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T Murayama

Publications and source records attributed to T Murayama.

At least 145 records · Page 8Linked to original sources

A ras homologue of Neurospora crassa regulates morphology.

In order to study the role of signal transduction pathways in the regulation of morphology in Neurospora crassa, we cloned and characterized a ras homologue, termed NC-ras2. The predicted protein product of this gene is composed of 229 amino acid residues and contains all the consensus sequences shared by the ras protein family. The gene is located in linkage group V. An NC-ras2 disruptant showed morphological characteristics very similar to those of the smco7 mutant, which also maps to linkage group V. Nucleotide sequence analysis revealed that the smco7 mutant harbored a single base deletion in the NC-ras2 gene, which is predicted to result in the truncation of the protein product. Introduction into the smco7 mutant of an NC-ras2 clone yielded stable transformants with a wild-type phenotype. The smco7 mutant exhibited very slow hyphal growth and the rate of conidial formation was approximately one two-hundredth of wild type. The smco7 mutation causes both the changes in the pattern of hyphal growth and the defects in cell wall synthesis. Both the diameter and the length of the apical compartment were shorter in the hyphae of the smco7 mutant. These results suggest that NC-ras2 is identical to smco7, and that the signal transduction pathway mediated by the NC-ras2 protein regulates the apical growth of hyphae, cell wall synthesis, and conidial formation in N. crassa.

Amino Acid Sequence↗

Effects of K+ channel blockers and K+ ionophore on isoprenaline-induced secretion of amylase from rat parotid acini.

The involvement of K+ channels in regulating secretion of amylase from isolated rat parotid acini was studied in conjunction with beta-adrenoceptor function. It was observed that increasing the concentration of extracellular K- or adding K+ channel blockers enhanced the secretion of amylase. Among several K+ channel blockers, tetraethylammonium, apamin and charybdotoxin were effective to enhance secretion by 48, 69 and 84%, respectively. Glibenclamide was without effect. A low concentration of isoprenaline (10(-7) M) enhanced secretion by 154% and its simultaneous application with tetraethylammonium gave a synergistic effect, producing 371% stimulation. Combination of tetraethylammonium and a low concentration of carbachol (10(-6) M) did not give the synergistic effect. Isoprenaline at the concentration of 10(-6) M enhanced secretion by 313% and this was reduced to 116% by 10(-5) M valinomycin, a K+ ionophore. Valinomycin was without effect on carbachol (10(-5) M)-induced secretion. Somatostatin (10(-5) M) and morphine (10(-4) M) also reduced isoprenaline-induced secretion of amylase. These results suggested the regulation of Ca(2+)-activated K+ channels by isoprenaline in amylase secretory processes in parotid acini.

Adrenergic beta-Agonists↗

Colon carcinoma glycoproteins carrying alpha 2,6-linked sialic acid reactive with Sambucus nigra agglutinin are not constitutively expressed in normal human colon mucosa and are distinct from sialyl-Tn antigen.

In human colon carcinoma, increased amounts of sialic acids have been found and correlated with tumor progression. Further, the degree of O-acetylation of sialic acid residues in normal mucosa is higher than in colon carcinoma. Thus, tumor-associated sialylated antigens may be constitutively expressed in O-acetylated form in normal mucosa unreactive with the respective monoclonal antibodies. We have earlier demonstrated a colon carcinoma-associated expression of alpha 2,6-linked sialic acid residues with the Sambucus nigra agglutinin (SNA). We report now that de-acetylation of normal and transitional colonic mucosa, in contrast to sialyl-Tn antigen, does not result in SNA binding. Further, the alpha 2,6-linked sialic acid recognized by SNA is distinct from that of sialyl-Tn antigen. This is confirmed by Northern blotting detecting transcripts for alpha 2,6 sialyltransferase of N-glycoproteins and measurement of activity for this sialyltransferase. Blot analysis by SNA of colon carcinoma cells revealed few reactive glycoproteins. Quantitative differences in lectin labeling and sialyltransferase activity were found in HCT116 colon carcinoma cell sub-lines. Our data suggest that SNA binding in human colon carcinoma is due to de novo expression of a specific sialic acid present on selected glycoproteins.

