• Title/Summary/Keyword: oxidative enzymes

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Partial Purification of Protein X from the Pyruvate Dehydrogenase Complex of Bovine Kidney

  • ;;;;Richard L. Veech
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.260-260
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    • 1994
  • Mammalian pyruvate dehydrogenase complex(PDC) enzyme consists of multiple oopies of three major oligomeric enzymes-El, E2 E3. And protein X is one of the enzymatic constituents which is tightly bound to E2 subunit This complex enzyme is responsible for the oxidative decarboxylation of pyruvate producing of acetyl CoA which is a key intermediate for the entry of carbohydrates into the TCA cycle for its complete metabolic conversion to CO$_2$. And the overall activity of the complex enzyme is regulated via covalent nodification of El subunit by a El specific phosphatase ad kinase. Protein X has lipoyl moiety that undergoes reduction and acetylation during ezymatic reaction and has been known h be involved in the binding of E3 subunit to E2 core and in the regulatory activity of kinase. The purification of protein X has not been achieved majorly because of its tight binding to E2 subunit The E2-protein X subcomplex was obtained by the established methods and the detachment of protein X from E2 was accomplished in the 0.1M borate buffer containing 150mM NaCl. During the storage of the subcomplex in frozen state at -70$^{\circ}C$, the E2 subunit was precipitated and the dissociated protein X was obtained by cntrifegation into the supernatant The verification of protein X was accomplished by (1)the migration on SDS-PAGE, (2)acetylation by 〔2$\^$-l4/C〕 pyruvate, and (3)internal amino acid sequence analysis of tryptic digested enzyme.

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Lipid-lowering and hepatoprotective effects of Vitis vinifera dried seeds on paracetamol-induced hepatotoxicity in rats

  • Almajwal, Ali Madi;Elsadek, Mohamed Farouk
    • Nutrition Research and Practice
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    • v.9 no.1
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    • pp.37-42
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    • 2015
  • BACKGROUND/OBJECTIVES: Red grape seeds as functional food are a good source of important bioactive components such as phenolics and antioxidants, which decrease oxidative stress that contributes to the pathogenesis of hepatotoxicity. The current study was conducted in order to evaluate the protective effect of red grape dried seeds (RGDS) on antioxidant properties, lipid metabolism, and liver and kidney functions of rats with paracetamol (750 mg/kg) induced hepatotoxicity. MATERIALS/METHODS: RGDS was added to the basal diet at 5, 10, and 20%. Thirty five adult male rats were assigned to five groups (n = 7) for a six-week feeding period; group (1) normal control, group (2) induced control, groups (3, 4, and 5) fed a diet with RGPS at different levels, 5, 10, and 20%, respectively. At the end of the feeding period, animals' blood and tissues were collected for estimation of serum lipid profile, serum liver, and kidney biomarkers. The protection was measured by detecting lipid peroxidation (LPO), glutathione (GSH), superoxide dismutase (SOD), Catalase (CAT) (in liver tissues), and liver histological examination. RESULTS: The results showed a significant (P < 0.05) decrease in levels of serum cholesterol, triglycerides, low density lipoprotein (LDL-C), and very low density lipoprotein (VLDL-C), with a significant increase in level of high density lipoprotein (HDL-C) for RGDS groups compared to induced control. Rats administered a diet containing RGDS levels produced significant (P < 0.05) hepatoprotection by decreasing the activities of liver enzymes, kidney parameters, and lipid peroxidation, while levels of GSH, SOD, and CAT were increased significantly to near the normal levels. CONCLUSION: The RGDS 20% group was more effective than others against hepatotoxicity of paracetamol, which may be attributed to RGDS total phenols and antioxidant contents, which were 1.438 mg and 1.231 mg, respectively.

Expression of Cu/Zn SOD Protein Is Suppressed in hsp 70.1 Knockout Mice

  • Choi, S-Mi;Park, Kyung-Ae;Lee, Hee-Joo;Park, Myoung-Sook;Lee, Joung-Hee;Park, Kyoung-Chan;Kim, Man-Ho;Lee, Seung-Hoon;Seo, Jeong-Sun;Yoon, Byung-Woo
    • BMB Reports
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    • v.38 no.1
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    • pp.111-114
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    • 2005
  • Heat shock proteins (HSPs) are known to protect cells from oxidative stress and other types of injuries. We previously reported the neuroprotective effect of HSP70 following cerebral ischemia and reperfusion using hsp 70.1 knockout (KO) mice. However, the precise role of HSP70 in neuroprotection has not been established yet. The purpose of this study was to investigate the relationship between HSP70 and antioxidant enzymes using hsp 70.1 KO mice. The activities of both SOD-1 and SOD-2 were significantly decreased in hsp 70.1 KO mice than in the wild type (WT) littermates. SOD-1 protein level in the hsp 70.1 KO mice was lower than that of WT. We speculate that HSP70 might be involved in regulation of expression of SOD-1 at the level of transcription or by post-transcriptional modification.

