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K Sagisaka

Publications and source records attributed to K Sagisaka.

At least 19 recordsLinked to original sources
PubMed1992

Methylated cytosine level in human liver DNA does not decline in aging process.

In order to ascertain a generality of the age-dependent decrease in DNA methylation level among different mammalian species, methylated cytosine contents in human liver and spleen DNA at different ages have been determined using high performance liquid chromatography (HPLC). Unexpectedly, the liver DNA revealed no appreciable decline with age while the spleen DNA showed a slight reduction. It indicates that a decrease of methylation level in genomic DNA is not a common denominator of age-related changes in mammals.

Adolescent↗
PubMed1992

Suppression of ischemia-reperfusion injury by liposomal superoxide dismutase in rats subjected to tourniquet shock.

To investigate the role of oxygen-derived free radicals in the pathogenesis of tourniquet shock, the authors present an experimental animal model. Two groups of rats were fastened with rubber tubes on both thighs (1.5 kg/cm2) for 6 h under pentobarbital anaesthesia. One group was administered liposomal superoxide dismutase (L-SOD 30,000 U/kg body weight), and the other liposome as a control 3 h prior to tourniquet removal. No rats in the control group (n = 20) survived more than 24 h after reperfusion, whereas 55% of animals treated with L-SOD (n = 20) survived for 24 h or more, and two recovered completely (P less than 0.005). Blood samples were obtained from the abdominal aorta after laparotomy of anaesthetized rats of both groups at different time intervals. Changes in the hematocrit value and blood urea nitrogen during the early periods after reperfusion were attenuated by prior administration of L-SOD, and the total plasma SOD activity of the control animals decreased promptly and continuously throughout the experimental period. This experimental model was very useful to study the pathogenesis of tourniquet shock with respect to reproducibility, induction of the shock stages and mortality. It is thought that oxygen-free radicals are involved in the induction of tourniquet shock, and L-SOD was, to a certain extent, effective against reperfusion injury in the early stages of shock.

Alanine Transaminase↗
PubMed1992

Blood grouping of minute samples using monoclonal anti-A,B antibody.

The availability of a monoclonal anti-A,B antibody (AB-E6E2) for blood grouping of blood and saliva stains was investigated. The antibody, which was reacted with A, B and AB antigens but not with O antigen, was produced from BALB/c mouse-mouse hybridoma raised to group AB red blood cells. AB-E6E2 had heat-elutability enough to detect ABO antigens in blood and saliva stains by absorption-elution test, whereas it had extremely weak double combining potency in mixed agglutination test. Application of elution-ELISA method will enable the detection of smaller amount of ABH antigens. The antibody, therefore, would be useful to segregate group O minute samples from the other blood group ones.

ABO Blood-Group System↗
PubMed1991

Application of single-locus hypervariable region DNA probes to deficiency cases in paternity testing.

Seven DNA probes which recognize single-locus hypervariable region (HVR) were applied to a paternity test in which the putative father and his wife were deceased. Three legitimate children, an illegitimate child and her mother were available for analysis. The cumulative paternity index of the illegitimate child derived from 15 conventional blood group markers was 18.71 and from 7 DNA probes 92,572.08, that is, 4,948 times higher than the former. Thus the DNA analyses gave nearly conclusive evidence that the putative father was the biological father of the child. The application of highly discriminating polymorphisms of DNA which recognize single HVR loci is considered to be extremely informative in cases of disputed parentage.

Child↗
PubMed1991

[Paternity test with single locus DNA probes].

