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H Inouye

Publications and source records attributed to H Inouye.

At least 73 records · Page 4Linked to original sources
PubMed1984

New X-ray spacings from central myelinated tissue.

New X-ray reflections have been detected from myelinated tissue of the mammalian C.N.S. Diffraction patterns from unfixed optic nerves of mouse, rat, guinea-pig and rabbit, and unfixed corpus callosum of calf were recorded during X-ray exposures of 1-6 days. The equatorial pattern was typical of lamellar myelin of the C.N.S.; however, the meridional pattern showed new features: a strong, sharp intensity maximum at 30.4 +/- 0.4 A (N = 11) spacing, and a weak, diffuse ring centered at 11.0 +/- 0.5 A (N = 5) spacing. The 30.4 A reflection was neither strictly arced like the equatorial reflections from lamellar myelin nor fully horizontal. Since the 30.4 A meridional reflection was not observed in patterns from myelinated nerve of the P.N.S., we suggest that this diffraction comes from the radial component, or interlamellar tight junctions, which is unique to mammalian C.N.S. myelinated tissue. The diffuse ring at 11.0 A probably comes from myelin protein.

Animals↗
PubMed1983

CD-resolved secondary structure of bovine plasma albumin in acid-induced isomerization.

Bovine plasma albumin (BPA) showed the acid-induced two-step transition, the N-F transition and acid-expansion. Changes in fractions of alpha-helix (f alpha), beta-form (f beta) and unordered form (fR) in the acid-induced isomerization of BPA were studied using the method of Chen et al. (1972) with two constraints: sigma fi = 1, 0 less than or equal to fi less than or equal to 1. pH-profiles of f alpha and fR showed the two-step change, one corresponding to the N-F transition and the other to the acid-expansion in 0.10 M KCl and in 0.02 M NaClO4. pH-profile of f beta showed one-step change, correlating to the later part (lower pH side) of the N-F transition. The N-F transition might thus involve the helix leads to beta and helix leads to coil transitions.

Animals↗
PubMed1983

Mutations that alter the signal sequence of alkaline phosphatase in Escherichia coli.

A phoA-lacZ gene fusion was used to isolate mutants altered in the alkaline phosphatase signal sequence. This was done by selecting Lac+ mutants from a phoA-lacZ fusion strain that produces a membrane-bound hybrid protein and is unable to grow on lactose. Two such mutant derivatives were characterized. The mutations lie within the phoA portion of the fused gene and cause internalization of the hybrid protein. When the mutations were genetically recombined into an otherwise wild-type phoA gene, they interfered with export of alkaline phosphatase to the periplasm. The mutant alkaline phosphatase protein was found instead in the cytoplasm in precursor form. DNA sequence analysis demonstrated that both mutations lead to amino acid alterations in the signal sequence of alkaline phosphatase.

Alkaline Phosphatase↗
PubMed1983

Analysis of regulation of phoB expression using a phoB-cat fusion.

The phoB gene, which encodes a positive control factor for a number of phosphate-regulated genes in Escherichia coli, was cloned into multicopy plasmid pBR322. A phoB-cat fusion that expressed chloramphenicol transacetylase from the phoB promoter was constructed. Studies of the expression of the phoB-cat fusion showed that the pattern of regulation of the phoB gene was similar to that of the phoA gene, the structural gene for alkaline phosphatase. The phoB gene was derepressed under conditions of phosphate starvation, was constitutively expressed in a phoR background, and required the phoM gene product for expression in a phoR strain. Finally, a functional phoB product was required for its own synthesis. Our results indicate either that phoA gene expression responds directly to the concentration of the phoB gene product in cells or that the phoA and phoB controlling elements are quite similar.

Acetyltransferases↗
PubMed1983

Immunogenetical study in patients with chronic glomerulonephritis.