The Mitogen-Activated Protein Kinase Signal Transduction Pathways in Alternaria Species

  • Xu, Houjuan;Xu, Xiaoxue;Wang, Yu-Jun;Bajpai, Vivek K.;Huang, Lisha;Chen, Yongfang;Baek, Kwang-Hyun
    • The Plant Pathology Journal
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    • v.28 no.3
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    • pp.227-238
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    • 2012
  • Mitogen-activated protein kinase (MAPK) cascades are conserved signaling modules in the eukaryotic cells. They are involved in many major cell processes in fungi such as stress responses, vegetative growth, pathogenicity, secondary metabolism and cell wall integrity. In this review, we summarized the advances of research on the MAPK signaling pathways in Alternaria species. As major phytopathogenic fungi, Alternaria species reduce crop production. In contrast to the five MAPK pathways known in yeast, only three MAPK pathways as Fus3/Kss1-type, Hog1-type, and Slt2-type have been characterized in Alternaria. The Fus3/Kss1-type MAPK pathway participates in regulation of vegetative growth, conidiation, production of some cell-wall-degrading enzymes and pathogenicity. The Hog1-type pathway is involved in osmotic and oxidative stress, fungicides susceptibility and pathogenicity. The Slt2-type MAP kinases play an important role on maintaining cell wall integrity, pathogenicity and conidiation. Although recent advances on the MAPK pathways in Alternaria spp. reveal many important features on the pathogenicity, there are many unsolved problems regarding to the unknown MAP kinase cascade components and network among other major signal transduction. Considering the economic loss induced by Alternaria spp., more researches on the MAPK pathways will need to control the Alternaria diseases.

Alteration in Pyridine Nucleotide Status in Cells as an Adaptive Response to Water Stress in Rice (Oryza sativa L.) Seedlings

  • Boo, Yong-Chool;Jung, Jin
    • Applied Biological Chemistry
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    • v.41 no.4
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    • pp.228-234
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    • 1998
  • An adaptive measure of photosynthetic cells to a condition identified with a reduction of cellular energy charge, caused by water deficit-induced impairment of photosynthetic ATP production, was investigated using hydroponically cultured rice seedlings. Water stress treatment of the seedlings resulted in a marked decrease in cellular ATP level, a significant increase in the content of NAD(H) and concurrent decrease in that of NADP(H) in shoots, which accompanied a decrease in the activity of NAD kinase (EC 2.7.1.23) that specifically converts NAD(H) to NADP(H). The decline in the enzyme activity was particularly evident in the $Ca^{2+}/calmodulin-dependent$ kinase, the major form of NAD kinase in plants, whereas the level of active calmodulin remained unchanged during water deficit. The ratio of $NADP^+$ to NADPH was maintained nearly constant and no increases were seen in the level of $H_2O_2$ and the activities of $superoxide/H_2O_2-detoxifying$ enzymes in shoots stress-treated for two days. Based on these results, it may be suggested that rice plants take a strategy to cope with an adverse situation of limited photophosphorylation created by water deficit in that cells facilitate ATP production through glycolysis and oxidative phosphorylation; in doing so, rice cells suppress NAD kinase activity, consequently up-sizing the NAD(H) pool at the expense of the NADP(H) pool. Several parameters associated with the stress symptoms are also of implicative that there is no overproduction of superoxide radical or the related active oxygen at least in rice seedlings.

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Protection by Sunghyangchungisan against Oxidative Endothelial Cell Injury (배양(培養)된 혈관(血管) 내피세포(內皮細胞)에서 산화성(酸化性) 세포(細胞) 손상(損傷)에 미치는 성향정기산(星香正氣散)의 보호(保護) 효과(效果))

