Forensic analysis of hallucinogenic fungi: a DNA-based approach

Hallucinogenic fungi synthesize two controlled substances, psilocin and psilocybin. Possession of the fungal species that contain these compounds is a criminal offence in North America. Some related species that are morphologically similar, do not contain the controlled substances. Therefore, unambiguous identification of fungi to the species level is critical in determining if a mushroom is illegal. We investigate a phylogenetic approach for the identification of species that contain the psychoactive compounds. We analyzed 35 North American specimens representing seven different genera of hallucinogenic and non-hallucinogenic mushrooms. We amplified and sequenced the internal transcribed spacer region of the rDNA (ITS-1) and a 5' portion of the nuclear large ribosomal subunit of rRNA (nLSU rRNA or 28S). ITS-1 locus sequence data was highly variable and produced a phylogenetic resolution that was not consistent with morphological identifications. In contrast, the nLSU rRNA data clustered isolates from the same species and separated hallucinogen containing and non-hallucinogen containing isolates into distinct clades. With this information, we propose an approach that combines the specificity of PCR detection and the resolving power of phylogenetic analysis to efficiently and unambiguously identify hallucinogenic fungal specimens for legal purposes.     

Characterization and haplotype analysis of 10 novel Y-STR loci in Chinese Han population

In this study, we analyzed allelic sequences of 10 novel Y-specific STR loci, DYS454, DYS510, DYS513, DYS520, DYS542, DYS544, DYS552, DYS561, DYS587 and DYS593, surveyed the distribution of haplotypes in a Chinese Han population. Extracted DNA was amplified with PCR, followed by a horizontal non-denaturing polyacrylamide gel electrophoresis with discontinuous buffer system. Purified alleles were sequenced on DNA sequencer (ABI Model 377) to verify the number of motif repeats. The number of alleles observed at each locus ranged from 3 to 8, yielding 102 haplotypes in 103 unrelated males samples. The allele diversity values for each locus ranged from 0.2099 (DYS544) to 0.7523 (DYS552). The haplotype diversity using all these loci was 0.9998. Our study revealed that they were valuable Y-specific markers for forensic applications.     

Fatal firearm wounds: a clinicopathologic study

Firearm-related injuries are a leading cause of morbidity and mortality in South Africa. This retrospective autopsy study included all firearm-related fatalities seen at a central mortuary in Durban. These cases had initially been assessed by a clinician before death, from July 1998 to June 1999.A total of 322 cases of fatal gunshot wounds were reviewed. Medical records and postmortem findings were compared by data capture onto a spreadsheet. Twenty-eight cases were excluded because of surgical alteration of the wounds, making autopsy evaluation impossible. In 50.7% of cases, the number of wounds was correctly identified and in 55.1% the position was correctly identified. In only 36.1% of cases were the entry and exit wounds correctly identified. The total number of missed wounds was 504 (48.3% of cases). The majority of missed wounds were on the upper limbs (25%), with the anterior trunk (21.8%) and lower limbs (18.5%) making up the remainder. This study demonstrates that physicians in South Africa do not adequately document and interpret gunshot wounds, which may influence the treatment of the patient and have medicolegal consequences. Emphasis should be placed on undergraduate and postgraduate teaching for physicians to correctly identify, interpret, and document firearm wounds.     

Munchausen syndrome by proxy (MSBP): an extreme form of child abuse with a special forensic challenge

Munchausen syndrome by proxy (MSBP) is a special form of child abuse in which an adult repeatedly produces symptoms of illness in a person under his/her care. In most cases the perpetrators are mothers who repeatedly and in different ways produce or feign symptoms of illness in their children in order to obtain medical treatment for them. MSBP is thus a special form of child abuse that is also of importance in the field of forensic medicine and a particular challenge to the medicolegal expert. We report two cases of poisoning with different substances (clozapine and clonidine) detected by toxicological investigations at our Department of Legal Medicine. The relevance of the problem for the medicolegal expert and the importance of an interdisciplinary co-operation are pointed out.     

STR genotyping and mtDNA sequencing of latent fingerprint on paper

A systematic study was conducted to investigate whether DNA can be successfully extracted from latent fingerprints deposited on ordinary paper and analysed using short tandem repeat profiling and mitochondrial DNA sequencing. In order to evaluate the performance of latent fingerprint analysis in a criminal case, experiments with varying conditions were carried out to improve our understanding of low copy number (LCN) DNA typing. After optimising the extraction methods to achieve increased sensitivity, the examination of touched paper can routinely yield the STR profile of the individual who has touched it. A fingerprint can therefore be considered as a potential source of DNA for genetic identification. Nevertheless, the findings of our "after enhancement experiment" (using chemically or physically pre-treated fingerprints), and our "mixture experiment" (using fingerprints from three to four people on the same sheet of paper) help to define the limitations of the low copy number PCR technique in forensic casework.     

