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Biomedical subjects

S Masaki

Publications and source records attributed to S Masaki.

At least 19 recordsLinked to original sources

Differential contribution of the FcR gamma chain to the surface expression of the T cell receptor among T cells localized in epithelia: analysis of FcR gamma-deficient mice.

The function of the Fc receptors gamma chain (FcR gamma) for the expression of the T cell receptor (TCR) complex and for T cell development, especially for T cells localized in epithelia, was investigated by analyzing FcR gamma-deficient mice. In wild-type mice, CD8 alpha alpha + beta -TCR alpha beta + T cells of intestinal intraepithelial lymphocytes (i-IEL) utilized CD3 zeta homodimers and zeta-FcR gamma heterodimers, whereas CD8 alpha alpha + beta -TCR gamma delta + i-IEL used zeta-FcR gamma and FcR gamma homodimers in the TCR complex. On the other hand, these T cells in FcR gamma-deficient mice contained only zeta homodimers. The surface expression of the TCR complex was reduced in CD8 alpha alpha + beta -i-IEL and dendritic epidermal T cells (DETC) in these mice, whereas the development of these T cells was normal. The degree of reduction appeared to depend on the expression level of FcR gamma. In contrast to these populations, TCR gamma delta + intraepithelial T cells in reproductive organs (r-IEL) were dramatically decreased, suggesting that the development of r-IEL is FcR gamma-dependent, probably due to the predominant usage of FcR gamma homodimers in the TCR complex. These results indicate that the FcR gamma chain contributes differently to the TCR expression and to the development of T cells localized in epithelia.

Animals

The gene Bfsp1 for the lens fiber cell beaded-filament structural protein CP94 maps to mouse chromosome 2.

Bfsp1 is a mouse gene encoding the lens beaded-filament structural protein CP94. By use of rat CP94 cDNA as a probe, Southern hybridization of mouse DNAs prepared from the interspecific backcross progeny between Mus molossinus-mit and C57BL/6J strains was carried out. The results indicated that Bfsp1 maps at position 58 (+/- 6) on chromosome 2, in the vicinity of the blind-sterile (bs) locus.

Animals

Apoptotic degeneration in the arhinencephalic brain of the mouse mutant Pdn/Pdn.

The homozygotes of a mouse strain with genetic polydactyly (Polydactyly Nagoya, Pdn) exhibit arhinencephaly and various brain malformations. In the present experiment, abnormal apoptotic degeneration in the arhinencephalic brain of Pdn/Pdn embryos and newborns was investigated immunohistochemically and by molecular genetic techniques. Polyclonal antibody against single-stranded DNA detected the nuclei of programmed dying cells (apoptotic cells) specifically in the interdigital necrotic zone of the normal mouse limb plate on day 14 of gestation. We used this antibody against single-stranded DNA to investigate the apoptotic degeneration in Pdn/Pdn brain. Abnormal apoptosis was observed in the infralimbic cortical plate, hypothalamus and periventricular thalamus on day 0 after birth in Pdn/Pdn brains. The TRPM-2 gene, which has been considered to mediate apoptosis, was expressed in the developing normal and Pdn/Pdn brains. TRPM-2 gene expressions in the brain stem and cerebellum of arhinencephalic Pdn/Pdn fetuses and newborns were higher than those of +/+ littermates. From these facts, it was suggested that the abnormal apoptosis caused a large amount of cell loss in the arhinencephalic mouse brain, and this cell loss induced the expansion of the ventricle, followed by the hydrocephaly.

Animals

cDNA sequence analysis of CP94: rat lens fiber cell beaded-filament structural protein shows homology to cytokeratins.

To study the molecular structure of the gene responsible for a lens fiber cell beaded-filament structural protein of 94kDa (CP94), we isolated its specific cDNA from a rat lens cDNA library by use of anti-mouse CP94 antiserum. The expressed fusion protein kept the epitopes specific against anti-chick CP97 as well as anti-mouse CP94 antibody, and the size was estimated as 190-200kDa, indicating that the cDNA insert of the clone seemed to encode a polypeptide with 80-90kDa in appearance. Northern analysis indicated that CP94 mRNA is expressed only in the lens, and not in the brain, skin, heart, kidney, lung, and liver, and the size was estimated to 2.1-2.3kb. In a lens of inherited microphthalmic mouse, Elo, a trace amount of mRNA with the size closely similar to that of rat mRNA was observed. The entire compiled sequence (1,873bp) showed an open reading frame covering the sequence of 533 amino acids totalling 58,857Da. No sequence homologous to the entire CP94 was found among the entries of any nucleotide and amino acid sequence databases; but with respect to a limited amino acid sequence of N-side region of CP94, a significant homology with cytokeratins was found.

