Suitability of specific soft tissue swabs for the forensic identification of highly decomposed bodies.

When decomposition of a recovered body is fairly advanced, identification based on common morphologic features is often impossible. In these cases, short tandem repeat (STR) marker genotyping has established itself as a convenient and reliable alternative. However, at very progressed stages of decomposition, postmortem tissue putrefaction processes can decrease DNA yields considerably. Hence, not all types of tissue are equally suitable for successful STR marker-based postmortem identification. Bone or dental material is often analysed in corpses with advanced decompositional changes. However, processing of these materials is very elaborate and time and resource consuming. We have therefore focused on the suitableness of various types of soft tissue swabs, where DNA extraction is easier and faster. By sampling 28 bodies at various stages of decomposition, we evaluated the suitability of different tissues for genotyping at varying degrees of physical decay. This was achieved by a systematic classification of the sampled bodies by morphological scoring and subsequent analysis of multiple tissue swabs of the aortic wall, urinary bladder wall, brain, liver, oral mucosa and skeletal muscle. In summary, we found variable degrees of suitability of different types of soft tissue swabs for DNA-based identification. Swabs of the aortic wall, the urinary bladder wall and brain tissue yielded the best results - in descending order - even at advanced levels of decay.

Age estimation based on aspartic acid racemization in dentine: what about caries-affected teeth?

Age estimation based on aspartic acid racemization (AAR) in dentine is one of the most precise methods in adult age. Caries induces protein degradation and may have an impact on the kinetics of AAR in dentine. We systematically examined standardized prepared dentine samples from caries-affected teeth to clarify the question, if caries-affected teeth should not be used for age estimation based on AAR at all, or if the analysis of dentine samples from such teeth may be useful after removal of the caries-affected tissue according to clinical standards. Our results suggest that caries may lead to an extensive protein degradation even in macroscopically healthy-appearing dentine samples from caries-affected teeth and may significantly affect the precision of age estimation. To ensure the quality of age estimation based on AAR in forensic practice, we recommend using dentine samples from healthy teeth. If only caries-affected teeth are available, dentine samples from at least two teeth from the same individual should be analyzed as it seems unlikely that caries-induced protein degradation occurred with identical kinetics in two different teeth. In any case, results of the analysis of caries-affected teeth must be interpreted with caution.

Age estimation based on aspartic acid racemization in human sclera.

Age estimation based on racemization of aspartic acid residues (AAR) in permanent proteins has been established in forensic medicine for years. While dentine is the tissue of choice for this molecular method of age estimation, teeth are not always available which leads to the need to identify other suitable tissues. We examined the suitability of total tissue samples of human sclera for the estimation of age at death. Sixty-five samples of scleral tissue were analyzed. The samples were hydrolyzed and after derivatization, the extent of aspartic acid racemization was determined by gas chromatography. The degree of AAR increased with age. In samples from younger individuals, the correlation of age and D-aspartic acid content was closer than in samples from older individuals. The age-dependent racemization in total tissue samples proves that permanent or at least long-living proteins are present in scleral tissue. The correlation of AAR in human sclera and age at death is close enough to serve as basis for age estimation. However, the precision of age estimation by this method is lower than that of age estimation based on the analysis of dentine which is due to molecular inhomogeneities of total tissue samples of sclera. Nevertheless, the approach may serve as a valuable alternative or addition in exceptional cases.

Age estimation of decomposed bodies based on a combined arteriosclerotic index.

Several methods exist for the estimation of the age at the time of death, ranging from mere visual inspection to costly laboratory examinations. The "combined arteriosclerotic index (CAI)" is considered to be a suitable low-budget tool for undecayed corpses. It defines the ratio between diameter and longitudinal pre-strain of the abdominal aorta. Its applicability in cases of decomposed corpses has not been studied yet. We examined whether it is a valid parameter in putrefied bodies as well and whether there is a correlation between CAI and the stage of decomposition. In conclusion the CAI becomes less accurate with increasing putrefaction. Nonetheless, even in case of high-grade putrefaction it remains a useful tool for instant age estimation which should be followed by the application of methods with higher accuracy.

Age of collagen in intracranial saccular aneurysms.

BACKGROUND AND PURPOSE: The chronological development and natural history of cerebral aneurysms (CAs) remain incompletely understood. We used (14)C birth dating of a main constituent of CAs, that is, collagen type I, as an indicator for biosynthesis and turnover of collagen in CAs in relation to human cerebral arteries to investigate this further. METHODS: Forty-six ruptured and unruptured CA samples from 43 patients and 10 cadaveric human cerebral arteries were obtained. The age of collagen, extracted and purified from excised CAs, was estimated using (14)C birth dating and correlated with CA and patient characteristics, including the history of risk factors associated with atherosclerosis and potentially aneurysm growth and rupture. RESULTS: Nearly all CA samples contained collagen type I, which was <5 years old, irrespective of patient age, aneurysm size, morphology, or rupture status. However, CAs from patients with a history of risk factors (smoking or hypertension) contained significantly younger collagen than CAs from patients with no risk factors (mean, 1.6±1.2 versus 3.9±3.3 years, respectively; P=0.012). CAs and cerebral arteries did not share a dominant structural protein, such as collagen type I, which would allow comparison of their collagen turnover. CONCLUSIONS: The abundant amount of relatively young collagen type I in CAs suggests that there is an ongoing collagen remodeling in aneurysms, which is significantly more rapid in patients with risk factors. These findings challenge the concept that CAs are present for decades and that they undergo only sporadic episodes of structural change.

