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Biomedical subjects

Y Miyake

Publications and source records attributed to Y Miyake.

At least 379 records · Page 21Linked to original sources
PubMed1991

Cardiovascular responses to norepinephrine and arginine vasopressin infusion in chronically catheterized fetal lambs.

Fetal cardiovascular responses to infused norepinephrine (NE) and arginine vasopressin (AVP) were compared in bilaterally vagotomized and chronically catheterized (control) fetal lambs. In the control fetuses, NE infusion was characterized during the first five minutes by a decreased fetal heart rate (FHR) and increased mean arterial blood pressure (MABP) and by a preejection period (PEP) followed by a gradual return to the original values. NE infusion in vagotomized fetuses was followed by tachycardia, decreased PEP without the initial prolongation and an MABP response similar to that in the controls. AVP in control fetuses caused a persistent decrease in the FHR and increase in MABP and PEP. Vagotomy affected only the FHR response: the decrease was persistent but smaller. AVP produced persistent fetal cardiovascular responses regardless of vagotomy; however, the biphasic response FHR pattern, suggestive of NE tachyphylaxis, was eliminated by vagotomy.

Animals↗
PubMed1991

[Late asthmatic response by recruitment of homocytotropic antibody from circulating blood to the airway wall in guinea pig].

Many reports have been published about the late asthmatic response (LAR) of animal model. But most of the animals used were sensitized actively and it is considered generally that it is impossible to generate LAR in passively sensitized animals, especially in guinea pig. About the mechanism of LAR, most discussion are focused on airway hyperreactivity, inflammation and chemical mediators. But little is known about the direct initiator of the late phase bronchoconstriction. Instead of the circulating antibody which is generated by antibody producing cells in actively sensitized animals, anti-serum was administered to guinea pigs intravenously as passive sensitization fifteen minutes prior to antigen inhalation. Four to eight hours after antigen inhalation, we investigated bronchoconstriction. Eight hours after antigen challenge, bronchoalveolar lavage (BAL) were performed. In the BAL fluid, macrophages decreased and neutrophils increased significantly. These results suggest that LAR can occur without airway inflammation and the inflammation might be the result of antigen-antibody reaction at the airway wall, and that the direct initiator of late phase bronchoconstriction might be the recruitment of homocytotropic antibody from circulating blood to the airway wall.

Animals↗
PubMed1990

Cloning and expression of a cDNA encoding mouse kidney D-amino acid oxidase.

A cDNA encoding D-amino acid oxidase (DAO;EC1.4.3.3) has been isolated from a BALB/c mouse kidney cDNA library by hybridization with the cDNA for the porcine enzyme. Analysis of the nucleotide (nt) sequence of the clone revealed that it has a 1647-nt sequence with a 5'-terminal untranslated region of 68 nt, an open reading frame of 1035 nt that encodes 345 amino acids (aa), and a 3'-terminal untranslated region of 544 nt that contains the polyadenylation signal sequence, ATTAAA. The deduced aa sequence showed 77 and 78% aa identity with the porcine and human enzymes, respectively. Two catalytically important aa residues, Tyr228 and His307, of the porcine enzyme, were both conserved in these three species. RNA blot hybridization analysis indicated that a DAO mRNA, of 2 kb, exists in mouse kidney and brain, but not liver. Synthesis of a functional mouse enzyme in Escherichia coli was achieved through the use of a vector constructed to insert the coding sequence of the mouse DAO cDNA downstream from the tac promoter of plasmid pKK223-3, which was designed so as to contain the lac repressor gene inducible by isopropyl-beta-D-thiogalactopyranoside. Immunoblot analysis confirmed the synthesis and induction of the mouse DAO protein, and the molecular size of the recombinant mouse DAO was found to be identical to that of the mouse kidney enzyme. Moreover, the maximum activity of the mouse recombinant DAO was estimated to be comparable with that of the porcine DAO synthesized in E. coli cells.

Amino Acid Sequence↗
PubMed1990

Antifungal drugs affect adherence of Candida albicans to acrylic surfaces by changing the zeta-potential of fungal cells.

