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C Miyamoto

Publications and source records attributed to C Miyamoto.

At least 37 records · Page 2Linked to original sources
PubMed1997

A homoserine lactone autoinducer regulates virulence of an insect-pathogenic bacterium, Xenorhabdus nematophilus (Enterobacteriaceae).

N-beta-Hydroxybutanoyl homoserine lactone (HBHL), the autoinducer of the luminescent system of Vibrio harveyi, has been identified as the first small compound to restore virulence to avirulent mutants of Xenorhabdus nematophilus. HBHL stimulated the level of lipase activity excreted by avirulent X. nematophilus and lowered the phenoloxidase activity in the hemolymph of insects infected with X. nematophilus, parameters that are both associated with insect pathogenesis. Moreover, mortality of the insects infected with avirulent X. nematophilus was restored upon injection with HBHL. Chloroform extraction of medium conditioned with wild-type but not avirulent X. nematophilus led to the isolation of a compound with the same chromatographic mobility as HBHL as well as the ability to stimulate the luminescence of a dim autoinducer-dependent mutant of V. harveyi. Transfer of the V. harveyi lux operon into avirulent and wild-type X. nematophilus generated dim and bright luminescent strains, respectively, which responded to HBHL and an agonist and antagonist in a manner analogous to their effects on the luminescence of dim autoinducer-deficient and bright wild-type strains of V. harveyi, indicating that similar HBHL-dependent regulatory systems exist in these two bacterial species.

Animals↗
PubMed1996

A point mutation within each of two ATP-binding motifs inactivates the functions of elongation factor 3.

We have investigated how point mutations in the two ATP-binding motifs (G(463)PNGCGK(469)ST and G(701)PNGAGK(707)ST) of elongation factor 3 (EF-3) affect ribosome-activated ATPase activity of EF-3, polyphenylalanine synthesis, and growth of Saccharomyces cerevisiae. The point mutation impaired the ribosome-activated ATPase activity of EF-3, when glycine(463 and 701) and lysine(469 and 707) were replaced with valine and arginine, respectively. Thus, each glycine and lysine residue in both ATP-binding motifs is indispensable for EF-3's binding with ATP and the ensuing generation of ribosome-activated ATPase activity. Additionally, the mutant EF-3s did not catalyze polyphenylalanine synthesis in vitro when each glycine(463 and 701) was replaced with valine. The mutant EF-3s did not support cell growth in TEF3-disrupted S. cerevisiae, when each lysine(469 and 707) and glycine(463) was replaced with arginine and valine, respectively. Thus, each of the two ATP-binding motifs of EF-3 is indispensable for the ribosome-activated ATPase activity of EF-3, which is required for protein synthesis and cell growth in S. cerevisiae.

Adenosine Triphosphatases↗
PubMed1996

In vitro evaluation of iodine-125-labeled monoclonal antibody (MAb 425) in human high-grade glioma cells.

Human high-grade glioma cell lines (A1207, U-87MG, U-373MG, and F39) with high levels of epidermal growth factor receptor (EGF-R) expression were incubated for 2-48 h with 1 microCi/ml of the EGF-R-specific 125I-MAb 425 and measured for surface-bound, cytoplasmic, and nuclear radioactivity. The A1207 and U-373MG cell lines showed the highest surface-bound radioactivity with 215.9 +/- 8.7 nCi (30 h) and 287.8 +/- 23.2 nCi (24 h)/10(6) cells, respectively, whereas the U-87MG and the F39 cell lines bound significantly less antibody (48.8 +/- 5.4 nCi [48 h] and 31.1 +/- 0.7 nCi [24 h]). Surface-bound antibody was efficiently internalized into the cytoplasm. The U-373MG, U-87MG, and A1207 cell lines achieved 19.8% +/- 2.1 internalization of the surface-bound antibody in contrast to > 40% for the F39 cell line. Only the A1207 cell line showed significant nuclear radioactivity. There was no correlation between the reported EGF-R number and amount of antibody bound or internalized. We conclude that binding and uptake of the 125I-MAb 425 is specific for human glioma cells and shows saturation kinetics independent of receptor density.

Antibodies, Monoclonal↗
PubMed1996

Mycoplasma hyorhinis infection levels in lungs of piglets with porcine reproductive and respiratory syndrome (PRRS).