Acetylation↗

Inhibition of protein kinase C-dependent noradrenaline release by wortmannin in PC12 cells.

Wortmannin is a specific phosphatidylinositol 3-kinase inhibitor, and it inhibits secretion in many cell types. Previously we reported that high K+ and external ATP stimulated a Ca2+ influx and [3H]-noradrenaline ([3H]NA) release from rat pheochromocytoma cell line 12 (PC12) cells in the presence of extracellular CaCl2. Addition of phorbol 12-myristate 13-acetate (PMA) stimulated [3H]NA release by itself and enhanced the maximal responses of high K+ and ATP. In this study, we investigated the effects of wortmannin on NA release from PC12 cells. Wortmannin inhibited the [3H]NA release induced by high K+ and ATP, and the stimulatory effects of PMA, in a dose-dependent manner. Wortmannin caused 50% inhibition of high K+-induced [3H]NA release at a concentration of 2.78+/-0.68 microM (n = 5). The increased cytosolic free Ca2+ concentrations ([Ca2+]i), induced by ATP, were not inhibited by wortmannin. Wortmannin inhibited PMA-induced phosphorylation of a 80-kDa protein in the cytosol fraction of PC12 cells. Calphostin C, a specific protein kinase C inhibitor, also inhibited high K+-, ATP-, and PMA-induced NA release, and the phosphorylation of the 80-kDa protein induced by PMA. Mastoparan, an amphiphilic tetradecapeptide from wasp venom, stimulated NA release in the presence or absence of extracellular CaCl2. Neither wortmannin nor calphostin C inhibited the NA release induced by mastoparan. These findings suggest that wortmannin inhibits the PKC-dependent pathway, not [Ca2+]i mobilization, resulting in the inhibition of NA release from PC12 cells.

Adenosine Triphosphate↗

Transesophageal Doppler echocardiographic assessment of systolic and diastolic coronary blood flow velocities at baseline and during adenosine triphosphate-induced coronary vasodilation in chronic aortic regurgitation.

Few reports exist on the changes in systolic and diastolic coronary flow velocities (CFVs) at baseline and during coronary vasodilation in patients with chronic aortic regurgitation (AR). We examined the left anterior descending CFVs in 21 patients with AR (11 patients with mild AR and 10 patients with moderate to severe AR), 9 patients without AR (no AR group), and 6 patients who had undergone surgery for moderate to severe AR (postoperation group) with transesophageal Doppler echocardiography. Adenosine triphosphate (ATP) was infused into a peripheral right arm vein at four different doses (35, 70, 100, and 140 micrograms/kg/min). Coronary flow velocity response in systole and diastole was calculated as the ratio of systolic peak and mean and diastolic peak and mean CFVs during maximal ATP infusion to those at baseline. The systolic peak and mean CFVs and the diastolic peak and mean CFVs at baseline were significantly increased in the moderate to severe group compared with those in the other groups (p < 0.05, respectively). Systolic and diastolic CFVs were significantly increased during ATP infusions in the four groups. No significant differences of systolic and diastolic CFVs were observed among the four groups during maximal ATP infusion. The coronary flow velocity response calculated from the peak and mean diastolic CFVs were significantly decreased in the moderate to severe group (1.6 +/- 0.3 and 1.7 +/- 0.4) compared with those in the other three groups (3.6 +/- 0.7 and 3.2 +/- 1.1 in the no AR group, 2.6 +/- 0.6 and 2.5 +/- 0.4 in the mild group, and 2.5 +/- 0.7 and 2.4 +/- 0.6 in the postoperation group) (p < 0.05, respectively). In conclusion, the systolic and diastolic left CFVs at baseline appeared to be significantly increased in patients with moderate to severe chronic AR. However, the velocities during coronary vasodilation by ATP were equal to those in other groups, resulting in a decrease of coronary flow velocity response in systole and diastole.

