Search PubMed⌕ Search

Biomedical subjects

M Amano

Publications and source records attributed to M Amano.

At least 91 records · Page 5Linked to original sources

Evidence for the involvement of endogenous aspartate in the mediation of carotid chemoreceptor reflexes in the rostral ventrolateral medulla of the rat.

2-Amino-5-phosphonovalerate (AP5; 153 pmol) injected into the rostral ventrolateral medulla (RVLM) inhibited pressor responses induced by carotid chemoreceptor stimulation. AP5 also inhibited pressor responses to aspartate (0.75 nmol) but not to glutamate (0.53 nmol) similarly injected. High K+ (50 mM) released endogenous aspartate and glutamate in a Ca2+-dependent manner from the RVLM. Chemoreceptor stimulation caused a release of aspartate but not of glutamate in the RVLM, and sinus nerve denervation abolished the release of aspartate. Increases in blood pressure induced by intravenous phenylephrine did not release aspartate. These results support the hypothesis that endogenous aspartate in the rat RVLM is involved in the mediation of chemoreceptor reflexes.

2-Amino-5-phosphonovalerate↗

Domain-specific phosphorylation of vimentin and glial fibrillary acidic protein by PKN.

PKN is a serine/threonine protein kinase with a catalytic domain homologous to the protein kinase C family and unique N-terminal leucine zipper-like sequences. Using analyses with the yeast two-hybrid system and in vitro binding assay, we found that the regulatory domain of PKN interacted with vimentin. We then examined whether PKN would phosphorylate vimentin in vitro. Vimentin proved to be an excellent substrate for PKN, and the phosphorylation of vimentin by PKN occurred in the head domain with the result of a nearly complete inhibition of its filament formation in vitro. Similar results were also obtained with another type III intermediate filament protein, glial fibrillary acidic protein (GFAP). These results raise the possibility that PKN may regulate filament structures of vimentin and GFAP by domain-specific phosphorylation.

Animals↗

Rho-associated kinase directly induces smooth muscle contraction through myosin light chain phosphorylation.

Small GTPase Rho plays pivotal roles in the Ca2+ sensitization of smooth muscle. However, the GTP-bound active form of Rho failed to exert Ca2+-sensitizing effects in extensively Triton X-100-permeabilized smooth muscle preparations, due to the loss of the important diffusible cofactor (Gong, M. C., Iizuka, K., Nixon, G. , Browne, J. P., Hall, A., Eccleston, J. F., Sugai, M., Kobayashi, S. , Somlyo, A. V., and Somlyo, A. P. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 1340-1345). Here we demonstrate the contractile effects of Rho-associated kinase (Rho-kinase), recently identified as a putative target of Rho, on the Triton X-100-permeabilized smooth muscle of rabbit portal vein. Introduction of the constitutively active form of Rho-kinase into the cytosol of Triton X-100-permeabilized smooth muscle provoked a contraction and a proportional increase in levels of monophosphorylation of myosin light chain in both the presence and the absence of cytosolic Ca2+. These effects of constitutively active Rho-kinase were wortmannin (a potent myosin light chain kinase inhibitor)-insensitive. Immunoblot analysis revealed that the amount of native Rho-kinase was markedly lower in Triton X-100-permeabilized tissue than in intact tissue. Our results demonstrate that Rho-kinase directly modulates smooth muscle contraction through myosin light chain phosphorylation, independently of the Ca2+-calmodulin-dependent myosin light chain kinase pathway.

Androstadienes↗

Phosphorylation of glial fibrillary acidic protein at the same sites by cleavage furrow kinase and Rho-associated kinase.