The infection levels of Mycoplasma hyorhinis, M. hyopneumoniae and M. hyosynoviae in the lung of piglets were examined in relation to porcine reproductive and respiratory syndrome (PRRS). These animals consisted of 43 PRRS piglets with PRRS, 2 piglets infected with PRRS virus but symptom-free, and 10 control piglets free of PRRS virus and its antibody. M. hyorhinis was isolated from 40 of the 43 PRRS piglets, from 1 of the 2 latent infected piglets and from 3 of the 10 control piglets. The number of M. hyorhinis isolated from the lungs of PRRS piglets was more than 10(5) CFU/g, but those isolated from the latent infected piglets and the control piglets were less than 10(3) CFU/g. In addition to this, Haemophilus parasuis and Pasteurella spp. were frequently isolated from the piglets with PRRS (51.2% and 25.6%, respectively). On the other hand, M. hyopneumoniae was isolated from only 4 of 55 piglets tested, and M. hyosynoviae was not isolated. M. hyorhinis was also detected directly in the lung emulsion samples from almost all the PRRS piglets using a polymerase chain reaction-based method.

Animals↗
PubMed1996

A simple preparation of mycoplasmal DNA template for PCR from biological samples using effective surfactants.

To prepare mycoplasmal DNA template for PCR from biological samples rapidly and easily, surfactants which can solubilize cell membranes effectively were investigated. 3-[3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) was considered an effective surfactant. This surfactant could solubilize mycoplasma cell membrane without suppressing the polymerase reaction. In addition, proteinase K treatment played an important role in preparing mycoplasmal DNA template from a simulated biological sample. It was therefore considered that a combination of proteinase K- and CHAPS- added lysis buffer would be more useful in preparing mycoplasmal DNA template. We could detect PCR products by using the lysis buffer with a simulated lung emulsion sample containing mycoplasma organisms at 10(4) CFU per gram.

Animals↗
PubMed1995

Characterization and gene cloning of 1,3-beta-D-glucan synthase from Saccharomyces cerevisiae.

1,3-beta-D-Glucan synthase of Saccharomyces cerevisiae was solubilized and purified up to 700-fold by product entrapment. The specific activity of the partially purified enzyme was around 4 mumol glucose incorporated.min-1.mg protein-1. In SDS/PAGE, enrichment of a 200-kDa protein was clearly observed in parallel with the increase in specific activity. mAbs that could immunoprecipitate the 1,3-beta-D-glucan synthase activity were isolated, and some of them also recognized this 200-kDa protein in the Western blot. Internal amino acid sequences of this 200-kDa protein were determined after lysyl endopeptidase digestion. With the information of these amino acid sequences, we cloned two genes, GSC1 and GSC2 (glucan synthase of S. cerevisiae 1 and 2), which are very similar to each other (88% at the amino acid level); hydropathy profiles of both proteins suggest that these genes encode integral membrane proteins which can be assumed to have approximately 16 transmembrane domains. Disruption of each gene was not lethal, but disruption of both genes was lethal. The 1,3-beta-D-glucan synthase activities of membrane and partially purified enzyme of gsc1::URA3 cells were significantly lower than those of the wild-type and gsc2::LEU2 cells.

Amino Acid Sequence↗
PubMed1995

Separable binding sites for the natural agonist endothelin-1 and the non-peptide antagonist bosentan on human endothelin-A receptors.

A three-dimensional model for the transmembrane domains of human endothelin-A receptor was built using structural information from bacteriorhodopsin and sequence alignment to other guanine-nucleotide-binding regulatory(G) protein-coupled receptors. Based on this model, 18 amino acids located at the inside of the receptor were mutated and analyzed for binding of the natural ligand endothelin-1 and bosentan, a recently described potent orally active endothelin antagonist [Clozel, M., Breu, V., Gray, G., Kalina, B., Löffler, B.-M., Burri, K., Cassal, J.-M., Hirth, G., Müller, M., Neidhart, W. & Ramuz, H. (1994) Pharmacological characterization of bosentan, a new potent orally active nonpeptide endothelin receptor antagonist, J. Pharmacol. Exp. Ther. 270, 228-235]. Mutation of Gly97, Lys140, Lys159, Gln165 and Phe315, located in transmembrane region 1, 2, 3, 3, and 6, respectively, caused reduced specific binding of 125I-labelled endothelin-1, despite an expression level similar to wild-type endothelin-A receptor. Mutation of Tyr263, Arg326 and Asp351 preserved endothelin-1 binding but caused reduced binding of bosentan. These amino acids, located on transmembrane regions 5, 6 and 7, respectively, are conserved among endothelin-A and endothelin-B receptors but not in other G-protein-coupled receptors. These observations demonstrate a dissociation of the binding site for the peptidic natural agonist endothelin-1 and the synthetic non-peptide antagonist bosentan. They provide the molecular basis for bosentan being a specific antagonist for both, endothelin-A as well as endothelin-B receptors and may in combination with studies on structure/activity relationship support the design of novel and more potent endothelin receptor antagonists.

