• Title/Summary/Keyword: pH dependent

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MMP and TIMP production in periodontal ligament fibroblasts stimulated by Prevotella nigrescens lipopolysaccharide (Prevotella nigrescens lipopolysaccharide로 자극한 치주인대 섬유아세포에서 기질금속단백분해효소와 단백분해효소억제제의 생성 양상에 대한 연구)

  • Yang, Won-Kyung;Lee, Woo-Cheol;Kim, Mi-Ri;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.30 no.5
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    • pp.372-384
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    • 2005
  • The purpose of this study was to monitor the secretion of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) by human periodontal ligament (PDL) fibroblasts stimulated with Prevotella nigrescens lipopolysaccharide (LPS), and to examine the effect of calcium hydroxide treatment on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. PDL fibroblasts were stimulated by the LPS (0, 0.1, 1, 10 ${\mu}g/ml$) or LPS (10 ${\mu}g/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days, for various periods of time (12, 24, 48 h). Immunoprecipitation were performed for protein level analysis of MMP-1 MMP-2 and TIMP-1. Total RNA was isolated and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 mRNA. According to this study, the results were as follows: 1. The p개duction of MMP-1 by stimulation with P. nigrescens LPS increased in time-dependent manner, and showed maximum value at 48 h in both protein and mRNA level. But there was no dose-dependent increas. 2. MMP-2 production time-dependently increased when stimulated with 1 and 10 ${\mu}g/ml$LPS, but there was no dose-dependent increase. 3. TIMP-1 p개duction increased to 24 h, but decreased at 48 h. It increased when stimulated with 0.1 and 1${\mu}g/ml$, but suppressed at 10 ${\mu}g/ml$ .4. P. nigrescens LPS pretreated with $Ca(OH)_2$ markedly downregulated MMP-1 gene expression.

Establishment of an Assay for P2X7 Receptor-Mediated Cell Death

  • Lee, Song-Yi;Jo, Sooyeon;Lee, Ga Eun;Jeong, Lak Shin;Kim, Yong-Chul;Park, Chul-Seung
    • Molecules and Cells
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    • v.22 no.2
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    • pp.198-202
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    • 2006
  • The $P2X_7$ receptor, an ATP-gated cation channel, induces cell death in immune cells and is involved in neurodegenerative diseases. Although the receptor plays various roles in these diseases, the cellular mechanisms involved are poorly understood and antagonists are limited. Here, the development of a cell-based assay for human $P2X_7$ receptor is reported. We established permanent lines of HEK 293 cells expressing a high level of $hP2X_7$ receptor. Functional activity of the $hP2X_7$ receptor was confirmed by whole-cell patch recording of ATP-induced ion currents. Prolonged exposure to ATP resulted in death of the $hP2X_7$-expressing HEK 293 cells and this cell death could be quantified. Two known $P2X_7$ antagonists, PPADS and KN-62, blocked ATP-induced death in a concentration-dependent manner. Thus, this assay can be used to screen for new antagonists of $hP2X_7$ receptors.

Polymerization of ADP-Ribose Pyrophosphatase: Conversion Mechanism of $Mg^{2+}-Dependent$ ADP-Ribose Pyrophosphatase into $Mg^{2+}-Independent$ Form

  • Kim, Dae-Ki;Kim, Jong-Hyun;Song, Eun-Kyung;Han, Myung-Kwan;Kim, Jong-Suk
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.826-831
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    • 2003
  • ADP-ribose pyrophosphatase (ADPRase) hydrolyzes ADP-ribose (ADPR) into AMP and ribose-5'-phosphate. It is classified into two groups, $Mg^{2+}$-dependent and $Mg^{2+}$-independent ADPRase, depending on its $Mg^{2+}$requirement. Here, we purified $Mg^{2+}$-dependent ADPRase from rabbit liver and examined what factors affect $Mg^{2+}$ requirement. The purified enzyme showed a single band with the molecular weight of 34 kDa on SDS-PAGE both in the presence and absence of 2-mercaptoethanol. The molecular weight of the native enzyme calculated by gel filtration was 68 kDa, indicating that ADPRase is a dimer made up of two identical subunits. $Mg^{2+}$-dependent ADPRase with the highest ADPR affinity had a $K_m$ of 160$\pm$10 $\mu$M and a pH optimum of around pH 9.5. Treatment of the purified ADPRase with heated cytosol fractions at 37$^{\circ}C$ for 3 h caused some changes in the chemical properties of the enzyme, including an increase in molecular weight, a decrease in solubility, and a loss of $Mg^{2+}$-dependency. The molecular weight of the cytosol-treated ADPRase measured by gel filtration was over 420 kDa, suggesting, for the first time, that ADPRase could be polymerized by undefined cytoplasmic factors, and that polymerization is accompanied by changes in the solubility and metal ion dependency of the enzyme.