Site- and phosphorylation state-specific antibodies are useful to analyze spatiotemporal distribution of site-specific phosphorylation of target proteins in vivo. Using several polyclonal and monoclonal antibodies that can specifically recognize four phosphorylated sites on glial fibrillary acidic protein (GFAP), we have previously reported that Thr-7, Ser-13, and Ser-34 on this intermediate filament protein are phosphorylated at the cleavage furrow during cytokinesis. This observation suggests that there exists a protein kinase named cleavage furrow kinase specifically activated at metaphase-anaphase transition (Matsuoka, Y., Nishizawa, K., Yano, T., Shibata, M., Ando, S., Takahashi, T., and Inagaki, M. (1992) EMBO J. 11, 2895-2902; Sekimata, M., Tsujimura, K., Tanaka, J., Takeuchi, Y., Inagaki, N., and Inagaki, M. (1996) J. Cell Biol. 132, 635-641). Here we report that GFAP is phosphorylated specifically at Thr-7, Ser-13, and Ser-34 by Rho-associated kinase (Rho-kinase), which binds to the small GTPase Rho in its GTP-bound active form. The kinase activity of Rho-kinase toward GFAP is dramatically stimulated by guanosine 5'-(3-O-thio)-triphosphate-bound RhoA. Furthermore, the phosphorylation of GFAP by Rho-kinase results in a nearly complete inhibition of its filament formation in vitro. The possibility that Rho-kinase is a candidate for cleavage furrow kinase is discussed.

Antibodies↗

Granulocyte colony-stimulating factor-induced comobilization of CD4- CD8- T cells and hematopoietic progenitor cells (CD34+) in the blood of normal donors.

The feasibility of transplantation of HLA-matched hematopoietic progenitor cells from the blood of normal donors given granulocyte colony-stimulating factor (G-CSF) has been reported recently. In the current study, the changes in T-cell subsets as well as CD34+ cells were determined in one blood volume leukapheresis products of six normal individuals given G-CSF. Examination of the T-cell subsets in the leukapheresis products showed three different patterns: one in which a discrete population of CD4- CD8- alphabeta T cells was found in addition to the typical CD4+ and CD8+ T cells in the unfractionated as well as in high- and low-density cells; a second in which the discrete population of CD4- CD8- alphabeta T cells was predominant only in the low-density fractions; and a third in which a discrete population of CD4- CD8- T cells was not observed. The median yield of CD4- CD8- T cells was about fourfold to fivefold higher than the calculated number present in one blood volume (5L) from normal individuals. The ratios of CD34+ cells to CD4+ and CD8+ T cells, and of CD4- CD8- T cells to CD4+ and CD8+ T cells, were highest in the low-density fractions. These fractions suppressed the mixed leukocyte, and may ameliorate graft-versus-host disease as compared with unfractionated cells.

Adult↗

Phenotypes and invariant alpha beta TCR expression of peripheral V alpha 14+ NK T cells.

A novel subset of peripheral T cells, peripheral NK T cells, is found to be a major population comprising 5% of splenic T and 40% of bone marrow T cells. The majority of peripheral NK T cells are characterized by the expression of an invariant TCR-alpha encoded by V alpha 14/J alpha 281 with a one nucleotide N region. Moreover, a specific reduction of V alpha 14+ NK T cells has been demonstrated to be tightly associated with various autoimmune diseases, indicating their decisive role in autoimmune disease development. In this study, we investigated the phenotypes of peripheral V alpha 14+ NK T cells and their TCR-beta repertoire. Peripheral V alpha 14+ NK T cells, comprise two populations, i.e., small and large sized cells, at an equal frequency, belonged to the CD4- CD8- fraction, and are heat stable antigen(bright), macrophage-1bright, B220bright, CD45RBdim, and Mel-14dim, but CD5-, distinct from thymic NK T cells. TCR-beta analysis clearly showed that peripheral V alpha 14+ NK T cells utilized two to three dominant invariant TCR-beta, such as V beta 8.2 D beta J beta 2.5/V beta 7 D beta J beta 2.1 in the spleen and liver, V beta 8.2 D beta J beta 2.5/V beta 8.3 D beta J beta 2.2/V beta 7 D beta J beta 2.6 in the bone marrow, and V beta 7 D beta J beta 2.1/V beta 3 D beta J beta 1.2 in intestinal intraepithelial lymphocytes. Judging from the unusual surface phenotypes, such as heat stable antigen, macrophage-1, B220, CD45RBdim, and Mel-14dim, which are known to be T cell activation markers, peripheral V alpha 14+ NK T cells may always be activated under physiologic conditions, resulting in the oligoclonal expansion of V alpha 14+ NK T cells with different invariant TCR-beta in different peripheral organs. The unique features of V alpha 14+ NK T cells are discussed.

