• Title/Summary/Keyword: ATP level

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Nucleotide Triphosphates Inhibit the Degradation of Unfolded Proteins by HslV Peptidase

  • Lee, Jung Wook;Park, Eunyong;Bang, Oksun;Eom, Soo-Hyun;Cheong, Gang-Won;Chung, Chin Ha;Seol, Jae Hong
    • Molecules and Cells
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    • v.23 no.2
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    • pp.252-257
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    • 2007
  • Escherichia coli HslVU is an ATP-dependent protease consisting of two heat shock proteins, the HslU ATPase and HslV peptidase. In the reconstituted enzyme, HslU stimulates the proteolytic activity of HslV by one to two orders of magnitude, while HslV increases the rate of ATP hydrolysis by HslU several-fold. Here we show that HslV alone can efficiently degrade certain unfolded proteins, such as unfolded lactalbumin and lysozyme prepared by complete reduction of disulfide bonds, but not their native forms. Furthermore, HslV alone cleaved a lactalbumin fragment sandwiched by two thioredoxin molecules, indicating that it can hydrolyze the internal peptide bonds of lactalbumin. Surprisingly, ATP inhibited the degradation of unfolded proteins by HslV. This inhibitory effect of ATP was markedly diminished by substitution of the Arg86 residue located in the apical pore of HslV with Gly, suggesting that interaction of ATP with the Arg residue blocks access of unfolded proteins to the proteolytic chamber of HslV. These results suggest that uncomplexed HslV is inactive under normal conditions, but may can degrade unfolded proteins when the ATP level is low, as it is during carbon starvation.

Wheat phytase potentially protects HT-29 cells from inflammatory nucleotides-induced cytotoxicity

  • Jeongmin An;Jaiesoon Cho
    • Animal Bioscience
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    • v.36 no.10
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    • pp.1604-1611
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    • 2023
  • Objective: The aim of this study was to investigate the protective effect of wheat phytase as a structural decomposer of inflammatory nucleotides, extracellular adenosine triphosphate (ATP), and uridine diphosphate (UDP) on HT-29 cells. Methods: Phosphatase activities of wheat phytase against ATP and UDP was investigated in the presence or absence of inhibitors such as L-phenylalanine and L-homoarginine using a Pi Color Lock gold phosphate detection kit. Viability of HT-29 cells exposed to intact- or dephosphorylated-nucleotides was analyzed with an EZ-CYTOX kit. Secretion levels of pro-inflammatory cytokines (IL-6 and IL-8) in HT-29 cells exposed to substrate treated with or without wheat phytase were measured with enzyme-linked immunosorbent assay kits. Activation of caspase-3 in HT-29 cells treated with intact ATP or dephosphorylated-ATP was investigated using a colorimetric assay kit. Results: Wheat phytase dephosphorylated both nucleotides, ATP and UDP, in a dose-dependent manner. Regardless of the presence or absence of enzyme inhibitors (L-phenylalanine and L-homoarginine), wheat phytase dephosphorylated UDP. Only L-phenylalanine inhibited the dephosphorylation of ATP by wheat phytase. However, the level of inhibition was less than 10%. Wheat phytase significantly enhanced the viability of HT-29 cells against ATP- and UDP-induced cytotoxicity. Interleukin (IL)-8 released from HT-29 cells with nucleotides dephosphorylated by wheat phytase was higher than that released from HT-29 cells with intact nucleotides. Moreover, the release of IL-6 was strongly induced from HT-29 cells with UDP dephosphorylated by wheat phytase. HT-29 cells with ATP degraded by wheat phytase showed significantly (13%) lower activity of caspase-3 than HT-29 cells with intact ATP. Conclusion: Wheat phytase can be a candidate for veterinary medicine to prevent cell death in animals. In this context, wheat phytase beyond its nutritional aspects might be a novel and promising tool for promoting growth and function of intestinal epithelial cells under luminal ATP and UDP surge in the gut.

Nitric Oxide-cGMP-Protein Kinase G Pathway Contributes to Cardioprotective Effects of ATP-Sensitive $K^+$ Channels in Rat Hearts