Amino Acid Sequence↗
PubMed1995

Ligand binding domain of the human endothelin-B subtype receptor.

We have employed both protein chemical and molecular biological approaches to determine the ligand binding domain of the endothelin-B subtype (ETB) receptor. The human ETB receptor purified from human placenta by using affinity chromatography was cross-linked with 125I-labeled endothelin-1 (ET-1) and then incubated in the presence of trypsin or thermolysin under nondenaturing conditions. The N-terminal amino acid sequence of the radiolabeled polypeptide encompassed approximately 115 amino acid residues starting from Ile85 of the human ETB receptor. This was confirmed by experiments in which the binding activity of endothelin-1 to various chimeric endothelin receptors was monitored in the presence and absence of competitive endothelin receptor antagonists such as BQ-123 and bosentan. The region from Ile138 to Ile197 (60 amino acid residues) of the ETB receptor was found to interact with both antagonists. Therefore, this sequence was determined to be the ligand binding domain. In addition, we found that part of the N-terminal domain in close proximity to the first transmembrane region was required for the ligand binding activity of the ETB receptor, and the 12 amino acid residues from Ser390 to Leu401 at the proximal cytoplasmic tail are perhaps necessary to maintain the ligand binding site in active form. The cysteine rich region from residue 400 to residue 403 in the C-terminus of the ETB receptor is involved in coupling of the guanine nucleotide-binding regulatory protein for ET-1-induced signal transduction.

Amino Acid Sequence↗
PubMed1994

Thallium imaging for brain tumors with results measured by a semiquantitative index and correlated with histopathology.

BACKGROUND: The optimal management of patients with brain tumors requires knowledge of the tumor characteristics upon presentation and the discovery of recurrence after therapy. Thallium-201 (Tl-201) chloride has shown varying uptake in tumors, depending on their viability and the type and degree of malignancy. This study explores the diagnostic potential of thallium imaging in patients with brain tumors. METHODS: Forty-three Tl-201 single photon emission computed tomographic scintigrams were performed on 40 patients with intracranial neoplasms, nearly equally divided between patients with no prior treatment and patients who had prior treatment and were suspected to have recurrent tumor and/or radiation necrosis. A thallium tumor index was calculated as the ratio of counts for a region of interest drawn in the lesion area and its mirror image in normal brain tissue. A two-tailed Student's t test was performed to compare the thallium index and histopathologic findings. RESULTS: A value of 1.5 for the thallium tumor index allowed for the best correlation between the prediction of malignancy and the histopathologic results. In the pretreatment group, a thallium tumor index greater than 1.5 correlated with high grade malignancy, and less than 1.5 correlated with either a well differentiated astrocytoma or benign cyst. In the posttreatment group, a thallium tumor index greater than 1.5 correlated with recurrent and/or residual malignant tumor. CONCLUSIONS: For those patients undergoing initial evaluation, the thallium study can help in the differential diagnosis of an intracranial mass lesion and offers confirmation of results of biopsy. For those patients who already have received treatment, the study can be used to detect recurrent or residual tumor.

Adult↗
PubMed1994

Identification of a region of the human endothelin ETA receptor required for interaction with bosentan.

Bosentan (Ro 47-0203, 4-tert-butyl-N-[6-(2-hydroxy-ethoxy)-5-(2- methoxy-phenoxy)-2,2'-bipyrimidin-4-yl]-benzenesulfonamide) is a new non-peptidic mixed antagonist of endothelin receptors whose binding activity was two orders higher for the endothelin ETA receptor than that for the endothelin ETB receptor. To identify which region of the human endothelin ETA receptor interacts with bosentan, we created various chimeric endothelin receptors containing domains from the endothelin ETA and ETB receptors in Chinese hamster ovary cells and studied the effect of bosentan on the binding of endothelin-1 to the chimeric receptors. We found that the chimeric endothelin ETB receptor containing domains from the endothelin ETA receptor, the second extracellular region including the proximal transmembrane region (B-region) revealed an affinity toward bosentan which was similar to that of the endothelin ETA receptor. In contrast, the chimeric endothelin ETA receptor, containing the B-region of the endothelin ETB receptor, reduced the binding affinity to the level of the endothelin ETB receptor. Since bosentan competes with endothelin-1 for binding to the endothelin ETA receptor, this receptor antagonist seems to interact with the (140-144) KLLAG sequence located at the carboxylterminus of the second transmembrane region of the endothelin ETA receptor, required for the natural ligand binding.

