STR analysis of artificially degraded DNA-results of a collaborative European exercise

Degradation of human DNA extracted from forensic stains is, in most cases, the result of a natural process due to the exposure of the stain samples to the environment. Experiences with degraded DNA from casework samples show that every sample may exhibit different properties in this respect, and that it is difficult to systematically assess the performance of routinely used typing systems for the analysis of degraded DNA samples. Using a batch of artificially degraded DNA with an average fragment size of approx. 200 bp a collaborative exercise was carried out among 38 forensic laboratories from 17 European countries. The results were assessed according to correct allele detection, peak height and balance as well as the occurrence of artefacts. A number of common problems were identified based on these results such as strong peak imbalance in heterozygous genotypes for the larger short tandem repeat (STR) fragments after increased PCR cycle numbers, artefact signals and allelic drop-out. Based on the observations, strategies are discussed to overcome these problems. The strategies include careful balancing of the amount of template DNA and the PCR cycle numbers, the reaction volume and the amount of Taq polymerase. Furthermore, a careful evaluation of the results of the fragment analysis and of automated allele calling is necessary to identify the correct alleles and avoid artefacts.     

„Spielarten des Kapitalismus“ und institutionelle Komplementaritäten in der Makroökonomie — Eine empirische Analyse

Der Beitrag geht der Frage nach, wie national unterschiedliche politökonomische Institutionen die Leistungskraft entwickelter Marktwirtschaften beeinflussen und ob institutionelle Komplementaritäten in der Makroökonomie existieren. Der Ansatz der ?Spielarten des Kapitalismus“ behauptet, dass es diese Komplementaritäten gibt und sich die Länder entsprechend dem in den ökonomischen Sphären vorherrschenden Modus der Koordination in klare Gruppen unterscheiden lassen. In dem Artikel werden die Kernaussagen des Ansatzes einer Reihe von empirischen Tests unterzogen, die auf einer zusammenfassenden Analyse einer Reihe von Fallbeispielen beruht, Die empirischen Ergebnisse bestätigen die Annahmen in überzeugender Weise. Dieliberalen undkoordinierten Marktwirtschaften variieren systematisch entsprechend dem relativen Verhältnis zwischen marktförmiger und strategischer Koordination. Institutionelle Komplementaritäten finden sich in beiden Typen, und sie bringen gesamtwirtschaftliche Leistungszuwächse hervor. Abschließend werden die Muster des institutionellen Wandels in den entwickelten politischen Ökonomien analysiert und einige politische Implikationen der Befunde diskutiert.     

Evaluation of ephedrine, pseudoephedrine and phenylpropanolamine concentrations in human urine samples and a comparison of the specificity of DRI amphetamines and Abuscreen online (KIMS) amphetamines screening immunoassays

The purpose of this study was to evaluate the ability of two amphetamine class screening reagents to exclude ephedrine (EPH), pseudoephedrine (PSEPH), and phenylpropanolamine (PPA) from falsely producing positive immunoassay screening results. The study also sought to characterize the prevalence and concentration distributions of EPH, PSEPH, and PPA in samples that produced positive amphetamine screening results. Approximately 27,400 randomly collected human urine samples from Navy and Marine Corps members were simultaneously screened for amphetamines using the DRI and Abuscreen online immunoassays at a cutoff concentration of 500 ng/mL. All samples that screened positive were confirmed for amphetamine (AMP), methamphetamine (MTH), 3,4-Methylenedioxyamphetamine (MDA), 3,4-Methylenedioxymethamphetamine (MDMA), EPH, PSEPH, and PPA by gas chromatography/mass spectrometry (GC/MS). The DRI AMP immunoassay identified 1,104 presumptive amphetamine positive samples, of which only 1.99% confirmed positive for the presence of AMP, MTH, MDA, or MDMA. In contrast, the online AMP reagent identified 317 presumptive amphetamine positives with a confirmation rate for AMP, MTH, MDA, or MDMA of 7.94%. The presence of EPH, PSEPH, or PPA was confirmed in 833 of the 1,104 samples that failed to confirm positive for AMP, MTH, MDA, or MDMA; all of the 833 samples contained PSEPH. When compared to the entire screened sample set, PSEPH was present in approximately 3%, EPH in 0.9%, and PPA in 0.8% of the samples. The results indicate that cross reactivities for EPH, PSEPH, and PPA are greater than reported by the manufacturer of these reagents. The distribution of concentrations indicates that very large concentrations of EPH, PSEPH, and PPA are common.     

