Identification of exhumed remains of fire tragedy victims using conventional methods and autosomal/Y-chromosomal short tandem repeat DNA profiling

In a fire tragedy in Manila in December 1998, one of the worst tragic incidents which resulted in the reported death of 23 children, identity could not be established initially resulting in the burial of still unidentified bodies. Underscoring the importance of identifying each of the human remains, the bodies were exhumed 3 months after the tragedy. We describe here our work, which was the first national case handled by local laboratories wherein conventional and molecular-based techniques were successfully applied in forensic identification. The study reports analysis of DNA obtained from skeletal remains exposed to conditions of burning, burial, and exhumation. DNA typing methods using autosomal and Y-chromosomal short tandem repeat (Y-STR) markers reinforced postmortem examinations using conventional identification techniques. The strategy resulted in the identification of 18 out of the 21 human remains analyzed, overcoming challenges encountered due to the absence of established procedures for the recovery of mass disaster remains. There was incomplete antemortem information to match the postmortem data obtained from the remains of 3 female child victims. Two victims were readily identified due to the availability of antemortem tissues. In the absence of this biologic material, parentage testing was performed using reference blood samples collected from parents and relatives. Data on patrilineal lineage based on common Y-STR haplotypes augmented autosomal DNA typing, particularly in deficiency cases.     

Genetic variability of 16 Y-chromosome STRs in a sample from Equatorial Guinea (Central Africa)

Nine Y-STR loci from the "minimal haplotype" included in Y-STR Haplotype Reference Databases (YHRD) together with eight additional Y-STRs (DYS437, DYS438, DYS439, DYS460, DYS461, GATA C4, GATA H4 and GATA A10) were analyzed in a sample of 101 males from Equatorial Guinea living in Madrid. Haplotype and allelic frequencies were calculated and genetic diversities were estimated for each genetic system as well as for the whole haplotype. An unexpected high frequency (6%) of intermediate alleles (13.2 and 14.2) was found in DYS385. For DYS19, two alleles were found in one sample. Another sample failed to amplify with DYS393 primers using either PowerPlex Y System (Promega Corporation) or the Y-PLEXtrade mark 12 (Reliagene, New Orleans, LA) commercial kits. Comparison between Equatorial Guinea and another African population (Mozambique; South East Coast) revealed a significant pairwise Phi(st) value between them (Phi(st)=0.03309; P=0.00000).     

Y-chromosomal STR haplotypes in three ethnic groups and one cosmopolitan population from Tunisia

The 11 Y-chromosomal short tandem repeats (STRs) included in the Promega Corporation PowerPlex Y System (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385, DYS437, DYS438 and DYS439) were typed in three ethnic groups ("Andalusians", Berber and Arab) and one cosmopolitan population (Tunis) from Tunisia, summing up 247 individuals, and 139 different haplotypes. Focusing the analysis on the seven Y-STRs of the YHRD Minimal Haplotype Core (DYS385 excepted), "Andalusians" showed no differences from the Cosmopolitan and the Arab samples previously published (our Arab sample presented an extremely low haplotype diversity), but were different from the Berbers. The Berbers from Tunisia were not different from those from Morocco.     

Mitochondrial DNA error prophylaxis: assessing the causes of errors in the GEP'02-03 proficiency testing trial

A qualitative and quantitative analytical method was developed and validated for the determination of 49 licit and illicit drugs in oral fluid. Small oral fluid samples, volume 1mL, were collected from volunteers using a modified Omni-Sal device and the analytes were extracted from an oral fluid/buffer mixture using a single Bond Elut Certify solid phase extraction cartridge. Liquid chromatography-tandem mass spectrometry (LC-MS-MS) and gas chromatography-repetitive full scan mass spectrometry (GC-MS) were used in parallel to analyze the extracts for the targeted drugs. Extracts were analyzed by GC-MS in their underivatized form and as their pentafluoropropionyl derivatives. Deuterated internal standards were used for quantification of drugs of abuse by LC-MS-MS to minimize matrix effects. Methadone-d(9) and tumoxetine were used as the internal standards for quantification of non-derivatized and derivatized analytes respectively by GC-MS. Linearity was demonstrated over the range 5-200 ng/mL and limits of detection were less than 4 ng/mL for each drug analyzed. The method demonstrated acceptable recoveries for most of the analytes and good intra- and inter-day precision. Acquisition of data by repetitive full scan GC-MS allows the addition of further analytes to the target menu.     