HLA-A, B and DR typing was performed on a total of 236 patients with chronic glomerulonephritis (CGN) which were diagnosed by renal biopsies evaluated by light microscopy, direct immunofluorescent staining and whenever indicated electron microscopy. The interrelationship between the HLA system and the histopathologic classification of CGN was examined. There were 20 patients with benign recurrent hematuria (BRH), 82 with IgA nephropathy (IgA-N), 24 with proliferative glomerulonephritis (PGN), 47 with minimal change nephrotic syndrome (MCNS), 4 with membranous nephropathy (MN), 43 with membranoproliferative glomerulonephritis (MPGN) and 16 with focal glomerulosclerosis (FGS). The following results were obtained: 1) Antigens from different loci were associated with different subgroups of CGN. There were three types of associations; a) HLA-A locus associated as found in PGN, b) HLA-B locus associated as found in BRH and c) HLA-DR locus associated as found in MCNS. IgA-N, MPGN and FGS seemed to be included in this group. 2) The association of HLA-A locus antigens with MCNS could be subdivided by age o onset suggesting that in MCNS HLA-A antigens are linked to disease expression. 3) A heterogeneity in IgA-N is suggested by its association with different HLA antigens; a) a HLA-B37 associated group, b) a HLA-DR4 or DYT associated group and c) a HLA-DEn (Dw 6.1) or B12 associated group. These data suggest differences in the mechanisms of HLA and disease association in each histopathologic subtype of CGN.

Adolescent↗
PubMed1982

HLA-D clusters associated with DR4 in the Japanese population.

A study of HLA-D clusters associated with DR4 was performed in the Japanese population. These clusters consist of DYT, DKT2, DB3, and Dw4. Forty-two Japanese typed as DR4 were investigated, and it was found that 17 (40.5%) were DYT, 7 (16.7%) DKT2, 7 (16.7%) DB3, and 4 (9.5%) Dw4.

HLA-DR4 Antigen↗
PubMed1982

Circular dichroic and fluoropolarimetric studies on tryptophyl residues in acid-induced isomerization of bovine plasma albumin.

The acid-induced isomerization (the N-F transition) and expansion of bovine plasma albumin were studied by measuring circular dichroic spectra and fluorescence polarization of tryptophyl residues. Decreases in the magnitude of ellipticities at 208, 222, 262 and 268 nm were observed in the N-F transition and acid-expansion. However, increases in the magnitude of ellipticities at 295-300 nm observed in the initial part of the N-F transition exactly correlated with the increase of rotational relaxation time of tryptophyl side chains obtained by fluorescence polarization measurement.

Animals↗
PubMed1982

Conformational changes of bovine plasma albumin prior to the salting-out of protein in concentrated salt solution.

By working at very low protein concentration (ca. 0.003%), it is possible to measure tryptophyl fluorescence intensity at 350 nm (F350) of bovine plasma albumin (BPA) as a function of pH under precipitating conditions (acidic concentrated salt solutions). Under such conditions, distinct changes in F350 were seen before the starting of precipitation of BPA and no further changes in F350 over the precipitating pH range. Comparison of pH-profiles monitored by F350 with those by solubility in the presence of various salts at various concentrations indicated that the change of solubility is observed after definite changes in conformation of the protein.

Animals↗
PubMed1982

Signal sequence of alkaline phosphatase of Escherichia coli.

The amino acid sequence of the signal sequence of phoA was determined by DNA sequencing by using the dideoxy chain termination technique (Sanger et al., Proc. Natl. Acad. Sci. U.S.A. 74:5463-5467, 1977). The template used was single-stranded DNA obtained from M13 on f1 phage derivatives carrying phoA, constructed by in vitro recombination. The results confirm the sequence of the first five amino acids determined by Sarthy et al. (J. Bacteriol. 139:932-939, 1979) and extend the sequence in the same reading frame into the amino terminal region of the mature alkaline phosphatase (Bradshaw et al., Proc. Natl. Acad. Sci. U.S.A., 78:3473-3477, 1981). As was predicted (Inouye and Beckwith, Proc. Natl. Acad. Sci. U.S.A. 74:1440-1444, 1977), the signal sequence was highly hydrophobic. The alteration of DNA sequence was identified for a promoter mutation that results in the expression of phoA independent of the positive control gene phoB and in insensitivity to high phosphate.

Alkaline Phosphatase↗
PubMed1981

Cloning and restriction mapping of the alkaline phosphatase structural gene (phoA) of Escherichia coli and generation of deletion mutants in vitro.

The structural gene for alkaline phosphatase (phoA) of Escherichia coli was cloned into the PstI site of pBR322, from a transducing bacteriophage, lambda p(phoA-proC). The restriction map of the plasmid was established. Based upon this information, several phoA deletion plasmids as well as a smaller phoA+ plasmid were constructed. The genetic map and restriction map were correlated by recombination analysis. Cells carrying one of the phoA+ plasmids overproduce alkaline phosphatase 10-fold upon phosphate limitation. However, both regulation and processing of the enzyme were found to be normal.

