Markus A. Rothschild

In vitro phase I metabolism of the synthetic cannabimimetic JWH-018

A potent synthetic cannabinoid receptor agonist, JHW-018, was recently detected as one of the most prominent active agents in abusively used incenses such as Spice and other herbal blends. The high pharmacological and addictive potency of JWH-018 highlights the importance of elucidating the metabolism of JWH-018, without which a meaningful insight into its pharmacokinetics and its toxicity would not be possible. In the present study, the cytochrome P450 phase I metabolites of JWH-018 were investigated, after in vitro incubation of the drug with human liver microsomes, followed by liquid chromatography–tandem mass spectrometry analysis. This revealed monohydroxylation of the naphthalene ring system, the indole moiety, and the alkyl side chain. In addition, observations were made of dihydroxylation of the naphthalene ring system, and the indole moiety, or as result of a combination of monohydroxylations of both the naphthalene ring system and the indole moiety or the alkyl side chain, or a combination of monohydroxylations of both the indole ring system and the alkyl side chain. There is also evidence of trihydroxylation at different locations of the hydroxyl groups in the molecule. Furthermore, dehydration of the alkyl side chain, in combination with both monohydroxylation and dihydroxylation as well as arene oxidation of the naphthalene ring system, combined with both monohydroxylation and dihydroxylation at different sites of oxidation were found. N-dealkylation also in combination with both monohydroxylation and dihydrodiol formation of the N-dealkylated metabolite was detected. Finally, a metabolite was found carboxylated at the alkyl side chain.

Complex suicides by self-incineration

Among the few cases of suicidal self-incineration primary or secondary complex suicides are a rarity. Accordingly the number of reports on this subject in the forensic literature is small. In a retrospective analysis of the Freiburg and Berlin autopsy material, four cases were found within an observation period of 11 years. These are described together with seven other cases reported in the literature. Altogether most of these suicides are primary combinations, especially with falls from a height. Other combinations include hanging or the infliction of stabs, cuts or gunshot injuries. The sex and age distribution is similar to that found for suicide by self-incineration alone.

Blood stain pattern analysis

Bloodstain pattern analysis (BPA) refers to the collection, categorization and interpretation of the shape and distribution of bloodstains connected with a crime. These kinds of stains occur in a considerable proportion of homicide cases. They offer extensive information and are an important part of a functional, medically and scientifically based reconstruction of a crime. The following groups of patterns can essentially be distinguished: dripped and splashed blood, projected blood, impact patterns, cast-off stains, expirated and transferred bloodstains. A highly qualified analysis can help to estimate facts concerning the location, quality and intensity of an external force. A sequence of events may be recognized, and detailed questions connected with the reconstruction of the crime might be answered. In some cases, BPA helps to distinguish between accident, homicide and suicide or to identify bloodstains originating from a perpetrator. BPA is based on systematic training, a visit to the crime scene or alternatively good photographic documentation, and an understanding and knowledge of autopsy findings or statements made by the perpetrator and/or victim. A BPA working group has been established within the German Society of Legal Medicine aiming to put the knowledge and practical applications of this subdiscipline of forensic science on a wider basis.

Expression of heat-shock protein 70 (Hsp70) in the respiratory tract and lungs of fire victims

Immunohistochemical investigation of the respiratory tract and lungs of 63 fire victims revealed a statistically significant enhanced expression of heat-shock protein 70 (Hsp70) in the epiglottis, the trachea, and the main and the peripheral bronchi compared with a control group. In the fire victims, a strong expression of Hsp70 was discernible not only particularly in the vessels but also in seromucous secretory cells, ciliated epithelial cells, smooth muscle cells, and alveolar cells. The results suggest a vital or supravital reaction due to the inhalation of hot fire fumes.