Amino Acid Sequence

A molecular genetic linkage map of mouse chromosome 10, including the Myb, S100b, Pah, Sl, and Ifg genes.

Restriction endonuclease fragment length variations (RFLV) on mouse chromosome 10 were detected in four genes, namely, the Myb protooncogene (Myb) and the genes for S100 beta protein (S100b), phenylalanine hydroxylase (Pah), and interferon-gamma (Ifg). RFLV were found in restriction patterns generated with BamHI for Myb, in those generated with BglII for S100b, in those generated with EcoRV for Pah, and in those generated with TaqI for Ifg. A multipoint backcross was carried out by the mating (129/Sv-Sl/+ x MOL-MIT)F1 x 129/SvJ(-)+/+. The Sl mutation has phenotypic effects which include deficiencies in pigment cells, germ cells, and blood cells. The following order of genes was derived from the results of the multipoint backcross, with distances between genes in parentheses: centromere--Myb--(34.9 cM)--S100b--(8.5 cM)--Pah--(8.5 cM)--Sl--(12.3 cM)--Ifg--telomere. Most laboratory strains and two strains of Mus musculus domesticus of wild origin carry the Myba, S100a, Paha, and Ifga alleles. In contrast, a strain of M. m. musculus, two strains of M. m. yamashinai, and two strains of M. m. molossinus carry the Mybb, S100b, Pahb, and Ifgb alleles. Other strains of wild origin carry various combinations of these alleles.

Alleles

Antiallergic effect of epinastine (WAL 801 CL) on immediate hypersensitivity reactions: (I). Elucidation of the mechanism for histamine release inhibition.

Epinastine caused an inhibition of histamine release from rat peritoneal mast cells induced by both antigen-antibody reaction and compound 48/80. Epinastine was similarly effective in inhibiting compound 48/80-induced histamine release not only from isolated rat peritoneal mast cells but also from rat mesenterial pieces. Also, histamine release from lung pieces obtained from actively sensitized guinea pigs after exposure to antigen challenge was markedly inhibited by epinastine. The drug was effective in inhibiting not only Ca2+ uptake into lung mast cells in actively sensitized guinea pigs but also Ca2+ release from the intracellular Ca store of rat peritoneal mast cells exposed to both compound 48/80 and substance P. No significant changes were observed in phosphodiesterase activity in rat peritoneal mast cells treated with epinastine, while adenylate cyclase activity was augmented by epinastine. Epinastine has no inhibitory effect on histamine release induced by Ca2+ or IP3 from permeabilized mast cells. However, the drug significantly and dose-dependently suppressed calmodulin activity suggesting that histamine release inhibition due to epinastine may be partly attributable to Ca(2+)-calmodulin dependent process(es). The drug caused no visible changes in thermodynamic behavior of lipids, either in order parameter or in differential scanning calorimetry, indicating that the drug has no influence on membrane fluidity.

Adenylyl Cyclases

Defect of a fiber cell-specific 94-kDa protein in the lens of inherited microphthalmic mutant mouse Elo.

Deficiency in a 94,000-dalton protein in the non-crystallin fraction from the Elo mouse lens was shown. To perform further investigations, we raised an antibody against the 94,000-dalton protein isolated from normal mouse lens. Western blot analysis with the antibody indicated that the protein was only present in the lens and not in the brain, lung, heart, liver, and kidney. In the lens, it was unique to the cortex and nucleus fractions, not being present in the epithelial cells. Furthermore, it was observed in the water-soluble fraction as well as in the urea-soluble fraction. The antibody weakly but clearly reacted with the chick CP97 lens peptide, a fiber cell-specific protein, and anti-CP97 antibody also reacted with the 94,000-dalton protein. From these results, we concluded that the protein corresponds to CP97 cytoskeletal protein in the mouse lens. The protein was deficient in the lenses from Elo mice, but microphthalmic lenses from CTA mice contained a normal level.

Animals

Effect of Tween 80 on formation of the superficial L1 layer of the Mycobacterium avium-Mycobacterium intracellulare complex.