Amino Acid Sequence↗

Formation of actin stress fibers and focal adhesions enhanced by Rho-kinase.

The small guanosine triphosphatase (GTPase) Rho is implicated in the formation of stress fibers and focal adhesions in fibroblasts stimulated by extracellular signals such as lysophosphatidic acid (LPA). Rho-kinase is activated by Rho and may mediate some biological effects of Rho. Microinjection of the catalytic domain of Rho-kinase into serum-starved Swiss 3T3 cells induced the formation of stress fibers and focal adhesions, whereas microinjection of the inactive catalytic domain, the Rho-binding domain, or the pleckstrin-homology domain inhibited the LPA-induced formation of stress fibers and focal adhesions. Thus, Rho-kinase appears to mediate signals from Rho and to induce the formation of stress fibers and focal adhesions.

3T3 Cells↗

Activation of salmon GnRH mRNA expression prior to differentiation of precocious males in Masu salmon.

Changes in salmon gonadotropin-releasing hormone (sGnRH) mRNA levels in the brain of underyearling male masu salmon, Oncorhynchus masou, were investigated to clarify sGnRH participation in differentiation of precocious males. Fish were immature with low gonadosomatic index (GSI < 0.100%) on April 16 and May 7. On June 11, fish were differentiated into two groups: future precocious males and immature males. Fish in the former group had high GSI (0.203%) with proliferation of primary spermatocytes in the testis, while the latter had low GSI (0.055%) with primary spermatogonia. sGnRH mRNA levels (the number of neurons expressing sGnRH mRNA and the total number of silver grains per unit area) in the ventral telencephalon (VT) and in the VT plus preoptic area (POA) already increased in May before the differentiation of precocious males. GSI was positively correlated with sGnRH mRNA levels in the VT and in the VT plus POA in April, and with mRNA levels in the POA in May. Activated sGnRH synthesis is concluded to be related to the appearance of precocious males in masu salmon.

Animals↗

Bioaccumulation profiles of polychlorinated biphenyls including coplanar congeners and possible toxicological implications in Baikal seal (Phoca sibirica).

Isomer specific concentrations of individual polychlorinated biphenyls (PCBs) including toxic non-ortho (IUPAC 77, 126, 169), mono-ortho (105, 118, 156) and di-ortho (137, 138, 153, 180) coplanar congeners were determined in the blubber of 40 Baikal seals (Phoca sibirica) and as their fish diet collected from Lake Baikal, Siberia. Residue levels of total PCBs in Baikal seals were noticeably high and comparable to those reported for seals from the North Sea, suggesting the recent usage of this compound in the watershed of Lake Baikal. Non-, mono-, and di-ortho coplanar congeners were also detected in Baikal seals and fish. An approach to estimate bioaccumulation profiles of PCB congeners revealed that the non-ortho PCBs, IUPAC 77, 126 and 169 seemed to be less persistent than other congeners. Furthermore, selective biotransformation of PCB congeners having either meta-para vicinal H atoms or both adjacent chlorinated meta-para and ortho-meta positions has been suggested. Comparison of 2,3,7,8-TCDD toxic equivalents (TEQ) of non-, mono- and di-ortho coplanar congeners in Baikal seals with those for other marine mammals suggested higher enrichment of mono-ortho congeners, particularly IUPAC 105 and 118, which contributed significantly to the total TEQs in Baikal seals. Results imply that the TCDD-like toxicity is relatively serious in Baikal seals, because of the enrichment of these toxic PCB congeners in tissues.

Journal Article↗

The T cell activation factor NF-ATc positively regulates HIV-1 replication and gene expression in T cells.

Clinical deterioration in human immunodeficiency virus type 1 (HIV-1) infection is associated with increased levels of viral replication and burden in the peripheral blood and lymphoid organs. T cell activation and ensuing cellular gene activation can be critical for HIV-1 replication. The hypothesis that the nuclear factor of activated T cells (NF-AT) may influence HIV-1 replication is therefore compelling given the tight correlation of HIV-1 transcriptional induction to T cell activation. We report that certain NF-AT(Rel) family members productively bind the kappaB regulatory elements, synergize with NF-kappaB and Tat in transcriptional activation of HIV-1, and enhance HIV-1 replication in T cells. These results link regulatory factors critical to T cell commitment directly to HIV-1 replication.