FINEX: a Probabilistic Expert System for forensic identification

A series of recent papers have shown how to formulate complex problems of forensic DNA identification inference, such as occur in disputed paternity or criminal identification cases, in terms of Probabilistic Expert Systems (PESs). However, at the present time, general purpose PES software is not particularly well suited to the repetitive tasks of: specifying an appropriate set of marker networks for a specific problem; for editing the many local conditional probability tables; and combining evidence from several genetic markers to evaluate likelihoods. Here, I describe a user-friendly prototype software tool called FINEX developed both to automate such tasks and also to evaluate likelihoods of interest. Ease of use is achieved by a graphical specification language that enables a user to quickly specify a range of forensic DNA problems. I describe the algorithms by which FINEX converts the user input in the graphical specification language and data on observed markers to the Bayesian networks used in PES.     

Bite mark documentation and analysis: the forensic 3D/CAD supported photogrammetry approach

Bite mark identification is based on the individuality of a dentition, which is used to match a bite mark to a suspected perpetrator. This matching is based on a tooth-by-tooth and arch-to-arch comparison utilising parameters of size, shape and alignment. The most common method used to analyse bite mark are carried out in 2D space. That means that the 3D information is preserved only two dimensionally with distortions. This paper presents a new 3D documentation, analysis and visualisation approach based on forensic 3D/CAD supported photogrammetry (FPHG) and the use of a 3D surface scanner. Our photogrammetric approach and the used visualisation method is, to the best to our knowledge, the first 3D approach for bite mark analysis in an actual case. The documentation has no distortion artifacts as can be found with standard photography. All the data are documented with a metric 3D measurement, orientation and subsequent analysis in 3D space. Beside the metrical analysis between bite mark and cast, it is possible using our method to utilise the topographical 3D feature of each individual tooth. This means that the 3D features of the biting surfaces and edges of each teeth are respected which is--as shown in our case--very important especially in the front teeth which have the first contact to the skin. Based upon the 3D detailed representation of the cast with the 3D topographic characteristics of the teeth, the interaction with the 3D documented skin can be visualised and analysed on the computer screen.     

Third molar root development in relation to chronological age: a large sample sized retrospective study

The aim of the present investigation, which is a continuation and extension of a previous published pilot study [Forensic Sci. Int. 129 (2002) 110], was to calculate the chronological age of an individual based on the dental developmental stages of third molars (so-called wisdom teeth). The evaluated material consisted of 2513 orthopantomograms (OPG's) of patients all of Belgian Caucasian origin between 15.7 and 23.3 years. The sample of OPG's was scored by two observers who were calibrated for intra- and interobserver reliability by means of kappa statistics. The kappa statistics revealed a strong agreement between the intra- and interobserver measurements. Further statistical analysis was performed in order to obtain multiple regression formulae for dental age calculation with the chronological age as the independent variable and the third molar developmental stages as dependent variables. Both for males and females a significant Pearson correlation coefficient was noticed for contralateral third molars, a smaller less significant one for the antimeres. For the application of the regression formulae one has to take into consideration the gender, the location and number of wisdom teeth. Probabilities for an individual to be older than 18 years were also calculated in case of fully developed wisdom teeth.     

The time-dependent expression of fibronectin,MRP8, MRP14 and defensin in surgically treated human skin wounds

Fibronectin, MRP8, MRP14 and defensin were detected immunohistochemically in 46 surgically treated, fresh (hours old) human wounds and in wounds of 13 individuals who died immediately from fatal trauma (airplane crashes or train rollovers). In immediate fatal trauma, it was not possible to detect fibronectin outside of bleeding areas nor could inflammatory cells be visualized in the interstitium using MRP8, MRP14, or defensin antibodies. Fibronectin staining could be regularly demonstrated in wounds at least 20 min-old. Granulocytic infiltrates limited to the perivasal space could be detected 20-30 min after infliction of the wound expressing MRP8, MRP14 and defensin. It was also possible to detect fibronectin networks and MRP8-, MRP14-, and defensin-positive granulocytes and macrophages in particular wounds up to 30 days-old. No differences between the expression of MRP8 and MRP14 could be demonstrated in the wounds, the majority of which were only several hours old. As wound age increased, the number of defensin-positive granulocytes detected decreased. The immunohistochemical detection of fibronectin is a useful way to demonstrate vitality in fresh wounds, beginning about 20-30 min post-trauma. However, detection of MRP8, MRP14 and defensin provides no advantage over the routine histological detection of granulocytes and macrophages in wounds under 1-2 days old.