Novel paternity testing by distinguishing parental alleles at a VNTR locus in the differentially methylated region upstream of the human H19 gene

Conventional PCR-based genotyping is useful for forensic testing but cannot be used to determine parental origins of alleles in DNA specimens. Here we describe a novel method of combined conventional genotyping and PIA typing (parentally imprinted allele typing) at a minisatellite region upstream from the H19 locus. The PIA typing uses two sets of primers and DNA digested with methylation-sensitive Hha I enzyme. The first amplification produces only the methylated fragment of paternal H19 allele, and the second detects polymorphism in the minisatellite. Hence, this distinguishes paternal and maternal alleles by difference in the DNA methylation. Furthermore, the polymorphism in this polymorphic locus was examined using 199 unrelated Japanese and 171 unrelated Germans, their polymorphism information content being 0.671 and 0.705, respectively. Feasibility of this typing is demonstrated for six families, and the usefulness is shown by application to paternity testing.     

Lower limits of detection of synchrotron radiation high-energy X-ray fluorescence spectrometry and its possibility for the forensic application for discrimination of glass fragments

Synchrotron radiation high-energy X-ray fluorescence (SR-XRF) analysis utilizing 75.5keV X-ray radiation from beam-line BL37XU at Super Photon Ring 8GeV (SPring-8), a third-generation synchrotron facility, was found to have advantages for forensic discrimination of glass samples. The lower limits of detection (LLD) for calibration curves were at the picogram level for Ba, Ce, and Sm and at the 10pg level for Sr, Zr, Sn, and Hf. The spectrum of NIST SRM 612 glass reference material demonstrated K-line peaks of 31 elements including rare-earth elements, and the relative standard deviations (R.S.D.) of all the measured elements except Ca were less than 9.7%. Fragments of collected sheet glass were used as samples for investigating the application of this technique to forensic analysis. Several trace elements such as Pb, Rb, Sr, Zr, La, Ce, and Hf were detected in the spectra of the samples, and these elements could be used as indexes to characterize the glass samples. But the "lower limits of detection (LLD)" of each element were not examined enough. In this report, these limits by synchrotron radiation X-ray spectrometry were clarified. By these results, this technique should provide an effective approach to the nondestructive discrimination of small glass fragments in the field of forensic science.     

Toxicological analysis of thiamylal in biological materials by gas chromatography/mass spectrometry

A reliable and sensitive method to analyze thiamylal in biological materials was developed, using gas chromatography/mass spectrometry (GC/MS). A quantitative determination was made by use of mass fragmentography with the lower detection limit of 0.01 microgram/g. Thiopental was used as the internal standard. Distribution of the drug in the blood and body tissues of rats was examined. The method was then used to detect thiamylal in tissues from an autopsied patient and concentration of this drug in the body materials was evaluated, from medico-legal aspects.     

Determination of ABO blood grouping from human oral squamous epithelium by the highly sensitive immunohistochemical staining method EnVision+

Using the highly sensitive immunohistochemical staining method EnVision+, which employs a dextran polymer reagent for the secondary antibody, the detection of the ABH antigens was attempted in the oral squamous epithelium. This new technique uses monoclonal antibody as a primary antibody and it takes about three hours for staining. The time is much shorter than conventional absorption-elution testing or absorption-inhibition testing for the determination of ABO blood grouping. Secretor saliva samples were stained at strong intensity by the antibody, which corresponded to its blood group and anti-H. On the one hand, nonsecretor saliva samples were stained at strong intensity only by the antibody that corresponded to its blood group, and at weak intensity only by anti-H. Since human oral squamous epithelium antigens were stained specifically by this method, we can examine the ABO blood group of saliva samples and perform cytodiagnosis at the same time. Our research suggested that the EnVision+ Method is a useful technique for ABO blood grouping of saliva in forensic cases.     

Comparison of Catalytic and Immunological Amylase Tests for Identifying of Saliva from Degraded Samples

The stability of salivary α‐amylase is a critical factor in both catalytic and immunological method‐based forensic saliva identification. This study aimed to assess the sensitivity of catalytic and immunological tests on degraded saliva samples. Degraded saliva stains were prepared by microbial decomposition using humid soil. Salivary α‐amylase activity was catalytically detected both qualitatively and quantitatively using the Phadebas^® amylase test. As immunological methods, we conducted qualitative and quantitative tests using the RSID?‐saliva test and ELISA, respectively. Salivary α‐amylase activity of degraded samples (incubated at 37°C for 12 h) was significantly lower than that of controls in the quantitative tests. All the degraded samples obtained by the humid soil produced negative results in the Phadebas^® tests, but showed positive results in the RSID?‐saliva test and ELISA. These results suggest that immunological tests are effective for testing degraded saliva samples that have lost their enzymatic activity.     

The use of moxibustion scars in the identification of individuals in forensic medicine

The use of moxibustion today does not only take place in Eastern Asia, but recently also in Europe and the United States. Moxibustion scars are often seen in autopsy cases in Japan. However, no reports of the use of moxibustion in autopsy cases have been reported. This paper reports on an autopsy case in which moxibustion scars were used to extrapolate on the past symptoms of the deceased, date of therapy and the location of the moxibustion clinic which administered the moxibustion.     

Trust in the leadership of governors and participatory governance in Tokyo Metropolitan Government

This article examines the relationship between the trust in leadership of political leaders and citizen participation by analysing data on Tokyo residents. Among the four variables we used to designate types of, or attitudes towards, public participation, only actual participation has a positive influence on trust. The normative recognition of participation is associated with a critical attitude towards the government. As the unclear needs of citizens have the greatest impact on trust, citizens’ trust is built through relationships between citizens and not between citizens and their government. The importance of positive actions from the local government such as public officials and political leaders that stimulate citizen awareness to the point where citizens take interest in public administration, practice participation, and enlarge participation opportunities, is increasing. We also need to consider the limitations of citizen ability to understand public issues and reflection of their needs, and the limitations of participatory governance on the decision-making process.