Heinz-Dieter Wehner

Delimitation of the time of death by immunohistochemical detection of insulin in pancreatic β-cells

To improve the possibilities to delimitate the time of death after longer laytime, it was examined if this is possible by immunohistochemical insulin detection. The results show that in our examination material, the pancreatic beta-cells of up to 12-day-old corpses produce a positive immunoreaction towards insulin in all cases, whereas none of the corpses older than 30 days show such a reaction. This means that in case of a negative immunoreaction, the time of death can be assumed to lie more than 12 days before the autopsy. The fact that a negative immunoreaction occurs consistently after 30 days leads to the conclusion that when insulin has been stained in a specimen, the death of the respective person must lie a maximum of 29 days earlier, whereby these time-limits may change in considerably different surrounding conditions.

The elimination kinetics of methanol and the influence of ethanol

SummaryFour male subjects aged between 20 and 29 years were given intravenous injections of methanol at a dosage of 10 mg per kg body weight, once without prior administration of ethanol, and once after oral ingestion of 0.3 g ethanol per kg body weight. The serum methanol concentration was monitored over the next 5 h (after methanol administration alone) and 6–7 h (after methanol administration following ethanol ingestion). The elimination of methanol administered alone was found to follow first-order kinetics with a rate constant for the elimination phase of 0.475-0.259 h−1, corresponding to an elimination half-life of 1.8-3.0h. When ethanol was also administered methanol oxidation was found to be completely blocked until the blood ethanol concentration had fallen to 0.2 g/kg. When the ethanol concentration had dropped to zero, methanol elimination followed exactly the same course as that observed in the experiment without prior administration of ethanol (k: 0.378-0.231 h−1;t/12: 1.5–2.7 h).ZusammenfassungVier männliche Probanden im Alter zwischen 20 Jahren und 29 Jahren wurden 10 mg Methanol pro Kilogramm Körpergewicht intravenös injiziert. Die Methanolapplikationen erfolgten einmal bei Ethanol-Nüchternheit, einmal nach vorangegangener oraler Zufuhr von 0,3 g Ethanol pro Kilogramm Körpergewicht. Der Verlauf der Serum-Methanolkonzentrationskurve wurde über 5 Stunden (alleinige Methanolzufuhr) bzw. 6 bis 7 Stunden (zusätzliche Ethanolzufuhr) beobachtet. Nach alleiniger Methanolapplikation folgte die Methanol-Elimination einer Funktion erster Ordnung mit terminalen Dispositionskonstanten zwischen 0,475h−1 und 0,259h−1, entsprechend Eliminations-Halbwertszeiten von 1.8 h bis 3,0 h. Nach gleichzeitiger Ethanolzufuhr wurde die Methanol-Oxydation bis herab zu Ethanolkonzentrationen von 0,2 g/kg vollständig blockiert. Nach Erreichen der Ethanol-Nüchternheit zeigte die Methanol-Elimination keine Unterschiede im Vergleich zu den Versuchen ohne gleichzeitige Ethanolbelastung (k: 0.378h−1 bis 0,231−1;t½: 1,5h bis 2,7h).

Delimitation of the time of death by immunohistochemical detection of thyroglobulin

To improve the possibilities to delimit the time of death after longer laytime it was examined if this is possible by immunohistochemical detection of thyroglobulin. The results show that in our examination material the colloid and the follicular cells of the thyroid glands of up to 5-day-old corpses produce a positive immunoreaction towards thyroglobulin in all cases whereas none of the corpses older than 13 days show such a reaction. This means that in case of a negative immunoreaction the time of death can be assumed to lie more than 6 days before the autopsy. The fact that a negative immunoreaction occurs consistently after 13 days leads to the conclusion that when thyroglobulin has been stained in a specimen, the death of the respective person must lie a maximum of 12 days earlier, whereby these time-limits may change in considerably different surrounding conditions.

Percentile charts to determine the duration of child abuse by chronic malnutrition

Longstanding quantitative or qualitative under-supply of nutrition leads to weight loss and, in children, to stagnation of growth and thus to stunted growth. A comparison of the expected growth, according to percentile growth curves, with the actual body size, gives an indication as to the period of time in which malnutrition took place. The moment in which the growth curve bends off and leaves the norm is to be interpreted as the earliest begin, the moment in which the attained growth would have been achieved as the latest begin of the nutritional impairment.

