The identification of a victim using the DGGE method for trace deposits collected on adhesive film

The denaturant gradient gel electrophoresis (DGGE) method was used in order to simultaneously estimate the genotypes of different factors in a gel plate consisting of one sheet. A genotype analysis of the blood groups (MN, Duffy, Kidd type) and serotype (Gc system) was carried out. DNA samples were extracted from trace deposits which were transferred on adhesive film from a blood trace obtained from a car tire after a fatal car accident. The reference DNA was prepared from the victim's blood. The PCR amplification fragments were amplified from the gene which controlled each blood group. The primers were designed in order to analyze the genotypes with one to three base substitutions in the amplification product. The denaturant concentration limit of the gel for the DGGE method to detect each genotype of the blood groups (MN, Duffy, Kidd type and Gc system) and other conditions of electrophoresis were performed according to previously methods.The each genotype of the blood groups and the Gc system were all simultaneously distinguished in one plate.     

Glucose phosphate isomerase variants in human bloodstains and seminal stains

Glucose phosphate isomerase (GPI) variants occurring in human red cells were also demonstrated in human semen. Phenotyping was possible from bloodstains of 6 weeks storage and seminal stains of 12 weeks storage. The GPI system may be a supplemental tool for medicolegal individualization of seminal stains.     

Chromophoric labeling of cannabinoids with 4-dimethylaminoazobenzene-4'-sulfonyl chloride

A simple and sensitive assay for the cannabinoids is presented using a dabsylation procedure. Dabsyl derivatives of delta 9-tetrahydrocannabinol (delta 9-THC) and cannabinol (CBN) were prepared by reacting with 4-dimethylaminoazobenzene-4'-sulfonyl chloride (dabsyl chloride) in acetone in the presence of sodium carbonate-sodium bicarbonate buffer (pH 10). Crystalline dabsylcannabinoids gave intense absorption in the visible region. With these derivatives, analysis by thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC) were tested. These techniques gave good separation and nanogram detection of dabsyl-THC and -CBN by using n-hexane-ethyl acetate-diethylamine (20:5:1) for TLC and MeOH--H2O (95:5) at 450 nm for HPLC.     

Components of paint thinner in body fluids clearly detected using the salting-out technique

For a more sensitive detection of paint thinner components in body fluids, we made use of a salting-out technique, with sodium chloride added to blood samples followed by gas chromatography, using the headspace method. The detection of ethyl acetate and isobutanol was considerably enhanced using these approaches.     

A very slow BF variant (S085) detected in a Japanese population

Using high-voltage agarose gel electrophoresis and immunofixation a very slow BF variant was detected in a Japanese person living in Yamanashi district. The family analysis suggested the hereditary occurrence of a new allele, BF*S085.     

A Case of Sudden Infant Death Due to Incomplete Kawasaki Disease

Although Kawasaki disease (KD) is a self‐limiting disease, it may cause sudden cardiac death. Diagnosis of KD is principally based on clinical signs; however, some infant cases do not meet the criteria. Such cases are identified as incomplete KD. The sudden death risk in incomplete KD cases is similar to conventional KD. In our 5‐month‐old case, he had been admitted to a hospital for a fever and suppuration at the site of Bacille de Calmette et Guerin (BCG) vaccination. However, after discharge from the hospital, his C‐reactive protein (CRP) levels declined, he got indisposed and died suddenly. A medico‐legal autopsy revealed myocarditis, coronaritis, platelet‐aggregated emboli in coronary arteries, and myocardial degeneration, suggesting that the fatal myocardial infarction was due to thrombus emboli in the coronary arteries. Forensic pathologists therefore should pay attention to the cardiac pathology originated from incomplete KD as a potential cause in cases of sudden infant death.     

Accuracy and Usefulness of the AVOXimeter 4000 as Routine Analysis of Carboxyhemoglobin

The measurement of blood carboxyhemoglobin (CO‐Hb) is important to determine the cause of death. The AVOXimeter 4000 (AVOX), a portable CO‐oximeter, has the advantages of a low purchase price and operating cost, ease of operation, and rapid results. Little information is available on the usefulness of AVOX in the forensic sample, and the previous study investigated only six samples. Therefore, in this study, we confirmed the usefulness of the AVOX through a comparison of its re s ults with data previously obtained using the double wavelength spectrophotometric method in autopsies. Regression analysis was performed between CO‐Hb levels measured by the AVOX and those measured by the conventional double wavelength spectrophotometric method in postmortem blood samples: a significant correlation was observed. This study suggests the usefulness of the AVOX to analyze postmortem blood, and the AVOX is suitable for routine forensic analysis and can be applied at the crime scene.     

A New Method for Human ABO Genotyping Using a Universal Reporter Primer System

Abstract: We developed a new method for forensic ABO genotyping based on a universal reporter primer (URP) system. This allows for the simultaneous detection of six single nucleotide polymorphism (SNP) sites in the ABO gene (nucleotide positions 261, 297, 526, 703, 796, and 803). This URP system provides obvious peaks, ranging from 82 to 151 bp in length. ABO genotypes were classified and successfully genotyped by our method, including minor alleles that may cause a discrepancy between the genetic data and serological phenotypes. Full profiles were identified using as little as 0.1 ng (0.05 ng/reaction) of standard K562 and 9947A DNA. Moreover, the success rate of genotyping from a URP system was much higher than that from a conventional primer extension method in degraded DNA. This method enables simple and rapid detection of multiple SNP sites on human ABO genes and is highly specific and sensitive when using limited and degraded DNA.