Seven kinds of DNA probes recognizing hypervariable DNA loci were applied to 28 cases of paternity test, involving two cases in which the putative fathers had died. The combinations of probe and restriction enzyme are as follows; MR24/1-HinfI, 3'Globin-PvuII, Ha-ras-PvuII, Mucin-PvuII, D2S44 (pYNH24)-MspI, D17S30 (pYNZ22)-MspI, D1S57 (pYNZ2)-RsaI. The reported number of the alleles are 37, 39, 5, 10, 33, 15, and 5, respectively. Those probes lie on different chromosomes except D1S57 on 1p and Mucin on 1q21. Exclusion probability (EP) and paternity index (PI) were calculated from the allele frequencies in Japanese population reported by Yokoi et al. Cumulative EP from 7 DNA probes was 0.999932, and cumulative PI ranged from 7.3 X 10(6) to 947. Also, cumulative EP from 17 kinds of conventional blood group markers (CBGM) was 0.9776, and cumulative PI ranged from 1290 to 0.11. Total EP from 7 DNA probes and CBGM was 0.999998478. Cumulative PI from 7 DNA probes were 5 to 2,000,000 times higher than that from CBGM. The single locus hypervariable DNA polymorphisms are considered to be informative for paternity test.

DNA Probes↗
PubMed1990

Binding of thyroid hormones to human hemoglobin and localization of the binding site.

Radiolabeled thyroid hormones were allowed to bind to erythrocyte cytosol and the complex was fractionated by Sephadex G-100 or by high-performance liquid chromatography (HPLC). On Sephadex G-100, four radioactive peaks (P1-P4) were obtained, whereas HPLC gave only three radioactive peaks (P1-P3). Chromatographic studies with human adult Hb and non-Hb cytosol protein fractions, which had been reacted with radiolabeled thyroid hormones, and immune precipitation with specific antisera for the hormones, confirmed that the first peak of Sephadex G-100 radioactivity was a mixture of Hb and non-Hb proteins, while the second peak was Hb. The third peak was free 125I and the fourth peak was unbound 125I-T3 or 125I-T4. The third peak of HPLC was confirmed to be a mixture of free 125I and unbound radiolabeled thyroid hormones. Scatchard analysis of the interaction between T4 and apo-Hb, and the alpha- and beta-chains of human Hb suggested the presence of the specific binding site(s) for the hormone. Interaction between T4 and synthesized peptides, which constitute the heme pocket of the beta-chain of Hb (beta 61-75, beta 71-85, beta 81-95), indicated that the T4 binding site of Hb resides within the heme-binding cavity. It is concluded that human erythrocyte cytosol does not contain "receptor" for thyroid hormones and cannot be a model for studying functions of cytosol "receptor" for the hormones; rather, it contains binding protein with large binding capacity, including Hb and non-Hb proteins, which possibly constitute a large reservoir for the hormone in blood.

Binding Sites↗
PubMed1990

Hypervariable polymorphic VNTR loci for parentage testing and individual identification.

Three kinds of variable number of tandem repeat DNA probes (VNTR: pYNZ22, pYNH24, and pYNZ2) showing hypervariable polymorphisms were studied. Allelic frequencies and their confidence intervals among Japanese individuals were obtained. Co-dominant segregation of the polymorphism was confirmed in family studies. Two a priori probabilities were calculated for each VNTR locus: exclusion probabilities for an alleged father/mother/child trio and for an alleged parent/child duo, and probabilities of matching of genotyped two unrelated individuals or two siblings. Availability as well as highly discriminating polymorphic pattern of VNTR loci makes it potentially very useful for forensic and human genetic purposes.

Alleles↗
PubMed1990

Investigation of paternity establishing without the putative father using hypervariable DNA probes.

Seven kinds of DNA probes which recognize hypervariable loci were applied for paternity test. The putative father was decreased and unavailable for the test. The two legitimate children and their mother (the deceased's wife) and the four illegitimate children and their mother (the deceased's kept mistress) were available for analysis. Paternity index of four illegitimate child was investigated. Allelic frequencies and their confidence intervals among unrelated Japanese individuals were previously reported from our laboratory, and co-dominant segregation of the polymorphism was confirmed in family studies. Cumulative paternity indices of four illegitimate children from 16 kinds of standard blood group markers were 165, 42, 0.09, and 36, respectively. On the other hand, cumulative paternity indices from 7 kinds of DNA probes are 2,363, 4,685, 57,678, and 54,994, respectively, which are 14, 113, 640, 864, and 1,509 times higher than that from standard blood group markers. The DNA analyses gave nearly conclusive evidence that the putative father was the biological father of the children. Especially, the paternity relation of the third illegitimate child could not be established without the DNA analyses. Accordingly, DNA polymorphism is considered to be informative enough for paternity test.