The effects of Tween 80 supplementation of liquid culture medium on the formation of the superficial L1 layer of the Mycobacterium avium-Mycobacterium intracellulare complex (MAC) were examined by serological and scanning electron microscopic experiments. Specific antiserum to the glycopeptidolipids on the L1 layer of M. avium S-139, made in a rabbit, was used for seroagglutination reactions with antigens prepared from strain S-139 grown in medium supplemented with various levels of Tween 80 (0, 0.05, 0.5, 5, and 50 mg/ml). The agglutination titers gradually decreased as the concentration of Tween 80 rose. Scanning electron microscopy showed that the fibrillar materials consisting mainly of glycopeptidolipids on the L1 layer of strain S-139 also disappeared with increases in the concentration of Tween 80. In addition, there was no obvious correlation between (i) the plasmid DNAs and serotypes of MAC and (ii) formation of the L1 layer of MAC. It is therefore concluded that Tween 80 used to supplement liquid culture medium affects formation of the L1 layer, which has been considered to be one of the pathogenic factors of MAC.

Animals

Comparative study of the adverse effects of various radiographic contrast media, including ioversol, a new low-osmolarity medium. II. The complement system and endothelial cells.

The effects of ioversol, iohexol, iopamidol and meglumine sodium amidotrizoate (MSA) on the complement system and endothelial cells were investigated. The protein bindings of the radiographic contrast media (RCM), each tested with guinea pig plasma, were less than 1%. When guinea pig serum was incubated with any of the RCM, activation of the complement system, which leads to hemolysis, was not influenced by the nonionic agents, ioversol, iohexol or iopamidol. However, MSA, an ionic agent, significantly reduced hemolytic activity at 370 mgI/ml. Perfusion of the abdominal aorta with nonionic agents did not elicit significant endothelial damage; ioversol induced the least damage among the nonionic RCM, while MSA caused remarkable endothelial damage. Although MSA caused a marked release of endothelin-1 from cultured endothelial cells obtained from porcine aorta, nonionic RCM did not induce significant endothelin-1 release; no influence was elicited by ioversol and iohexol caused a weak suppression, while iopamidol had the opposite effect. These results indicate that ioversol could be used as a safe contrast medium in intravascular administration.

Animals

Sex differences in cognitive abilities: a cross-cultural perspective.

Studies in Western cultures have indicated significant sex differences in certain cognitive abilities. To determine whether similar differences occur in a non-Western culture, this study administered a cross-linguistic battery of tests to high school students in Japan and America. In both cultures, girls averaged significantly higher scores on a Story Recall test, the Digit-Symbol test and a Word Fluency test whereas boys achieved significantly higher scores on a Mental Rotation test. The analysis of standardized test scores further indicated that the size of the sex difference was culture-independent in three out of these four cases. These results are discussed in the context of the GESCHWIND and GALABURDA [Cerebral Lateralization, Biological Mechanisms, Associations and Pathology, Bradford Books, Cambridge, Massachusetts] account of the contribution of testosterone to left-right asymmetries in early cerebral development.

Adolescent

Effect of Tween 80 on the growth of Mycobacterium avium complex.

The effect of Tween 80 on the growth of Mycobacterium avium complex (MAC) in liquid culture condition was investigated. Observation of the colony-forming units (CFU) and the morphology of MAC with transmission and scanning electron microscopy showed that Tween 80 at 0.05% in the medium (ca. 0.5 mg/ml) had bacteriostatic action and caused cell elongation. Tween 80 at 0.5% or more in the medium (ca. 5 mg/ml) reduced the quantity of MAC glycolipids and also led to false positive or positive results in biochemical tests for mycobacterial identification using nitrate reductase, urease, or arylsulfatase. To determine whether or not surfactants could reduce the MAC permeability barrier, the minimal inhibitory concentration (MIC) of antituberculosis drugs on MAC was determined in liquid medium with or without several kinds of surfactants including Tween 80. Five surfactants including Tween 80 increased the activity of antituberculosis drugs to MAC. These findings suggest that Tween 80 acts directly on the cell wall of MAC.

Antitubercular Agents

Restriction fragment length variations and chromosome mapping of two mouse metallothionein genes, Mt-1 and Mt-2.

Restriction endonuclease fragment length variations (RFLVs) were found through the use of cDNA probes for metallothionein genes 1 (Mt-1) and 2 (Mt-2) in the mouse. RFLVs were detected in restriction patterns generated by BglII and XbaI in the Mt-1 gene and by PvuII in the Mt-2 gene. All laboratory strains carry the Mt-1a and Mt-2a alleles. Among strains of wild origin, some Western European subspecies (Mus mus domesticus and M. m. brevirostris) also carry the Mt-1a and Mt-2a alleles. In contrast, a European subspecies (M. m. musculus) and the great majority of subspecies from East Asian countries (M. m. molossinus, Chinese mice of wild origin, and M. m. yamashinai) carry the Mt-1b and Mt-2b alleles. A domesticus strain from Bulgaria and two castaneus strains from Thailand and Philippines carry the intermediate combination of Mt-1b and Mt-2a alleles. Using the RFLVs, we mapped the Mt-1 and Mt-2 genes on chromosome 8, and they appear to be very closely linked since no recombination was observed between them in any of the mice examined. Data from three-point cross tests showed that the recombination frequencies are 4.31% between Os and Mt, 15.52% between Mt and Prt-2, and 19.83% between Os and Prt-2. The gene order of Os-Mt-1,Mt-2-Prt-2 has been confirmed.

