vienna between January 1st, 1995 and December 31st, 1997 were compared to investigate whether they represent two different subgroups of the drug-taking community. Analysis indicated that drug-related emergencies were significantly younger and that the proportion of females was higher than amongst the fatalities. In addition, emergencies were more likely than fatalities to occur in private residences than public places. No significant association between the number of contacts with the Vienna Ambulance Service and the fatal outcome of drug use could be substantiated. The results of this study indicate that drug-related emergencies and drug-related deaths represent two different subpopulations of the drug-taking community. Therefore, different strategies of prevention are considered.

Determination of glucitollysine for the quantitation of non-enzymatic glucosylation by ion exchange chromatography and reverse phase liquid chromatography.

A specific and sensitive method for the quantitative determination of the stable, reduced glucose-lysine adduct, glucitollysine (GL), in plasma protein samples is described. The method uses standard amino acid ion exchange chromatography followed by reverse phase high performance liquid chromatography after derivatisation of GL to a fluorescent product. Moreover, GL was characterised and identified in plasma samples by means of mass spectroscopy. GL measured in plasma samples of eleven type I diabetics and two healthy controls showed a significant linear correlation to concomitantly determined haemoglobin AI and glucosylated plasma proteins, but did not correlate with plasma glucose levels. This method allows the estimation of non-enzymatic glucosylation in biological samples with a high degree of specificity and sensitivity down to the low nanogram range.

Prediction of gas chromatographic retention indices of a diverse set of toxicologically relevant compounds.

For a set of 846 organic compounds, relevant in forensic analytical chemistry, with highly diverse chemical structures, the gas chromatographic Kovats retention indices have been quantitatively modeled by using a large set of molecular descriptors generated by software Dragon. Best and very similar performances for prediction have been obtained by a partial least squares regression (PLS) model using all considered 529 descriptors, and a multiple linear regression (MLR) model using only 15 descriptors obtained by a stepwise feature selection. The standard deviations of the prediction errors (SEP), were estimated in four experiments with differently distributed training and prediction sets. For the best models SEP is about 80 retention index units, corresponding to 2.1-7.2% of the covered retention index interval of 1110-3870. The molecular properties known to be relevant for GC retention data, such as molecular size, branching and polar functional groups are well covered by the selected 15 descriptors. The developed models support the identification of substances in forensic analytical work by GC-MS in cases the retention data for candidate structures are not available.

Systematic toxicological analysis: computer-assisted identification of poisons in biological materials.

A new software was developed to improve the chances for identification of a "general unknown" in complex biological materials. To achieve this goal, the total ion current chromatogram was simplified by filtering the acquired mass spectra via an automated subtraction procedure, which removed mass spectra originating from the sample matrix, as well as interfering substances from the extraction procedure. It could be shown that this tool emphasizes mass spectra of exceptional compounds, and therefore provides the forensic toxicologist with further evidence-even in cases where mass spectral data of the unknown compound are not available in "standard" spectral libraries.

Detection of mustard gas bis(2-chloroethyl)-sulfide in urine.

Extraction procedure and detection of bis(2-chloroethyl)sulfide from urine samples of two patients are described using gas chromatography-mass spectrometry. Concentrations are found in the low ppb-range. The result is discussed in connection with the reactivity of this substance and the corresponding metabolites.

Detection of bis(2-chlorethyl)-sulfide (Yperite) in urine by high resolution gas chromatography-mass spectrometry.

Bis(2-chlorethyl)-sulfide (Yperite) could be detected by GC/MS in urine samples, concentration ranging from 1 to 30 ppb. These results were supported by high resolution mass spectral data.

Automatic screening in postmortem toxicology.