Adenosine Triphosphate↗

Protein kinase A-dependent IL-6 production induced by calcitonin in human glioblastoma A172 cells.

In human glioblastoma A172 cells, interleukin-6 (IL-6) production was induced by interleukin-1 beta (IL-1 beta) and dibutyryl cyclic AMP. These cells have been shown to induce IL-6 production via a cAMP-protein kinase A system. Since calcitonin (CT) and calcitonin gene-related peptide (CGRP) are known to increase cAMP accumulation in murine and rat astrocytes, we examined whether these neuropeptides induced IL-6 production in A172 cells. Human CT and human CGRP increased IL-6 production and cAMP accumulation in a dose-dependent manner. A specific protein kinase A inhibitor, H-89, inhibited both CT- and CGRP-induced IL-6 production. CT and CGRP have been shown to cross-react with each other. To exclude the possibility of this cross-reactivity, we studied the additive effects of CT and CGRP and the inhibitory effects of specific inhibitors. When 100 nM CT was added, cAMP accumulation stimulated by 10 nM CGRP (the maximal dose) was increased. CGRP (8-37), a specific CGRP receptor inhibitor, inhibited cAMP accumulation and IL-6 production induced by CGRP, but did not inhibit these effects when they were induced by CT. Salmon CT (8-32), a specific inhibitor of the CT receptor, inhibited cAMP accumulation induced by CT, but did not inhibit the effect induced by CGRP. These results demonstrated that CT can induce IL-6 production via cAMP accumulation and the effects of CT are mediated via its own receptors.

Calcitonin↗

Specificity of polyclonal antibodies raised against a novel 24,25-dihydroxyvitamin D3-bovine serum albumin conjugant linked through the C-11alpha or C-3 position.

Novel hapten-carrier conjugants were prepared by coupling 11 alpha-hemiglutaryloxy-(24R)-24,25-dihydroxyvitamin D3 or (24R)-24,25-dihydroxyvitamin D3 [24,25(OH)2D3] 3-hemiglutarate with bovine serum albumin (BSA), to obtain an antibody with high specificity and affinity for use in 24,25(OH)2D3 immunoassay. The polyclonal antibodies showing high titre were each elicited in three or four rabbits against these two conjugants; the antibodies obtained from the former and the latter conjugants were expressed as Ab11 and Ab3, respectively. These had a much higher affinity for 24,25(OH)2D3 than that of the vitamin D binding protein (DBP). Specificity of the antibodies was investigated by crossreactivities with 11 related compounds in a radioimmunoassay (RIA) system. The Abll well discriminated the 1 alpha-hydroxylated metabolites such as 1,24,25(OH)3D3 (< or = 0.69%) and 1,25(OH)2D3 (< or = 0.25%), but significantly crossreacted with some side chain modified compounds such as (24S)-24,25-dihydroxyvitamin D3 [24S,25(OH)2D3] (> or = 67%), 25(OH)D3 (> or = 14%) and 25,26(OH)2D3 (> or = 23%). On the other hand, the Ab3 showed only negligible crossreactivities with the compounds having a different side chain structure such as 24S,25(OH)2D3 (< or = 3.0%), 25(OH)D3 (< 0.3%) and 25,26(OH)2D3 (< or = 0.53%). A significant crossreaction was found only with 1,24,25(OH)3D3 (> or = 68%). These results demonstrated that the Ab3 are promising for developing an immunoassay system which is much more specific and sensitive than conventional competitive protein binding assays based on DBP.

24,25-Dihydroxyvitamin D 3↗

Inhibition of inducible nitric oxide synthase expression by endothelin in rat glial cells prepared from the neonatal rat brain.