Alleles

Linkage analysis of the mutation locus in the eye lens obsolescence (Elo) mouse.

To map precisely the mutation locus of eye lens obsolescence (Elo) on mouse chromosome 1, subsequent linkage analysis was achieved using backcross mating between 129/SvSl-Elo (Elo/+) and 129/SvSl (+/+). Mouse genomic DNAs from 17 strains including the Elo mutant mouse were first digested with several restriction enzymes and analyzed by hybridization using gamma 2- and gamma 4-crystallin cDNAs as probes. Restriction endonuclease DraI showed distinct RFLP patterns in both cases. When gamma 2-crystallin cDNA was used as the probe, two strong bands were observed at 4.0 and 2.4 kb in the majority of strains, but the former fragment shifted to the 3.4-kb position in 129/SvSl-Elo (Elo/Elo) and CFO. The polymorphism between 4.0- and 3.4-kb fragments corresponded to the gamma 1-crystallin locus (Cryg-1), and that of the 2.4-kb one, to the gamma 2-crystallin locus (Cryg-2). Mouse DNAs were also analyzed by hybridization using gamma 4-crystallin cDNA (Cryg-4). In this case, 3.4- and 3.0-kb fragments were observed in Elo and wild-type mice, respectively. The backcross offsprings were analyzed with respect to Elo, Idh-1, Cryg-1, and Cryg-4 loci. Among 223 mice analyzed, recombination between Elo and Idh-1 loci was observed in three offsprings; and that between Cryg-1 or Cryg-4 and Idh-1 loci, in one offspring. No recombination occurred between Cryg-1 and Cryg-4 alleles.

Alleles

Plasmid profiles of Mycobacterium avium complex isolated from swine.

Twenty strains of Mycobacterium avium complex (MAC) isolated from swine and five strains from humans were examined for drug susceptibility and plasmid content. Four strains of swine origin and two strains of human origin harbored plasmid DNAs differing in molecular weights. No relationship between plasmid contents and drug resistance was observed. Southern DNA-DNA hybridization showed that small plasmids from swine MAC strains were homologous to those from human origin at the nucleotide level.

Animals

New antibiotics, resorcinomycins A and B: antibacterial activity of resorcinomycin A against mycobacteria in vitro.

Resorcinomycin A, N-[(S)-alpha-guanidino-3,5-dihydroxy-4-isopropylphenylacetyl]glyci ne, S-RSM-A, a new antibiotic produced by Streptoverticillium roseoverticillatum, has an antibacterial spectrum directed towards mycobacterial species. It is not active against Gram-negative and Gram-positive bacteria, except mycobacteria and weakly active against mycoplasmas. In vitro activities of the S-, R- and S,R-isomers of RSM-A against atypical mycobacterial strains were compared with those of streptomycin (SM) and kanamycin (KM) by 2-fold agar dilution methods using Middlebrook 7H10 agar medium. Although R-RSM-A and S,R-RSM-A were comparable or inferior to both antibiotics, the antimycobacterial activity of S-RSM-A was superior to that of SM and KM.

Anti-Bacterial Agents

Structural study of immunoaffinity-purified DNA polymerase alpha-DNA primase complex from calf thymus.

The DNA polymerase alpha-DNA primase complex was purified over 17,000-fold to near homogeneity from calf thymus using an immunoaffinity column. Sodium dodecyl sulfate gel electrophoresis revealed three polypeptides with molecular weights of 140, 50 and 47 kDa, in a ratio of 1:2:0.25. The complex showed a sedimentation coefficient of 9.7 S, a Stokes radius of 56 A and a native molecular weight of 250-260 kDa. Taken together, the data suggest that the calf thymus dNA polymerase alpha-DNA primase complex is essentially a heterotrimer of large (140 kDa) and small (50 kDa) subunits in a ratio of 1:2, with a globular conformation. Electron-microscopic studies of the complex revealed a spherical particle of 120 A in diameter, in agreement with the physiochemical results. The binding of the complex to DNA was also demonstrated.

Animals