Alkaline Phosphatase↗
PubMed1981

HLA-D/DR linked immune response to Candida allergen: an in vivo and in vitro study in Japanese.

Immune response to Candida allergen in man was studied using the lymphocyte reaction in vitro and the skin test in vivo. One hundred and ten normal healthy Japanese were typed for HLA-A, B, D and DR. It was found that HLA-B7, Dw1 and DR1 were strongly associated with high responsiveness and Den and Mt2 with low responsiveness. In a family study, the inheritance of high responsiveness was observed. In the reaction patterns of HLA-D homozygote and heterozygote cells, it was found that Dw1/DR1 and Dw12/DR2 were associated with a high responders and that DYT/DR4 and DEn (6.1)/DR- were associated with low responders. It is considered that the Dw1/DRl gene might be more dominant as an immune response; the Ir gene when compared to a gene such as the DYT, and the EDn (6.1) as an immune suppressive; Is gene. A genetic factor can be implicated in the secondary response (but not the primary response) to Candida allergen. Therefore, the existence of HLA-linked Ir and/or Is gene to Candida allergen is discussed.

Adult↗
PubMed1981

Role of macrophages in T lymphocyte response to Candida allergen in man with special reference to HLA-D and DR.

Activation of naturally sensitized human T lymphocytes to Candida allergen was studied using three HLA-D and DR heterozygote Japanese cells (Dw1.DR1/DYT.DR4, Dw12.DR2/DYT.DR4, DYT.DR4/DEn.DR blank) and four HLA-D and DR homozygote cells (Dw1.DR1, Dw12.DR2, DYT.DR4, DEn.DR blank). In vitro activation of T lymphocytes to Candida allergen was found to require the presence of autologous or allogeneic compatible HLA-Dw1.DR1 and Dw12.DR2 macrophages.

Antigens, Fungal↗
PubMed1980

Cloned primed-lymphocyte-test reagents in the dissection of HLA-D.

Human T lymphocytes obtained as blasts on day 4 from a primary mixed leukocyte culture (MLC) were cloned in the presence of T cell growth factor (TCGF) and feeder cells. Parameters important in producing higher-specific-activity TCGF were evaluated; irradiation of the responding cells as well as removal of adherent cells or inclusion of indomethacin in the culture was important. In addition, the presence of an irradiated lymphoblastoid cell line (LCL) cell in the TCGF-producing system enhanced activity in the supernate. The long-term maintenance of progeny from clones was achieved by utilizing the LCL autologous with either the responding or sensitizing cells from the initial MLC as feeder cells. Under those conditions, clones could be expanded for 7 or more wk with the maintenance of PLT reactivity. Had all the cells in each clone been maintained for the full 7 wk, more than 1 X 10(10) cells could have been developed in each clone. The cloned reagents provide a higher degree of antigen-specific reactivity than do normal PLT cells. It is to be anticipated that as the requirements for cloning are made more stringent, including the recloning of the cells, these reagents will aid greatly in the dissection of the complexity attendant to HLA-D.

Clone Cells↗
PubMed1980

Long-term maintenance of "cloned" human PLT cells in TCGF with LCL cells as a feeder layer.

The long-term maintenance of T cells "cloned" by limiting dilution in TCGF was enhanced by the use of irradiated autologous lymphoblastoid cell line (LCL) cells as well as irradiated LCL cells of the individual to which the T cells were originally primed. It was possible to obtain more than 1 X 10(12) cells from a "clone" seeded at one cell per well. Some of the clones tested express primed LD-typing activity.

Cell Division↗
PubMed1980

TCGF production for cloning and growth of functional human T lymphocytes.

In an effort to increase the potency of T cell growth factor (TCGF), several variables were examined for their effects on the production of TCGF. The following manipulations enhanced the potency of TCGF: first, the removal of adherent cells and addition of indomethacin to the producing cultures; second, irradiation with 1000 rads of the cells used to produce TCGF; and, third, the addition of Epstein-Barr virus transformed lymphoblastoid (LCL) cells. It was also noted that the addition of irradiated feeder cells increased the efficiency of limiting dilution cloning.

Cell Division↗