• Title/Summary/Keyword: thiol

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Reactive Oxygen Species and Nitrogen Species Differentially Regulate Neuronal Excitability in Rat Spinal Substantia Gelatinosa Neurons

  • Lee, Hae In;Park, A-Reum;Chun, Sang Woo
    • International Journal of Oral Biology
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    • v.39 no.4
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    • pp.229-236
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    • 2014
  • Reactive oxygen species (ROS) and nitrogen species (RNS) are implicated in cellular signaling processes and as a cause of oxidative stress. Recent studies indicate that ROS and RNS are important signaling molecules involved in nociceptive transmission. Xanthine oxidase (XO) system is a well-known system for superoxide anions ($O{_2}^{{\cdot}_-}$) generation, and sodium nitroprusside (SNP) is a representative nitric oxide (NO) donor. Patch clamp recording in spinal slices was used to investigate the role of $O{_2}^{{\cdot}_-}$ and NO on substantia gelatinosa (SG) neuronal excitability. Application of xanthine and xanthine oxidase (X/XO) compound induced membrane depolarization. Low concentration SNP ($10{\mu}M$) induced depolarization of the membrane, whereas high concentration SNP (1 mM) evoked membrane hyperpolarization. These responses were significantly decreased by pretreatment with phenyl N-tert-butylnitrone (PBN; nonspecific ROS and RNS scavenger). Addition of thapsigargin to an external calcium free solution for blocking synaptic transmission, led to significantly decreased X/XO-induced responses. Additionally, X/XO and SNP-induced responses were unchanged in the presence of intracellular applied PBN, indicative of the involvement of presynaptic action. Inclusion of GDP-${\beta}$-S or suramin (G protein inhibitors) in the patch pipette decreased SNP-induced responses, whereas it failed to decrease X/XO-induced responses. Pretreatment with n-ethylmaleimide (NEM; thiol-alkylating agent) decreased the effects of SNP, suggesting that these responses were mediated by direct oxidation of channel protein, whereas X/XO-induced responses were unchanged. These data suggested that ROS and RNS play distinct roles in the regulation of the membrane excitability of SG neurons related to the pain transmission.

Bacillus stearothermophilus 에서 부분 정제한 Cytosine Deaminase 의 특성

  • 장영채;이경형;김성영;조윤래;김종규
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.305-309
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    • 1992
  • Cytosine deaminase (EC 3.5.4.1) from BaciNus stc~urorhermophilus was partially purified 7.2-fold with an overall yield of 52.7%. The partially purified enzyme deiiminated cytosine only.but not 5-methylcytosine and 5-fluorocytosine. The apparent Michaclis constant. Km valuefor cytosine was 5.9 mM. The enzyme was relatively stable in the range of pH 4.0 to 7.0.furthermore extremely thermo-stable : more than 75'X) of the activity was remained afterheating at 80$^{\circ}$C for I0 min at pH 6.5. The enzyme had a pH optimum at around pH7.0 to 7.5. and temperature optimum at 35 to 31$^{\circ}$C. And the activation energ (En value)determined from an Arrhenius plot was 26 Kcal/mol. The enzyme activity was stronglyinhibited by heavy metal ions such as Cd", Hg". Cut' at 1 mM, anJ by o-phenanthroline,and p-chloromcrcuribcnzoate at I mM. But the enrymc activity was activatetl increased byGMP, and CMP at 1 mM.ased by GMP, and CMP at 1 mM.

