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Biomedical subjects

Y Kawamoto

Publications and source records attributed to Y Kawamoto.

At least 109 records · Page 6Linked to original sources

Genetic profile of alloxan-induced diabetes-susceptible mice (ALS) and-resistant mice (ALR).

Two inbred strains from a foundation stock derived from Crj: CD-1 (ICR) mice were established after more than 20 generations of full-sib mating, and by simultaneous selective breeding for developing and not developing diabetes after alloxan administration (45 mg/kg in males, 47 mg/kg in females). To elucidate the genetic background of the two inbred strains, i. e., alloxan-induced diabetes-susceptible (ALS) strain and alloxan-induced diabetes-resistant (ALR) strain, their biochemical genetic markers and immunogenetic markers were examined. 1) For both strains, investigation of biochemical genetic markers at 19 loci and immunogenetic markers at 11 loci revealed no variation in any gene within the same strain, showing to be homogeneous, thus indicating establishment of the inbred strains. 2) The two strains differed from each other at 2 loci of biochemical genetic markers and at 5 loci of immunogenetic markers. 3) The ALS and ALR strains can be regarded as new inbred strains derived from ICR mice. 4) The results show that the marker genes of the two strains are different at 7 loci, but it remains unclear whether or not these genes are involved in the difference between the two strains in susceptibility to alloxan.

Alloxan↗

[Genetic parameters of body weight, muscle weight and skeletal characteristics in Japanese quail].

Heritability, phenotypic and genetic correlations of body weight, muscle weight and skeletal characteristics of Japanese quail males at 8 weeks of age were investigated to obtain basal information on breeding and on genetic monitoring by morphometrical methods. For this study, 221 male progenies were used. Measurements were taken on body weight, muscle weight and four kind of skeletal characteristics (skeletal weight, skeletal length, skeletal width and skeletal height). Heritability estimates were 0.67 for body weight, 0.36 to 0.56 for muscle weight, 0.79 to 0.94 for skeletal weight, 0.35 to 0.77 for skeletal length, 0.17 to 0.32 for skeletal width and 0.41 to 0.84 for skeletal height, respectively. The phenotypic and genetic correlations were found to be highly positive between body weight, muscle weight and skeletal length. A high degree of genetic correlation was observed between femur length, ossa cruris length and ossa metatarsalia length. These results suggest that the body weight, muscle weight and skeletal characteristics can be improved by breeding and that these genetically determined skeletal characteristics might serve for strain identification and genetic monitoring in Japanese quail.

Animals↗

Bronchial reactivities in guinea pigs to acetylcholine or histamine exposure.

The bronchial reactivities in Hartley guinea pigs to acetylcholine (ACh) or histamine (Hist) were investigated, and the following results obtained; 1. Eight-week-old animals were exposed to ACh and Hist. A significant relationship was observed between the concentrations of the chemicals and the time needed to produce falling down (TNPFD) due to the asthmatic reaction to ACh and Hist. 2. Eight-week-old animals were exposed to 0.1% ACh and 0.05% Hist, for which the mean TNPFD +/- standard error were 377 +/- 33 sec and 122 +/- 5 sec respectivity. However, no difference in reactivity between male and female animals was noted. 3. Eight- and 9-week-old animals were exposed to 0.01% ACh and 0.025% Hist. A positive correlation was observed (r = 0.736, p less than 0.01) between the TNPFD for ACh and that for Hist. 4. Growing animals from 2 weeks to 20 weeks old were exposed to 0.08% ACh and 0.025% Hist. After inhalation of both chemicals, 6-week-old animals showed the greatest prolongation of mean TNPFD (lowest sensitivity). 5. Eight-week-old animals were exposed to 0.08% ACh and 0.025% Hist. With both of these chemicals, a positive correlation was observed between TNPFD and dose threshold (ACh r = 0.886, p less than 0.001; Hist r = 0.891, p less than 0.001).

Acetylcholine↗

Diversity of surface protein antigen A gene in the chromosomal DNA of Streptococcus sobrinus.

A recombinant plasmid, pYA724, containing an 8.45-kb DNA fragment encoding surface protein antigen A (spaA) from Streptococcus sobrinus 6715 was used to examine the DNA homology of the spaA gene with chromosomal DNA of Streptococcus sobrinus B13N. Restriction endonuclease BamHI-digested pYA724 DNA was 32P radio-labeled by nick-translation, and a Southern blot DNA-DNA hybridization experiment was carried out. pYA724 DNA hybridized with a BamHI fragment of B13N chromosomal DNA, and the size of this fragment was larger than 8.45 kb. The chromosomal DNA was digested with BamHI-Pst I, BamHI-Hind III, and BamHI-Sal I, and analyzed by Southern blot DNA hybridization. pYA724 DNA hybridized with different sizes and numbers of DNA fragments of B13N chromosomal DNA; however, the sizes were not identical to those of internal fragments expected from the pYA724 restriction map. SpaA gene clone derivative plasmids, pYA745 and pYA756, were digested with EcoRI and Hinc II, respectively, and electrophoresed together with the digested B13N chromosomal DNA on the same gel; then Southern blot analysis was carried out. Fragments with sizes corresponding to the EcoRI or HincII internal fragments of pYA745 and pYA756 were not observed among the B13N fragments. These data indicate that serotype d S. sobrinus B13N has DNA homologous with the spaA gene from serotype g strain 6715, although within the same biotype strain has a diversity of arrangement within the chromosome.