Amino Acid Sequence↗
PubMed1994

Identification of specific regions of the human endothelin-B receptor required for high affinity binding with endothelin-3.

To investigate the endothelin-3 (ET-3) binding region of the endothelin-B (ETB) receptor, we have transiently produced various chimeric endothelin receptors in transfected Chinese hamster ovary cells. Using 125I-ET-1 as the radioactive ligand in the displacement experiment, the replacement of both the second and third extracellular regions including the flanking transmembranes of the ETB receptor with the corresponding domains of the endothelin-A (ETA) receptor, increased the apparent Ki value for ET-3 from 5 x 10(-11) M to 10(-8) M. The affinity of this chimeric receptor, ETB-BC, for ET-3 was about two orders lower than ETB yet one order higher than ETA. Previously we have reported the involvement of Lys-140 located in the C-terminus of the second transmembrane region of the ETA receptor for ET-1 binding (Eur. J. Biochem., 220, 37-43, 1994). To assess the importance of the corresponding Lys-161 of the ETB receptor in ET-3 binding, we have replaced it with Ile in the ETB receptor. The mutant receptor had a 5.6-fold reduction in its affinity for ET-3, but its affinity for ET-1 remained similar. These results demonstrate that Lys-161 of the receptor is important for high affinity binding with ET-3 which, in part, confers the non-selective binding characteristics of the ETB receptor for ET isopeptides.

Amino Acid Sequence↗
PubMed1994

Identification of a ligand-binding site of the human endothelin-A receptor and specific regions required for ligand selectivity.

To investigate the ligand-binding site of the human endothelin-A-receptor subtype (ETA), we have produced various chimeric and mutated receptors in chinese hamster ovary cells. The substitution of Lys140 with Ile located in the C-terminus of the second transmembrane region caused a 13-fold reduction in affinity for endothelin-1 (ET-1) and 3.6-fold lower Bmax than those values for the original receptor. Correspondingly, the mutated ETA receptor with the Lys140-->Ile substitution failed to induce an increase in the intracellular calcium concentration in the presence of 1 nM ET-1. Thus, the Lys140 in the ETA receptor is important in ligand binding. ETA and ETB receptors possess the ET isopeptides selective and non-selective binding activities, respectively. Displacement experiments and the binding of 125I-ET-3 to various chimera receptors demonstrated that both the third and fourth extracellular regions, including the flanking transmembrane regions, are responsible for the ligand-binding selectivity of the ETA receptor.

Amino Acid Sequence↗
PubMed1994

Indium-111-labeled anti-EGFr-425 scintigraphy in the detection of malignant gliomas.

BACKGROUND: The monoclonal antibody anti-epidermal growth factor receptor (EGFr) antibody-425, against the epidermal growth factor receptor, has the potential to bind specifically to gliomas and not normal brain tissue. A prospective study was conducted (1986-1988) to evaluate the use of Indium-111 (111In)-labeled anti-EGFr-425 in the localization of gliomas before radioimmunotherapy with Iodine-125 (125I)-labeled anti-EGFr-425. METHODS: Twenty-eight patients with intracranial neoplasms were injected intravenously with an average dose of 2.2 mCi 111In-labeled anti-EGFr-425. Planar and single-photon emission computed tomography scans were performed after 48 and 72 hours. Control studies also were performed in two cases with 111In-labeled Co 17-1A (an antibody to colorectal cancer) and in one case with unlabeled 111In chloride. RESULTS: The immunoscintigraphic findings were generally in good agreement with computerized tomographic findings. The definitive diagnosis was established by biopsy findings: 23 gliomas (1 Grade I, 5 Grade II, 6 Grade III, and 11 Grade IV), 1 meningioma, and 4 metastatic lesions. The localization of gliomas with 111In-labeled anti-EGF-425 had a sensitivity of 0.96, a specificity of 0.60 and an accuracy of 0.90. CONCLUSION: Immunoscintigraphy with 111-In labeled anti-EGFr-425 can be useful in the management of malignant gliomas, especially before radioimmunotherapy with 125I-labeled anti-EGFr-425.