Medicolegal aspects of witnessed suicide due to gunshot to the head. II. Legal medicine aspects and examination of the firing hand

The second part of the paper on suicides by gunshots to the head in the presence of witnesses focuses on relevant morphological autopsy findings such as entrance site, signs of close range or contact shots, bullet path etc. and also discusses selected aspects of ascertaining gunshot residues. For the identification of the shooter an integral medicolegal assessment of all the facts including the investigation results and the autopsy findings is essential. However, the morphological findings alone do not allow safe diagnosis, as for example in a homicide the temporal region, which was affected in all our cases, may have been deliberately chosen by the perpetrator as a localization typical of suicide. Thus methods to ascertain gunshot residues on the firing hand (by means of adhesive films and the polyvinyl-alcohol collection method--PVAL) are of great practical importance. In seven cases adhesive films and/or the polyvinyl-alcohol collection method were used. In one case the gunshot residues (GSR) were analysed by means of tape lifts and subsequent scanning electron microscopy (SEM). It was found that especially the combined application of topographical (adhesive tape/PVAL) and cumulative (SEM) methods allowed for the doubtless identification of the shooter, thus usually confirming the suicide. By the example of one case it is demonstrated that without the immediate collection of evidence at the scene objective reconstruction of the event becomes impossible. On the basis of the reported cases recommendations are finally given for a differentiated approach in the medicolegal evaluation of alleged witnessed suicide by gunshot (to the head).     

Development of a finger printing device for use on a mobile robot

In this paper, we describe the development of a device for fuming fingerprints with cyanoacrylate (Super Glue) to enable police tactical units to obtain fingerprint evidence from suspicious packages using a remote-controlled robot. Through a series of initial experiments and preliminary designs, we show that effective cyanoacylate fuming requires sufficient heat, humidity, and airflow. This work led to the development of a final working prototype, called robot accessory for fuming fingerprint evidence (RAFFE), which is currently being field tested by the Calgary Police Service.     

Trace analysis of peroxide explosives by high performance liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry (HPLC-APCI-MS/MS) for forensic applications

An HPLC-APCI-MS(/MS) method for the (trace) analysis of the most commonly encountered peroxide explosives, hexamethylenetriperoxidediamine (HMTD) and triacetonetriperoxide (TATP), has been developed. With this method, HMTD and TATP have been analyzed in the same run. (Pseudo-)molecular ions of these peroxides have been obtained as base peak under the same condition. A series of product ions was produced from these pseudo-molecular ions ([HMTD - 1]+ and [TATP + NH4]+) in the MS/MS analysis. We also pioneered in showing that a TATP molecular ion [TATP + H]+ can be observed with HPLC-MS/MS. The limit of detection for HMTD and TATP was 0.26 and 3.3 ng, respectively, on column by HPLC-MS in the Full Scan mode and 0.08 and 0.8, respectively, by HPLC-APCI-MS/MS in Selected Reaction Monitoring (single mass unit) mode. The method presented has been applied successfully for the identification of peroxides in the bulk solid state (powder sample), as well as in post-blast extracts originating from a forensic case. For the post-blast extracts, the use of tandem MS has been shown clearly to be of crucial importance for the identification and detection of the peroxide explosives.     

A genetic basis for anomalous band patterns encountered during DNA STR profiling

Since 1995 the Forensic Science Service (FSS) has carried out DNA profiling of reference samples for the UK National DNA Database and in forensic casework using two multiplex STR profiling systems. During this period, profiles with anomalous banding patterns, although comparatively rare, have been encountered regularly. The FSS has collected instances of triallelic patterns and aberrant diallelic patterns. A systematic examination of these patterns has provided insight into their underlying genetic cause. The triallelic patterns could be classified into two types based on the relative intensities of their component alleles. In the Type 1 pattern the alleles were of uneven intensity, whereas in the Type 2 pattern, all three alleles were of even intensity. Evidence is presented that the more frequent Type 1 pattern is the result of somatic mutation at a heterozygous locus, and the Type 2 pattern is the result of a localized chromosomal rearrangement at a heterozygous locus. Directly from the Type 1 pattern, it was possible to deduce the size difference between the progenitor and mutated allele. All mutational changes were found to be multiples of four nucleotides, suggesting the loss or addition of one or more tetrameric repeat units. Aberrant diallelic patterns were identified by analysts due to an unexpectedly large difference in intensity between alleles at a heterozygous locus. While some of these diallelic patterns are likely caused by the same genetic phenomena described above occurring at a homozygous locus, others are demonstrated to be caused by a mutation in the primer binding sequence, leading to a reduction in amplification efficiency of one allele. It is concluded that based on a visual inspection of a profile, it is possible to infer a likely genetic basis directly from the triallelic pattern. By contrast, the aberrant diallelic patterns can be due to any one of a number of possible genetic effects.     

Y-SNP typing of U.S. African American and Caucasian samples using allele-specific hybridization and primer extension

Multiplex analysis of genetic markers has become increasingly important in a number of fields, including DNA diagnostics and human identity testing. Two methods for examination of single nucleotide polymorphisms (SNPs) with a potential for a high degree of multiplex analysis of markers are primer extension with fluorescence detection, and allele-specific hybridization using flow cytometry. In this paper, we examined 50 different SNPs on the Y-chromosome using three primer extension multiplexes and five hybridization multiplex assays. For certain loci, the allele-specific hybridization method exhibited sizable background signal from the absent alternate allele. However, 100% concordance (>2000 alleles) was observed in ten markers that were typed using both methods. A total of 18 unique haplogroups out of a possible 45 were observed in a group of 229 U.S. African American and Caucasian males with the majority of samples being assigned into 2 of the 18 haplogroups.