Delimitation of the time of death by immunohistochemical detection of calcitonin

To improve the possibilities of delimitating the time of death after longer laytime it was examined if this is possible by immunohistochemical detection of calcitonin. The results show that in our examination material the c-cells of the thyroid glands of up to 4-day-old corpses produce a positive immunoreaction towards calcitonin in all cases whereas none of the corpses older than 13 days show such a reaction. This means that in the case of a negative immunoreaction the time of death can be assumed to lie >4 days before the autopsy. The fact that a negative immunoreaction occurred consistently after 13 days leads to the conclusion that when calcitonin has been stained in a specimen, the death of the respective person must lie a maximum of 12 days earlier, whereby these time-limits may change in considerably different surrounding conditions.

Immunohistochemical detection of methadone in the human brain

To develop a method of detecting methadone in the human brain by immunohistochemistry, brain tissue of frontal cortex, cerebellum, hippocampus, basal ganglions and brain stem from victims of a lethal methadone overdose was examined. The staining was performed with a monoclonal anti-methadone antibody from the mouse, originally developed for immunochemichal purposes (ELISA). With the help of the DAKO((R)) Catalyzed Signal Amplification (CSA) System, a specific positive immunoreaction was obtained in the neurons of the frontal cortex and hippocampus, as compared with specimen from deaths without exposition to methadone. Thus, along with metamphetamine, phenobarbital, morphine and insulin, immunohistochemical detection is also possible for methadone and the intake of this medicament can now be proven morphologically.

Demonstration of morphine in ganglion cells of the hippocampus from victims of heroin overdose by means of anti-morphine antiserum

Abstract To investigate the topography of morphine distribution in the human brain, a method has been developed to detect morphine immunohistochemically. In this study hippocampus tissue from victims of heroin overdose (blood morphine concentrations 220 ng/g–1500 ng/g; 6-MAM positive urine sample), known for its high concentration of μ-opiate receptors was used. The immunohistochemical staining was performed with an anti-morphine antiserum originally developed for radio-immuno-assays. In comparison with control specimens from cases of sudden death without morphine exposition or a history of heroin abuse, the brains from victims of heroin overdose showed selectively stained ganglion cells, axons and dendrites, suggesting a massive concentration of morphine in the neuronal structures.

Insulin- or morphine-injection? Immunohistochemical contribution to the elucidation of a case

Two autopsy cases of an elderly couple who died on the same day will be used to underline the importance of immunohistochemistry of forensic practice. At first unexplainable injection marks on the upper arms of the corpses and the possibility of a closely related physician injecting insulin and certifying a natural death made it important, considering suspect insulin concentrations in the blood, to exclude insulin injections in these marks. Further, the statement that morphine had been injected for the analgesia of tumour pains, was reinforced by immunohistochemistry.

Immunocytochemical examinations of biological traces on expanding bullets (QD–PEP)

When a crime victim has been injured with several different objects, it is of central importance for the forensic investigation to be able to show which object caused which injury, especially if one of the injuries was lethal. In cases of bullet penetration wounds it is often not possible to find such evidence. However, immunocytochemical investigations can accurately match a victims injury to a particular bullet path through the body. In cases where expanding bullets have been used and the heart or liver has been struck by a projectile, it can be shown that the cells remaining on the bullet stem from those particular organs. In this case the specific cytological evidence was established by means of marking heart- and liver-specific tissue proteins with appropriate antibodies (cardiac troponin I and HepPar 1) followed by disclosure with an appropriate chromogen. Thus, in principle, cells can be used as evidence after being extracted from the projectiles by either damp cotton-wool swabs or adhesive trace evidence tape. Because of the specificity of the used immunocytochemical antibodies, finding evidence of an antigen on a particular projectile proves that it was the object that injured the organs.

Immunhistochemischer Morphinnachweis an der subkutanen Injektionsstelle

ZusammenfassungMorphin ist neben Insulin und Heparin eines der am häufigsten subkutan applizierten Medikamente. Finden sich im Rahmen einer gerichtsmedizinischen Obduktion subkutane Injektionsstellen, ist es von großem Wert die injizierte Substanz bestimmen zu können. Das vorliegende immunhistochemische Verfahren zeigt, dass eine morphinspezifische positive Immunreaktion des subkutanen Fettgewebes einer subkutanen Morphininjektion zuzuordnen ist und dass des Weiteren ein verbreiteter immunpositiver interstitieller Raum zwischen den subkutanen Lipozyten auf die Intravitalität der Morphininjektion hindeutet.AbstractMorphine apart of insulin and heparin is one of the most frequently subcutaneous applied medicaments. If subcutaneous injection sites are found within the framework of a forensic medical autopsy, it is of big value to be able to differentiate which substance was injected at this point. The present immunohistochemical study points, that the subcutaneous injection of morphine can be detected by the morphine-specific positive immunoreaction of the subcutaneous fatty tissue and that furthermore a widened intercellular space between the subcutaneous lipocytes points to the intravitality of the injection.