The effect of sub-inhibitory concentrations of antifungal drugs on the adherence of Candida albicans to acrylic surfaces was investigated. Among five antifungals tested, azalomycin F and aculeacin A significantly enhanced the adherence. The zeta-potential of fungal cells was affected by antifungal drugs, whereas no significant change in cell surface hydrophobicity was observed. The relationship obtained between the change in the adherence and that in zeta-potential suggests that the enhanced adherence was caused by decreased electric repulsive forces.

Acrylates↗
PubMed1990

Molecular cloning and sequence analysis of a cDNA encoding a porcine kidney renin-binding protein.

A complementary DNA encoding a renin-binding protein (RnBP) has been isolated from a porcine kidney cDNA library by immunological screening of in vitro translation products from the cDNAs. Analysis of the nucleotide sequence of the clone revealed a 1,342-nucleotide sequence with a 5'-terminal untranslated region of 52 nucleotides, an open reading frame of 1,206 nucleotides that encodes 402 amino acids, and a 3'-terminal untranslated region of 84 nucleotides that contains the polyadenylation signal sequence, AATAAA. The predicted amino acid sequence contains no hydrophobic amino-terminal sequence and does not show significant homology to those of other identified proteins. The in vitro translated RnBP was found to have the same molecular weight, 42,000, as that of the purified RnBP from porcine kidney on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and it formed a complex with renin purified from porcine kidney, which indicates that the cDNA encodes a functional RnBP without a propeptide sequence. The RnBP cDNA probe hybridized to a 1.5-kilobase mRNA in kidney, liver, adrenal, and pituitary glands, the amount being much greater in kidney than in the other tissues. Southern blot analysis showed the presence of a unique gene for RnBP in the porcine genome.

Amino Acid Sequence↗
PubMed1990

Parathyroid hormone-related protein is a possible autocrine growth inhibitor for lymphocytes.

Adult T-cell leukemia (ATL)-related cells have the ability to produce a newly-isolated calcium-regulating protein, parathyroid hormone-related protein (PTHrP). The present study revealed that lectin-stimulated normal lymphocytes produce immunoreactive (IR)-PTHrP. When the T-cell-enriched fraction was purified from normal lymphocytes and then treated with lectin, a similar amount of IR-PTHrP was detected, suggesting that IR-PTHrP is an actual product of T-lymphocytes. A biologically active fragment of PTHrP, PTHrP(1-34), suppressed DNA synthesis in lectin-stimulated lymphocytes at concentrations greater than 50 pg/mL; the same concentration range of IR-PTHrP detected in the cultured media of lectin-stimulated lymphocytes. Therefore, it is reasonable to postulate that PTHrP is a cytokine inhibiting the cellular growth of normal lymphocytes.

Cell Division↗
PubMed1990

A triazine dye, cibacron blue 3G-A induces Staphylococcus aureus to form giant clusters.

Cibacron blue 3G-A (CB), one of the triazinyl dyes commonly used as a ligand for affinity chromatography, induced staphylococci to form giant clusters without affecting cell viability. It was demonstrated that the cluster formation was not a mechanical aggregation of cells by CB, but a consequence of its biological effect on staphylococcal growth. Clusters induced by CB did not form a regular arrangement, and were different from clusters induced by SDS or polyanethole sulfonate.

Cell Division↗
PubMed1990

Parathyroid hormone-related protein in tumor tissues obtained from patients with humoral hypercalcemia of malignancy.

We assessed the relationship between the parathyroid hormone-related protein (PTHrP) and the development of humoral hypercalcemia of malignancy (i.e., hypercalcemia due to the production by solid tumors of hypercalcemic factors) by assaying tumor extracts from hypercalcemic and normocalcemic patients with cancer for immunoreactive PTHrP contents. Immunoreactive PTHrP was demonstrated in extracts of 21 of 22 tumor tissues obtained from 22 patients with humoral hypercalcemia of malignancy. Immunoreactive PTHrP was rarely seen in tumor tissue extracts obtained from 34 normocalcemic patients with cancer and two hypercalcemic patients with cancer with severe bone metastases. Gel filtration studies of tumor extracts revealed a molecular-size heterogeneity of immunoreactive PTHrP. These radioimmunoassay data indicate a close relationship between detection of PTHrP in tumor tissues and the development of humoral hypercalcemia of malignancy.

