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Biomedical subjects

R Mukoyama

Publications and source records attributed to R Mukoyama.

6 recordsLinked to original sources
PubMed1998

Gender determination from dental pulp by using capillary gel electrophoresis of amelogenin locus.

Gender of dental pulp DNA was determined by the capillary gel electrophoresis (CGE) method with amelogenin locus as a marker. The analysis of the male sample resulted in two peaks, a Y-locus standard sample was added and the CGE method repeated. This resulted in an amplification of the second peak and judging from the difference between the allele sizes of the X and Y loci, it was confirmed that the first peak was for the X and the second for the Y locus. The female sample was then analyzed and one peak was observed. As before the X-locus standard sample was added to this sample and resulted in an amplification of the peak, thus verifying it to be the X locus. The CGE method was conducted for 10 male and 10 female samples and double peaks verified the male sample, while one peak verified the female sample. Gender was thus correctly determined in all cases.

Amelogenin↗
PubMed1998

[Repeat structure of DYS389 locus].

In the DYS389 locus, tetranucleotide repeat units (CTGT and CTAT) connect complicatedly, and repeat sequences to show polymorphism are found in two sections. Because non-repetitive sequence including the same sequence as forward primer exists among these repeat sequences, a pair of primers for the DYS389 locus give two amplification products (DYS389 I and DYS389 II), both of which are polymorphic. These polymorphisms come from the differences in the number of CTGT and CTAT repeats. DNA was extracted from the dental pulp of 68 males and saliva of 48 males, and amplified for the DYS389 locus. And then the length polymorphisms and the repeat structures were examined. As results of denaturing polyacrylamide gel electrophoresis, alleles 8-12 appeared in the DYS389 I locus, while alleles 24 29 appeared in the DYS389 II locus, with alleles 11 and 27 showing the most frequent. As results of sequencing of many samples, we confirmed that the repeat structures of the DYS389 I and II alleles were (CTGT)3 (CTAT)r and (CTGT)p (CTAT)q (CTGT)3 (CTAT)r, respectively. DYS389 I alleles were related to the repeat numbers of r (r = 8-12), and DYS389 II alleles to the repeat numbers of p + q + r (p = 4.5; q = 11-14; r = 8-12). Even the same DNA size of DYS389 II alleles had different repeat structures, indicating that DYS389 is effective for the purposes of individual identification and paternity testing.

Alleles↗
PubMed1995

[Application of enzyme-linked immunosorbent assay (ELISA) to the medico-legal identification].

Medico-legal identification of saliva stains and bloodstains was performed by an enzyme-linked immunosorbent assay (ELISA) using a horseradish peroxidase conjugate in combination with the use of monoclonal antibodies. Activity of alpha-amylase in the stains was measured for an identification of saliva using an anti-human amylase antibody, and secretory IgA was detected for a species identification using an anti-IgA antibody. ABO and Lewis blood group antigens were detected using anti-A, anti-B, anti-H, anti-Lea and anti-Leb antibodies. After the solubilization of ABH antigens in blood stains with octyl-beta-D-glucose solution, ABO typing has been performed using fully automated system composed of a unit for blood group reaction based on ELISA and a unit for reading of result based on photometric analysis.

ABO Blood-Group System↗
PubMed1995

[Sex determination from dental calculus by polymerase chain reaction (PCR)].

We studied sex determination from dental calculus by the PCR method using primers which recognize DYZ3 region of Y-chromosome and DXZ1 of X-chromosome. The amounts of DNA obtained from 1.7-25.0 mg of dental calculus samples ranged 0.6-26.4 micrograms, and the majority of DNA was low molecular weight DNA. After PCR amplification, Y and X products were observed by the method of polyacrylamide gel electrophoresis. Both Y and X amplification products were observed in 26 out of 29 male samples, while, Y products was not detected in two samples and neither Y nor X product was detected in one sample. In 17 out of 20 female samples, only X product was observed, while, Y and X products were observed in three samples. The minimum amount of DNA for sex determination was 3 pg. Sex determination using DNA in dental calculus will be quite useful for forensic application because it can be done without destruction of morphological characteristics of teeth.

DNA↗
PubMed1989

Determination of ABO blood groups from saliva and saliva stains by an indirect enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies.

The detection of A, B and H blood group substances (ABH-BGS) in saliva and in saliva stains has been investigated quantitatively by an indirect ELISA using a horseradish peroxidase conjugate in combination with the use of monoclonal antibodies. Through this method, the reaction specificity to BGS in the saliva was very high and its detection sensitivity was found to be approximately 1,000 times greater than has been achieved in a hemagglutination-inhibition test. The monoclonal anti-A and anti-B reagents reacting with both secretor and non-secretor saliva in a hemagglutination-inhibition test and in this ELISA method were selected from among commercial monoclonal antibodies. However, no monoclonal anti-H reagent was found to react with non-secretor saliva. The BGS level was determined by the use of calibration curves of A, B and H standard BGS from human gastric mucosa and was expressed in units, based on the inhibition titer of the standard BGS. In 230 saliva samples, ABH-BGS were detectable, except for H BGS in non-secretor saliva. The BGS levels in saliva stains experimentally prepared were found to be approximately proportional to the levels in the original saliva. As for actual and aged stains, it was possible to detect BGS in most cigarette butts and in aged stains, however, such detection proved impossible in saliva samples from postage stamp.

ABO Blood-Group System↗