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Effect of Chelation with Calcium Disodium EDTA on Haemato-biochemical and Trace Mineral Profile in Blood from Lead Exposed Calves

  • Patra, R.C.;Swarup, D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.8
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    • pp.1130-1134
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    • 2005
  • An experiment was performed using 20 calves of about one-month old to investigate the effect of chelation therapy with calcium disodium ethylenediaminetetraacetate ($CaNa_2$EDTA) alone or along with antioxidant $\alpha$-tocopherol in lead loaded calves on blood trace minerals, erythrocytic sulfahydryl groups and some haematobiochemical parameters. Fifteen calves were given lead orally at a daily dose of 7.5 mg of 99% pure lead acetate/kg body weight for 28 days. Then the lead was withdrawn on day 28 and the calves were randomly divided into three groups. Each group of five animals was either treated with $CaNa_2$EDTA alone at the dose rate of 110 mg/kg body weight in two divided doses for 4 days or along with $\alpha$-tocopherol at the dose rate of 100 mg/kg body weight orally daily for 7 days, keeping the remaining five calves as lead-exposed untreated controls. Blood samples were collected at the end of the lead exposure (day 0) and thereafter on day 2, 4, 7 and 10 from the start of the chelation treatment. The treatment with EDTA alone led to slow but non-significant improvement in blood copper level, but incorporation of antioxidant $\alpha$-tocopherol in chelation therapy resulted in its significant decline, as recorded on day 7-post treatment. Withdrawal of lead or treatment with $CaNa_2$EDTA alone or along with $\alpha$-tocopherol enhanced the erythrocytic thiol contents and the levels of T-SH and P-SH became statistically (p<0.05) comparable to those of lead-exposed controls by day 7 and 4, respectively. There was no significant (p>0.05) change in serum urea, creatinine, total protein and albumin levels between the treatment groups. It is concluded from the present investigation that treatment with $CaNa_2$EDTA at the present dose rate is safe to be used for chelation in lead loaded calves.

Antimicrobial Effect of 2-Phenylethynyl-Butyltellurium in Escherichia coli and Its Association with Oxidative Stress

  • Pinheiro, Franciane Cabral;Bortolotto, Vandreza Cardoso;Araujo, Stifani Machado;Poetini, Marcia Rosula;Sehn, Carla Pohl;Neto, Jose S.S.;Zeni, Gilson;Prigol, Marina
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1209-1216
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    • 2018
  • This study aimed to evaluate the antimicrobial activity of 2-phenylethynyl-butyltellurium (PEBT) in Escherichia coli and the relation to its pro-oxidant effect. For this, we carried out the disk diffusion test, minimum inhibitory concentration (MIC) assay, and survival curve analysis. We also measured the level of extracellular reactive oxygen species (ROS), activity of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), and level of non-protein thiols (NPSH). PEBT at 1.28 and 0.128 mg/disk exhibited antimicrobial capability in the disk diffusion test, with an MIC value of 1.92 mg/ml, whereas PEBT at 0.96, 1.92, and 3.84 mg/ml inhibited bacterial growth after a 9-h exposure. PEBT at 3.84, 1.92, and 0.96 mg/ml increased extracellular ROS production, decreased the intracellular NPSH level, and reduced the SOD and CAT activities. Glutathione or ascorbic acid in the medium protected the bacterial cells from the antimicrobial effect of PEBT. In conclusion, PEBT exhibited antimicrobial activity against E. coli, involving the generation of ROS, oxidation of NPSH, and reduction of the antioxidant defenses in the bacterial cells.

Effect of Terminalia chebula on Physiological Activity in Mice (가자(Terminalia chebula) 추출물이 마우스의 생리활성에 미치는 영향)

  • 박종옥;이승은
    • Journal of Life Science
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    • v.14 no.1
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    • pp.148-153
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    • 2004
  • In this study, we investigated the effect of water, extract of Terminalia Chebula (TC) on physiological activity in mice. TC water extract showed hemagglutination against several different types of red blood cells. $LD_{50}$ of TC extract was 390 mg/kg (po). Treatment of TC water extract orally administered 200, 300 mg/kg daily for one week. Hepatic cytosolic enzymes, xanthine oxidase and aldehyde oxidase activities were significantly increased comparison with normal group. Treatment of TC water extract increased hepatic malondialdehyde (MDA) formation, and reduced glutathione content. We also found that the decreased activities of glutathione S-transferase and glutathione reductase but was not affected activities of $\gamma$-glutamylcysteine synthetase after treatment of TC water extract. These results suggested that increase of the hepatic lipid peroxide is caused by glutathione reduction.