In primary cultured rat glial cells, a combination of inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) stimulates production of nitrite via expression of the inducible form of nitric oxide synthase (iNOS). In these cells, simultaneous addition of endothelin (ET) decreased iNOS expression and nitrite accumulation induced by TNF-alpha/IL-1beta. The inhibitory effect of ET on TNF-alpha/IL-1beta-stimulated iNOS expression appears to be mediated by ET(B) receptors, because (1) both ET-1 and ET-3 inhibited the effects of TNF-alpha/IL-1beta on iNOS expression and nitrite accumulation, (2) a selective ET(B) receptor agonist, Suc-[Glu(9),Ala(11,15)]-ET-1 (8-21) (IRL1620), decreased the effects of TNF-alpha/IL-1beta, and (3) a selective ET(B) receptor antagonist, N-cis-2,6-dimethylpiperidinocarbonyl-L-gamma-methylleucyl-D- 1-methoxycarbonyltryptophanyl-D-norleucine, abolished the inhibitory effects of ETs and IRL1620. Incubation of glial cells with lipopolysaccharide (LPS) caused an increase in iNOS expression. Simultaneous addition of ET-3 decreased the effects of LPS (10 and 100 ng/ml) on iNOS expression. Furthermore, cyclic AMP-elevating agents (dibutyryl cyclic AMP and forskolin) inhibited TNF-alpha/IL-1beta-induced and LPS-induced iNOS expression and nitrite accumulation. These findings suggest that ETs can decrease TNF-alpha/IL-1beta-induced and LPS-induced iNOS expression via ET(B) receptors and that cyclic AMP may be involved in this process.

Animals↗

Involvement of L-type-like amino acid transporters in S-nitrosocysteine-stimulated noradrenaline release in the rat hippocampus.

Nitrogen oxides, such as nitric oxide, have been shown to regulate neuronal functions, including neurotransmitter release. We investigated the effect of S-nitroso-L-cysteine (SNC) on noradrenaline (NA) release in the rat hippocampus in vivo and in vitro. SNC stimulated [3H]NA release from prelabeled hippocampal slices in a dose-dependent manner. SNC stimulated endogenous NA release within 30 min to almost five times the basal level in vivo (microdialysis in freely moving rats). In a Na+-containing Tyrode's buffer, SNC-stimulated [3H]NA release was inhibited 30% by the coaddition of L-leucine. In the Na+-free, choline-containing buffer, SNC-stimulated [3H]NA release, which was similar to that in the Na+-containing buffer, was inhibited markedly by L-leucine, L-alanine, L-methionine, L-phenylalanine, and L-tyrosine. The effects of the other amino acids examined were smaller or very limited. The effect of L-leucine was stronger than that of D-leucine. A specific inhibitor of the L-type amino acid transporter, 2-aminobicyclo[2.2.1]-heptane-2-carboxylate (BCH), inhibited the effects of SNC on [3H]NA release in the Na+-free buffer. Uptake of L-[3H]leucine into the slices in the Na+-free buffer was inhibited by SNC, BCH, and L-phenylalanine, but not by L-lysine. The effect of SNC on cyclic GMP accumulation was not inhibited by L-leucine, although SNC stimulated cyclic GMP accumulation at concentrations up to 25 microM, much less than the concentration that stimulates NA release. These findings suggest that SNC is incorporated into rat hippocampus via the L-type-like amino acid transporter, at least in Na+-free conditions, and that SNC stimulates NA release in vivo and in vitro in a cyclic GMP-independent manner.

Amino Acid Transport Systems↗

Aldosterone-producing adrenocortical carcinoma metastases found seven years after adrenalectomy.

