A sandwich enzyme-linked immunosorbent assay for ABO blood typing of semen by using anti-p 84 monoclonal antibody as a marker of blood group substance in semen

A blood group substance (BGS), a protein with ABH antigenic activity, was isolated from human seminal plasma and designated as p 84 (Sato, 1995). We have developed a method for determining the ABO blood type of semen by performing a sandwich enzyme-linked immunosorbent assay (ELISA) in which p 84 is captured with an anti-p 84 monoclonal antibody, and evaluated the specificity and sensitivity of this method. Although BGS activity was detected in semen sensitively by this method, it was not detected in saliva, urine, breast milk, blood or vaginal secretions. Since the concentration of p 84 in semen was independent of the secretion status, the status can be determined as non-secretor when p 84 but not BGS activity was detected. To determine the stability of BGS activity on p 84, dried stains of semen on filter paper were kept at 4, 26, and 37 degrees C for 8 months, 2 years and 1 month, respectively, and their BGS activities were examined. After 8 months at 4 degrees C, over 60% of the original BGS activity was recovered from the stain. The activity could be detected even from a square as small as 0.25 by 0.25 cm. After 1 month at 37 degrees C and 2 years at 26 degrees C, 31 and 20% of the BGS activity, respectively, still remained. It could be detected from the pieces of 1.0 by 1.0 cm and 0.5 by 0.5 cm squares, kept for 1 month at 37 degrees C and 2 years at 26 degrees C, respectively. Finally, semen was mixed with saliva or blood at varying volumetric ratios and used for the sources of dried stains. The BGS activity of p 84 could be detected in the stains until the ratio between semen and saliva or blood reached 1:4. We conclude that this sandwich ELISA offers a more sensitive and specific method for determining the ABO blood type of semen samples obtained from sexual assault victims than existing methods, such as the conventional absorption-elution and classical hemagglutination-inhibition tests.     

Population data on the X chromosome short tandem repeat locus HumHPRTB in two regions of Germany

This report contains the results of two population studies on the X chromosome STR HumHPRTB carried out in a Northern and a Southern region of Germany. The numbers of unrelated individuals were 443 and 335, respectively. Eight alleles (alleles 9 to 16) were found. In female individuals 29 different genotypes were encountered. In German populations the HumHPRTB STR was characterized by the following data: PIC = 0.750; HET = 0.769: MEC = 0.556. Allele distribution met the Hardy-Weinberg expectations. The Northern and Southern populations did not show any significant differences.     

Fatal aspiration of sardine fry in a patient with lung cancer

We report a fatal case of death due to unusual aspiration of sardine fry in an elderly Japanese man with lung cancer. The cause of death was sudden respiratory arrest while eating. Autopsy revealed peculiar materials with cell nests and pigmented particles, together with striated muscle and skin, in the ectatic bronchioles of the left lower lobe. Serial histologic sections suggested that the structures observed were the eyeballs of small animals that appeared to have been inhaled. The patient had habitually eaten sardine fry and rice gruel, which were also detected in the gastric contents. Therefore, the eyes were considered to be those of the fry, which is a popular food item in Japan. This was confirmed by histologic examination of fry that were obtained commercially.     

A study on the use of photopolymer reflex plate in the forgery of Chinese seals

This study compares the identifying characteristics between the impressions made with original Chinese seals and photopolymer replicate seals, and between the impressions made with two photopolymer replicate seals that originate from one single negative. Various seal materials common to the Chinese community such as cow's horns and ivory are employed. Other than the common factors such as different incident angle upon imprinting or different degree of inking that constitute the natural variation in impressions, the physical changes of the seal materials with temperature will be considered. This study concludes that photopolymer replicate seal impressions can be differentiated from the originals in respect to print density and stroke morphology, but it is very difficult to differentiate between two photopolymer replicate seal impressions with the same origin. This study also proposes a simple method for the examination of forged Chinese seal impressions.     

A density gradient technique for use in forensic soil analysis

A high-density aqueous salt solution for the preparation of density gradients is presented. It has been used successfully by the authors in forensic soil analysis. It has a density range that allows for the separation of a soil specimen's heavy mineral components. It has no odor or toxic fumes, which eliminates the need to use a hood during preparation, and is far superior to the organic liquids normally used to prepare density gradients. This liquid should cause many forensic scientists to reexamine their attitudes towards using density gradients in forensic soil casework.     

Spectroscopic determination of skin viability. A predictor of postmortem interval

We have demonstrated that skin viability decreases at a measurable rate following death in an animal model. The decreased skin viability was measured by fluorescein diacetate and ethidium bromide using fluorescence emission spectroscopy. There is significant decrease of the fluorescence intensity of the fluorescein diacetate assay between the 1-4 h, the 6-24 h, and the >40 h time points postmortem. For times between 6-24 h and >40 h postmortem the ethidium bromide assay showed consistent and significant increases in signal. The fluorescence measurements in this study showed that under the experimental conditions the time of death could be determined for <4, 6-24, and >40 hapotmotrem. The application of these assays in the field will require further study of the environmental factors.     

Unexpected childhood death due to hemolytic uremic syndrome

Two cases of unexpected childhood death due to hemolytic uremic syndrome are reported. A 21-month-old girl who was discovered dead in bed following a short illness was found at autopsy to have overwhelming sepsis resulting from transmural colitis. Escherichia coli serotype 0157A was isolated from the intestine, and renal changes of hemolytic uremic syndrome were found. A 4-year-old girl died suddenly in hospital from intracranial hemorrhage while being treated for hemolytic uremic syndrome-related renal failure. Culture of urine and feces grew verocytotoxin producing E. coli. These cases demonstrate that hemolytic uremic syndrome may be a rare cause of unexpected childhood death and that the diagnosis may not be established prior to autopsy. Postmortem culture of tissues and fluids in cases of suspected sepsis in children may be essential in establishing this diagnosis, because histologic evaluation may be compromised by profound sepsis and tissue putrefaction. Accuracy in diagnosis may have significant public health and medicolegal consequences.     

The Origin and Outcome of an Amicus Brief in Kumho Tire v. Carmichael (1999)

Law and Human Behavior -     

Discrepancy between CYP2D6 phenotype and genotype derived from post-mortem dextromethorphan blood level

OBJECTIVE: To describe the death of a toddler after a therapeutic dose of dextromethorphan and its investigation. STUDY DESIGN: Case report, cytochrome P450 phenotype and genotype determination in the victim and post-mortem drug redistribution study performed in rats. RESULTS: A 20-month Asian male who received 3 mg of dextromethorphan once at 09:00 h and again at 22:00 h was found dead at 04:35 h. Post-mortem examination showed signs of early bronchopneumonia (bacterial cultures were negative); dextromethorphan and dextrorphan blood concentrations taken from the heart cavity were 500 ng/ml (1. 84 micromol/l) and 200 ng/ml (0.78 micromol/l), respectively. Despite the dextromethorphan level being almost 100-fold higher than expected after therapeutic doses, intentional or unintentional overdose was extremely unlikely; other potential causes were investigated. Post-mortem drug redistribution study performed in rats showed that dextromethorphan does not undergo extensive redistribution after death (6+/-5-fold increase) and could not explain the observed dextromethorphan level. The dextromethorphan/dextrorphan concentration ratio of 2.5 found in this toddler was compatible with a slow CYP2D6 metabolizer phenotype. However, the toddler exhibited a fast metabolizer genotype. Potential reasons for this discrepancy are discussed. CONCLUSION: CYP450 phenotypes derived from post-mortem blood levels should be interpreted with caution and preferably confirmed by a genotype analysis.