Adult↗
PubMed1990

Degradation peptides of secretin after storage in acid and neutral aqueous solutions.

During the storage of secretin in acid and neutral aqueous solutions, five degradation peptides (A1, A2, A3, A4, A5) and one degradation peptide (N1) were produced, respectively. They were isolated in pure form by HPLC, and the intramolecular structures were studied by a combination of amino acid analysis, enzymatic digestions, HPLC, and Fab-mass spectroscopy. Although the degradation peptides are composed of the same amino acids as secretin after acid hydrolysis (except A1 and A4 which are cleavage products S16-27 and S4-27, respectively), reversed-phase HPLC analysis of their digestive fragments with trypsin and alpha-chymotrypsin are different from those of secretin. By Fab-mass spectroscopy, the m/z values for the S1-6 fragments obtained from secretin, A2, and A3 were 663, 663, and 645, respectively. When S1-6 from A2 was treated with aminopeptidase M, a fragment obtained was identical with the synthetic beta-aspartyl3 S3-6, as determined by HPLC. The A2 and N1 peptides are completely the same based on various chemical analyses. The A3 peptide can also be rapidly degraded to secretin and beta-aspartyl3 secretin. Consequently, A1 and A4 are concluded to be the cleavage peptides of secretin, S16-27 and S4-27, respectively, A2 and N1 are concluded to be beta-aspartyl3 secretin, and A3 is concluded to be aspartoyl3 secretin.

Amino Acid Sequence↗
PubMed1990

Mechanism and kinetics of secretion degradation in aqueous solutions.

In order to clarify the mechanism of secretin degradation in aqueous solutions, the formation of degradation products from secretin, aspartoyl3 secretin and beta-aspartyl3 secretin was investigated; the stabilities of these three peptides were investigated as well. Aspartoyl3 secretion and beta-aspartyl3 secretin, degradation peptides produced during the storage of secretin in aqueous solutions, were isolated by preparative reversed-phase HPLC (RP-HPLC). The amounts of secretin and its two degradation peptides resulting from storage of secretin in various buffer solutions (pH 2.3 to 10.0, mu = 0.5 M, 60 degrees C) were determined by analytical RP-HPLC. Secretin and the isolated degradation peptides were stored separately in various aqueous buffer solutions resulting in the degradation of each peptide. A mixture of secretin and its degradation or cleavage peptides was formed in each solution. The observed degradation rates for each peptide approximately followed first-order kinetics. The pH-rate profiles for conversion of secretin and beta-aspartyl3 secretin were similar, while that for aspartoyl3 secretin was different from these two. Aspartoyl3 secretin was more stable than the others at pH 2.3 to 4.0, but it was easily degraded between pH 5.0 and 10.0. Investigation of aspartoyl3 secretin degradation showed that its degradation was related to the pH value of the solution, and that hydroxide ion catalyzes the ring opening of the aspartoyl peptide. Secretin was most stable in pH 7.0 buffer solution and more stable in acidic solutions than in alkaline solutions. Secretion was mainly degraded through the following pathways: cleavage peptides reversible secretin in equilibrium aspartoyl peptide in equilibrium beta-aspartyl peptide vector cleavage peptides.

Buffers↗
PubMed1990

Sustained release of insulin by double-layered implant using poly(D,L-lactic acid).

This report describes the advantage of double-layered implants using low molecular weight poly(DL-lactic acid) in the sustained release of insulin. The double-layered implant consisted of a polymer matrix containing insulin and a polylactic acid layer which was coated partially on one of the surfaces of the insulin:polymer matrix. The double-layered implants were compared with single-matrix implants from the standpoint of the in vitro dissolution test and in vivo performance. In vitro release rates were controlled by changing the amount of poly(DL-lactic acid) used in the polymer layer. In an in vivo test using diabetic animals, the double-layered implants provided a sustained release of insulin for 19 d, as judged by the changes in blood glucose levels and serum insulin levels after the subcutaneous implantation.

