Hideo Ishizu

Can microorganisms produce alcohol in body cavities of a living person?: a case report.

Unusual endogenous ethanol production in intraabdominal bloody fluid of an individual who was stabbed in the abdomen and who developed peritonitis after a peritoneotomy is discussed. In the intraabdominal bloody fluid, 2.45 mg/g ethanol and 0.079 mg/g n-propanol were detected. The level of ethanol in the heart blood was about 1 mg/g. The level of n-propanol indicates that a large quantity of ethanol was produced endogenously in the intraabdominal bloody fluid. In an animal experiment in which rats were injected with 20 mL of 10% glucose mixed 5:1 with a presumed volume of rat blood into the abdominal cavity after injury of the small intestine to allow enterobacteria to spread into the cavity, a significant quantity of ethanol was produced in the administered fluid while the animals were alive. The antemortem ethanol production in the intraabdominal bloody fluid of the victim might have been caused by the microorganisms responsible for the peritonitis after the operation.

Typing of Y chromosome single nucleotide polymorphisms in a Japanese population by a multiplexed single nucleotide primer extension reaction

We have developed a new method for typing single nucleotide polymorphisms (SNPs) on the human Y chromosome based on a multiplexed single nucleotide primer extension. This method has the advantage that several SNPs are typed rapidly and simultaneously. We examined 15 different SNP loci on Y chromosome, M9, M105, M122, M125, M128, M130, SRY465, IMS-JST006241, IMS-JST006841, IMS-JST002611, IMS-JST003305, IMS-JST008425, IMS-JST021354, IMS-JST021355 and IMS-JST055457, in 159 Japanese males. From the typing results of these 15 loci, we found 13 haplotypes. Gene diversity for each locus ranged from 0.025 to 0.486 and the haplotype diversity was estimated to be 0.838. This method could be readily applied for personal identification and paternity testing.

Sudden unexpected death due to rupture of the stomach

We report a case of sudden unexpected death due to rupture of the stomach. A 49-year-old man was found dead in a public lavatory. Autopsy findings revealed two rupture wounds measuring 14 cm and 6 cm located in the fundus of stomach at the side of the greater curvature despite of any superficial injury. The deceased had an ulcer in the lesser curvature of stomach, and dilation in this area was expected to be impaired. Under this condition, excessive over-eating resulting in over-extension of the stomach wall at the greater curvature was speculated to have caused stomach rupture.

Automated Quantitation of Polyamines by Improved Cation-Exchange High-Performance Liquid Chromatography Using a Pump Equipped With a Plunger Washing System

Abstract An improved method for simple, precise, and sensitive quantitation of polyamines[putrescine, spermidine, spermine, cadaver-ine(in part) and 1, 6-diaminohexane] by cation-exchange high performance liquid chromatography is described. Postcolumn deriv-atization using the o-phthalaldehyde method was employed to detect them. Analysis took 45 minutes and the minimum detectable amount of each polyamine was 10 – 20 pmol/μ1. An isocratic pump equipped with a plunger washing system and a mobile phase of pH 5.3 are the advantages of this improved method. Consequently, an operator can carry out routine and automated polyamine analysis without any hesitation regarding maintenance of the pump or column damage due to crystal precipitation from the mobile phase on or in the pump, which has been a problem previously encountered with cation-exchange resin. Reproducibility of the day to day precision and duplicate determination, and simultaneous reproducibility of the standard mixture were calculated (CV<2.2%).

Simultaneous determination of nitrazepam and its metabolites in urine by micellar electrokinetic capillary chromatography

We applied micellar electrokinetic capillary chromatography to simultaneous separation and determination of nitrazepam and its major metabolites, 7-aminonitrazepam and 7-acetamidonitrazepam, in spiked urine. Prior to electrophoresis, the three compounds were successfully extracted from the spiked urine with commercial disposable solid-phase cartridges. The optimum running buffer for the separation was prepared by combining 85 parts of 60 mM sodium dodecyl sulphate-6 mM phosphate-borate, adjusted to pH 8.5, with 15 parts of methanol. The separation order, completed within 25 min, was 7-aminonitrazepam > 7-acetamidonitrazepam > nitrazepam, at an applied potential of 20 kV. We obtained reproducible electropherograms in successive repetitions, and few other peaks or interferences appeared in the electropherogram. The detection limits of the three compounds were 50-100 pg (0.1-0.2 microgram/ml of analyte in spiked urine), and the recoveries were 78.9-100.8% for 1 microgram/ml and 84.1-100.3% for 5 micrograms/ml. The application of this method to forensic or clinical samples is demonstrated.

