Examination of seminal stain by HPLC assay of phenolphthalein.

Quantitative high performance liquid chromatography (HPLC) to detect semen was investigated in this study. Briefly, 1cm of a gauze thread with a seminal stain was soaked in the reaction mixture (phenolphthalein diphosphate tetrasodium dissolved in acetate buffer) for 5-10 min, and the supernatant was analyzed by HPLC with a spectrophotometric detector. Phenolphthalein was liberated from the reagent in the presence of acid phosphatase, and the liberated phenolphthalein was detected objectively and was unaffected by blood contamination.

Identification of animal species using the partial sequences in the mitochondrial 16S rRNA gene.

This study investigated a PCR direct sequencing method for species identification by analyzing partial sequences in the mitochondrial 16S rRNA genes of many animal species amplified with universal primers. Samples from 182 vertebrates and 103 invertebrates were analyzed, and the sequences could be obtained in 182 and 72 species, respectively. The sequence divergence was sufficient to identify the species at the level of genus with the aid of the GenBank database and the BLAST tool.

The proof of flat-line scalp EEGs of brain dead patients by an automatic EEG analysis system.

Scalp electroencephalograms (EEGs) of brain dead patients are macroscopically flat under 7 or 10 microV/mm electroencephalograph sensitivity, but significant noises are detected in EEGs under 2 microV/mm sensitivity, interfering with the analysis. EEGs of 20 brain dead patients (17-76 years old) were therefore analyzed quantitatively as equivalent electric potentials in frequency bands delta, theta, alpha and beta using the automatic EEG analysis system developed in Kansai Medical University. The equivalent electric potentials in each band were about or less than 1 microV, which is used as a criterion of judgment of flat-line EEGs or brain death.

Detection of single nucleotide polymorphism in the FUT2 gene by temperature gradient gel electrophoresis.

A simple and rapid analysis system for single nucleotide polymorphisms (SNPs) was investigated for the FUT2 gene using the temperature gradient gel electrophoresis (TGGE) method. The 426-bp or 259-bp FUT2 fragments were amplified from heterozygous samples using primers, and the heteroduplex and homoduplex bands were detected by TGGE. The FUT2 fragments amplified from homozygous samples were denatured and re-annealed with a known sequence fragment, forming heteroduplex bands which were analyzed by TGGE.

Simple colorimetric semiquantitation method of hippuric acid in urine for demonstration of toluene abuse.

A semiquantitative method for hippuric acid in toluene sniffers' urine was explored by modifying the method by Tomokuni and Ogata. The color of sample urine mixed with pyridine and benzenesulfonyl chloride was yellow, and became reddish by addition of distilled water. Using the color chart, the concentration of hippuric acid could be semiquantitated in a few minutes.

Mung bean nuclease treatment improves Se genotyping by allele-specific inverse polymerase chain reaction amplification.

The allele-specific inverse polymerase chain reaction (PCR) technique, which has been explored to detect two linked polymorphic regions simultaneously, was applied to genotype the Se system. The major alleles of the Se system in Japanese are Se, sej defined by a single nucleotide substitution in the Se allele, and se(fus) generated by recombination between the Sec1 and FUT2 genes. The first PCR products using gene-specific primers were self-ligated, and each allele was detected by the second inverse-PCR using allele-specific primers.

Se genotyping following allele-specific polymerase chain reaction amplification.

The polymorphism of the Sec2 gene, which determines Se blood type, has been reported. This study presents an Se genotyping system by the allele-specific polymerase chain reaction amplification method. The Se, sej and se(fus) alleles were amplified using allele-specific primers.