The impact of total body water on breath alcohol calculations.

Due to a legislative amendment in Austria to determine breath alcohol (BrAC) instead of blood alcohol (BAC) in connection with traffic offences, many results of blood alcohol calculations were simply converted using distinct conversion factors. In Austria, the transformation of BAC to BrAC was carried out by using a factor of 1:2000, which, however, is commonly known to be too low. Noticing the great demand for a calculation method that is not exclusively based on blood alcohol, a formula for calculating breath alcohol based on blood alcohol was published in 1989, but in which the body surface area (BSA) was considered the most important influencing variable. In order to refine this new method, a liquor intake experiment was conducted combined with measurements of total body water (TBW) as an additional variable, using hand to foot bioelectrical impedance assessment (BIA). The test group comprised 37 men and 40 women to evaluate the accuracy of TBW and BSA as an individual parameter for alcohol concentration. The correlation coefficient of BrAC with TBW was constantly higher than with BSA (maximum = 0.921 at 1 h and 45 min after cessation of alcohol intake). These results are valid for both men and women as well as in a gender independent calculation. Hence, for an accurate back calculation of BrAC adjusted values of eliminations rates had to be found. This study describes mean elimination rates of BrAC for both men (0.065 ± 0.011 mg/L h-1) and women (0.074 ± 0.017 mg/L h-1). As previously shown women displayed a significantly higher elimination rate than men (p = 0.006).

Does the prior application of the field kit bullet hole testing kit 3 on a suspected bullet hole bias the analysis of atomic absorption spectrophotometry?

Forensic ballistics is the study of bullet trajectory and consists of determining gunshot residue (GSR) to identify bullet holes. Among several highly sensitive methods, atomic absorption spectrophotometry (AAS) is employed to analyze GSR in the laboratory. However, it is sometimes necessary to identify bullet holes immediately at a crime scene. The purpose of this examination was to determine whether the use of the field test Bullet Hole Testing Kit 3 (BTK3) on a suspected bullet hole would influence the outcome of AAS-analysis: Three commonly encountered firearms (Glock17, Tokarev, and Colt) were fired at skin, wood, and cloth. AAS-analysis was performed with and without previous BTK3 application. The results clearly indicate that there is no significant interaction on the grounds of BTK3 use (BTK3 vs. no-BTK3 [kit_nokit] [Pb: p = 0.1309; Sb: p = 0.9111], material*kit_nokit [Pb: p = 0.5960; Sb: p = 0.9930], distance*kit_nokit [Pb: p = 0.4014; Sb: p = 0.9184], and firearm type*kit_nokit [Pb: p = 0.9662; Sb: p = 0.9885]); hence, applying this field kit does not falsify later AAS outcomes.

Establishment and characterization of a primary and a metastatic melanoma cell line from Grey horses.

The Grey horse phenotype, caused by a 4.6 kb duplication in Syntaxin 17, is strongly associated with high incidence of melanoma. In contrast to most human melanomas with an early onset of metastasis, the Grey horse melanomas have an extended period of benign growth, after which 50% or more eventually undergo progression and may metastasize. In efforts to define changes occurring during Grey horse melanoma progression, we established an in vitro model comprised of two cell lines, HoMel-L1 and HoMel-A1, representing a primary and a metastatic stage of the melanoma, respectively. The cell lines were examined for their growth and morphological characteristics, in vitro and in vivo oncogenic potential, chromosome numbers, and expression of melanocytic antigens and tumor suppressors. Both cell lines exhibited malignant characteristics; however, the metastatic HoMel-A1 showed a more aggressive phenotype characterized by higher proliferation rates, invasiveness, and a stronger tumorigenic potential both in vitro and in vivo. HoMel-A1 displayed a near-haploid karyotype, whereas HoMel-L1 was near-diploid. The cell lines expressed melanocytic lineage markers such as TYR, TRP1, MITF, PMEL, ASIP, MC1R, POMC, and KIT. The tumor suppressor p53 was strongly expressed in both cell lines, while the tumor suppressors p16 and PTEN were absent in HoMel-A1, potentially implicating significance of these pathways in the melanoma progression. This in vitro model system will not only aid in understanding of the Grey horse melanoma pathogenesis, but also in unraveling the steps during melanoma progression in general as well as being an invaluable tool for development of new therapeutic strategies.

Micromorphological changes in cardiac tissue of drug-related deaths with emphasis on chronic illicit opioid abuse.

