• Title/Summary/Keyword: specific inhibitor

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The Role and Localization of Nitric Oxide Synthase in Neurogenic Inflammation of the Rat Airways (백서의 기도 선경성 염증에서 산화질소 합성효소(Nitric Oxide Synthase)의 역할과 분포)

  • Shim, Jae-Jeong;Lee, Sang-Yub;Lee, Sang-Hwa;Suh, Jung-Kyung;Kim, Chul-Hwan;Cho, Jae-Youn;In, Kwang-Ho;Yoo, Seo-Hwa;Kang, Kyung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.3
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    • pp.420-433
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    • 1996
  • Background : There have been many debates about the effects of nitric oxide on the neurogenic inflammation. The role of nitric oxide in the neurogenic inflammation of airways will be required a better understanding of the localization and types of nitirc oxide synthase(NOS) activity in the neurogenic inflammation of airways. Method : To investigate the role of nitric oxide in airway neurogenic inflammation, 1) the effects of neurokinin receptor antagonist (FK224) and nitric oxide synthase inhibitor, $N^{\omega}$-nitro-L-arginine (L-NNA) on plasma extravastion were evaluated in four groups of Sprague-Dawley rats ; sham operation group(sham NANC group), electrical vagal stimulation group(NANC2 group), intravenous pretreatment groups with FK224 (1mg/kg ; FK224 group), and L-NNA(1mg/kg ; L-NNA group) 15 minutes before vagal NANC stimulation. 2) NOS activity in trachea with neurogenic inflammation was localized by immunohistochemical stain. Immunohistochemical stain was performed by antibodies specific for inflammatory cells(iNOS), brain(bNOS), and endothelium (eNOS) on trachea obtained from sham NANC, NANC2, and FK224 groups. Results : The results are that plasma extravsation in neurogenic inflammation of rat airways was inhibited by FK224, but enhanced by L-NNA pretreatment(P<0.05). There was significantly increased infiltration of inflammatory cells in subepithelium of neurogenic inflammatory trachea, but the reduction of subepithelial infiltration of inflammatory cells was observed after pretreatment with FK224(P<0.05). Immunostaining with anti-iNOS antibody showed strong reactivity only in infiltrated inflammatory cells in neurogenic rat trachea, and these iNOS reactivity was reduced by pretreatment with FK224. bNOS immunoreactivity was significantly increased only in the nerves both of neurogenic inflammatory and FK224 pretreated trachea compared with sham NANC trachea(p<0.05). eNOS immunoreactivity was not significant change in endothelium in neurogenic inflammation of rat trachea. Conclusion : These results suggest that nitric oxide released from iNOS in infiltrated inflammatory cells has main role in neurogenic inflammation of rat trachea. The presence of bNOS immunoreactivity in the nerves indicates that nitric oxide may be released from the nerves in rat trachea with neurogenic inflammation.

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Inhibitory Effect of Camp Antagonist and Pka Inhibitors, and Stimulatory Effect of Adenylate Cyclase Agonist on Cathepsin K Processing in Cultured Mouse Osteoclasts (cAMP 길항제와 PKA 억제제 및 Adenylate Cyclase 촉진제의 백서 파골세포에서 Cathepsin K 생성에 대한 효과)

  • Shim, Youn-Soo
    • Journal of dental hygiene science
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    • v.6 no.1
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    • pp.1-9
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    • 2006
  • Cathepsin K (cat K) is the major cysteine protease expressed in osteoclasts and was thought to play a key role in matrix degradation during bone resorption. It was shown that the intracellular maturation of cat K was prevented by the cAMP antagonist, Rp-cAMP, and the protein kinase A (PKA) inhibitors of KT5720 and H89. In contrast, forskolin, a adenylate cyclase agonist, rather induced Cat K processing and maturation in osteoclasts. Furthermore, to determine whether cat K processing and maturation signaling involves protein kinase C (PKC), mouse total bone cells were treated with calphostin C, a specific inhibitor of PKC, however, no effect was observed, indicating that calphostin C did not affect to osteoclast-mediated cat K processing and maturation. Thus, it is indicated that the cAMP-PKA signaling pathway regulates cat K maturation in osteoclasts. Since secreted proenzymes have the potential to reenter the cell via M6P receptor, to prevent this possibility, it was tested cAMP antagonist Rp-cAMP and the PKA inhibitors KT5720 and H89 in the absence or presence of M6P. Inhibition of cat K processing by Rp-cAMP, KT5720, or H89 was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of Rp-cAMP, KT5720 and H89. These dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of cat K processing.

