Pulmonary edema in fatal heroin overdose: immunohistological investigations with IgE, collagen IV and laminin - no increase of defects of alveolar-capillary membranes

Pulmonary edema complicating heroin overdosage is a well recognized entity and regarded as the major mechanism contributing to death in heroin addicts. It's pathogenesis is unknown, several mechanisms are discussed: hypoxia-induced increase of pulmonary capillary permeability, depressed myocardial contractility, centrally induced respiratory depression, primary toxic effects on the alveolar capillaries and acute anaphylactic shock. The present study included opiate-related deaths (n=23) and a control group of sudden cardiovascular deaths (n=12) to verify the hypothesis, that defects of the alveolar capillary membranes and/or an acute anaphylactic reaction leads to pulmonary congestion, edema and hemorrhages. Lung specimens were obtained from these 35 autopsies of persons autopsied in the Institute of Forensic Medicine, University of Bonn, in 1997 and 1998. All specimens were examined with hematoxylin-eosin, prussian blue and investigated with immunohistological methods using primary antibodies against collagen IV, laminin and IgE. Defects of the basal laminae of the alveoli were found, demonstrated by laminin and collagen IV, and the number of IgE-positive cells was counted in both groups. There was an increased but not significant number of IgE-positive cells in the heroin-group and defects of the epithelial and endothelial basal laminae were found in both groups without significant differences.     

Numerical density of delta-opioid receptor expressing neurons in the frontal cortex of drug-related fatalities

In animal experiment and in cell culture, chronic morphine treatment has been followed by a reduction as well as an increase of the delta-opioid receptor (OR) number. The present postmortem morphometric study of morphine-related fatalities of drug addicts (n=12, 22-35 years old, with blood unconjugated morphine levels from 27.1 to 407 ng/ml, m.v. 176.9 ng/ml) versus a non-addicted control group (n=13, 10-44 years old) is intended to examine whether chronic opiate exposure also affects the numerical density of deltaOR expressing neurons in the human neocortex (area 10 according to Brodmann (Vergleichende Lokalisationslehre der Grosshirnrinde (1909) Johann Ambrosius Barth, Leipzig)). For the immunohistochemical procedure, vibratome sections (100 microm) were incubated with a monoclonal antibody against the deltaOR diluted 1:100, and immunoreactive sites were visualized using an immunoperoxidase protocol. The numerical densities of OR expressing and Nissl-stained neurons were assessed morphometrically (camera lucida drawings). In both collectives, the anti deltaOR immunoreactivity was predominantly localized in pyramidal neurons of layers (L) II/III and V as well as in round and ovoid neurons of L VI. In the drug-related fatalities, the density of neurons expressing deltaOR protein amounted for 2515+/-240/mm(3), in the control group for 2616+/-204/mm(3), thus displaying no statistically significant difference. These findings go along with the binding behavior of opioid ligands in postmortem brains of heroin addicts revealing similar receptor densities and affinities in the control subjects and addicts.     

Duration of the fire and degree of charring of a burned cadaver]

A 37-year-old woman was found dead severely charred in the bedroom of her house. The autopsy revealed as cause of death a strangulation of the neck. On the basis of the decomposition of the body the experts were asked for a statement on the duration of burning. There seems to be only little medicolegal experience on the duration of burning to a defined state of decomposition. Based on observations during cremations published in the literature and experimental burnings a burning time of approximately 20 minutes (after temperatures of more than 700 degrees C had been reached) is supposed to be sufficient to produce the present state of the body.     

Forensic case profiling aspects on multiple homicides from the Cologne-Bonn metropolitan area 1985-2000

The medicolegal and subsequent criminologic interpretation of forensic and pathological findings in cases of homicide makes up an important tool of case profiling. In a retrospective study of 26 cases of "multiple homicides" involving 31 perpetrators (30 males, 1 female, mean age 33.5 years) and 73 victims (33 males, 40 females, mean age 36 years, 68 fatalities, 5 survivors), autopsy reports and prosecution authorities' files were investigated with regard to individual characteristics of victims and offenders, circumstances as well as mode of commitment. The major aim of this study was to comprehensively elucidate and characterise relevant forensic and criminologic features, which may gain importance for forensic case profiling. Forty-six victims were found in the close social environment of the perpetrator and 45 homicides were committed either in the victim's, the perpetrator's or the shared domicile. The main motives included concealment of a crime (n=13), personal conflicts/domestic arguments (n=7) and greed (n=12). The relevant injuries with regard to the cause of death were attributable to sharp force (n=13), blunt force (n=7), gunshot wounds (n=24), ligature strangulation (n=3), smothering (n=5), fire/carbon monoxide (n=4) and combined impacts (n=11). In 15 cases, so called defence injuries were found. In 5 victims a post-mortem blood alcohol concentration >1.5 g/l was determined. In six perpetrators, a severe psychiatric impairment of juridical responsibility was ascertained (Section 20 German criminal code, n=2, psychosis; Section 21 German criminal code, n=4, acute alcohol intoxication). As far as conviction data were available, 27 crimes were juridically assessed as murder, 12 as manslaughter and one as bodily harm with fatal consequences.     