  • Lee Dong-Uhn;Kim Young-Kyun
    • Herbal Formula Science
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    • v.8 no.1
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    • pp.147-167
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    • 2000
  • Reactive oxygen species (ROS) play an important role in the pathogenesis of a variety of life threatening conditions such as atherosclerosis, myocardial infarction and cerebral stroke. In this study, the effect of Sunghyangchungisan (SHCS) as a cytoproctant against ROS-induced cell injury was studied by investigating its effect on $H_{2}O_2-induced$ cell injury in cultured endothelial cells derived from the human umbilical vein. SHCS effectively proteced the cells against $H_{2}O_2-induced$ injury determined by trypan blue exclusion ability and lactate dehydrogenase (LDH) release. The effect of SHCS was concentration-dependent and the concentrations to inhibit by 50% the cell death and LDH release were $0.9{\pm}0.1$ and $1.2{\pm}0.1\;mg/ml$, respectively. In addition, SHCS effectively protected the cells against t-butylhydroperoside- and menadione-Induced injury as well. SHCS inhibited lipid peroxidation determined by malondialdehyde production. SHCS exerted as an effective scavenger of ROS produced by exposing the cells to $H_{2}O_2$ The activities of the intracellular ROS scavenging enzymes such as superoxide dismutase, catalase and glutathione peroxidase were not Influenced by SHCS.These results indicate that SHCS might exert as an effective cytoprotectant against ROS-induced cell injury. Further intensive studies would provide us insights into mechanisms of the pharmacological actions of SHCS.

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Isolation of Lactic Acid Bacteria Showing Antioxidative and Probiotic Activities from Kimchi and Infant Feces

  • Ji, Keunho;Jang, Na Young;Kim, Young Tae
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1568-1577
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    • 2015
  • The purpose of this study was to investigate lactic acid bacteria with antioxidative and probiotic activities isolated from Korean healthy infant feces and kimchi. Isolates A1, A2, S1, S2, and S3 were assigned to Lactobacillus sp. and isolates A3, A4, E1, E2, E3, and E4 were assigned to Leuconostoc sp. on the basis of their physiological properties and 16S ribosomal DNA sequence analysis. Most strains were confirmed as safe bioresources through nonhemolytic activities and non-production of harmful enzymes such as β-glucosidase, β-glucuronidase and tryptophanase. The 11 isolates showed different resistance to acid and bile acids. In addition, they exhibited antibacterial activity against foodborne bacteria, especially Bacillus cereus, Listeria monocytogenes, and Escherichia coli. Furthermore, all strains showed significantly high levels of hydrophobicity. The antioxidant effects of culture filtrates of the 11 strains included 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging capacity, 2.2'-azino-bis (2-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation scavenging activity, and superoxide dismutase activity. The results revealed that most of the culture filtrates have effective scavenging activity for DPPH and ABTS radicals. All strains appeared to have effective superoxide dismutase activity. In conclusion, the isolated strains A1, A3, S1, and S3 have significant probiotic activities applicable to the development of functional foods and health-related products. These strains might also contribute to preventing and controlling several diseases associated with oxidative stress, when used as probiotics.

Effect of Rhodiola Sachalinensis Administration and Endurance Exercise on Insulin Sensitivity and Expression of Proteins Related with Glucose Transport in Skeletal Muscle of Obese Bucker Rat (홍경천 섭취와 운동수행이 비만 쥐의 인슐린 민감도와 골격근내 당수송 관련 단백질 발현에 미치는 영향)

  • Oh Jae-Keun;Shin Young-Oh;Jung Hee-Jung;Lee Jung-Eun
    • Journal of Nutrition and Health
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    • v.39 no.4
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    • pp.323-330
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    • 2006
  • Peripheral insulin resistance in obese/type II diabetes animals results from an impairment of insulin-stimulated glucose uptake into skeletal muscle. Insulin stimulate the translocation of GLUT4 from intracellular location to the plasma membrane. Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) is implicated in mediation of fusion of GLUT4-containing vesicle with the plasma membrane. Present study investigated regulatory effects of Rhodiola sachalinensis administration and exercise training on the expression of GLUT4 protein and SNAREs protein in skeletal muscles of obese Zucker rats. Experimental animals were randomly assigned into one of five groups ; lean control(LN), obese control(OB), exercise-treated(EXE), Rhodiola sachalinensis-treated(Rho), combine of Rho & EXE (Rho-EXE). All animals of exercise training (EXE, Rho-EXE) performed treadmill running for 8 weeks, and animals of Rho groups (Rho, Rho-EXE) were dosed daily by gastric gavage during the same period. After experiment, blood were taken for analyses of glucose, insulin, and lipids levels. Mitochondrial oxidative enzyme (citrate synthase, CS ; $\beta$-hydroxyacyl-CoA dehydrogenase, $\beta$-HAD) activity were analysed. Skeletal muscles were dissected out for analyses of proteins (GLUT4, VAMP2, syntaxin4, SNAP23). Results are as follows. Exercise and/or Rhodiola sachalinensis administration significantly reduced body weight and improved blood lipids (TG, FFA), and increased insulin sensitivity. Endurance exercise significantly increased the activity of mitochondrial enzymes and the expression of GLUT4 protein, however, administration of Rhodiola sachalinensis did not affect them. The effect of exercise and/or Rhodiola sachalinensis administration on the expression of SNARE proteins was unclear. Our study suggested that improvement insulin sensitivity by exercise and/or Rhodiola sachalinensis administration in obese Zucker rats is independent of expression of SNARE proteins.