Antigens, Bacterial↗

A population-genetic study of crab-eating macaques (Macaca fascicularis) on the island of Angaur, Palau, Micronesia.

Blood protein polymorphisms of an introduced population of the crab-eating macaques on Angaur Island, Palau, Micronesia, were examined electrophoretically to assess genetic variability. Results showed a high degree of genetic heterozygosity and some distinctive features in the genetic constitution of this island population. Negative evidence is presented regarding ancestry from a single pair of macaques. Their origin is discussed in relation to the genetic structure of the present population.

Alleles↗

[Reaction of various experimental animals to exposure to acetylcholine or histamine, and morphological observations of bronchial smooth muscle].

We investigated the sensitivity of the airway of various experimental animals to acetylcholine (ACh) and histamine (Hist). The experimental animals were exposed to 0.1% ACh or 0.05% Hist. Guinea pigs and rabbits exhibited an asthmatic reaction to both chemicals, but rabbits seemed to have a milder reaction than guinea pigs to both chemicals. Mice, rats and hamsters showed no reaction. We then performed a morphological study of the airways of 5 species in order to clarify the reason for their different reactivities to ACh and Hist. In the morphological study, abundant smooth muscle could be seen in the terminal bronchioles and respiratory bronchioles of guinea pigs and rabbits. In contrast, animals of other species had little smooth muscle in either site. Mice had no respiratory bronchioles. Consequently, we concluded that there is a high correlation between sensitivity to ACh and Hist and the extent of smooth muscle distribution around the airway.

Acetylcholine↗

Viscoelasticity of fibrinogen solution and of blood during coagulation studied by a new damped oscillation rheometer.

The behavior of a newly developed damped oscillation type rheometer was analyzed for fibrinogen solution and blood during coagulation. This rheometer consists of a cylindrical tube suspended from a torsion wire, that is filled with liquid to be tested. The logarithmic damping factor (LDF) during coagulation for blood and fibrinogen solution was obtained by this rheometer, which was closely related to the changes of viscosity and/or viscoelasticity of the blood sample. The slight increase of LDF prior to the rapid decrease was observed for blood. The increase of LDF would be reflected in the formation of the aggregation structure of red blood cells (rouleaux network) prior to the formation of fibrin network. The value of LDF for fibrinogen solution sharply increased and then decreased through a maximum value with the progress of coagulation, although the change of LDF was remarkably dependent on the fibrinogen concentration. The initial increase in LDF for fibrinogen solution was considered to be due to the formation of small clots in the solution. The decrease in LDF after attaining a maximum value is ascribed to the formation of fully developed fibrin network. The maximum value of LDF during coagulation for fibrinogen solution is higher than that for blood. The behavior was compared with that for non-biological fluids such as viscosity standard liquids and polyvinyl alcohol solution. From those data, it was concluded that the higher value of LDF than that for Newtonian liquids was due to the formation of aggregation structure or inhomogeneous fine clots in the liquid, which was accompanied with the appearance of the elasticity.

Blood Coagulation↗

Serological survey and virus isolation of simian T-cell leukemia/T-lymphotropic virus type I (STLV-I) in non-human primates in their native countries.

Infection with a simian retrovirus (STLV-I) closely related to human T-lymphotropic virus type I (HTLV-I) was investigated in non-human primates living in their native countries in Africa and Asia. Serum antibodies cross-reacting with HTLV-I antigens were detected in 85 of 567 non-human primates of 30 species. Seropositive animals were found among African green monkeys, olive baboons, Sykes' monkeys, mandrills and patas monkeys in several countries in Africa, and cynomolgus monkeys, Celebes macaques and siamangs in Indonesia. The frequency of seropositivity was much higher in adult than in young African green monkeys, cynomolgus monkeys and Celebes macaques. STLV-Is were isolated by establishing II lines of virus-producing lymphoid cells in the presence of interleukin-2 from 5 species of seropositive non-human primates, i.e. the African green monkey, Sykes' monkey, Celebes macaque, cynomolgus monkey and siamang. All these cell lines had T-cell markers and Tac antigen, and the cell lines from the African green monkey and Sykes' monkeys were Leu2a+ while those from other species were Leu3a+. These cell lines expressed viral antigens reacting with human sera from adult T-cell leukemia (ATL) patients and monoclonal antibodies (MAbs) against p19 and p24 of HTLV-I core proteins, and produced virus particles having RNA-dependent DNA polymerase activity. Cellular DNAs from these cell lines contained provirus sequences homologous to HTLV-I, shown by Southern blot hybridization. The restriction patterns of these provirus genomes were different from those of HTLV-I and were also dissimilar in the different species.