We report a case of metastatic adrenocortical carcinoma detected 7 years after adrenalectomy. A 52-year-old woman, who had undergone adrenalectomy for an aldosterone-producing adrenocortical carcinoma at age 45, was found on examination by computerized tomography to have enlarged paraaortic lymph nodes. These nodes were surgically resected, and the histological diagnosis from the resected tissue was metastatic adrenocortical carcinoma. The patient has now survived for more than 9 years following the original adrenalectomy. Evidence suggests that this was a slow-growing tumor, because the primary tumor was sharply demarcated and the metastases were found 7 years after the original operation. We believe that aggressive surgical resection of metastatic lesions could lead to prolonged survival in patients with adrenocortical carcinomas of this type.

Adrenal Cortex Neoplasms↗

Carcinosarcoma in the region of the female urethra.

A 61-year-old woman with acute urinary retention was found to have a carcinosarcoma in the region of the urethra. Evaluation of the computed tomogram suggested a urethral tumor, which was resected by a transperineal approach. She received local radiotherapy after surgery, and is alive at 1-year follow-up with a tumor metastasis to the pelvis.

Carcinosarcoma↗

Sonographically-detected impalpable testicular cancer with retroperitoneal bulky metastases: a case report.

We report a case of a sonographically-detected impalpable embryonal cell carcinoma with bulky retroperitoneal metastases. A 34-year-old man, who presented with left flank pain, was presumed to have an extragonadal retroperitoneal germ cell tumor. Scrotal sonography revealed a hypoechoic lesion, 7 mm in diameter, which was histologically diagnosed as a primary embryonal cell carcinoma. Evidence suggested that the primary tumor had grown slowly, as the tumor was well encapsulated. This case suggests that some extragonadal germ cell tumors arise from a primary testicular cancers, and that successful treatment of these tumors should include consideration that they may have arisen as a primary testicular mass.

Adult↗

Theiler's murine encephalomyelitis virus subgroup strain-specific infection in a murine macrophage-like cell line.

We compared infection of a murine macrophage-like cell line, J774-1, with two Theiler's murine encephalomyelitis virus subgroup strains. The GDVII strain, which is highly virulent and produces acute polioencephalomyelitis in mice, did not actively replicate in J774-1 cells, although there was a significant inhibition in cellular protein synthesis. In contrast, the DA strain, which is less virulent and causes demyelination with a persistent virus infection, productively infected J774-1 cells; however, there was less virus produced than in BHK-21 cells, and there was little if any cellular protein shutoff. These in vitro data may provide some explanation for the biological activities that are observed between both subgroup strains.

Actins↗

Human cytomegalovirus induces interleukin-8 production by a human monocytic cell line, THP-1, through acting concurrently on AP-1- and NF-kappaB-binding sites of the interleukin-8 gene.

Cytomegalovirus (CMV) infection induced interleukin-8 (IL-8) gene transcription in a human monocytic cell line, THP-1 cells, leading to IL-8 secretion. The functional analysis of the IL-8 gene revealed that both AP-1- and NF-kappaB factor-binding elements were involved in conferring the responsiveness to CMV. Moreover, electrophoretic mobility shift assays demonstrated that CMV induced the formation of NF-kappaB and AP-1 complexes. These results suggest that CMV activates these transcriptional factors, resulting in IL-8 gene expression.

Animals↗

Yield of computed tomography and bronchoscopy for the diagnosis of Mycobacterium avium complex pulmonary disease.

Mycobacterium avium complex (MAC) pulmonary disease with nodules and bronchiectasis is increasing. But the usefulness of computed tomography (CT) and bronchoscopy for diagnosis and the significance of MAC isolation from respiratory secretions are still unclear. For a 4-yr period, we prospectively examined the role of bronchoscopy with bronchial washing and transbronchial lung biopsy in 26 patients who had clusters of small nodules in the periphery of the lung associated with ectatic changes of the draining bronchi on the CT scan. None of them was infected with human immunodeficiency virus. Thirteen of the 26 patients (50%) had cultures positive for MAC, six in the sputum and 13 in the bronchial washing. Epithelioid granuloma was demonstrated in eight of 13 patients with culture-positive MAC and in two of 13 patients in whom MAC was culture-negative. Rapidly growing mycobacteria were cultured in the two patients. Seven of the eight biopsy-positive patients received treatment and responded by sputum conversion and/or radiographic improvement. We found that the CT finding was a useful clue to suspect MAC pulmonary disease and that the bronchial washing was more sensitive than the routine expectorated sputum for MAC isolation. Demonstration of granuloma in more than half of the MAC-positive patients would suggest that MAC may have invaded the lung tissue rather than colonized in the airways.