DNA↗
PubMed1990

Haptoglobin typing of human bloodstains using a specific DNA probe.

The stability of DNA in human bloodstains and various post mortem tissues has been investigated. High molecular weight (HMW) DNA was usually recovered from dried bloodstains, even those up to a few years old, but very rapid degradation was found to occur post mortem in the liver, pancreas, spleen and kidney. Other tissues such as the heart, thyroid and skeletal muscle were found to give a reasonable yield of HMW DNA during the first few days after death. The feasibility of using DNA extracted from forensic bloodstain specimens for the detection of DNA polymorphisms was explored using a human haptoglobin (Hp) alpha chain specific probe. Using HindIII and XbaI digests the Hp genotypes Hp2, Hp1F and Hp1S were distinguished by Southern blot analysis in DNA prepared from 1 cm2 bloodstains up to 15-18 months old.

Autoradiography↗
PubMed1990

Individual identification of human bloodstains investigated with hypervariable DNA loci.

Seven kinds of hypervariable DNA probes, which recognize hypervariable loci, were applied for individual identification of human bloodstains. Allelic frequencies and their confidence intervals among unrelated Japanese individuals were previously reported from our laboratory, and codominant segregation of the polymorphism was confirmed in family studies. One-month-old bloodstains from ten individuals were investigated in the present study. The probability of matching was calculated using the database among Japanese population. Cumulative probability of matching from 7 kinds of hypervariable DNA probes was 1.1 x 10(-11), which was 4.5 x 10(9) times higher than that from 6 kinds of common blood group markers such as ABO, MN, Rh-Hr, P1, Hp and PGM1. Accordingly, DNA polymorphism is considered to be informative enough for individual identification of bloodstains.

Alleles↗
PubMed1990

Hypervariable regions of DNA for parentage testing and individual identification.

Four kinds of DNA probes that recognize hypervariable regions (HVR) were studied for parentage testing and individual identification. Allele frequencies and their confidence intervals among unrelated Japanese individuals were obtained. Codominant segregation of the polymorphism was confirmed in family studies. Two a priori probabilities were calculated for each HVR locus: the exclusion probabilities for an alleged father/mother/child trio and for an alleged parent/child duo, and the probabilities of matching of genotypes of two unrelated individuals or two siblings. The ease of availability of the probes and their highly discriminating polymorphic patterns mean they could be very useful for forensic purposes.

Alleles↗
PubMed1989

Human identification and sex determination of dental pulp, bone marrow and blood stains with a recombinant DNA probe.

Recombinant DNA hybridizing specifically to a 300 nucleotide repeat DNA sequence (BLUR8) of human specificity and to human repeat DNA sequence (pHY10) on the Y chromosome was used for human identification and sex determination of degraded DNA samples of blood stains, dental pulp, and bone marrow. This radioactive technique enabled reliable and sensitive human and sex determination from blood stains that were more than 80 years old. Less than 1 piece of 0.5 cm length thread of blood stain was enough for both tests. DNA from relatively fresh dental pulp and bone marrow was clearly identified. The human identification test, which could recognize up to 0.3 ng DNA correctly, was 3 to 5 times more sensitive than the sex determination test.

Adolescent↗
PubMed1989

Hypervariable polymorphism of autosomal origin detected by the Y-chromosome derived probe, pHY10.

A recombinant DNA probe (pHY10) hybridizing specifically to human DNA family DYZ1, 3,000 copies of which are present on the long arm of the Y chromosome, was used for probing human genome DNA digested with various restriction enzymes. To our surprise, the probe detected a hypervariable polymorphism of autosomal origin in human DNA when digested with TaqI. None of other 12 restriction enzymes revealed polymorphic patterns. Codominant segregation of the polymorphism was established in family studies. This probe has been widely used in the detection of the Y chromosome. Its ease of availability as well as highly discriminating polymorphic pattern makes it potentially very useful for forensic and human genetic purposes.

DNA Probes↗
PubMed1989

[Some aspects of the ABH blood group from the standpoint of personal identification].

