Profenofos metabolites in human poisoning

Profenofos and its metabolites were determined in a case of fatal poisoning. Little profenofos and large amounts of metabolites were detected by gas chromatography/flame photometric detection in the acid extracts of blood and urine after methylation with diazomethane. Four major metabolites containing phosphorus were identified with the synthesized metabolites, namely, despropylated profenofos, desethylated profenofos and des-S-propylated profenofos, respectively. 4-Bromo-2-chlorophenol (BCP), an aryl moiety of profenofos, was also determined in blood and urine with high performance liquid chromatograph (HPLC) as free or conjugated metabolites.     

Highly sensitive HLA-DNA typing from formalin-fixed and paraffin-embedded tissue samples

Nucleotide sequences have been determined for more than 1700 different alleles at the core of the human leukocyte antigen (HLA) system. The highly polymorphic character of these genes affects adaptive immune response and is also useful for forensic applications. HLA typing from formalin-fixed and paraffin-embedded tissue provides abundant useful information for both clinical settings and forensic investigations. This study, which investigated the potential use of DNA from formalin-fixed and paraffin-embedded tissue samples in an HLA PCR sequence-specific primer and probe (SPP) system, showed that tissue fixed in formalin for less than 3 days and embedded in paraffin can serve as a useful source of DNA for PCR-SPP typing kits.     

Thymus and adrenal glands in elder abuse

Endogenous glucocorticoid-induced thymic involution is generally considered to be an important finding for determining child abuse. The present study investigated the weight of the thymus and the adrenal glands in elder abuse cases to identify a potential marker for elder abuse. There was no significant difference in the thymus and the adrenal weight between elder abuse and control cases. However, the elder abuse cases in which the duration of abuse was less than 3 months showed a significant increase in the adrenal weight in comparison to control cases. In such cases, histopathological findings showed a loss of intracellular light granules from the zona fasciculata, which might indicate a loss of cholesterol due to the overproduction of glucocorticoid. These results might imply that the elderly, who were maltreated for less than 3 months, were in the early phase of a long-term stress state during which stress-induced overproduction of glucocorticoid was observed in adrenal glands as indicated by Selye. Our results suggest that an increase in adrenal weight may be a potential marker for elder abuse of relatively short periods, especially less than a few months.     

Analysis of AmpFlSTR MiniFiler™ loci and its forensic application

The allele frequencies of eight MiniFiler™ loci have been analyzed in 101 Japanese individuals living in Kanagawa with informed consent by means of ABI 310 Genetic Analyzer. A total of 7 alleles for D13S317, 8 alleles for D7S820, 11 alleles for D2S1338, 11 alleles for D21S11, 5 alleles for D16S539, 14 alleles for D18S51, 8 alleles for CSF1PO, and 13 alleles for FGA were observed. The polymorphic profiles of these MiniFiler™ loci in the present study were essentially the same as those obtained by using the AmpFlSTR^® Identifiler^® PCR Amplification kit. The combined matching probability of eight MiniFiler™ loci and cumulative probability of paternity exclusion were estimated as 1.97 × 10⁻¹⁰ and 0.9996, respectively. The MiniFiler™ kit was useful for individual identification in forensic analysis.     

Preventing elderly suicide through primary care by community-based screening for depression in rural Japan

Depression is a major cause of suicide among the elderly. Few previous community-based interventions against depression have reduced the suicide rate. This study aims to evaluate outcomes of a community-based program to prevent suicide among the elderly using a quasi-experimental design with a neighboring reference group. The program, including depression screening with follow-up and health education through primary care and public health nursing, was implemented for 10 years in Matsudai town, a rural area of Japan (population 6,015; suicide rate per 10(5) [65-year-olds] for males 290.6, and for females 361.3). Changes in the suicide risk were estimated by the incidence rate ratio (IRR). The female risk of completing suicide in the intervention area was reduced by 70% (age-adjusted IRR: 0.30; 95% CI: 0.14-0.67), while there was no change in the risk for males in the intervention area nor for males or females in the reference area (Kawanishi town: population 9,425; elderly suicide rate for males 212.2, females 151.9). A ratio of the female IRR in the intervention area to that in its prefecture was also estimated at 0.45 (95% CI: 0.19-0.97), showing that the reduction of suicide risk in the intervention area was greater than the historical trend. A community intervention against suicide using management of depression with nonpsychiatric, primary health care would be effective for elderly females, but not males.     

Determination of phenotypes of esterases (Set) in fresh saliva and saliva stains by disc electrophoresis and the distribution of Set phenotypes in the Japanese population

A rapid and simple disc electrophoretic technique for the determination of saliva esterase (Set) types is described. The frequency of Set types was F 16.4%, FS 49.8% and S 33.8%. The estimated gene frequency of Set-1F was 0.413 and of Set-1S was 0.587. An accurate determination of Set types was possible in 5 – 7-week-old saliva stains. This system may be useful in medicolegal applications.     

Development of Multiple Assays using 46 SNPs for Comprehensive mtDNA Haplogrouping and Application to Highly Degraded DNA

Six multiplex PCR systems using single‐base extension reactions to analyze 46 mitochondrial DNA (mtDNA)‐coding region single nucleotide polymorphisms (SNPs) that define 42 haplogroups, that is, 24 major mtDNA haplogroups and 18 subclades, were devised. To improve the usefulness of the established systems for the analysis of degraded DNA samples, novel primers to render amplicons with sizes <150 bp were designed. By applying these systems to 214 Japanese individuals, 24 different haplogroups (power of discrimination = 93.4%) were found. To assess the effectiveness of our systems in grouping degraded DNA, an ancient bone sample of a Jomon skeleton was analyzed and then classified as haplogroup N9b. We conclude that the present systems are powerful screening tools for major haplogroups of mtDNA in addition to the prevalent subhaplogroups in the Japanese population and that these systems are capable of analyzing highly degraded DNA samples in forensic studies.     

Rapid and simple sex determination method from dental pulp by loop-mediated isothermal amplification

Sex determination from dental pulp DNA was examined by loop-mediated isothermal amplification (LAMP) method. Amelogenin locus was analyzed for sex determination. A set of four specially designed primers was prepared based on database from Gene Bank, and loop primers were designed to shorten the analysis time. Analysis was performed using 32 dental pulp DNA samples removal from permanent teeth stored at room temperature for 1–25 years after extraction. The X allele was detected in approximately 32 min with real-time turbidimeter and the Y allele was detected in approximately 34 min. Analysis time was reduced to half when using loop primers. Visual detection was also possible as the amplified product showed white turbidity. Sex determination by LAMP method was rapid and simple, and it should prove useful in unknown bodies of mass disasters.