Adult↗

Muscarinic acetylcholine receptors on rat thymocytes: their possible involvement in DNA fragmentation.

Several studies have shown that the nervous (and hormonal) system controls immune functions. In the present study, we examined the presence of muscarinic acetylcholine receptors and the effect of carbachol on DNA fragmentation in adult rat thymocytes. Rat thymocytes possessed high affinity binding sites for the muscarinic antagonist [3H]3-quinuclidinyl benzilate (QNB). The average number of binding sites per cells was 3000, and the equilibrium dissociation constant of [3H]QNB on intact cell was approximately 80 nM. The binding was inhibited by an M1- and M3-selective antagonist, 4-diphenylacetoxy-N-methylpiperidine methiodine (4-DAMP). Hydrocortisone (100 mg/kg, s.c.) treatment of rats for 2 days prior to sacrifice increased the average number of [3H]QNB binding sites on thymocytes by 82 +/- 33%. The gel electrophoresis of DNA extracted from carbachol-treated thymocytes revealed a ladder pattern typical of intranucleosomal fragmentation. The addition of oxotremorine-M also induced DNA fragmentation and the effects of muscarinic agonists were inhibited by the addition of atropine or 4-DAMP. The results suggest the existence of muscarinic receptors and the possible involvement in apoptosis in thymocytes.

Animals↗

Effects of homologous atrial, brain, and C-type natriuretic peptides on isolated heart and blood vessels of bullfrog.

The cardiovascular effects of homologous natriuretic peptides (fNPs) were examined in the bullfrog, Rana catesbeiana. Synthetic bullfrog atrial natriuretic (fANP), brain natriuretic (fBNP) and C-type natriuretic peptides (fCNP I and fCNP II), were tested in vitro and compared under the same experimental conditions. All frog NPs produced a significant, and concentration-dependent, reduction in tension (relaxant effect) in the isolated dorsal aorta. Frog CNP II exhibited similar vasorelaxation profile as that of fANP, while fBNP and fCNP I had lower activity than fANP. Frog NPs inhibited norepinephrine induced contraction and fCNP II was most potent. In isolated preparations of atrium and ventricle, fCNP I and II produced a significant, and concentration-dependent, reduction in tension, but neither fANP nor fBNP produced any significant effects. Frog CNP II is most potent among fNPs in relation to reduce the cardiac output. Chronotropic responses of the heart to administrations of NPs were insignificant. The present results for the first time showed that fNPs play roles in the control of cardiovascular system, both blood vessels and heart, in the bullfrog.

Animals↗

Direct action of melatonin on hen ovarian granulosa cells to lower responsiveness to luteinizing hormone.

The presence of a melatonin-binding component in the membrane fraction of the ovarian granulosa cells of hens that has characteristics of a receptor such as high affinity, limited capacity, reversible binding, and binding specificity was demonstrated by radioligand binding assays. When the cells were incubated in vitro with various concentrations of luteinizing hormone (LH) in the presence and absence of melatonin (1 and 5 nM), the dose-response curve for the LH-stimulated progesterone production was shifted to a higher concentration of LH: ED50 of LH was approximately two times greater in the presence of melatonin. The results suggest a direct action of melatonin on the ovarian granulosa cells of the hen to lower the responsiveness to LH for progesterone production.

Animals↗