The systematic analysis of postmortem samples is one of the most challenging tasks in forensic toxicology. For determining cause of death, analysis of different tissues can be indispensable. Automation of these analyses would increase reproducibility and therefore lead to more reliable and comparable results. Recent developments in analytical toxicology and the availability of automation devices for various analytical stages, such as sampling, preliminary testing, sample extraction, chromatographic separation, identification, and data processing are examined and discussed. At present only parts of the analytical procedure have been automated-however, the goal should be the integration of these parts into a single, continuous process. Currently, only one "fully-automated" procedure for the comprehensive screening of blood and urine (excluding sample pretreatment, which remains separate) has been published. But it can be expected that automation of analytical procedures in forensic toxicology will indeed progress, even with regard to the very complex screening of postmortem samples.

Suicide of a body packer.

The smuggling of illicit drugs by means of body packing has become a common problem at European airports. Europe is considered to be the fastest growing market for cocaine worldwide, and the air route is the most frequently used method of trafficking cocaine. Smuggling illicit drugs by use of body packing is considered to be a high toxicological hazard because of the risk of leakage or a package bursting. We report about the first case of suicide of a body packer by re-ingesting the content of excreted cocaine packages. The consequence of this case is that the death scene investigation and autopsy assessments in case of a body packer's death should always consider the possibility of re-ingested packages. Detention personnel should be instructed by forensic and criminalistic experts to take preventive measures.

Identification of the general unknown. Application of mass selective detectors in forensic toxicology.

One of the basic aims of forensic toxicology is the identification of previously unknown drugs and poisons. This is frequently achieved using the combination of gas chromatography and benchtop quadrupole or ion trap mass spectrometers. The influence of matrix loading on the mass spectral quality was tested, and it was found that a realistic amount of matrix changed the pattern of the spectra obtained by the ion trap mass spectrometer. Disturbed mass spectra led to unsuitable suggestions from the library search and thus rendered the identification of the "general unknown" more difficult. On the other hand, higher selectivity and lower detection limits favored the ion trap technology for target analysis because of the capability of MS-MS.

Gas chromatographic analysis of illicit heroin samples.

Morphine and 6-acetylmorphine in illicit heroin samples can be easily determined by propionylation in aqueous solution (disodium hydrogen phosphate buffer). Heroin, 3-propionylmorphine and 6-acetyl-3-propionylmorphine are separated on methyl-phenylpolysiloxane liquid phases (5% phenyl content). The analytical procedure and the corresponding statistical data are presented together with the retention indices of typical street heroin concomitant drugs. The procedure can also be applied to the analysis of opium preparations and very dilute heroin (morphine) samples.

General unknown screening in postmortem tissue and blood samples: a semi-automatic solid-phase extraction using polystyrene resins followed by liquid-liquid extraction.

The identification of general unknown poisons in complex biological materials like postmortem blood and tissue is a great challenge for the forensic toxicologist. Therefore, a screening procedure utilizing a semi-automatic work-up with an ASPEC system was developed. A broad range of different compounds can be isolated by using non-selective and generally applicable organic polymeric sorbents such as OASIS HLB or Isolute 101. Because colloidal solutions were applied to these sorbents, the denaturation of proteins, which can result in an irreversible loss of significant compounds by adsorption and occlusion, could be avoided. Because of the process of micellar chromatography followed by liquid-liquid extraction of the crude extract, very clean fractions were obtained from such complex matrices as postmortem blood, liver, and brain samples. High recoveries (72-100%) and good day-to-day relative standard deviations (1-17%) could be achieved with both polymeric sorbents. The procedure paves the way for the identification of general unknown poisons in target organs and is therefore a useful tool in the field of forensic toxicology.

Drugs and driving in Vienna, Austria.

Drugs that affect the central nervous system are generally assumed to have the potential to impair driving ability. In a retrospective survey, police files and the results of toxicological urine analysis from drivers suspected of driving under the influence of drugs in Vienna from 1993 to 1996 were investigated. Decisive for police intervention was "unsafe driving" (swerving, hesitating, going too slowly, etc.), driving at high speed within the city limits, driving through red lights or stop signs, and driving at night without lights. In one-fifth of the cases drivers caused a traffic accident. Casting suspicion on driving under influence of drugs was mainly caused by impaired coordination of movements, bloodshot eyes, slurred speech, drowsiness, conspicuous behavior, and changed pupils. In the majority of the study population more than two symptoms were recorded by police. In 94% of the cases police suspicions could be confirmed by toxicological urine analysis.