AIMS: The main intention of this retrospective study was to investigate whether chronic illicit drug abuse, especially the intravenous use of opioids (heroin), could potentially trigger the development of myocardial fibrosis in drug addicts. DESIGN: A retrospective case-control study was performed using myocardial tissue samples from both drug-related deaths (DRD) with verifiable opioid abuse and non-drug-related deaths in the same age group. SETTING: Department of Forensic Medicine, Medical University of Vienna, Austria (1993-94). PARTICIPANTS: Myocardial specimens were retrieved from 76 deceased intravenous opioid users and compared to those of 23 deceased non-drug users. MEASUREMENTS: Drug quantification was carried out using the enzyme-multiplied immunoassay technique (EMIT), followed by [gas chromatography-mass spectrometry (GC-MS), MAT 112(®) ], and analysed using the Integrator 3390A by Hewlett Packard(®) and LABCOM.1 computer (MSS-G.G.). The amount of fibrous connective tissue (FCT) in the myocardium was determined by using the morphometric software LUCIA Net version 1.16.2(©) , Laboratory Imaging, with NIS Elements 3.0(®) . FINDINGS: Drug analysis revealed that 67.11% were polydrug users and the same proportion was classified as heroin addicts (6-monoacetylmorphine, 6-MAM)-32.89% were users of pure heroin. In 76.32% of DRD cases, codeine was detected. Only 2.63% consumed cocaine. The mean morphine concentrations were 389.03 ng/g in the cerebellum and 275.52 ng/g in the medulla oblongata, respectively. Morphometric analysis exhibited a strong correlation between DRD and myocardial fibrosis. The mean proportion of FCT content in the drug group was 7.6 ± 2.9% (females: 6.30 ± 2.19%; males: 7.91 ± 3.01%) in contrast to 5.2 ± 1.7% (females: 4.45 ± 1.23%; males: 5.50 ± 1.78%) in the control group, indicating a significant difference (P = 0.0012), and a significant difference in the amount of FCT between females and males (P = 0.0383). There was no significant interaction of age and FCT (P = 0.8472). CONCLUSIONS: There is a long-term risk of cardiac dysfunction following chronic illicit drug abuse with opioids as a principal component. Regular cardiological examination of patients receiving substitution treatment with morphine is strongly recommended.

Copy number expansion of the STX17 duplication in melanoma tissue from Grey horses.

BACKGROUND: Greying with age in horses is an autosomal dominant trait, associated with loss of hair pigmentation, melanoma and vitiligo-like depigmentation. We recently identified a 4.6 kb duplication in STX17 to be associated with the phenotype. The aims of this study were to investigate if the duplication in Grey horses shows copy number variation and to exclude that any other polymorphism is uniquely associated with the Grey mutation. RESULTS: We found little evidence for copy number expansion of the duplicated sequence in blood DNA from Grey horses. In contrast, clear evidence for copy number expansions was indicated in five out of eight tested melanoma tissues or melanoma cell lines. A tendency of a higher copy number in aggressive tumours was also found. Massively parallel resequencing of the ~350 kb Grey haplotype did not reveal any additional mutations perfectly associated with the phenotype, confirming the duplication as the true causative mutation. We identified three SNP alleles that were present in a subset of Grey haplotypes within the 350 kb region that shows complete linkage disequilibrium with the causative mutation. Thus, these three nucleotide substitutions must have occurred subsequent to the duplication, consistent with our interpretation that the Grey mutation arose more than 2,000 years before present. CONCLUSIONS: These results suggest that the mutation acts as a melanoma-driving regulatory element. The elucidation of the mechanistic features of the duplication will be of considerable interest for the characterization of these horse melanomas as well as for the field of human melanoma research.

A cis-acting regulatory mutation causes premature hair graying and susceptibility to melanoma in the horse.

In horses, graying with age is an autosomal dominant trait associated with a high incidence of melanoma and vitiligo-like depigmentation. Here we show that the Gray phenotype is caused by a 4.6-kb duplication in intron 6 of STX17 (syntaxin-17) that constitutes a cis-acting regulatory mutation. Both STX17 and the neighboring NR4A3 gene are overexpressed in melanomas from Gray horses. Gray horses carrying a loss-of-function mutation in ASIP (agouti signaling protein) had a higher incidence of melanoma, implying that increased melanocortin-1 receptor signaling promotes melanoma development in Gray horses. The Gray horse provides a notable example of how humans have cherry-picked mutations with favorable phenotypic effects in domestic animals.