Nitrite Scavenging Effect of Maillard Reaction Products Derived from Glucose-Amino Acids (Glucose-아미노산계 Maillard 반응생성물의 아질산염 소거작용)

  • Kim, Seon-Bong;Lee, Dong-Ho;Yeum, Dong-Min;Park, Jin-Woo;Do, Jung-Roung;Park, Yeung-Ho
    • Korean Journal of Food Science and Technology
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    • v.20 no.3
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    • pp.453-458
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    • 1988
  • This research was carried out to investigate the effects of Maillard reaction products and nondialyzable melanoidins on the nitrite-scavenging. Nitrite-scavenging reactions were done at the different pH conditions(pH 1.2, 4.2 and 6.0). Maillard reaction products and nondialyzable melanoidins, produced from the glucose-amino acids(lys., gly., arg., his.)model systems, had a great of nitrite-scavenging effects. Nitrite-scavenging effects of Maillard reaction products and nondialyzable melanoidins were also pH dependent, being higher at pH 1.2 and lower at pH 6.0. By the treatment of Maillard reaction products and nondialyzable melanoidins with sodium borohydride, nitrite-scavenging effects were remarkably decreased at pH 1.2.

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Effect of salicylic acid and its analogues on stomatal closing in Commelina communis L. (닭의장풀의 기공닫힘에 미치는 살리실릭산과 그 유사물의 효과)

  • Joon-Sang Lee
    • Journal of Environmental Science International
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    • v.4 no.4
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    • pp.15-15
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    • 1995
  • ABA and SA showed different effect on stomatal closing on same condition. The addition of 1 M salicylic acid to fully opened stomata resulted in a significant reductionn of 22 % in stomatal aperture. However, 1 mM ABA reduced 73 % of stomatal aperture. The light absorption spectra of the salicylic acid solution showed that SA was degraded within 1 hour. Therefore, SA solution was resupplied % the detached epidermis every 30 min. during incubation and it was found that even at 10 $mu$M SA induced stomatal closing significantly. Its effect was also greatly pH dependent. The reduction of stomatal aperture caused by 1 mM SA was most effective at lower pH (pH 7.2, 5 %: pH 6.2, 40 %; pH 5.2, 78 %). Therefore, if SA was properly treated to the epidermal strips in the medium, the effects of SA on stomatal closing were similar with those of ABA.

Effect of salicylic acid and its analogues on stomatal closing in Commelina communis L. (닭의장풀의 기공닫힘에 미치는 살리실릭산과 그 유사물의 효과)

  • 이준상
    • Journal of Environmental Science International
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    • v.4 no.4
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    • pp.317-321
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    • 1995
  • ABA and SA showed different effect on stomatal closing on same condition. The addition of 1 M salicylic acid to fully opened stomata resulted in a significant reductionn of 22 % in stomatal aperture. However, 1 mM ABA reduced 73 % of stomatal aperture. The light absorption spectra of the salicylic acid solution showed that SA was degraded within 1 hour. Therefore, SA solution was resupplied % the detached epidermis every 30 min. during incubation and it was found that even at 10 $\mu$M SA induced stomatal closing significantly. Its effect was also greatly pH dependent. The reduction of stomatal aperture caused by 1 mM SA was most effective at lower pH (pH 7.2, 5 %: pH 6.2, 40 %; pH 5.2, 78 %). Therefore, if SA was properly treated to the epidermal strips in the medium, the effects of SA on stomatal closing were similar with those of ABA.

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Kinetic Studies on the Mechanism of Hydrolysis of 4'-[N-(9-Acridinyl)]-1'-(N-methanesulfonyl)-3'-methoxyquinonediimide (4'-[N-(9-Acridinyl)]-1'-(N-methanesulfonyl)-3'-methoxyquinonediimide의 가수분해 반응메카니즘에 관한 반응속도론적 연구)

  • Kim, Tae Rin;Chung, Dong In;Pyun, Sang Yong
    • Journal of the Korean Chemical Society
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    • v.40 no.12
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    • pp.733-740
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    • 1996
  • The rate constants for the hydrolysis of 4'-[N-(9-acridinyl)]-1'-(N-methanesulfonyl)-3'-methoxyquinonediimide(AMQD) were determined by ultraviolet visible spectrophotometer in water at $25^{\circ}C.$ The rate equation which could be applied over wide pH ranges were obtained. On the basis of pH-rate profile, Bronsted plot, hydrolysis product analysis, general base catalysis and substituent effect, the plausible hydrolysis mechanism was proposed: Below pH 3.00, the hydrolysis reaction was proceeded by the attack of water to 4'-position of quinonoid after protonation at nitrogen of acridinyl and between pH 3.00 and 9.00, the addition of water and hydroxide occurred competitively. However, above pH 9.00, the rate constants were dependent upon only the concentration of hydroxide ion.