Cells, Cultured↗

Association between ion fluxes and defense responses in pea and cowpea tissues.

The glycopeptide elicitor from a pea pathogen, Mycosphaerella pinodes, induced rapid alkalinization and increases in levels of Na+ and K+ ions in the extracellular solution upon contact with pea and cowpea tissues. The presence of monensin, nigericin, lidocaine, quinidine or phenytoin together with the elicitor markedly inhibited these changes, whereas the presence of valinomycin, gramicidin D, tetraethylammonium, CsCl and aminopyridine did not. The production of phytoalexins in pea and cowpea tissues was also strongly inhibited by the simultaneous presence of the former reagents but not of the latter reagents. Inhibitory effects on the production of phytoalexins were diminished when monensin, nigericin or a Na(+)-channel blocker was applied 3 h after the start of treatment with elicitor. Furthermore, orthovanadate and neomycin, which suppress defense responses in both tissues, also inhibited the above mentioned changes. By contrast, the species-specific suppressor from M. pinodes inhibited the elicitor-induced release of Na+ and K+ ions from pea tissues, but, conversely, by itself it elicited either the defense response or the release of Na+ and K+ ions from cowpea tissues. The results indicate that these ion-related changes, in particular the efflux of Na+ and K+ ions, might be closely associated with the signal transduction system for defense responses at the tissue level.

Fabaceae↗

Adequacy of pacing rate during exercise in rate responsive ventricular pacing.

Our objective was to determine the adequate pacing rate during exercise in ventricular pacing by measuring exercise capacity, cardiac output, and sinus node activity. Eighteen patients with complete AV block and an implanted pacemaker underwent cardiopulmonary exercise tests under three randomized pacing rates: fixed rate pacing (VVI) at 60 beats/min and ventricular rate-responsive pacing (VVIR) programmed to attain a heart rate of about 110 beats/min or 130 beats/min (VVIR 110 and VVIR 130, respectively) at the end of exercise. Compared with VVI and VVIR 130, VVIR 110 was associated with an increased peak oxygen uptake (VVIR 110: 20.3 +/- 4.5 VVI: 16.9 +/- 3.1; P < 0.01; and VVIR 130: 19.0 +/- 4.1 mL/min per kg, respectively; P < 0.05 and a higher oxygen uptake at anaerobic threshold (15.3 +/- 2.7, 12.7 +/- 1.9; P < 0.01, and 14.6 +/- 2.6 mL/min per kg; P < 0.05). The atrial rate during exercise expressed as a percentage of the expected maximal heart rate was lower in VVIR 110 than in VVI or VVIR 130 (VVIR 110: 75.9% +/- 14.6% vs VVI: 90.6% +/- 12.8%; P < 0.01; VVIR 110 vs 130: 89.1% +/- 23.1% P < 0.05). There was no significant in cardiac output at peak exercise between VVIR 110 and VVIR 130. We conclude that a pacing rate for submaximal exercise of 110 beats/min may be preferable to that of 130 beats/min in respect to exercise capacity and sympathetic nerve activity.

Cardiac Output↗

Combination chemotherapy for malignant paraganglioma.

Treatment with a combination chemotherapeutic regimen consisting of cyclophosphamide, vincristine, and dacarbazine for malignant paraganglioma with hepatic metastasis is reported. A 51-year-old male presented with tumors in the retroperitoneal space and liver. The patient was diagnosed as having paraganglioma based on elevated levels of serum neuron-specific enolase, urinary catecholamine and vanillylmandelic acid, and on histological findings of the liver specimen. The patient was treated with this combination chemotherapy in repeated 21-day cycles. Temporary improvement in laboratory findings and a 20% reduction in the size of the hepatic masses were observed without severe adverse effects.

Antineoplastic Combined Chemotherapy Protocols↗

Normal contrast enhancement of extraocular muscles: fat-suppressed MR findings.