Suicidal chloroquine poisoning: clinical course, autopsy findings, and chemical analysis.

Suicidal overdose of chloroquine is rare. We present a 14-year-old girl who was admitted to our Emergency Department after intentional ingestion of 7.5 g of chloroquine base followed by cardiac and respiratory arrest. Despite early mechanical ventilation, cardiac support, and treatment with high doses of diazepam, the patient died. Laboratory investigation indicated disseminated intravascular coagulation confirmed by petechial hemorrhages in the leptomeninges at autopsy, a finding that has not previously been described. Postmortem tissue analysis demonstrated early drug distribution to the medulla oblongata and cerebellum.

[Pulmonary lipids in fatal burns (author's transl)]. / Uber das Verhalten der Lungenfette nach vitalen Verbrennungen.

The pulmonary lipids were investigated at autopsy in 22 deaths from burns (3 scalds among them) by means of histological examination, thin-layer chromatography and gas liquid chromatography. Leaving out of consideration 4 cases with concomitant mechanical trauma 7 out of 18 burned victims showed fat embolism of the lungs, the degree varying from slight (4) to medium (3). Cases surviving for a short period showed fat embolism more frequently than those with a rapidly fatal course. The extracted pulmonary lipids were separated by thin-layer chromatography into 5 fractions (phospholipids, cholesterol, fatty acids, triglycerides, cholesterol esters) and evaluated densitometrically. The triglyceride fraction was increased in cases of fat embolism after burns, as well as in control cases of post-traumatic fat embolism. For further lipid characterisation the fatty acids were determined by gas chromatography. Calculation of the amounts of the individual fatty acids showed that in cases of at least medium intensity the ratio of oleic acid/linoleic acid and oleic acid/stearic acid is shifted towards oleic acid.

[The umbilical cord blood lead concentration: a comparison between an urban and rural population (author's transl)]. / Bleigehalt des Nabelschnurblutes. Vergleich eines städtischen Kollektivs mit einem ländichen

The cord blood lead level was determined to 40 newborn infants. The values were significantly higher in the infants born to mothers resident in Vienna than in the infants born to the group of mothers from the rural surroundings of Eisenstadt. The overall mean blood level was 17.7+/-7.4 mug/100 g. Increasing pollution requires investigations concerning possible effects on the growing fetus and would serve as foundation for future trend assessment studies.

[Drug fatalities from the forensic medicine viewpoint. 10 years experiences of the Vienna Institute of Forensic Medicine]. / Drogentodesfälle aus gerichtsmedizinischer Sicht. 10 Jahre Erfahrungen aus dem Institut für Gerichtliche Medizin, Wien.

The problem of illicit drug abuse has of course not spared Austria. In fact, considering only the increase in the official figures on drug-related deaths over the past few years, this problem has dramatically worsened in Austria. Drug-related deaths represent the most serious consequence of substance abuse, but according to the Pompidou Group classification such deaths are only one of eight indicators in assessing the extent of drug abuse in a community. We analyzed reports on all postmortems classified as drug-related deaths according to the official definition of the Austrian Federal Ministry of Internal Affairs undertaken at the Institute of Forensic Medicine in Vienna between 1984 and 1993. The annual number of drug-related deaths increased nearly ten-fold during the study period. In about a third of the cases accidental single substance overdosage (of mainly a morphine-containing drug) was the cause of death. In the majority of the corpses more than one drug, including alcohol, was detected. During the past 10 years there was an evident trend towards a multisubstance abuse pattern. Since the plateau reached in 1988, the incidence of HIV-1 infection among decreased drug injectors has decreased.