DNA Commission of the International Society for Forensic Genetics (ISFG): recommendations regarding the role of forensic genetics for disaster victim identification (DVI).

The ISFG membership consists of scientists and medical professionals specialized in using genetic testing for kinship analysis and the individualization of biological material. This expertise makes the forensic geneticist a resource of advice to international and national organizations dealing with human identifications and causes many DNA laboratories to get involved in DVI tasks. The present recommendations are meant to educate more forensic geneticists about their potential involvement in mass fatality preparedness and possible DVI efforts, as well as to provide practical guidance for each of the laboratories' individual tasks. The idea to work on DNA-specific recommendations was born after a round table discussion dealing with the 2004 Tsunami disaster in south east Asia during the 21st congress of the International Society for Forensic Genetics on the Azores, Portugal, in September 2005. The ensuing discussion between scientists and pathologists that had been involved in the International Center in Khao Lak, Thailand, revealed the need for the scientific community to be better prepared to answer the local authorities' questions by formulating generally acceptable scientific standards for the most efficient use of DNA-based victim identification methods. These recommendations, as well as the many cited references, are intended to provide guidance on establishing preparedness for the forensic genetics laboratory, on collecting and storing ante-mortem and post-mortem samples suitable for DNA analysis, on DNA extraction and genetic typing strategies, on data management, and on issues related to the biostatistical interpretation and reporting of results.

STR loci Penta D and Penta E: Austrian Caucasian population data.

Allele frequencies for the short tandem repeat (STR) loci Penta D and Penta E, included in the PowerPlex 16 kit were obtained from a sample of 269 unrelated Austrian Caucasian individuals. Both loci met Hardy-Weinberg expectations.

Results of a collaborative study of the EDNAP group regarding the reproducibility and robustness of the Y-chromosome STRs DYS19, DYS389 I and II, DYS390 and DYS393 in a PCR pentaplex format.

A collaborative exercise was carried out by the European DNA Profiling Group (EDNAP) in the frame work of the STADNAP program, i.e. standardization of DNA profiling in Europe, in order to evaluate the performance of a Y-chromosome STR pentaplex, which includes the loci DYS19, DYS389 I and II, DYS390 and DYS393 and to determine whether uniformity of results could be achieved among different European laboratories. Laboratories were asked to analyze the five Y-STRs using singleplex and multiplex conditions in three bloodstains and one mixed stain (95% female and 5% male). All the laboratories reported the same results even for the mixed stain included in the exercise. This demonstrates the reproducibility and robustness of Y-chromosome STR typing even with multiplex formats and proves the usefulness of Y-STR systems for analyzing mixed stains with a male component.A total of 930 male samples from 10 different populations from Europe were also analysed for all the loci included in the pentaplex. Eight of these ten populations also included haplotype data. As for single gene analysis, haplotype diversity was higher in Germany and Italy and lower in Western European countries and Finland. Pairwise haplotype analysis shows the Finnish departure from the rest of the populations and a relatively homogeneity in the other European populations with F(ST) estimates lower than 0.05.UPGMA analysis shows an association of Western European population (Ireland, UK, Portugal and Galicia) on the one hand and central European populations on the other.

Population genetics of ten STR loci (AmpFlSTR SGM plus) in Austria.

A population study on the ten short tandem repeat (STR) loci D3S1358, VWA, D16S539, D2S1338, D8S1179, D21S11, D18S51, D19S433, TH01 and FGA was performed on 204 unrelated Austrian Caucasians. The DNA was amplified by multiplex PCR using the AmpF/STR SGM plus kit. All loci met Hardy-Weinberg expectations. The combined power of exclusion for the ten STR loci was 0.999976. The results show that these loci are very useful for forensic purposes.

Results of collaborative study regarding the standardization of the Y-linked STR system DYS385 by the European DNA Profiling (EDNAP) group.

Y-chromosome linked short tandem repeat (STR) loci are inherited as a closely linked haplotype, which appears to remain stable in a given paternal lineage over many generations. In forensic cases, Y-linked STRs are particularly useful for the identification of human remains as well as in rape cases with mixed male/female stain samples. DYS385 is derived from tandemly duplicated segments of the Y chromosome thus giving rise to two fragments of variable length which do not behave like alleles but genotypes. The European DNA Profiling (EDNAP) group has carried out a collaborative exercise among 14 participating laboratories using DYS385 for typing of five unknown bloodstains and a control sample. Furthermore, population data from eight different European countries with samples sizes between 91 and 150 male individuals were collected. The results confirm previous observations that DYS385 is one of the most informative Y-linked STR loci. It could also be demonstrated that reproducible results can be obtained independently from the electrophoretic separation and detection methods used. Thus DYS385 may serve as a useful complementation to the routinely used autosomal STR systems in special cases.