PURPOSE: To evaluate contrast enhancement of normal extraocular muscles in the orbit. METHODS: Noncontrast and contrast-enhanced T1-weighted images of the orbit were acquired using a fat-suppression MR technique in eight patients with no orbital disease. Contrast enhancement of 64 extraocular muscles was evaluated and compared with that of 16 temporal muscles. RESULTS: Compared with temporal muscles, all extraocular muscles markedly enhanced after administration of contrast material. CONCLUSIONS: Because normal extraocular muscles enhanced markedly with contrast material, more attention should be paid to these muscles when using contrast-enhanced, fat-suppressed T1-weighted MR imaging to evaluate pathologic conditions.

Adipose Tissue↗

[Uterine cervical carcinoma after radiotherapy: comparison between MR imaging and histopathological findings].

The findings of MRI and pathologic investigation were correlated in curatively irradiated uterine cervical carcinoma. Four patients having residual carcinoma diagnosed by biopsy underwent hysterectomy. MRI demonstrated the mass lesion in one patient with pathologic confirmation of massive viable cancer cells (case I). Of the other three patients, MRI demonstrated normal configuration of the uterine cervix. Cervical signal intensity, however, varied. Hyperintensity was noted in an area of the posterior wall on T2WI in case 2. The anterior wall of the case 2 and the other two cases showed hypointensity. Cervical specimens with normal intensity showed only a small number of degenerated cancer cells. On the other hand, pathologic examination of the posterior wall of the case 2 revealed both cancer cells with varying degrees of degeneration and necrotic tissues. Degeneration of cancer cells was stronger in the superficial layer than the deeper layer. Fibrosis, hemorrhage, granulation and hyalinization were hypointense on T2WI. T2 elongation reflected not only the residual tumor but the post-irradiation changes. Post-irradiated cervix with normal intensity indicated that only a small number of degenerated cancer cells may persist even if the biopsy was positive. We conclude that MRI is useful in evaluating tumor response to radiotherapy.

Carcinoma, Mucoepidermoid↗

[Usefulness of MR imaging in evaluating and predicting effect of neoadjuvant chemotherapy on uterine cervical carcinoma].

The purpose of this study was to assess the usefulness of MR imaging (MRI) in evaluating and predicting the effect of neoadjuvant chemotherapy on uterine cervical carcinoma. MRI was performed before and after neoadjuvant chemotherapy (NA-C) of systemic BOMP in 32 patients with cervical carcinoma. (1) In 8 patients who suffered from hysterectomy after NA-C, MR estimates of tumor diameter were within 5 mm of the surgical sample in all cases. In addition, MRI accurately evaluated parametrial state. (2) In 10 patients, the reduction rates after 2 courses of NA-C were compared with those after 3 courses, and no significant difference was found between them (p = 0.628). (3) Statistical analysis was performed to assess the correlation between the response to NA-C and MR findings before NA-C such as MR staging, tumor volume, signal intensity and contrast enhancement of tumor, body invasion, total cervical replacement, hydrometra, and ascites. Analysis of variance and multiregression analysis revealed that tumor volume (p = 0.012) and body invasion (p = 0.079) were important factors in predicting the effect of NA-C on cervical carcinoma. In conclusion, MRI is useful in evaluating and predicting the effect of NA-C on uterine cervical carcinoma.

Adult↗

[A case of local recurrence of rectal cancer that achieved a complete response with high-dose interstitial brachytherapy].

We report a case of local recurrence of rectal cancer that achieved complete response with high-dose interstitial brachytherapy. The patient was a forty-nine-year-old woman. In September 1995, she underwent abdominoperineal resection of rectum for rectal cancer (T4 N2 M0, moderately differentiated adenocarcinoma). In February 1996, she complained of genital bleeding. MRI examination revealed a mass of 70 x 22 x 20 mm in the pelvis, which was diagnosed to be recurrent rectal cancer with needle biopsy. Interstitial brachytherapy (60 Gy/twelve fractions/six days) was performed, and a complete response was achieved. One year after the treatment, no tumor was detected in the pelvis. No skin erosion, skin ulcer, urinary cystitis, nor ileitis occurred. High-dose interstitial brachytherapy delivers high-dose irradiation to a tumor while keeping the dose to the surrounding normal organs at a low level. This procedure may be an effective modality for local recurrence of rectal cancer.

Adenocarcinoma↗