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.     

Risk factors for sexual victimization among male and female homeless and runaway youth

Risk factors associated with the likelihood of being sexually victimized by a stranger or friend/acquaintance since being on the street was examined among 372 homeless and runaway youth. Young people were interviewed on the streets and in shelters by outreach workers using a systematic sampling strategy. Youth who engaged in more high-risk behaviors were expected to be at greater risk for sexual victimization by both known and unknown assailants. Results indicated that for females, running from home for the first time at an earlier age was associated with sexual victimization by both a stranger and friend/acquaintance. However, engaging in deviant subsistence strategies, survival sex, and grooming predicted being sexually victimized by a friend/acquaintance. For males, survival sex and grooming predicted stranger sexual victimization, whereas sexual orientation was associated with sexual victimization by a friend/acquaintance. Overall, 35% of the sample had been sexually victimized.     

Suicide with a butcher's bolt

The captive bolt pistol is an atypical firearm exclusively produced and used for butchery of breeding animals, but in some rare cases, it has been used as a lethal weapon for committing suicide by butchers, breeders and other people who have access to such weapons during their professional activities. This study describes the suicide committed by a butcher in the province of Udine (N. Italy) in 2001 who shot himself with his own captive bolt pistol that produced in the right temporal region a circular wound and a bone lesion of the temporal squama with a groove involving the frontal lobe; he died five days later of the fatal consequences of the meningo-encephalic lesions. The medico-legal issues implicated in this case, seen in the light of the data reported in the international literature, illustrate the difficulties faced when diagnosing these types of lesions, bearing in mind their rarity and peculiar nature, and introduce elements of differential diagnosis regarding lesions produced by similar weapons that lead to ascertainment of the event as accidental, suicidal, or homicidal.     

Ritual homicide during dissociative trance disorder

A case report is described of a man who developed a single period of recurrent episodes of dissociative trance disorder (DTD) during a family stress. The cultural environment was permeated by thoughts and beliefs in magic. During one trance state, the man performed an exorcism on the 6-month-old baby of his brother-in-law, and the baby died from cerebral hemorrhage. In the 5-year follow-up, no other episodes of DTD developed, and the man is serving an 18-year sentence.     

The importance of a urine sample in persons intoxicated with flunitrazepam--legal issues in a forensic psychiatric case study of a serial murderer

The sedative-hypnotic benzodiazepine flunitrazepam (FZ) is abused worldwide. The purpose of our study was to investigate violence and anterograde amnesia following intoxication with FZ, and how this was legally evaluated in forensic psychiatric investigations with the objective of drawing some conclusions about the importance of urine sample in a case of a suspected intoxication with FZ. The case was a 23-year-old male university student who, intoxicated with FZ (and possibly with other substances such as diazepam, amphetamines or cannabis), first stabbed an acquaintance and, 2 years later, two friends to death. The police investigation files, including video-typed interviews, the forensic psychiatric files, and also results from the forensic autopsy of the victims, were compared with the information obtained from the case. Only partial recovery from anterograde amnesia was shown during a period of several months. Some important new information is contained in this case report: a forensic analysis of blood sample instead of a urine sample, might lead to confusion during police investigation and forensic psychiatric assessment (FPA) of an FZ abuser, and in consequence wrong legal decisions. FZ, alone or combined with other substances, induces severe violence and is followed by anterograde amnesia. All cases of bizarre, unexpected aggression followed by anterograde amnesia should be assessed for abuse of FZ. A urine sample is needed in case of suspected FZ intoxication. The police need to be more aware of these issues, and they must recognise that they play a crucial role in an assessment procedure. Declaring FZ an illegal drug is strongly recommended.