Population data for 101 Austrian Caucasian mitochondrial DNA d-loop sequences: application of mtDNA sequence analysis to a forensic case.

The sequence of the two hypervariable segments of the mitochondrial DNA (mtDNA) control region was generated for 101 random Austrian Caucasians. A total of 86 different mtDNA sequences was observed, where 11 sequences were shared by more than 1 individual, 7 sequences were shared by 2 individuals and 4 sequences were shared by 3 individuals. One of the four most common mtDNA sequences in Austrians is also the most common sequence in both U.S. and British Caucasians, found in approximately 3.0% of Austrians, 4.0% of British, and 3.9% of U.S. Caucasians. Of the remaining three common Austrian sequences, one was not observed in either U.S. or British Caucasians. However, three British Caucasians exhibited a similar sequence type. Therefore, this particular cluster of sequence polymorphisms may represent a common "European" mtDNA sequence type. In general, Austrian Caucasians show little deviation from other Caucasian databases of European descent. Finally, mtDNA sequence analysis was applied to a forensic case, where hairs found at a crime scene matched the control hairs from the suspect.

Report of the European DNA Profiling Group (EDNAP)--an investigation of the hypervariable STR loci ACTBP2, APOAI1 and D11S554 and the compound loci D12S391 and D1S1656.

This paper describes the results of three collaborative exercises which continues the EDNAP theme to explore whether uniformity of DNA profiling results could be achieved between European laboratories using STRs. In an earlier exercise, complex hypervariable AAAG-repeat STR loci were investigated, but reproducibility was found to be poor because of the variation of techniques used by participating laboratories. In the exercise reported here, an internal allelic ladder composed of ACTBP2 and D11S554 fragments was distributed. This ladder was used to size ACTBP2 analysed by a "singleplex" PCR amplification and D11S554 combined with APOAI1 in a separate "duplex" reaction. Laboratories were asked to test 7 blood stains, one of which was a known control, and to report the results to the co-ordinating laboratory. The exercise demonstrated that ACTBP2 showed good reproducibility between laboratories, whereas further testing would be needed to validate APOAI1 and D11S554 for interlaboratory comparisons. In separate exercises, the simple loci D12S391 and D1S1656 were tested; both of these showed excellent reproducibility between laboratories.

Review of active compression-decompression cardiopulmonary resuscitation (ACD-CPR). Analysis of iatrogenic complications and their biomechanical explanation.

Our review takes a critical look at the active compression-decompression technique (ACD) for cardiopulmonary resuscitation (CPR). ACD-CPR was developed following a report of successful resuscitation performed by a medical amateur using a household plunger. The efficacy of the principle of active decompression has been demonstrated by animal and human studies. Potential iatrogenic complications from the CardioPump were evaluated only when large clinical trials were already underway. Our prospective analysis of autopsy patients and systematic randomised studies in corpses prove that ACD-CPR using the CardioPump considerably increases the rate of iatrogenic complications and especially of sternum fractures. The experimental use of the CardioPump in corpses and the analysis of a variety of different parameters, especially of the rubber cushion pads mounted in the silicone cup to prevent skin abrasions, revealed a statistically significant correlation between sternum fractures and female sex (P < 0.01) and usage of the rubber cushion pad (P = 0.045). Biomechanical studies showed that the transmission of forces from the CardioPump is greatly dependent on chest shape. The lower the sternum is sunken compared with the surrounding structures, the higher the force which is transmitted via the central area of the device onto the sternum. The rubber cushion pad shortens the distance between CardioPump and sternum by 5 mm and therefore increases the sternal loading. Sex differences in the shape of the sternum and especially the thickness may account for the significant correlation between sternum fractures and female sex.

Report of the European DNA profiling group (EDNAP): an investigation of the complex STR loci D21S11 and HUMFIBRA (FGA).

This paper describes a collaborative exercise which was intended to demonstrate whether uniformity of DNA profiling results could be achieved between European laboratories using two complex short tandem repeat (STR) loci. The loci D21S11 and HUMFIBRA (FGA) were chosen because they are commonly used by different European laboratories. D21S11 has approximately 14 common alleles (f > 0.001), whereas HUMFIBRA has 19 common alleles. Laboratories were asked to test seven blood stains, one of which was a known control, and to report the results to the coordinating laboratory. The exercise demonstrated that complex STRs were amenable to standardisation.