Estimating the probability of identity in a random dog population using 15 highly polymorphic canine STR markers

Dog DNA-profiling is becoming an important supplementary technology for the investigation of accident and crime, as dogs are intensely integrated in human social life. We investigated 15 highly polymorphic canine STR markers and two sex-related markers of 131 randomly selected dogs from the area around Innsbruck, Tyrol, Austria, which were co-amplified in three PCR multiplex reactions (ZUBECA6, FH2132, FH2087Ua, ZUBECA4, WILMSTF, PEZ15, PEZ6, FH2611, FH2087Ub, FH2054, PEZ12, PEZ2, FH2010, FH2079 and VWF.X). Linkage testing for our set of marker suggested no evidence for linkage between the loci. Heterozygosity (HET), polymorphism information content (PIC) and the probability of identity (P((ID)theoretical), P((ID)unbiased), P((ID)sib)) were calculated for each marker. The HET((exp))-values of the 15 markers lie between 0.6 (VWF.X) and 0.9 (ZUBECA6), P((ID)sib)-values were found to range between 0.49 (VWF.X) and 0.28 (ZUBECA6). Moreover, the P((ID)sib) was computed for sets of loci by sequentially adding single loci to estimate the information content and the usefulness of the selected marker sets for the identification of dogs. The estimated P((ID)sib) value of all 15 markers amounted to 8.5 x 10(-8). The presented estimations turned out to be a helpful approach for a reasonable choice of markers for the individualisation of dogs.     

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).     

Injuries in fatal cases of falls downstairs

Downstairs falls frequently occur within domestic environments and are mainly associated with elderly and intoxicated individuals, often feature multiple injuries on various parts of the body. In most cases it is not possible to determine the cause of the fall and/or death solely by means of external examination. In this retrospective study, which covers a period of 11 years, all cases of death which included a fall downstairs in their case history, were collected from the Forensic Institutes of the Universities of Bonn and Greifswald, Germany. Falls downstairs made up to 2% (166 cases) of all postmortem examinations carried out within this period. Interestingly, almost double of the amount of such falls applied to males as to females. The primary cause of death was cranio-cerebral trauma and the vast majority of skull injuries associated with falls downstairs were found above 'the hat brim line'. Injuries were also often found on several other parts of the body at once. Nineteen of the 116 examined individuals exhibited agonal injuries. In these cases, postmortem examination revealed pre-existing disease or intoxication to be the cause of death and thus, cause of the fall. The injury pattern only allows a tendency towards vital or agonal incident as a conclusion.     

Area-specific increased density of mu-opioid receptor immunoreactive neurons in the cerebral cortex of drug-related fatalities

In animal experiments and in cell culture, chronic morphine treatment has been followed by "up-regulation" as well as "down-regulation" of the mu-opioid receptor (OR) number. The present postmortem morphometric study of morphine-related fatalities of drug-addicts (n=13, 20-35 years old, with blood unconjugated morphine levels from 27.1 ng/ml to 458 ng/ml, m.v. 198.5 ng/ml) versus a non-addicted control group (n=13, 10-44 years old) was intended to examine, whether chronic opiate exposure affects the numerical density of mu-OR expressing neurons in the human neocortex (areas 11, 24 and 25 according to Brodmann). For the immunohistochemical procedure, vibratome sections (100 microm) were incubated with a monoclonal antibody against the mu-OR, diluted 1:100, and immunolabelled sites were visualized using an immunoperoxidase protocol. The numerical densities of OR immunoreactive neuronal profiles and Nissl-stained central profiles were assessed morphometrically (camera lucida-drawings). In both groups, the anti-mu-OR-immunoreactivity was mainly localized in pyramidal neurons of layers (L) II/III and V and in multiform neurons of L VI. In the areas 24 and 25, the density of the immunoreactive neuronal profiles did not display a significant difference between the two examined groups. In the area 11, however, the number of immunolabelled neuronal profiles amounted to 2777+/-206 mm(3) in the drug-related fatalities and to 2320+/-124 mm(3) in the control group and thus was significantly increased.     

Automated headspace solid-phase dynamic extraction for the determination of cannabinoids in hair samples

This article describes a fully automated procedure for detecting cannabinoids in human hair samples. The procedure uses alkaline hydrolysis and headspace solid-phase dynamic extraction (HS-SPDE), followed by on-coating derivatization and gas chromatography-mass spectrometry (GC-MS). SPDE is a further development of solid-phase microextraction (SPME), based on an inside needle capillary absorption trap. It uses a hollow needle with an internal coating of polydimethylsiloxane as extraction and pre-concentration medium.Ten mg of hair were washed with deionised water, petroleum ether and dichloromethane. After adding deuterated internal standards, the sample was hydrolyzed with sodium hydroxide and directly submitted to HS-SPDE. After absorption of analytes for an on-coating derivatization procedure, the SPDE-needle was directly placed into the headspace of a second vial containing N-methyl-N-trimethylsilyl-trifluoroacetamide before GC-MS analysis. The limit of detection was 0.14 ng/mg for Delta(9)-tetrahydrocannabinol, 0.09 ng/mg for cannabidiol, and 0.12ng/mg for cannabinol. Absolute recoveries were in the range of 0.6 to 8.4%. Linearity was verified over a range from 0.2 to 20 ng/mg, with coefficients of correlation between 0.998 and 0.999. Intra- and inter-day precision were determined at two different concentrations and resulted in ranges between 2.3 and 6.0% (intra-day) and 3.3 and 7.6% (inter-day). Compared with conventional methods of hair analysis, this automated HS-SPDE-GC-MS procedure is substantially faster. It is easy to perform without using solvents and with minimal sample quantities, and it yields the same sensitivity and reproducibility. Compared to SPME, we found a higher extraction rate, coupled with a faster automated operation and greater stability of the device.