Carbamate-Based Surface Reactions for Release of Amine Molecules from Electroactive Self-Assembled Monolayers

  • Hong, Dae-Wha;Kang, Kyung-Tae;Hong, Seok-Pyo;Shon, Hyun-Kyong;Lee, Tae-Geol;Choi, In-Sung S.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.08a
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    • pp.208-209
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    • 2011
  • In this work, we developed self-assembled monolayers (SAMs) of alkanethiols on gold that can release amine groups, when an electrical potential was applied to the gold. The strategy was based on the introduction of the electroactive carbamate group, which underwent the two-electron oxidation with simultaneous release of the amine molecules, to alkanethiols. The synthesis of the designed thiol compounds was achieved by coupling isocyanate-containing compound with hydroquinone. The electroactive thiols were mixed with hydroxyl-containing alkanethiol [$HS(CH_2)_{11}OH$] to form mixed monolayers, and cyclic votammetry was used for the characterization of the release. The mixed SAMs showed a first oxidation peak at +540 mV (versus Ag/AgCl reference electrode), demonstrating irreversible conversion from carbamate to hydroqinone with simultaneous release of the amine groups. The second and third cycles showed typical reversible redox reaction of hydroquinone and quione: the oxidation and reduction occurred at +290 mV and -110 mV, respectively. The measurement of ToF-SIMS further indicates that electrochemical-assisted chemical reaction successfully released amine groups. This new SAM-based electrochemistry would be applicable for direct release of biologically active molecules that contain amine groups.

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Glutathione (GSH) Response as a Metabolic Biomarker to Benzo(α)pyrene and Aroclor 1254 Exposure in the Pacific Oyster Crassostrea gigas

  • Jo, Qtae;Choy, Eun-Jung;Lee, Yong-Hwa;Ko, Sung-Jung;Song, Young-Chae;Kim, Myoung-Jin;Hwang, Eung-Ju
    • Fisheries and Aquatic Sciences
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    • v.9 no.4
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    • pp.140-145
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    • 2006
  • We measured activities of the ubiquitous tripeptide non-protein thiol (L-${\gamma}$-glutamyl-L-cysteinyl-glycine), glutathione (GSH), which is believed to playa fundamental role in detoxifying xenobiotics in biological systems, as a metabolic biomarker for benzo(${\alpha}$)pyrene and Aroclor 1254 exposure in the Pacific oyster Crassostrea gigas. Reproductive oysters were exposed to the pollutants for 50 days by the algal vectoring method in which the oysters were fed with concentrated standard algal foods grown in culture media containing Aroclor 1254 (0, 5, 50, 500 ng/g) or benzo(${\alpha}$)pyrene (0, 10, 100, 1,000 ng/g). Both pollutants induced maternal GSH activities in 10 days in a dosage-dependent manner (p<0.05), although Aroclor 1254 was stronger. The pollutant-driven GSH elevation persisted for 20 to 30 days depending on the pollutants and concentrations. Thereafter, a drastic decline in the GSH activity was observed due to metabolic failure, after which the oyster GSH remained at low levels throughout the remainder of the experiment. The pollutant exposures influenced maternal reproductive output in terms of fertilization, hatching, and morphology. These results imply that changes in activity of the GST-catalyzing molecule can be used as an oyster biomarker for Aroclor 1254 and benzo(${\alpha}$)pyrene exposure.