  • Cuong, Cang Van;Kim, Na-Ri;Cho, Hee-Cheol;Kim, Eui-Yong;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.2
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    • pp.95-100
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    • 2004
  • Ischemic preconditioning (IPC) has been accepted as a heart protection phenomenon against ischemia and reperfusion (I/R) injury. The activation of ATP-sensitive potassium $(K_{ATP})$ channels and the release of myocardial nitric oxide (NO) induced by IPC were demonstrated as the triggers or mediators of IPC. A common action mechanism of NO is a direct or indirect increase in tissue cGMP content. Furthermore, cGMP has also been shown to contribute cardiac protective effect to reduce heart I/R-induced infarction. The present investigation tested the hypothesis that $K_{ATP}$ channels attenuate DNA strand breaks and oxidative damage in an in vitro model of I/R utilizing rat ventricular myocytes. We estimated DNA strand breaks and oxidative damage by mean of single cell gel electrophoresis with endonuclease III cutting sites (comet assay). In the I/R model, the level of DNA damage increased massively. Preconditioning with a single 5-min anoxia, diazoxide $(100\;{\mu}M)$, SNAP $(300\;{\mu}M)$ and 8-(4-Chlorophenylthio)-guanosine-3',5'-cyclic monophosphate (8-pCPT-cGMP) $(100\;{\mu}M)$ followed by 15 min reoxygenation reduced DNA damage level against subsequent 30 min anoxia and 60 min reoxygenation. These protective effects were blocked by the concomitant presence of glibenclamide $(50\;{\mu}M)$, 5-hydroxydecanoate (5-HD) $(100\;{\mu}M)$ and 8-(4-Chlorophenylthio)-guanosine-3',5'-cyclic monophosphate, Rp-isomer (Rp-8-pCPT-cGMP) $(100\;{\mu}M)$. These results suggest that NO-cGMP-protein kinase G (PKG) pathway contributes to cardioprotective effect of $K_{ATP}$ channels in rat ventricular myocytes.

An Allocation of Safety Integrity Level to Inductive Loop type Train Control System (유도루프식 열차제어시스템 안전무결성등급 할당)

  • Ryou, Sung-Kyun;Park, Jae-Young;Yun, Hak-Sun
    • The Journal of the Korea institute of electronic communication sciences
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    • v.8 no.12
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    • pp.1905-1910
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    • 2013
  • This paper demonstrates the result of Safety Integrity Level (SIL) allocation for IL-type Train Control System(IL-TCS), by applying the semi-quantitative approach. IL-type TCS is defined in this paper as the set of Hardware and Software ATS equipment, Track-side ATP equipment, On-board ATP equipment, Track-side ATO equipment, On-board ATO equipment. SIL allocation is performed for these constituent subsystems of TCS. Based on three principles of the semi-quantitative method, the SIL allocation process is performed for the subsystems composing TCS.

A study of eatablishing ETCS Level 1 to speed up conventional lines (기존선 속도향상을 위한 ETCS Level 1 구축방안 연구)

  • Yoon, Yong-Ki;Jeong, Rag-Gyo;Kim, Chae-Duck
    • Proceedings of the KSR Conference
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    • 2011.05a
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    • pp.666-670
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    • 2011
  • ATS(Automatic Train Stop)s are is effected by track environment, signal distance and train braking distance. So onboard signaling systems such as ATP(Automatic Train Protection) are installed on conventional lines by some train operation companies. With the mixed signalling, the onboard signaling system is overlaid on a conventional line with ATS, it is possible to run the line with conventional trains and ATP trains and to use the advantages of ATP(higher speed or more trains on the line). This paper includes guidelines a mixed signalling(ETCS and ATS) system architecture, operation concepts and infill balise installation.

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Effect of Carbon Monoxide Intoxication on the Change in Contents of Cerebral Energy Metabolites of Rats (흰쥐에서의 일산화탄소(一酸化炭素) 중독(中毒)이 뇌(腦)에너지 대사(代謝) 관련물질(關聯物質) 함량변화(含量變化)에 미치는 영향)

  • Yun, Jae-Soon;Choi, Shin-Kyu
    • YAKHAK HOEJI
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    • v.33 no.3
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    • pp.149-155
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    • 1989
  • To predict the influence of carbon monoxide poisonining on cerebral energy metabolism, rats were exposed to 5000 ppm environment for 30 minutes. Carboxyhemoglobin (HBCO) saturation rate in this condition was 72% equally in male and female rats. Cerebral cortex in the rats showed lower level of ATP, glucose, creatine phosphate and higher level of lactate, pyruvate by anaerobic glycolysis. As for the levels of ATP, creatine phsphate and glucose, the cerebral cortex contents of them were larger in female rats of estrus than in male rats, whereas there was no difference between sexes in the levels of pyruvate and lactate. According to time passage from CO intoxication, the mode of changes in cerebral energy metabolite contents was similar in both sexes.