Age Factors↗

Inactivation of influenza B virus by normal guinea-pig serum.

Normal guinea-pig serum (GPS) lacking detectable antiviral antibody efficiently neutralized the infectivity of influenza B virus grown in chick embryos or MDCK cells. The inhibitor was heat-labile and sensitive to trypsin digestion. This beta-like inhibitor required Ca2+ and the complement components C1 and C4 for its activity. In contrast, GPS did not inactivate influenza A virus. Influenza B virus from which the neuraminidase activity of the spikes on the viral envelope had been eliminated by trypsin digestion was also inactivated to a level comparable to untreated virus. Complement component C1 alone bound directly to influenza B virus and inhibited its haemagglutinin activity. We suggest that the beta-like inhibitor in GPS is a component of the classical complement pathway which is triggered by the protein moiety of influenza B virus haemagglutinin, leading to virus neutralization.

Animals↗

Substitutions of cysteine residues of Escherichia coli heat-stable enterotoxin by oligonucleotide-directed mutagenesis.

The Escherichia coli 18-amino-acid, heat-stable enterotoxin STp has six cysteine residues linked intramolecularly by three disulfide bonds. These disulfide bonds are important for toxic activity, but the precise role of each bond is not clear. We substituted cysteine residues of STp in vivo by oligonucleotide-directed site-specific mutagenesis to dissociate each disulfide bond and examined the biological activities of the resulting mutants. The Cys-6----Ala and Cys-17----Ala mutations caused a complete loss of toxic activity. The Cys-5----Ala, Cys-10----Ser, and Gly-16, Cys-17----Cys-16, Gly-17 mutations caused a large decrease in toxic activity. These results mean that all three disulfide bonds formed at fixed positions are required for full expression of the biological activity of STp. However, a weak but significant toxicity still remained after three mutations, Cys-5----Ala, Cys-10----Ser, and Gly-16, Cys-17----Cys-16, Gly-17. This indicates that STp has some flexibilities in its conformation to exert toxic activity and that the role of each disulfide bond exerting toxic activity is not quite the same.

Amino Acid Sequence↗

Rapid isolation of endogenous inhibitor of factor B.

An inhibitor of mouse factor B was purified from lysate of L cells by a human factor B-affinity column. The purified inhibitor was found to be homogeneous with a molecular weight of about 25,000 and identical to the 25K protein isolated from L cell-lysate by the coprecipitation method with anti mouse factor B. Hemolytic titration of the activity and PAGE-analysis of the inhibitor-factor B complex showed that 1 mol of inhibitor reacted with 1 mol of mouse factor B and inactivated its hemolytic activity.

Animals↗

Cytotoxicity and calcium-dependent antigen of Yersinia.

The relationship between invasiveness and calcium dependency was examined in various strains of Yersinia enterocolitica and Y. pseudotuberculosis by using established cell lines. Infection with calcium-dependent bacteria resulted in the formation of microvilli and the adherence of bacteria on the cell surface, and the adherent bacteria were ingested 1.5 hr after infection. Morphological changes in the cells became visible 2 to 3 hr after infection, and intracellular multiplication of the ingested bacteria was noted. When the cells were incubated with bacteria at 37 C for 1.5 hr and then at 25 C, however, the morphological changes in the infected cells were not observed. No isogenic strains that had lost calcium dependency for growth at 37 C were able to elicit the morphological changes in the cells, though they possessed the ability to adhere to and penetrate the cells. The antigen(s) supposedly related to cytotoxicity of the calcium-dependent Yersinia was sought by using antibodies prepared against calcium-dependent bacteria and then absorbed with calcium-independent bacteria and with calcium-independent bacterial cytosol. Double diffusion tests between the antisera and bacterial cytosol extracts revealed the presence of an antigen which was a cytoplasmic substance common to all calcium-dependent but not calcium-independent strains of Y. enterocolitica and Y. pseudotuberculosis.

Animals↗

Endogenous inhibitors of factor B.

proteins which were able to bind noncovalently with mouse factor B were found in cells that are nonsecretors of factor B such as mouse-established monocytic cells and L cells but not in peritoneal resident macrophages. These proteins were isolated from lysates of L cells and separated into four distinct proteins by preparative SDS-polyacrylamide gel electrophoresis, with molecular weights of 25K , 28K , 33K, and 35K . The individual proteins formed a complex with purified mouse factor B at a molecular ratio of 1: 1 and inhibited its hemolytic activity. Proteins 25K and 28K inhibited the hemolytic activity of an activated form of factor B combined with cobra venom factor as well as that of the native form. These inhibitors did not affect the hemolytic activity of the second component of complement in mouse serum. The inhibitory activity of the 25K protein was partially inhibited by antiserum raised against it in rabbits.

Animals↗