Martin Steinlechner

Species identification by means of the cytochrome b gene

Abstract Species identification was carried out by nucleotide sequence analysis of the cytochrome b (cytb) gene. The aim of the study was to identify biological specimens from diverse vertebrate animals by extracting and amplifying DNA from 44 different animal species covering the 5 major vertebrate groups (i.e. mammals, birds, reptiles, amphibians and fishes). The sequences derived were used to identify the biological origin of the samples by aligning to cytb gene sequence entries in nucleotide databases using the program BLAST. All sequences were submitted to the GenBank including new species which were not observed in the databases. The applicability of this method to the forensic field is demonstrated by simulated casework conditions where different types of samples including problematic specimens such as hair, bone samples, bristles and feathers were investigated to identify the species.

AGE DEPENDENT APOPTOSIS AND LOSS OF RHABDOSPHINCTER CELLS

PURPOSE To our knowledge the exact age dependent morphological and functional changes of the sphincter mechanism have not been investigated. Therefore, cell densities of the urethra and the urethral rhabdosphincter across various age groups, and the appearance of apoptosis were examined to explore the changes in these structures during the aging process. MATERIALS AND METHODS Specimens were obtained from 16 male and 7 female cadavers 5 weeks to 92 years old. Histological sections were taken from 3 different levels of the rhabdosphincter and urethra. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling method was used to detect apoptosis in the urethra and rhabdosphincter. In all specimens relative volume densities of the striated muscle fibers, apoptotic indexes and diameters of the rhabdosphincter and urethra were determined. RESULTS An age dependent increase of apoptosis of the striated muscle fibers of the rhabdosphincter led to a dramatic decrease in the number of striated muscle cells. In the 5-week-old neonate 87.6% and in the 91-year-old woman 34.2% of the rhabdosphincter consisted of striated muscle cells. Overall, a direct linear correlation between the age of the specimens and decrease in volume densities of the striated muscle cells was evident. CONCLUSIONS The dramatic decrease in the number of striated muscle cells in the rhabdosphincter of the elderly due to apoptosis represents the morphological basis for the high incidence of stress incontinence in this population.

Urinary incontinence in the elderly and age-dependent apoptosis of rhabdosphincter cells

With advancing age, a progressive and age-dependent decrease of the density of striated muscle cells can be observed in the rhabdosphincter. This continuous loss of striated muscle cells due to apoptosis may finally lead to urinary incontinence.

Efficient DNA database laboratory strategy for high through-put STR typing of reference samples.

DNA intelligence databases were installed successfully in various countries during the past few years. It is a general trend that laboratories performing STR analysis for DNA databases have to adjust to increased sample through-put, especially when dealing with a high number of reference samples. In contrast to routine forensic casework analysis, where samples of suspects and unknown samples are interpreted with regard to the specific circumstances of the case and are kept distinctly apart from other cases, DNA databases consist of single, primarily unlinked DNA profiles. Problems areas associated with the high number of anonymous DNA profiles are the risk of logistic errors, such as sample mix-up during the laboratory procedure, and the risk of typing errors during manual transcription of data and/or results. Thus, DNA databases clearly require new laboratory strategies to rise to the challenge. This paper presents an efficient automated laboratory strategy on the platform of a laboratory management information system (LIMS) with the Austrian DNA Intelligence Database as example. Two goals were tackled in particular: first, data safety by avoiding both manual interaction during critical laboratory steps (i.e. when DNA is transferred form one tube into another), and errors due to manual transcription of sample information and results. Secondly, efficient sample processing by automizing the laboratory procedure with the help of robotic instruments, thus, giving the DNA staff more time to analyze data.

Camphor ingestion for abortion (case report)

The case of a 16-year-old girl who ingested 30 g of camphor dissolved with 250 ml wine to induce abortion is reported. The girl started vomiting 45 min after ingestion, which may have saved her life. Camphor is present in a number of over-the-counter medications, mainly for external application, and is readily available in drugstores. The substance is highly toxic and rapid in onset. The reported human lethal dose is 50 to 500 mg kg-1. Camphor ingestion may lead to abortion because camphor crosses the placenta and fetuses lack the enzymes to hydroxylate and conjugate with glucuronic acid. The girl was charged with intended abortion.

EMPOP—the EDNAP mtDNA population database concept for a new generation, high-quality mtDNA database

Abstract The European DNA Profiling Group (EDNAP) MtDNA Population Database (EMPOP) is an international collaborative project between DNA laboratories performing mtDNA analysis and the DNA laboratory of the Institute of Legal Medicine (GMI) in Innsbruck, Austria. The goal is to set up a directly accessible mtDNA population database, which can be used in routine forensic casework for frequency investigations. Most forensic laboratories do not have the capacity to rapidly generate massive amounts of sequence data. Altogether, however, they dispose of a respectable body of data. Apart from a mutual, non-competitive exchange of molecular genetic techniques, every forensic institute can contribute to the EMPOP project with its familiarity with the individual technology and with the polymorphisms typical for the population inhabiting its surroundings. The EMPOP core laboratory in Innsbruck provides the bioinformatic infrastructure with which to analyze, transcribe, store, and share the data.

Male/female DNA mixtures: a challenge for Y-STR analysis

Abstract We present the optimisation of two Y-STR multiplexes for forensic casework applications especially for the analysis of mixtures of male and female DNA. The procedure involved: (1) a new design of the PCR primers for the loci DYS389, DYS390 and DYS391 in order to improve the PCR efficiency and to reduce the length of the amplification products, and (2) the addition of PCR Enhancer to the reaction mix, increasing the specificity of the method.

APOPTOSIS OF RHABDOSPHINCTER CELLS: THE MAIN CAUSE OF URINARY INCONTINENCE WITH ADVANCING AGE?

Reproduction Form B-1 InStitUtii I UNIVERSITY OF INNSBRUCK, INNSBRUCK, AUSTRIA I