Quantitative Mass Spectrometric Analysis of Mixed Self-Assembled Monolayers for Biochips

  • Son, Jin Gyeong;Shon, Hyun Kyong;Hong, Daewha;Choi, Changrok;Han, Sang Woo;Choi, Insung S.;Lee, Tae Geol
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.275-275
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    • 2013
  • Formation and characterization of self-assembled monolayers (SAMs) on various surfaces are the essential basis for many other applications, including molecular switches, biosensors, microfluidics, and fundamental studies in surfaces and interfaces. To improve the performance at these applications, it is a key to control the quantity of each molecule in various mixed SAMs on the surface. In this study, using mixed SAM of carbamate-based hydroquinone (HQ)-PhBr and11-mercaptoundecanol, the quantitative mass spectrometric method of mixed SAM was developed based on comparison study with XPS and FT-IR methods. In addition, our method was applied to another mixed SAM of biotinylated PEG alkane thiol and 11-mercaptoundecanol for verification purpose. Time-of-flight secondary mass spectrometry (ToF-SIMS) analysis was performed to identify and quantify each molecule of mixed SAM along with principal component analysis (PCA). Since there is no matrix effect in the X-ray photoelectron spectroscopy (XPS) and Fourier transform-infrared (FT-IR) techniques, we compared ToF-SIMS results with XPS and FT-IR results. Because PCA results from ToF-SIMS analysis are well matched with XPS and FT-IR results from both mixed SAMs, we are expecting that our method will be useful to identify and quantify each molecule in various mixed SAMs.

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Exploration of the Glycosyltransferase BmmGT1 from a Marine-Derived Bacillus Strain as a Potential Enzyme Tool for Compound Glycol-Diversification

  • Liu, Quanquan;Ren, Pengfei;Liu, Yang;Qin, Wen;Li, Huayue;Li, Wenli
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.931-937
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    • 2018
  • Glycosyltransferases (GTs) from microbes are an emerging and rich source for efficient glycol-transformation of natural/unnatural compounds. Here, we probed the catalytic capability and substrate promiscuity of BmmGT1 from marine-derived Bacillus methylotrophicus B-9987. The regioselectivity of BmmGT1 on macrolactin A (1) was explored by optimization of the reaction conditions, in which a series of O-glycosylated macrolactins (1a-1e) were generated, including two new di/tri-O-glucosyl analogs (1b and 1e). Furthermore, BmmGT1 was able to catalyze the glycosylation of the thiol (S-) or amine (N-) sites of phenolic compounds (2 and 3), leading to the generation of N- (2a) or S-glycosides (3a and 3b). The present study demonstrates that BmmGT1 could serve as a potential enzyme tool for O-, N-, or S-glycosyl structural diversification of compounds for drug discovery.

Cadmium-induced COX-2 Expression in Cerebrovascular Endothelial Cells (카드뮴이 뇌혈관 내피세포에서의 $PGE^2$ 및 COX-2 발현에 미치는 영향)

  • Park Dong-Hyun;Kim Young-Chae;Moon Chang-Kiu;Jung Yi-Sook;Baik Eun-Joo;Moon Chang-Hyun;Lee Soo-Hwan
    • Environmental Analysis Health and Toxicology
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    • v.21 no.3 s.54
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    • pp.275-282
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    • 2006
  • In order to get insight into the mechanism of cadmium (Cd)-induced brain injury, we investigated the effects of Cd on the induction of COX-2 in bEnd.3 mouse brain endothelial cells. Cd induced COX-2 expression and $PGE_2$ release, which were attenuated by thiol-reducing antioxidant N-acetylcysteine (NAC) indicating oxidative components might contribute to these events. Indeed, Cd increased cellular reactive oxygen species (ROS) level and DNA binding activity of nuclear factor-kB (NF-kB), an oxidative stress sensitive transcription factor. Cd-induced $PGE_2$ production and COX-2 expression were significantly attenuated by Bay 11 7082, a specific inhibitor of NF-kB and by SB203580, a specific inhibitor of p38 mitogen activated protein kinase (MAPK). These data suggest that Cd induces COX-2 expression through activation of NF-kB and p38 MAPK, the oxidative stress-sensitive signaling molecules, in brain endothelial cells.