Inhibitory Effects of Marine Natural Products on Melanogenesis in B16 Melanoma Cells (B16 멜라닌 세포에서 해양소재 추출물의 멜라닌 생성 저해 효과)

  • Lee, Chan;Jang, Jung-Hee;Ahn, Eun-Mi;Park, Chan-Ik
    • The Korea Journal of Herbology
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    • v.27 no.4
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    • pp.73-80
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    • 2012
  • Objectives : Under normal condition melanin protects the skin from extracellular stimuli including ultraviolet (UV)-induced oxidative skin damages, but excess production and accumulation of melanin can induce hyperpigmentation causing esthetic problems. Therefore, in this study we tried to search for natural skin whitening materials from marine natural resources. Methods : Water and ethanol extracts of marine natural resources were prepared from Porphyra thalli (PT), Laminariae thallus (LT), Ostreae concha (OC), Sargassum thallus (ST), Undaria thallus (UT), Codium thalli (CT), Enteromorpha thalli (ET), Syngnathoides biaculeatus (SB), and Hippocampus coronatus (Hc). Their effects against UVB and ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis were investigated based on melanin formation in B16 mouse melanoma cells. The mRNA and protein expression of enzymes involved in the melanogenic process were further examined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Results : Water extract of Ostreae concha (OCW/E) effectively inhibited UVB and ${\alpha}$-MSH-induced melanin production in B16 melanocytes, which seemed to be mediated by inhibition of mRNA expression of tyrosinase and tyrosinase-related protein 1 (TRP-1). In another experiment, ethanol extracts from Porphyra thalli (PTE/E), Laminariae thallus (LTE/E), Sargassum thallus (STE/E), Undaria thallus (UTE/E), Codium thalli (CTE/E), Syngnathoides biaculeatus (SBE/E), and Hippocampus coronatus (HcE/E) significantly suppressed UVB and ${\alpha}$-MSH-induced melanin formation. Furthermore, ethylacetate fraction isolated form LTE/E (LTE/EEt) decreased UVB and ${\alpha}$-MSH-elevated extracellular melanin levels via inhibition of tyrosinase protein expression. Conclutions : These results suggest that marine natural resources such as Porphyra thalli, Laminariae thallus, Ostreae concha, Sargassum thallus, Undaria thallus, Codium thalli, Syngnathoides biaculeatus and Hippocampus coronatus have anti-melanogenic effects, thereby exhibiting high potentials to be utilized as one of the ingredients for the development of new whitening functional cosmetics.

The Oxidative Effects of Benzo[a]pyrene in Rat Hepatocyte Primary Culture (랫드 간세포 일차배양에서 Benzo[a]pyrene의 산화 효과)

  • Im, Tae Jin
    • Journal of Environmental Science International
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    • v.13 no.4
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    • pp.413-420
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    • 2004
  • The objectives of present study were to investigate the effects of benzo[a]pyrene(BaP) on cytotoxicity, lipid peroxidation and antioxidant enzymes in rat hepatocyte primary culture. Primary cultures of rat hepatocytes were incubated for 24 hr, 48 hr or 72 hr in the presence of various concentrations (0, 10, 20, 30, 50 or 100 $\mu.$ M) of BaP. Cytotoxicity and cell viability were determined by measuring glutamic oxaloacetic transaminase(GOT) activity, lactate dehydrogenase(LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MIT) value. Lipid peroxidation was evaluated using thiobarbituric acid reactive substances(TBARS) assay. Effects on antioxidant system were determined by measuring glutathione peroxidase(GPx) activity, glutathione reductase(GR) activity and glutathione concentration. Activities of GOT and LDH, MTT value as well as TBARS concentration were not affected by up to 100 $\muM$ of BaP for 24 hr incubation. However, BaP at the concentration of 50 $\muM$ for 48 hr incubation or at the concentration of 30 $\muM$ for 72 hr incubation began to increase LDH activity and TBARS concentration but decrease MTT value, representing that BaP caused cytotoxicity and decreased cell viability in dose- and time-dependent manners. GPx activity began to be decreased by BaP at the concentration of 50 $\muM$ for 72 hr incubation. Whereas, GR activity began to be decreased by BaP at the concentration of 20 $\muM$ for 72 hr incubation. Glutathione concentration began to be decreased by BaP at the concentration of 20 $\muM$ for 72 hr incubation and was further reduced to 90% by 100 $\muM$ of BaP. These results demonstrate that BaP caused cytoctoxicity and decreased cell viability by increasing lipid peroxidation and decreasing glutathione concentration as well as activities of GPx and GR.