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ATP-Induced Histamine Release Is in Part Related to Phospholipase $A_2$-Mediated Arachidonic Acid Metabolism in Rat Peritoneal Mast Cells

  • Lee, Yun-Hye;Lee, Seung-Jun;Seo, Moo-Hyun;Kim, Chang-Jong;Sim, Sang-Soo
    • Archives of Pharmacal Research
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    • v.24 no.6
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    • pp.552-556
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    • 2001
  • Histamine and arachidonic acid (AA) release was measured using the P2-purinoceptor antaongists, phospholipase $A_2{\;}(PLA_2)$ and cyclooxygenase (COX)/lipoxygenase (LOX) inhibitors to determine whether or not ATP-induced histamine release is associated with arachidonic acid (AA) release in rat peritoneal mast cells. ATP increased histamine release in a dose dependent manner, whereas adenosine did not. PPADS (a selective P2X-purinoceptor antagonist) and suramin (a nonselective P2X,2Y-purinoceptor antagonist) inhibited ATP-induced histamine release in a dose dependent manner. However, RB-2 (a P2Y-purinoceptor antagonist) did not block ATP-induced histamine release. Manoalide and oleyloxyethyl phosphorylcholine (OPC), secretory PLA$_2$ inhibitors, also inhibited ATP-induced histamine release dose-dependently. Both COX inhibitors (ibuprofen and indomethacin) and LOX inhibitors (baicalein and caffeic acid) inhibited ATP-induced histamine in a dose dependent manner. ATP significantly increased [$^3H$]AA release by 54%. PPADS and suramin significantly inhibited ATP-induced [3H]Ph release by 81% and 39%, respectively. ATP-induced histamine release was significantly inhibited by a variety of protein kinase inhibitors, such as bisindolmaleimide, genistein, methyl 2,5-dihydroxycinnamate, W-7 and trifluoperazine. Overall, the results suggest that ATP-induced histamine release is in part related to the PLA2-mediated AA metabolism and P2X-purinoceptors.

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Z-Domain Frequency Dependent Network Equivalent for Electromagnetic Transient and Harmonic Assessment (전자기 과도현상 해석과 고조파 평가를 위한 Z영역 주파수 의존 등가시스템 개발)

  • Wang, Y.P.;Chong, H.H.;Kim, K.Y.;Lee, J.T.;Han, H.H.;An, B.C.;Jeon, Y.S.
    • Proceedings of the KIEE Conference
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    • 2006.07a
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    • pp.145-146
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    • 2006
  • The recent power systems are very complex and to model them completely is impractical for analysis of electromagnetic transient. Therefore areas outside the immediate area of interest must be represented by some form of Frequency Dependent Network Equivalent (FDNE). In this paper a method for developing Frequency Dependent Network Equivalent (FDNE) using Z-domain rational Function Fitting is presented and demonstrated. The FDNE is generated by Linearized Least Squares Fitting(LSF) of the frequency response of a Z-domain formulation. This Three-port FDNE have been applied to the test AC power system. The electromagnetic transient package PSCAD/EMTDC is used to assess the transient response of the Three-port FDNE developed under different condition. The study results have indicated the robustness and accuracy of Three-port FDNE for analisys of electromagnetic transient and harmonic assessment.

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Protective Effects of Samul-tang on ${H_2O_2}-induced$ Cell Apoptosis in Cultured Cardiomyoblast Cells ($H_2O_2$에 의한 배양심근세포고사에 미치는 사물탕의 방어효과)

  • 박종운;한상혁;김도환;문병순
    • The Journal of Korean Medicine
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    • v.22 no.4
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    • pp.58-68
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    • 2001
  • Objectives : This study was designed to investigate the protective mechanisms of Samul-tang (SMT) on $H_2O_2$-induced toxicity in H9c2 cardiomyoblast cells. Methods : The cultured cells were pretreated with SMT and exposed to $H_2O_2$. The cell damage was assessed by using MTT assay. Also, we used Hoechst staining, Western blotting analysis. Results : SMT significantly reduced both $H_2O_2$-induced cell death and chromatin fragmentation. The decrease of Bcl2 expression by $H_2O_2$ was inhibited by SMT. In addition, the increase of Bax expression was also inhibited by SMT. In particular, Fas expression, which is generally recognized as cell death inducing signal by Fas/FasL interaction, was markedly decreased by $H_2O_2$ in a time-dependent manner, whereas this decrease was completely prevented by SMT. The cotreatment of SMT and $H_2O_2$ in H9c2 cells also induced the phosphorylation of ERK in a time-dependent manner. Moreover, PD098059, a specific inhibitor of ERKl/2, attenuated the protective effect of SMT on $H_2O_2$-induced toxicity in H9c2 cardiomyoblast cells. Furthermore, the protective effect of SMT was significantly blocked by treatment of SB203580, a specific inhibitor of p38. Conclusions : Taken together, this study suggests that the protective effects of the water extract of SMT against oxidative damages may be mediated by the modulation of Bel2 and Bax expression via the regulation of ERK and p38 signaling pathway.

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