Adult↗
PubMed1994

Intracavitary brachytherapy for squamous cell carcinoma of the esophagus.

From September 1983 through January 1989, 8 patients with a diagnosis of squamous cell carcinoma of the esophagus AJCC stages I to III, were treated with intracavitary brachytherapy following external irradiation. Within 3 months after completion of treatment, there was radiographic evidence of complete response in 6 of the 8 patients. None of the responders recurred locally. All the patients have died, with a mean survival of 17 months. One patient died of a stroke 2 1/2 years after completion of treatments without clinical evidence of esophageal cancer. Treatment was well tolerated, with minimal acute toxicity, and one patient developed stricture at the site of tumor, which was successfully treated by dilatation. Addition of systemic chemotherapy to this regimen warrants investigation.

Aged↗
PubMed1994

Cloning of the Saccharomyces cerevisiae gene whose overexpression overcomes the effects of HM-1 killer toxin, which inhibits beta-glucan synthesis.

A gene whose overexpression can endow Saccharomyces cerevisiae cells with resistance to HM-1 killer toxin was cloned from an S. cerevisiae genomic library. This gene, designated HKR1 (Hansenula mrakii killer toxin-resistant gene 1), contains a 5.4-kb open reading frame. The predicted amino acid sequence of the protein specified by HKR1 indicates that the protein consists of 1,802 amino acids and is very rich in serine and threonine, which could serve as O-glycosylation sites. The protein also contains two hydrophobic domains at the N-terminal end and in the C-terminal half, which could function as a signal peptide and transmembrane domain, respectively. Hkr1p is found to contain an EF hand motif of the calcium-binding consensus sequence in the C-terminal cytoplasmic domain. Thus, Hkr1p is expected to be a calcium-binding, glycosylated type I membrane protein. Southern and Northern (RNA) analyses demonstrated that there is a single copy of the HKR1 gene in the S. cerevisiae genome, and the transcriptional level of HKR1 is extremely low. Gene disruption followed by tetrad analysis showed that HKR1 is an essential gene. Overexpression of the truncated HKR1 encoding the C-terminal half of Hkr1p made the cells more resistant to HM-1 killer toxin than the full-length HKR1 did, demonstrating that the C-terminal half of Hkr1p is essential for overcoming the effect of HM-1 killer toxin. Furthermore, overexpression of HKR1 increased the beta-glucan content in the cell wall without affecting in vitro beta-glucan synthase activity, suggesting that HKR1 regulates beta-glucan synthesis in vivo.

Amino Acid Sequence↗
PubMed1994

Chronic actinic dermatitis: a clinical and photobiological study in 6 Japanese patients.

Chronic actinic dermatitis (CAD) has distinct clinical features different from polymorphous light eruption (PLE). In order to clarify the difference between CAD and PLE, not only in clinical histories and findings but also in photobiological and histopathological reactions to phototests, we investigated 6 Japanese patients with CAD and performed provocative phototests which are our standardized methods in diagnosing PLE. On provocative phototests in CAD, pruritic papules were reproduced with smaller doses of UVB, at longer hours after irradiation (48-72 h) and they lasted for more days than in patients with PLE. Our study demonstrated that although milder cases of CAD and severe cases of PLE could not be distinguished clearly based on photobiological reactions alone, typical cases of CAD showed completely different provocative phototest results from those of PLE.

Adult↗
PubMed1994

Differential regulation of c-fos gene expression by two types of human endothelin receptor in Chinese hamster ovary cells.

To investigate the nuclear signalling pathway induced by endothelin (ET) isopeptides, we have established permanent Chinese hamster ovary (CHO) cell lines, CHO-ETA/fos-lacZ and CHO-ETB/fos-lacZ, that produce both a c-fos-beta-galactosidase fusion protein and either the type A or the type B human ET receptor. These cell lines permitted a colorimetric measurement of c-fos expression, which was induced by the signal transduction system with ET receptors and ET isopeptides. We found that the ET-1-dependent c-fos expression was so efficient that it could respond to low concentrations (even a physiological concentration) of ET-1. For example, CHO-ETA/fos-lacZ and CHO-ETB/fos-lacZ responded to ET concentrations of 5 x 10(-9) M and 5 x 10(-13) M respectively. Using this highly sensitive system, the H-7 sensitive protein kinase was found to be involved in signal transduction mediated by ETA, and also partly in the ETB-mediated pathway. These lines of evidence suggest that c-fos expression occurs through at least two different pathways, depending on the concentration of ET in plasma.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