Genetic analysis of sudden unexplained death: A multidisciplinary approach

Each year infants, children and young adults die suddenly and unexpectedly. In many cases the cause of death can be elucidated by medico-legal autopsy, however, a significant number of these cases remain unexplained despite a detailed postmortem investigation and are labeled as sudden unexplained death (SUD). Post-mortem genetic testing, so called molecular autopsy, revealed that primary arrhythmogenic disorders including long QT syndrome and catecholaminergic polymorphic ventricular tachycardia (CPVT) may account for a certain number of these cases. Because of the inheritance of these diseases, close relatives of the deceased may also at potential risk of carrying fatal cardiac disorders. Therefore, advanced diagnostic analyses, genetic counseling and interdisciplinary collaboration should be integral parts of clinical and forensic practice. In the present study, we performed mutation analyses of the major genes causing cardiac channelopathies in 15 SUD cases. In four cases we found putative pathogenic mutations in cardiac ion channel genes. Clinical and genetic examination of family members of SUD victims was also performed and affected family members were identified. This study demonstrates that molecular genetic screening needs to become an inherent part of the postmortem examination. This will enhance the ability of screening family members of SUD victims who may be at risk. The present data also illustrate that detection and follow up of familial cases of sudden death is challenging and requires a close multidisciplinary collaboration between different medical disciplines, with great responsibility for the forensic pathologist.

How does spinal canal decompression and dorsal stabilization affect segmental mobility? A biomechanical study

IntroductionWhen decompression of the lumbar spinal canal is performed, segmental stability might be affected. Exactly which anatomical structures can thereby be resected without interfering with stability, and when, respectively how, additional stabilization is essential, has not been adequately investigated so far. The present investigation describes kinetic changes in a surgically treated motion segment as well as in its adjacent segments.Material and methodsSegmental biomechanical examination of nine human lumbar cadaver spines (L1 to L5) was performed without preload in a spine-testing apparatus by means of a precise, ultrasound-guided measuring system. Thus, samples consisting of four free motion segments were made available. Besides measurements in the native (untreated) spine specimen further measurements were done after progressive resection of dorsal elements like lig. flavum, hemilaminectomy, laminectomy and facetectomy. The segment was then stabilised by means of a rigid system (ART®) and by means of a dynamic, transpedicularly fixed system (Dynesys®).ResultsFor the analysis, range of motion (ROM) values and separately viewed data of the respective direction of motion were considered in equal measure. A very high reproducibility of the individual measurements could be verified. In the sagittal and frontal plane, flavectomy and hemilaminectomy did not achieve any relevant change in the ROM in both directions. This applies to the segment operated on as well as to the adjacent segments examined. Resection of the facet likewise does not lead to any distinct increase of mobility in the operated segment as far as flexion and right/left bending is concerned. In extension a striking increase in mobility of more than 1° compared to the native value can be perceived in the operated segment. Stabilization with the rigid and dynamic system effect an almost equal reduction of flexion/extension and right/left bending. In the adjacent segments, a slightly higher mobility is to be noted for rigid stabilization than for dynamic stabilisation. A linear regression analysis shows that in flexion/extension monosegmental rigid stabilisation is compensated predominantly in the first cranial adjacent segment. In case of a dynamic stabilisation the compensation is distributed among the first and second cranial, and by 20% in the caudal adjacent segment.SummaryMonosegmental decompression of the lumbar spinal canal does not essentially destabilise the motion segment during in vitro conditions. Regarding rigid or dynamic stabilisation, the ROM does not differ within the operated segment, but the distribution of the compensatory movement is different.

A 21 marker insertion deletion polymorphism panel to study biogeographic ancestry

Insertion/deletion polymorphisms have recently received increased interest in the forensic genetics community. This class of markers combines the advantageous genetic properties of single nucleotide polymorphisms (i.e., low mutation rate, genetic stability, and short amplicon size) with the technical advantage of short tandem repeat markers (simple detection by fluorescence-labelled PCR and capillary electrophoresis). For a large number of indel markers significant differences in allele frequencies between the major populations have been reported, making this class of markers suitable for the analysis of biogeographic ancestry. We have developed a multiplex PCR assay designed to establish the biogeographic ancestry of forensic DNA samples based on insertion/deletion polymorphisms. A panel of 21 short indels with allele frequency differences between three major population groups (European, African and Asian) was selected to be incorporated into a single-tube multiplex PCR assay. The assay is highly sensitive, requiring less than 0.5 ng of genomic DNA for successful typing. Due to the short fragment lengths below 200 bp, the assay is ideally suited for the typing of challenging forensic genetic case work samples. A population genetic study has been performed proving the performance of the assay in inferring the ancestral population of individuals. The chosen 21 markers are sufficient to distinguish between three major global population groups. Furthermore, the assay design leaves room for an extension in order to cover additional population groups.