HLA-DQ alpha genotype and allele frequencies in an Austrian population.

The HLA-DQ alpha system is used to an increasing degree in forensic serology, for stain investigations, and paternity cases. Therefore samples which were taken from 85 unrelated persons living in Austria were examined. DNA was analyzed by polymerase chain reaction (PCR) amplification followed by hybridization to allele-specific oligonucleotide probes in a reversed dot-blot technique after extraction with phenol/chloroform. HLA-DQ alpha allele frequencies of German, Finnish, Dutch, British, Italian, Caucasian/USA and of Austrian populations were compared. Six common HLA-DQ alpha alleles are detectable. These alleles determine 21 possible genotypes. In our study 20 genotypes were obtained; the genotype 3/3 was not observed. The HLA-DQ alpha typing system using PCR-amplified DNA is a simple and rapid method for typing very small amounts of DNA and very old and/or degraded DNA. Therefore it is a helpful method in forensic casework.

Serious complications from active compression-decompression cardiopulmonary resuscitation.

Complications arising from techniques of cardiopulmonary resuscitation (CPR) were reviewed by analysing the autopsy protocols of 25 patients who died after standard (Std) CPR and 31 who died after active compression-decompression (ACD) CPR, 15 of them preceded by Std CPR. The results can be summarised as follows: After Std CPR (n = 25) rib fractures were detected in 28%, sternal fractures in 16%, and no injuries in 68%. After ACD-CPR (n = 16) rib fractures occurred in 68%, sternal fractures in 68% and no injuries in 25%. After ACD-CPR following Std CPR(n = 15) rib fractures were detected in 93%, sternal fractures in 93%, and no patients were without thoracic fracture. In two patients severe cardiac injuries occurred clearly attributable to CPR. In conclusion cardiopulmonary resuscitation by the ACD-technique caused rib and sternal fractures more often than Std CPR and has a higher risk for iatrogenic cardiac and possible fatal injury.

Lesions of the alar ligaments. In vivo and in vitro studies with magnetic resonance imaging.

STUDY DESIGN: This study analyzed anatomic characteristics of the alar ligaments and the possibility of imaging them with magnetic resonance imaging. Also determined was whether artificial ruptures of the alar ligament can be recognized experimentally. OBJECTIVE: To determine the ability of magnetic resonance imaging to visualize normal, torn, resected alar ligaments. SUMMARY OF BACKGROUND DATA: There are no studies about computed tomography or magnetic resonance imaging findings of alar ligaments and after anatomic sections. Direct visualization of the complete ligament is not possible for computed tomography. No precise diagnostic method for showing a ruptured alar ligaments has been described. Magnetic resonance imaging seems to be the method of choice for distinguishing between normal and pathologic soft tissue. METHODS: Fifteen specimens from accident victims underwent anatomic dissection. In addition, ligaments from three groups were examined: 1) eight volunteers, 2) seven patients, and 3) 17 fresh cadaveric specimen before anatomic exploratory dissection. In seven of these specimens, one ligament was cut to simulate an artificial disruption and magnetic resonance imaging was repeated. RESULTS: Lesions of the alar ligaments were found in four of 15 prepared specimens. Using magnetic resonance imaging, the alar ligaments could be identified in all volunteers, patients, and specimen except one. No ruptures were found in the 17 specimens. Of the seven resected specimens, all cuts could be demonstrated by magnetic resonance imaging. CONCLUSION: Magnetic resonance imaging is useful for showing lesions of the alar ligaments because of a high soft tissue contrast, plane independence imaging, possibility of functional scans, and secondary reconstruction from three-dimensional data sets.

Postmortem injuries inflicted by domestic golden hamster: morphological aspects and evidence by DNA typing.

A case of postmortem animal scavenging by a domestic golden hamster (Mesocricetus auratus) is presented. A 43-year-old woman, who was not under medical treatment, was found dead in her flat with the lower part of her body naked and her legs straddled. Her face showed extensive lesions of the soft tissues which the medical examiner interpreted as vital scalping injuries. The total findings at the scene suggested at first a sexual offence. On autopsy the face injuries were identified as postmortem defects by animal scavenging with the teeth marks typical of rodents. In fact, the deceased had kept in her flat a free-range golden hamster whose burrow contained numerous fingernail-sized pieces of skin, fatty and muscular tissue. By means of DNA typing it was proved that these pieces of tissue belonged to the deceased.