Identification of human skeletal muscle from a tissue fragment by detection of human myoglobin using a double-sandwich ELISA

A method for identifying human skeletal muscle by detection of human myoglobin using a double-sandwich ELISA was developed. When an extract was prepared from 0.1 g skeletal muscle homogenized with 10 ml PBS, this method was able to detect human myoglobin in extracts diluted 10(4)-fold. There was no difference in the detection limit between individuals or sites of origin of skeletal muscles. Species specificity was good and no cross reaction occurred with skeletal muscle from other animals except the gorilla. Our method could also discriminate between skeletal muscle and other organs or tissues except the heart. Human myoglobin could be detected in skeletal muscles under the following conditions: putrefied at room temperature for 5 months, dried at room temperature for 11 months, heated at 100 degrees C for 72 h and immersed in fresh water at room temperature for 6 days. Two practical cases to which this method was applied are presented.

Massive pulmonary embolization with cerebral tissue due to gunshot wound to the head.

Autopsy findings in a 22-year-old man who received a gunshot wound to the head and died at the scene are presented. Massive white-grey emboli were found in the pulmonary arteries, and these were identified as cerebral cortical or glial tissue by histological examination. It was thought that the momentary severe increase of intracranial pressure and simultaneous rupture of the superior sagittal sinus due to the kinetic energy of the bullet had caused the lacerated cerebral tissue to enter the blood stream.

Effects of cocaine administration route on the formation of cocaethylene in drinkers : an experiment using rats

The effects of cocaine administration route on the formation of cocaethylene, an active metabolite of cocaine produced in the presence of ethanol, were investigated using rats. When 20 mg/kg cocaine was administered into the stomach together with 2 g/kg ethanol, maximum liver concentrations of cocaine and cocaethylene, 8410 +/- 3600 and 1680 +/- 520 ng/g, respectively, were observed at 15 min. In other tissues the maximum levels of both the substances were attained in 30 min, but were much lower than those in the liver. Intramuscular administration of 20 mg/kg cocaine with 2 g/kg oral ethanol gave levels of liver cocaine as low as 103 +/- 29 to 150 +/- 35 ng/g, resulting in no detection of liver cocaethylene over the entire 180-min study period, although gradual accumulation of cocaethylene was observed in other tissues. The accumulation patterns of cocaine and cocaethylene in blood of alcohol-intoxicated rats after the i.m. administration of cocaine were similar to those in blood of drinkers after nasal insufflation of cocaine. Despite i.v. administration of 1 mg/kg cocaine with 2 g/kg oral ethanol, no detectable amounts of cocaethylene appeared in any tissues over the entire 60-min study period. The present findings are considered to be of importance in the fields of forensic and clinical toxicology for clarifying (1) the rate of in vivo cocaethylene formation and (2) the distribution of cocaine and cocaethylene in blood and tissues.

Histopathological changes of the hypophysis in malnutrition in elderly subjects

Major objectives in forensic gerontology are physical and mental disorders during aging. which can be caused by various factors involving nutrition and stress, often accompanied by dysfunction in the neuroendocrine systems including the hypophysis. The objective of the present study was to investigate the histopathological changes in the adenohypophysis in elderly subjects using autopsy materials. Hypophyses with a scaphoid shape (group S: 16 males and 4 females: mean age. 78.6 years) and a normal one (group C: 30 males and 20 females: mean age. 65.2 years) were compared. Incidence of the scaphoid-shaped hypophysis mildly increased with age, being 17% in the elderly over 65 years of age. The weight of the pituitary gland in group S (0.42 ± 0.1 g) was lower than that of group C (0.65 ± 0.2 g). The degree of fibrosis was higher in group S (31.6% ± 5.4%) than in group C (18.3% ± 6.3%). lmmunohistochemical staining showed no significant differences in the proportion of the ACTH cells and the TSH cells between the two groups (p>0.05). However, there was an increase in the proportion of gonadotrophs. prolactin cells, and S-100-containing cells in group S and a decrease in that of GH cells (p<0.05). These findings may be associated with reduced anabolic, gonadal and hepatic functions due to malnutrition.

Polymorphism of the D12S391 microsatellite in a Japanese population sample

Using the polymerase chain reaction (PCR), we studied the short tandem repeat (STR) polymorphism observed at the D12S391 locus. In 350 Japanese examined, 14 different alleles ranging from 209 bp to 261 bp were detected. Allele 18 (221 bp) showed the highest frequency at 0.30. Observed and expected values of respective genotypes satisfied the Hardy-Weinberg equilibrium (chi 2 = 24.08, P = 0.24, df = 20). In addition, 18 additional sequence structures (suballeles), were detected in this study. Within the suballeles, sequence variants, in which the initial repeat of (AGAT) was replaced with (AGGT), was found in five samples. It was found that the analysis of single-strand conformation polymorphism (SSCP) before sequence analysis was useful for distinguishing these suballeles.