International standards in cases of mass disaster victim identification (DVI)

Mass disasters with a large number of unknown victims are among the biggest challenges for the police and forensic disciplines. Historical events illustrate the development of different methods which can be used in such circumstances. The fire in the Ring-Theater of Vienna (Austria) in 1881 with 449 victims of which 284 were subsequently identified [1], or the fire in the Bazar de la Charite in Paris (France) with 126 victims in 1897 [2] are examples of the usefulness of identification procedures such as forensic autopsy, odontology, and finger printing. These two disasters from the nineteenth century were the beginning of modern identification processes in legal medicine [2]. In the following years several major accidents, especially after the introduction of regular civil air transportation services, demanded an effective diaster victim identification (DVI) system. Even today the identification of victims of mass disasters is still one of the most important tasks. Forensic experts are involved in different ways depending on the country of residence, and include forensic pathologists, forensic dentists, forensic anthropologists, forensic molecular biologists and other specialists. The organization of the identification process differs from country to country, and within the countries, depending on the historical and political structures and on the kind of disaster. The success of the identification essentially relies on the organization, the experience, and the documentation of findings. The communication between countries or states can be difficult, particularly in cases where victims have different nationalities. Therefore, the solution to such problems should be a standardized process. The recent natural disaster in Japan, triggered by a massive earth quake, also illustrates the need for standards in cases of radioactive contamination. Nobody has experiences in cases of simultaneous chemical, biological, or radioactive/nuclear (CBRNE) situations and assaults with a large number of contaminated bodies.

Immunohistochemical expression of fibronectin in the lungs of fire victims proves intravital reaction in fatal burns

Immunohistochemical studies about the presence of fibronectin in the lungs were performed in a group of 73 fire victims (63 cases of intravital and 10 cases of postmortem burn) as well as in an unselected control group of 55 individuals not exposed to fire before death. The cases of intravital burn showed a significantly stronger fibronectin expression than the control cases and the cases of postmortem burn. Fibronectin was mainly present in macrophages of the peribronchial lung parenchyma and, not associated with cells, in the matrix of peribronchial tissue. Our findings suggest that higher levels of fibronectin expression in the lung tissue of burn victims compared to fire-unrelated deaths may serve as an indicator of an early intravital inflammatory response to fire damage.

Examination of postmortem animal interference to human remains using cross-species multiplex PCR

Postmortem animal interference may be confused at first sight with injuries of vital origin, thus arousing suspicion of external violence preceding death. A reliable classification of the origin of such doubtful injuries is of crucial importance, a fact that is especially true for the investigation of suspected homicide and/or mammade body mutilation after death. In forensic pathology, the identification of injuries as caused by animals postmortem and the classification of a particular species as responsible for a specific injury pattern under question is usually done by forensic pathologists with vast practical experience and special knowledge of the appearance and morphology of tooth marks of carnivores and rodents, respectively. However, a molecular biological investigation of such wounds could provide genetic evidence that an injury pattern present on a corpse was truly caused postmortem by animal interference and thus support the pathologists expertise. For this purpose, we developed a panel of small species-specific short-tandem repeat systems (<150 bp) for animals typically involved in postmortem scavenging of human remains, such as dogs and cats as well as wild-living rodents (mice and rats) having possible access to death scenes inside apartments or buildings. A specific and sensitive cross-species multiplex polymerase chain reaction was then established including the species-specific animal markers, thus enabling the genetic identification of wounds caused postmortem by different animals on human remains.