[Population study in West Austria using DNA single locus probes]. / Populationsstudie in West-Osterreich unter Verwendung von DNA-Single-Locus-Sonden.

Allele frequency distribution of the single-locus system YNH 24 (n = 302), G3 (n = 251), MS 43A (n = 333), and MS 31 (n = 333) was determined in Western Austria. After digestion with Hinf I, electrophoresis and Southern blotting, the genomic DNA was hybridised with the probes YNH 24, G3, MS 43A, and MS 31. Blood samples were taken from 333 unrelated caucasians living in the area of Innsbruck, Tyrol, Austria. The fragment distribution was calculated for each of the 4 single-locus systems. A single fragment pattern, indicating homozygosity, was shown in 9.93% with YNH 24, 12.45% with G3, 9.61% with MS 43A, and 7.21% with MS 31; the corresponding heterozygosity rates were 90.07%, 87.55%, 90.39%, and 92.79%, respectively. Our data are compared with those from England [18], Germany [3], Greenland [7], and Denmark [14]. Discrimination indices (Table 2) were calculated and statistical parameters (Table 3) presented.

[RFLP analysis of single hairs]. / RFLP-Analyse von Einzelhaaren.

Investigations were performed with the root of one hair torn out 6 times in parallel. Yield of DNA was about the same with 6 extraction methods tested. Hairs stored at RT up to 8 weeks in non sterile environment showed the same result as deeply frozen hairs. Hairs of one test person could not be typed in repeated investigations, whereas hairs of another person could successfully be typed in every case (n > 100). 35 hairs of 66 samples investigated from 11 other persons could be typed.

Immunocytochemical typing of ABO blood groups in vaginal swabs partly contaminated with semen.

ABH typing by immunocytochemical method has been carried out on 163 selected vaginal swabs. 127 cases (78%) were determined correctly, 11 cases (7%) incorrectly and 25 cases (15%) could not be classified. Often, not all of the vaginal cells showed the expected positive staining, which was not counted as a false result. The incorrect results were not dependent on the secretor status, but 37% of the non-secretor cases could not be classified immunocytochemically, as compared with 12% of the secretors. In a second series, 61 vaginal swabs, dried on microscope slides, have been covered with semen from an A1 B secretor. Absorption of heterologous antigens by the vaginal epithelia could be demonstrated only after extremely long incubation with semen and extremely long incubation with the anti-A or anti-B antibodies. From the 163 "native" swabs, 17 gave a positive reaction with the acid phosphatase test, but only one false ABH result. A possible influence of bacteria upon the results is discussed. We believe that in practice, no faults in immunocytochemical ABH typing have to be expected, due to absorption of heterologous antigens.

DNA profiling of bloodstains on linen pretreated with remedies used for cleaning and maintaining clothes.

Linen was pretreated with 20 remedies for cleaning and maintaining clothes and 20 microliters blood was applied on each sample and dried. Restriction enzyme digest of bloodstain DNA was irregularly inhibited by highly concentrated residues of 2 detergents and a stain-remover and colour-brightener. An additional dialysis step to purify DNA (Gill 1987) reliably prevented disturbance. High molecular DNA was obtained in every case and bandshifts were not observed.

[Modification of DNA typing of blood stains by textile stain carriers]. / Beeinflussung der DNA-Typisierung von Blutspuren durch textile Spurenträger.

Samples of 20 microliters blood were applicated von 55 different textiles, containing all usual materials for clothes, straight from the fabric and after thorough washing. DNA profiling was influenced only by blue jeans and blue terry towel straight from the fabric; in some of these samples there was an inhibition of the restriction enzyme digest that could not be prevented by an additional dialysis step.

[ABH antigens on spermatozoa membrane--absorption or local synthesis?]. / ABH-Antigene auf der Spermatozoenmembran--Absorption oder lokale Synthese?

ABH antigens can be demonstrated on the membrane of spermatozoa from semen, but not on spermatozoa from the epididymis. It is discussed in the literature, if the antigens are absorbed from the antigen rich seminal fluid, or if they are synthesized on the membrane, according to a haploid gene expression. In order to answer this question, cleaned spermatozoa from semen and spermatozoa from tissue samples of epididymidis, ductus epididymidis and the prostate region were investigated using the immunoenzyme technique. It was found that spermatozoa were ABH marked only after contact with the secretions of the accessory glands. Results indicate, that the ABH antigens on the spermatozoa derive from the seminal fluid.