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Simultaneous Detection of Seven Phosphoproteins in a Single Lysate Sample during Oocyte Maturation Process (난자성숙 과정의 단일 시료에서 일곱 가지 인산화 단백질의 동시 분석 방법)

  • Yoon, Se-Jin;Kim, Yun-Sun;Kim, Kyeoung-Hwa;Yoon, Tae-Ki;Lee, Woo-Sik;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.3
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    • pp.187-197
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    • 2009
  • Objective: Phosphorylation and dephosphorylation of proteins are important in regulating cellular signaling pathways. Bead-based multiplex phosphorylation assay was conducted to detect the phosphorylation of seven proteins to maximize the information obtained from a single lysate of stage-specific mouse oocytes at a time. Methods: Cumulus-oocyte complexes (COCs) were cultured for 2 h, 8 h, and 16 h, respectively to address phosphorylation status of seven target proteins during oocyte maturation process. We analyzed the changes in phosphorylation at germinal vesicle (GV, 0 h), germinal vesicle breakdown (GVBD, 2 h), metaphase I (MI, 8 h), and metaphase II (MII, 16 h in vitro or in vivo) mouse oocytes by using Bio-Plex phosphoprotein assay system. We chose seven target proteins, namely, three mitogen-activated protein kinases (MAPKs), ERK1/2, JNK, and p38 MAPK, and other 4 well known signaling molecules, Akt, GSK-$3{\alpha}/{\beta}$, $I{\kappa}B{\alpha}$, and STAT3 to measure their phosphorylation status. Western blot analysis and kinase inhibitor treatment for ERK1/2, JNK, and Akt during in vitro maturation of oocytes were conducted for the confirmation. Results: Phosphorylation of ERK1/2, JNK, p38 MAPK and STAT3 was increased over 3 folds up to 20 folds, while phosphorylation of the other three signal molecules, Akt, GSK-$3{\alpha}/{\beta}$, and $I{\kapa}B{\alpha}$ was less than 3 folds. All of these results except for Akt were statistically significant (p<0.05). Conclusion: This is the first report on the new and valuable method measuring many phosphoproteins simultaneously in one minute sample such as oocyte lysates. All of the three MAPKs, ERK1/2, JNK, and p38 MAPK are involved in the process of mouse oocyte maturation. In addition, STAT3 might be important regulator of oocyte maturation, while Akt phosphorylation at Serine 473 may not be involved in the regulation of oocyte maturation.

Studies on Genetice of Blast Resistance in Rice L Inheritance of Resistance to Specific Races of Blast Fungus and Relationship between Their Resistance and II, VIII, XI and XII Linkage Groups in Some Rice Varieties (수도품종의 도열병 저항성 유전분석 제1보 특정 도열병 균계에 대한 저항성 품종들의 저항성 유전분리와 II, VIII, XI 및 XII번 연관과의 관계)

  • Chae, Y.A.;Park, S.Z.;Ha, S.B.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.26 no.1
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    • pp.32-39
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    • 1981
  • In order to study the genetic system of the blast resistant varieties, the conidial suspension of mutant races of T-2$^{+t}$, N-2$^{+t}$, C-8$^{+t}$ was inoculated at 4-5 leaf stage by injector for F_2 seedlings from the crosses between seven resistant varieties and four maker lines easily detectable at seedling stage. The results are summarized as follows; 1. The fertility of cross between Semi-dwaf testers and Indica resistant varieties except Carreon was about 74 percents. 2. The segregation modes of resistance varied with varieties and blast races. However, the resistance was expressed as dominance in all cases. Tetep, Tadukan and Carreon showed more complicated segregation for resistance than that of the bred lines. 3. For blast races used, four segregation ratios such as 3:1, 9:7, 13:3 and 37:27 were found in the Tatukan, Tetep, and IR747, and three segregation ratios such as 3:1, 13:3 and 15:1 in the Carreon, and two segregation ratios of 3:1 and 13:3 with Suweon 287, Suweon 288, and Iri342. 4. In the segregation of the resistance to the each races, the ratios of 3:1, 13:3, 15:1 were fitted to T-2$^{+t}$, and the ratios of 3:1, 13:3, 9:7 and 37:27 to N-2$^{+t}$ and C-8$^{+t}$. 5. Suweon 287, Suweon 288 and Iri342 carried one simple dominant gene and inhibitor gene was considered in some cross combinations. Meanwhile Tadukan, Tetep and IR747 seemed to carry one to three resistant genes, and in some cross combinations, the expression of these genes were simple dominant, inhibiting, duplicating and complimentary action. 6. Resistance genes to blast races, T-2$^{+t}$, N-2$^{+t}$ and C-8$^{+t}$ in the Tadukan, Tetep, Carreon, Suweon 287, Suweon 288 and Iri342 were found to be independent with the linkage group of II(lg), VIII(la), XI(bc), and XII(gl).bc), and XII(gl).

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