1. Preparation of red cell or saliva specific ABH antibodies To prepare agglutinins specific for red cell (anti-Hr, -Ar and -Br) or for saliva (anti-Hs, -As and -Bs), many animals were immunized with red cell or saliva. Then antisera were alternately absorbed with saliva or red cell. By these procedures, specific anti-Hr, -Ar, -Hs, -As and -Hs were prepared. Specificity of antisera to saliva was confirmed by latex agglutination which was adsorbed with saliva. 2. Quantitation of red cell or saliva specific antigen Quantitative analyses of Hs, As and Bs in saliva were performed by agglutination inhibition test, indicating that considerable individual difference in quantity of these antigens was observed. However, the difference was proved not to be genetically controlled. On the other hand, quantity of Hr or Ar antigens were distributed similarly among H, A, B and AB groups, or between A and AB groups, respectively. 3. Localization of Hr and Hs antigen in the salivary glands Distribution of the antigen was examined by ABC technique. Hs antigen was mainly located in the serous gland and slightly in the mucous gland and ductus. However, Hr was observed mainly in red cell and slightly in the ductus. H antigen detected with U. europaeus anti-H was distributed in all the glands. 4. Heterogeneity of Ulex europaeus anti-H (UE) UE is known to consist of two isolectins (UE-1 and UE-2) with different sugar specificity. Rabbit antiserum to UE-1 or UE-2 was prepared.(ABSTRACT TRUNCATED AT 250 WORDS)

ABO Blood-Group System↗
PubMed1979

Preparation of anti-alpha 2-macroglobulin using Canavalia lineata DC lectin for differentiating species-specificity of blood stains.

To differentiate species-specificity of blood stains, anti-alpha 2-macroglobulin was raised in rabbits against Canavalia lineata DC lectin-serum complex (LSC). Adsorption of anti-LSC with human lipoprotein resulted in antiserum specific for alpha 2-macroglobulin. It was confirmed by Ouchterlony test that the antiserum adsorbed successively with monkey serum or anti-LSC adsorbed directly with monkey serum reacted with only human serum but not with mammalian ones. Immunoelectrosyneresis and anti-LSC consumption test could identify species-specificity of blood stains kept for up to two years and for up to several years, respectively. It is indicated that anti-LSC is quite effective for differentiating species-specificity of blood stains.

Adsorption↗
PubMed1979

On the receptors of human red cells reacting with Phaseolus coccineus L. lectin.

In order to elucidate receptors of proteolytic enzyme-treated red cells which react with Phaseolus coccineus L. lectin, the receptors prepared by affinity chromatography were serologically investigated. P. coccineus lectin had high agglutinin activity for bromelin-, papain- and pronase-treated red cells but that for the cells treated with ficin and trypsin was relatively low. Analyses of chemical composition revealed that sialic acid of the receptors from normal red cells was considerably much as compared with that from the treated cells. On the contrary, the enzyme treatment did not affect particularly carbohydrate composition of the receptors. Disc electrophoresis showed that the patterns of receptors from red cells treated with bromelin or papain were different from those from the other cells. On two-dimensional immunoelectrophoresis, the receptor of trypsin-treated cells gave five precipitation lines against anti-stroma and that of papain-treated cells three lines, but any other receptors showed no line. These findings indicate that there are plural receptors for P. coccineus lectin in red cells treated with each of proteolytic enzymes and that the receptors from respective red cells have electrophoretically and serologically different property.

Chromatography, Affinity↗
PubMed1978

Fragmentation of human albumin with proteolytic enzymes and its antigenicity with special reference to human-specificity.

Human albumin prepared by the trichloroacetic acid method was treated with bromelin, ficin, papain, pronase or trypsin. Pronase- or trypsin-treated albumin showed four fragments on polyacrylamide disc electrophoresis. When rabbit antiserum to pronase-treated albumin was adsorbed with monkey albumin, antibody with human specificity was completely abolished, whereas anti-albumin serum adsorbed with pronase-treated albumin retained the antibody to human albumin. It was considered that antigenic site with human specificity in albumin structure was inactivated by pronase.

Antigens↗