Segmental analysis for cocaine and metabolites by HPLC in hair of suspected drug overdose cases

Hair samples of eight postmortem cases were analyzed in segments of 1 to 3 cm for cocaine, benzoylecgonine and cocaethylene. Samples were prepared for analysis by digestion in 0.1 M HCl and subsequent extraction with mixed-mode solid-phase extraction columns. Measurement was made by reversed-phase, narrow-bore HPLC and fluorescence detection using two laboratory-made internal standards. The concentrations were in the region of 0.29-316 ng/mg of hair for cocaine, 0.43-141 ng/mg of hair for benzoylecgonine and 0.93-1.83 ng/mg of hair for cocaethylene. All eight investigated cases had cocaine-positive segments. In six of the cases, all segments were positive, suggesting regular cocaine use and two showed in-between negative segments indicating an interruption or a change of the abuse intensity. The results showed a second, remarkable observation, i.e. enormous concentration differences (factor >150) for both cocaine and benzoylecgonine between the different subjects. Furthermore, interindividual cocaine/benzoylecgonine ratios ranged from 0.02 to 8.43. We believe these observations could in part be attributed to both some of the still existing limitations in the analytical approach(es), especially the mandatory hair washing steps, and in our still too limited knowledge of the hair incorporation processes. Nevertheless, in some cases, segmental analysis proved to be an important tool to distinguish, together with postmortem examination, deadly chronic abuse from single acute drug overdosage.     

Three-dimensional cranio-facial reconstruction in forensic identification: latest progress and new tendencies in the 21st century

Three-dimensional (3D) cranio-facial reconstruction can be useful in the identification of an unknown body. The progress in computer science and the improvement of medical imaging technologies during recent years had significant repercussions on this domain. New facial soft tissue depth data for children and adults have been obtained using ultrasound, CT-scans and radiographies. New guidelines for facial feature properties such as nose projection, eye protrusion or mouth width, have been suggested, but also older theories and "rules of thumbs" have been critically evaluated based on digital technology. New fast, flexible and objective 3D reconstruction computer programs are in full development. The research on craniofacial reconstruction since the beginning of the 21st century is presented, highlighting computer-aided 3D facial reconstruction. Employing the newer technologies and permanently evaluating and (re)questioning the obtained results will hopefully lead to more accurate reconstructions.     

Observations on dental structures when placed in contact with acids: experimental studies to aid identification processes

In a "mafia" crime case, a magistrate asked us whether it is possible to destroy a cadaver by immersing it in acids, and would it be possible to identify any residues. The aim of this study was to observe the behavior of teeth exposed to four kinds of acid solutions. The teeth were placed in plastic containers with 25 mL of acid and observed. The experiences showed that teeth are completely dissolved after 14 h of immersion in 37% solution of hydrochloric acid, while at 90h in 96% sulfuric acid, the destruction of the samples is still incomplete. In nitric acid the teeth undergo a complete dissolution in 12 h, and in 17 h in aqua regia (chloroazotic acid-hydrochloric/nitric acid 1:3). It was possible to recognize the characteristic morphological features of dental tissues and structures up until the advanced stages of degradation.     

Semi-automated ultrasound facial soft tissue depth registration: method and validation

A mobile and fast, semi-automatic ultrasound (US) system was developed for facial soft tissue depth registration. The system consists of an A-Scan ultrasound device connected to a portable PC with interfacing and controlling software. For 52 cephalometric landmarks, the system was tested for repeatability and accuracy by evaluating intra-observer agreement and comparing ultrasound and CT-scan results on 12 subjects planned for craniofacial surgery, respectively. A paired t-test evaluating repeatability of the ultrasound measurements showed 5.7% (n = 3) of the landmarks being significantly different (p < 0.01). US and CT-scan results showed significant differences (p < 0.01) using a Wilcoxon signed rank test analysis for 11.5% (n = 6) of the landmarks. This is attributed to a difference in the volunteer's head position between lying (CT) and sitting (US). Based on these tests, we conclude that the proposed registration system and measurement protocol allows relatively fast (52 landmarks/20 min), non-invasive, repeatable and accurate acquisition of facial soft tissue depth measurements.     

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.     

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.     

Differences in trauma symptoms and family functioning in intra-and extrafamilial sexually abused adolescents

This study investigated to what extent abuse-related symptoms and family functioning are related to intra- or extrafamilial sexual abuse. One hundred adolescents (12 to 18 years old) were recruited shortly after disclosure of the abuse. Information from the participants was obtained through self-report questionnaires and a semistructured interview. Fifty-three percent of the adolescents reported clinically significant symptoms. Data did not support the idea that intrafamilial sexually abused adolescents report more symptoms than extrafamilial sexually abused adolescents. Type of abuse did not account for the differences and variety of reported symptoms or for differences in family functioning. Family functioning-in particular, lack of cohesion- was an independent contributor to internalizing trauma-related problems.