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Evaluation of Hygienic Status of University Foodservice Operation using ATP bioluminescence Assay (ATP bioluminescence Assay를 이용한 대학 급식시설의 위생상태 평가에 관한 연구)

  • 박영숙
    • Korean journal of food and cookery science
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    • v.16 no.2
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    • pp.195-201
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    • 2000
  • An investigation was conducted to evaluate the hygienic status of university foodservice operation by using conventional swabbing technique plus standard plate count and ATP bioluminescence assay. The results of the study were as follows: 1) For all kitchen boards, knives, feeding trays, and dish towels tested, there was an overall agreement at 84.7% level between the results obtained using ATP bioluminescence and plate count when using a pass/fail cut-off of 3$\times$ control values for ATP assay and 40 CFU(colony forming unit)/㎠ for plate count. 2) The agrement rate between ATP assay and standard plate count was 87.5% for the samples before use, 29.2% for those during use, and 42.7% for those after cleaning and sanitizing. 3) The plate counts of three university foodservice operations for kitchen board, kitchen knife, feeding tray and dish towel were within the acceptable limits when tested before using. However, none of them were within the acceptable limits when tested during using and after cleaning and sanitizing. 4) Above results suggested that an immediate action needs to be taken to reduce the potential danger of cross-contamination and also effective sanitary control methods needs to be developed to improve the sanitary condition.

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DMI Display implemented as single layout based on ERTMS specification for Integrated Onboard Signaling System (통합 열차제어 신호시스템을 위한 단일 레이아웃의 ERTMS 기반 DMI 화면의 설계 및 구현)

  • Jung, Ji-Chan;Kim, Hyung-Hoon;Cho, Yong-Gee
    • Proceedings of the KSR Conference
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    • 2011.10a
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    • pp.251-254
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    • 2011
  • This paper describes the design and implementation of the DMI display based on ERTMS specification for the Integrated Onboard Signaling System (IOSS). IOSS is consists of ATS, ATC, and ATP based on ERTMS/ETCS Level 1. We propose an improved DMI display design which has single layout concept for the IOSS. When a train is in the ATS, ATC, or ATP railway section, it is protected by the ATS, ATC, or ATP equipment, respectively. Therefore, DMI should display the different information according to the each signaling system. We design DMI display implemented as single layout for ATP, ATS and ATC information. This single layout display has a well integrated look and feel using the ERTMS specification and layout. This is for the driver to perceive the information easily and not to experience inconvenience resulting from the sense of difference when the train crossed into the different signaling section. Finally, we show the implementation result following this concept.

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A Study on the Protective Effect and Its Mechanism of Zinc against Immuno-cytotoxicity of Methylmercury (유기수은의 세포면역독성과 이에 대한 아연의 방어효과 및 기전)

  • 고대하;염정호;오경재
    • Journal of Environmental Health Sciences
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    • v.27 no.2
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    • pp.82-91
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    • 2001
  • This study was carried out to elucidate the protective effect of zinc chloride(ZnCl$_2$) and its mechanism against the immuno-cytotoxicity of methylmercury chloide($CH_3$HgCl). This study was observed in the culture of EMT-6 cells which are originated from mammary adenocarcinoma of Balb/c mouse. Cytotoxicity of metals was measured by cell viability and NO$_2$$^{[-10]}$ , and mitochondrial function was evaluated by adenosine triphosohate (ATP) production. $CH_3$HgCl significantly decreased the sythesis of nitric oxide(NO), ATP and glutathione(GSH) in a dose-dependent manner. ZnCl$_2$ significantly increased the synthesis of GSH in a dose-dependent manner, but synthesis of NO and ATP were not changed. The immuno-cytotoxicity of $CH_3$HgCl was not fully protected when combined addition of ZnCl$_2$, whereas ZnCl$_2$ prior to addition of $CH_3$HgCl completly protected the Hg-induced immuno-cytotoxicity. Similarly, intracellular accumulation of mercury significantly decreased by ZnCl$_2$. Degree of diminution of intracellular mercury was larger in ZnCl$_2$ prior to addition of $CH_3$HgCl than in combined addition of ZnCl$_2$ and $CH_3$HgCl.. Dithiothreitol(DTT) or buthionine sulfoximine(BSO) addition at 50$\mu$M or less, which was not toxic to the cells, did not affect synthesis of NO and ATP. DTT increased intracellular GSH level and DTT pretreatment protected toxicity induced by $CH_3$HgCl as shown complete recover in the NO and ATP values. BSO decreased intracellular GSH level and BSO pretreatment exaggerated toxicity induced by $CH_3$HgCl as shown synergistic reduction in the NO and ATP values. These results indicated that the protective effects of zinc against immuno-cytotoxicity of methylmercury associated with increasing cellular level of GSH. Increased intracellular GSH transports methylmercury to out of cells. In accordance with intracellular level of mercury decreased, immuno-cytotoxicity of methylmercury decreased. These result also suggest that the protective mechanism of zinc against the mercury toxicity would be exerted in the immune system in vivo.

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