Estimation of age at death based on aspartic acid racemization in elastic cartilage of the epiglottis.

Age estimation based on aspartic acid racemization (AAR) has been applied successfully to various tissues. For routine uses, AAR is analyzed in dentine. For cases in which teeth are unavailable, analyzing AAR in purified elastin has been shown to be an alternative method. The suitability of elastic cartilage from the epiglottis as an elastin source for age estimation based on AAR was tested. A total of 65 tissue samples (cartilage) of epiglottis and 45 samples of elastin purified from the elastic cartilage of epiglottis samples were analyzed. While the D-aspartic acid content of total tissue samples increased with age only slowly, its increase with age in purified elastin samples was similar to that in purified elastin from other tissues. The relationship between the D-aspartic acid content and age was shown to be close enough for age estimation based on AAR in purified elastin from the elastic cartilage of the epiglottis, provided a sufficient quality of elastin purification. Age estimation based on AAR in purified elastin from the epiglottis might serve as a valuable alternative in cases in which other tissues (e.g., teeth) are unavailable.

[Influence of grave chamber systems on decomposition]. / Zum Einfluss von Grabkammersystemen auf die Dekomposition.

Grave chambers allow bodies to decompose independent of hydrogeological circumstances. We examined the degree of body decomposition in 11 grave chambers after burial times between 12.1 and 15.3 years (mean 13.0 years). In 8 of the examined 11 bodies, bones were separated due to complete loss of tissue continuity. In 3 cases, the graves were partly not properly constructed and tissue continuity was at least partially preserved. Biochemical estimation of the age at the time of death on the basis of six teeth gathered during the inspection of the grave chambers showed only one result that could not be explained by the standard deviation of the method used.

Retention rate and side effects in a prospective trial on hepatitis C treatment with pegylated interferon alpha-2a and ribavirin in opioid-dependent patients.

Hepatitis C viral (HCV) infection is present in 30 to 98% of intravenous drug users. Intravenous substance abuse represents the main route of HCV transmission in industrialized countries. A multi-centre, randomized, controlled, prospective study assessed sustained virological response (SVR), adverse events such as depressive episodes and retention rate of HCV treatment in opioid-dependent patients. Stabilized, opioid-dependent patients with chronic HCV infection (genotype 2 or 3) received pegylated interferon alpha-2a in combination with ribavirin 800 mg/day (Group A) or 400 mg/day (Group B). Participants were randomized, blocked and stratified by genotype and viral load. A standardized psychiatric assessment, Beck Depression Inventory (BDI) and Van Zerssen's list of complaints were administered at each study visit. In 31 months, 300 opioid-dependent patients were screened; 190 (63.3%) were hepatitis C antibody positive. According to study protocol, out of 75 'potential-to-treat' patients with genotype 2 or 3, 17 stable patients (22.6%) were included in the study. All participants completed the study. Significant haemoglobin decreases occurred in both Groups A (P = 0.001) and B (P = 0.011). All the patients had an end-of-treatment (week 24) HCV RNA negativity. Fifteen (88.2%) achieved SVR at week 48. Overall, 52.9% developed depressive symptoms during treatment. Because of the prompt initiation of antidepressant medication at first appearance of depressive symptoms, no severe depressive episodes occurred. Our data show a high retention rate and reliability, and good viral response for both treatments. Hepatitis C treatment in stable opioid-dependent patients was efficacious, suggesting that addiction clinics can offer antiviral therapy in combination with agonistic treatment as part of multi-disciplinary treatment.

Office-based treatment in opioid dependence: a critical survey of prescription practices for opioid maintenance medications and concomitant benzodiazepines in Vienna, Austria.

BACKGROUND: The success of maintenance treatment for opioid dependence in office-based settings is influenced by the extent of treatment coverage, the availability of effective medications and the capacity of general practitioners to prescribe opioids in adequate doses with a minimum of concomitant benzodiazepine prescriptions. METHODS: This study compares prescriptions for opioid maintenance and concomitant benzodiazepine from Viennese physicians in 2002 and 2005 using health insurance prescription records (n = 30,309). RESULTS: Between 2002 and 2005, the number of patients prescribed opioids more than doubled (ratio 1:2.3), slow-release oral morphine replaced methadone as the most frequently prescribed medication (57.1 vs. 23.4%; buprenorphine 19.5%), and the ratio of benzodiazepine to opioid prescriptions significantly declined (0.76:1 vs. 0.42:1). Many patients were prescribed concomitant benzodiazepines (27%), in some cases from a secondary physician. CONCLUSION: Increased utilization of opioid medications in office-based settings will facilitate better treatment coverage. However, safeguards are necessary to ensure that general practitioners have sufficient training and support to safely and appropriately provide treatment, including the reduction in concomitant benzodiazepine use.