Animals↗
PubMed1990

Macular oscillatory potentials in humans. Macular OPs.

Studies of the focal macular electroretinogram (ERG) have been made with special reference to oscillatory potentials (OPs) by using a fundus monitoring system in humans. Human macular OPs consist of 3 to 4 wavelets (mean peak interval, approximately 6.5 msec). The distribution of OPs in relation to those in a- and b-waves was studied. The amplitudes of a-waves, b-waves, and OPs of the upper macula were significantly larger than those of the lower macula. The distribution of OPs is relatively sparse in the fovea, becoming more dense than the a- and b-waves from the fovea toward the parafovea, and differing even more toward the perifovea. There was no statistical difference of amplitude in a- and b-waves between nasal and temporal macula. The amplitude of OPs in the temporal macula, however, was significantly larger than in the nasal macula. In some macular diseases, such as diabetic maculopathy, cystoid macular edema, or the convalescent stage of central serous chorioretinopathy, macular OPs were selectively reduced, leaving the a- and b-waves intact. Macular OPs can provide a new aspect of macular function and can be a sensitive indicator to assess that function in macular diseases.

Adult↗
PubMed1990

Purification of staphylococcal exfoliative toxin by high pressure liquid chromatography.

Exfoliative toxin (ET) isolated from a clinical strain of Staphylococcus aureus was purified to homogeneity, using a 3-step HPLC system. NH2-terminal 20 amino residues of purified ET was found to be identical with ETA of S. aureus TA (7), S. aureus TC16 (9) and S. aureus ZM (10), but stability of purified ET was completely different from that of ETA. This purification system gave a high yield of pure ET, which exhibited higher purity than specimens purified by more complicated and time-consuming procedures. It is useful for small-scale purification for the comparative study of ET and easy to scale up for preparative purification.

Amino Acid Sequence↗
PubMed1990

Gene expression of D-amino acid oxidase in rabbit kidney.

Although D-amino acid oxidase (DAO) [EC 1.4.3.3] activity in rabbit kidney extract was undetectable, protein immunoreactive toward rabbit anti-pig kidney DAO antiserum and RNAs that hybridized with fragments of human and pig DAO cDNAs were detected distinctly in the rabbit kidney. A cDNA clone, RD22, was isolated from the rabbit kidney cDNA library by hybridization with a fragment of human DAO cDNA. Analysis of the nucleotide sequence revealed a 2,018 nucleotide sequence encoding a protein consisted of 347 amino acids. The number of amino acid residues was identical with those of human and pig DAOs, and the amino acid sequence showed 80 and 83% identity with pig and human DAOs, respectively. RNAs that hybridized with RD22 DNA fragment also existed in rabbit kidney, and their sizes were the same as those of the RNAs detected with the human and pig DAO cDNA fragments. RD22-derived protein was hardly synthesized by an in vitro expression system. However, a cDNA fragment lacking most of the 5'-untranslated region and its mutants containing base changes around the initiation codon did direct protein synthesis. Moreover, the protein derived from the partial cDNA fragment containing a large part of the coding region sequence showed immunoreactivity toward anti-pig DAO antiserum. The results suggest that one of the causes of the very poor synthesis of DAO protein in rabbit kidney is translational suppression in the synthetic process.

Amino Acid Sequence↗
PubMed1990

Adherence of Staphylococcus aureus to cultured epidermal cells during differentiation.

The adherence of clinical isolates of Staphylococcus aureus to cultured mouse epidermal cells was studied. Adherence of the isolates to the cells varied from strain to strain. When epidermal cell differentiation was induced by raising the calcium concentration in the medium, three out of 10 strains tested adhered better to calcium-induced differentiated cells than to undifferentiated cells, and one strain demonstrated higher adherence to undifferentiated cells than to differentiated cells. No significant difference between the adherences to both types of epidermal cells was observed with the other six strains. No relationship was observed between adherence and surface hydrophobicity with bacterial cells. Lipoteichoic acid and N-acetyl sugars caused limited inhibition of adherence. The adherence assay method employed in this study is useful for investigating the effects of epidermal cell differentiation on bacterial adherence in vitro.

Animals↗