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        Comparison of Spectrophotometric, HPLC and Chemilumines­cence Methods for 3-Nitrotyrosine and Peroxynitrite Interaction

        Turan Nilufer Nermin,Ark Mustafa,Demiryurek Abdullah Tuncay The Pharmaceutical Society of Korea 2005 Archives of Pharmacal Research Vol.28 No.3

        We have studied the interaction of 3-nitrotyrosine with peroxynitrite using three different methods; chemiluminescence, spectrophotometry and HPLC. Peroxynitrite-induced luminol or lucigenin chemiluminescence were significantly decreased by 3-nitrotyrosine, in concentration­dependent manners. The intensity of the peroxynitrite spectrum was also markedly reduced in the presence of 3-nitrotyrosine in the spectrophometric assay. However, there was no attenuation of the 3-nitrotyrosine signal in the HPLC assay after mixing with peroxynitrite. The interaction of 3-nitrotyrosine and hypochlorous acid (HOCI) was also studied via the chemilumines-cence assay, where the HOCI-induced responses were markedly inhibited by 3-nitrotyrosine. These results suggest that caution should be taken when studying the levels or interactions of 3-nitrotyrosine.

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        Comparison of Spectrophotometric, HPLC and Chemiluminescence Methods for 3-Nitrotyrosine and Peroxynitrite Interaction

        Nilufer Nermin Turan,Mustafa Ark,Abdullah Tuncay Demiryurek 대한약학회 2005 Archives of Pharmacal Research Vol.28 No.3

        We have studied the interaction of 3-nitrotyrosine with peroxynitrite using three different methods; chemiluminescence, spectrophotometry and HPLC. Peroxynitrite-induced luminol or lucigenin chemiluminescence were significantly decreased by 3-nitrotyrosine, in concentrationdependent manners. The intensity of the peroxynitrite spectrum was also markedly reduced in the presence of 3-nitrotyrosine in the spectrophometric assay. However, there was no attenuation of the 3-nitrotyrosine signal in the HPLC assay after mixing with peroxynitrite. The interaction of 3-nitrotyrosine and hypochlorous acid (HOCl) was also studied via the hemiluminescence assay, where the HOCl-induced responses were markedly inhibited by 3-nitrotyrosine. These results suggest that caution should be taken when studying the levels or interactions of 3-nitrotyrosine.

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        Chronic ouabain treatment induces Rho kinase activation

        Aysun Ozdemir,Gürkan Şahan,Ayşegül Demirtas,Eda Aypar,Gürkan Gözübüyük,Nilüfer Nermin Turan,Mustafa Ark 대한약학회 2015 Archives of Pharmacal Research Vol.38 No.10

        Ouabain is an endogenous Na?/K?-ATPaseinhibitor whose chronic administration induces hypertension. Endogenous ouabain levels increase in humanessential hypertension. On the other hand, Rho/Rho kinase(ROCK) pathway has been implicated in various animalmodels of hypertension. In the current work, we evaluatedthe possible involvement of Rho kinase in ouabain-inducedhypertension. Ouabain was administered daily (20 lg/kg,i.p.) to Wistar rats for 6 weeks. After the ouabain treatment,we evaluated the possible changes in vascular responsesto KCl and phenylephrine alone and in thepresence of Rho kinase inhibitor Y27632. We also determinedthe expressions of ROCKs, Rho A and phosphorylationof myosin binding subunit of myosin light chainphosphatase (pMYPT) and activation of Rho A. Agonistinducedcontractions in the presence of Y27632 are significantlydecreased and Y27632-induced relaxations inaortas precontracted with phenylephrine are significantlyenhanced with the chronic treatment of ouabain. Althoughthe expressions of ROCK I and ROCK II remained unchanged,pMYPT expression was significantly increased inouabain-treated group. Moreover, Rho A expression andactivation were decreased after treatment with ouabain. Although Rho kinase expression did not change in aortas,increased basal Rho kinase activation may contribute to thedevelopment of ouabain-induced hypertension. Our currentdata present the first evidence that Rho kinase is involvedin the development of ouabain-induced hypertension inrats.

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