Cellular redox state is known to be perturbed during ischemia and that $Ca^{2+}$ and $K^2$ channels have been shown to have functional thiol groups. In this study, the properties of thiol redox modulation of the ATP-sensitive $K^2$ ($K_{ATP}$) channel were examined in rabbit ventricular myocytes. Rabbit ventricular myocytes were isolated using a Langendorff column for coronary perfusion and collagenase. Single-channel currents were measured in excised membrane patch configuration of patch-clamp technique. The thiol oxidizing agent 5,5'-dithio-bis-(2-nitro-benzoic acid) (DTNB) inhibited the channel activity, and the inhibitory effect of DTNB was reversed by dithiothreitol (disulfide reducing agent; DTT). DTT itself did not have any effect on the channel activity. However, in the patches excised from the metabolically compromised cells, DTT increased the channel activity. DTT had no effect on the inhibitory action by ATP, showing that thiol oxidation was not involved in the blocking mechanism of ATP. There were no statistical difference in the single channel conductance for the oxidized and reduced states of the channel. Analysis of the open and closed time distributions showed that DTNB had no effect on open and closed time distributions shorter than 4 ms. On the other hand, DTNB decreased the life time of bursts and increased the interburst interval. N-ethylmaleimide (NEM), a substance that reacts with thiol groups of cystein residues in proteins, induced irreversible closure of the channel. The thiol oxidizing agents (DTNB, NEM) inhibited of the $K_{ATP}$ channel only, when added to the cytoplasmic side. The results suggested that metabolism-induced changes in the thiol redox can also modulate $K_{ATP}$ channel activity and that a modulatory site of thiol redox may be located on the cytoplasmic side of the $K_{ATP}$ channel in rabbit ventricular myocytes.
Three kinds of macro-reticular bead-typed chelating resins having thiol groups were obtained from basic resins like poly(strene-co-divinylbenzene) (PSD) and poly(styrene-co-methyl methacrylate-co-divinylbenzene) (PSMD): the chelating resin (I) was prepared by chloromethylation of phenyl rings of PSD followed by thiolation using thiourea. The chelating resin (ll) was designed to provide enough space to chelate heavy metal ions; one chloromethyl group was obtained by chlorination of hydroxymethyl group provided by reduction of carboxylic ester group of PSMD and another chloromethyl group was obtained by direct chloromethylation of pendent phenyl group using chloromethyl methyl ether. Both of chloromethyl groups were thiolated by using thiourea. The chelating resin (III) was prepared by chlorosulfonation of phenyl rings of PSD followed by thiolation using sodium hydrosulfide. The adsorbtivity toward heavy metal ions was evaluated. The hydrophobic chelating resin (I) with thiol groups showed highly selective adsorption capacity f3r mercury ions. However, the chelating resin (II) with thiol groups showed mere effective adsorption capacity toward mercury ions than chelating resin (I) with thiol groups, and showed some adsorption capacity for other heavy metal ions like Cu$\^$2+/, Pb$\^$2+/, Cd$\^$2+/ and Cr$\^$3+/. On the other hand, the chelating resin (III) which have hydrophilic thiosulfonic acid groups was found to be effective adsorbents for some heavy metal ions such as Hg$\^$2+/, Cu$\^$2+/, Ni$\^$2+/, Co$\^$2+/, Cr$\^$3+/ and especially Cd$\^$2+/ and Pb$\^$2+/.
An experiment was carried out with 48 IVRI 2CQ rats 6-8 week old, weighing 50-100 g, to study the effect of lead exposure on antioxidant defense, lipid peroxide level, status of thiol groups and concentration of lead in the liver and kidneys at the end of the exposure and also after withdrawal of lead administration. Twenty four rats were given lead at a daily dose rate of 1 mg lead/2 ml of distilled water/kg body weight as lead acetate solution intraperitoneally for a period of 30 days. Another 24 control rats received 2 ml of sterile normal saline solution (0.85% NaCl)/kg body weight in an identical manner. A many-fold increase in concentration of lead was associated with a non-significant (p>0.05) decrease in the activities of superoxide dismutase (SOD) in the liver (27%) and kidneys (12%) and catalase in kidneys (22%). A significant (p<0.05) increase in lipid peroxide level was recorded in the liver (40%) compared with control values. There were significant (p<0.05) decreases in the total thiol and protein bound thiol contents in liver and an increase in non-protein bound thiol groups in the kidneys of lead exposed rats. During the 10 day observation period after withdrawal of lead administration, no significant change was observed with respect to any of the above parameters indicating that a 10 day withdrawal period was not enough for restoration of normality. It is concluded that the magnitude of response and the resultant changes in the lipid peroxide concentration, and the activities of SOD and catalase were not identical in the liver and kidneys of lead-exposed rats.
Microorganisms capable of utilizing diaminododecane containing amine groups diterminally were isolated from the soil by enrichment culture. One strain of these isolated strain, designated as EL-0112L, was selected for this study. The results of this study were as follows. 1. This isolated strain EL-0112L was identified as Microbacterium, from the results of morphological, cultural, and biochemical tests. This isolated strain was named temporarily Microbacterium sp. EL-0112L for convenience. 2. Microbacterium sp. EL-0112L was tested for ability to utilize different kinds of substitued alkanes containing cyan, amine, chloro, and thiol groups(monoterminally or diterminall substituted) as carbon source. Pentamethylenediamine, hexamethylenediamine, n-decane, laurylamine, and alkane derivatives containing cyan, chloro, and thiol groups were not utilized by Microbacterium sp. EL-0112L. 3. The alkane derivatives that did not serve as growth substrates were tested further in oxidation tests using resting cell preparation of Microbacterium sp. EL-0112 L. Alkane derivatives containing cyan, chloro, thiol groups, and n-decane were oxidized by Microbacterium sp. EL-0112 L. It is possible that this isolated strain is also able to degrade their substituted counterparts since they are structually similar to diaminododecane. The remarkable substrates that were being oxidized were dichlorodecane, and 1-dodecanethiol. Microbacterium sp. EL- 0112L could not oxidize pentamethylenediamine, and hexamethylenediamine. 4. The metabolic products formed from diaminododecane by Microbacterium sp. EL-0112 L were acid compound containing carboxyl group and not containing amine group. On the thin layer chromatography, Rf values of these metabolic products were different from that of the product formed by Corynebacterium sp. EL-0112L. These results suggested the specificity of diaminododecane as carbon source.
Proceedings of the Korean Vacuum Society Conference
/
2011.08a
/
pp.197-197
/
2011
In this study, in contrast with cases in which Scanning Tunneling Microscopy (STM) tip-induced reactions were instigated by the tunneling electrons, the local electric field, or the mechanical force between a tip and a surface, we found that the tungsten oxide (WO3) covered tungsten (W) tip of a STM acted as a chemical catalyst for the S-H dissociative adsorption of phenylthiol and 1-octanethiol onto a Ge(100) surface. By varying the distance between the tip and the surface, the degree of the tip-catalyzed adsorption could be controlled. We have found that the thiol head-group is the critical functional group for this catalysis and the catalytic material is the WO3 layer of the tip. After removing the WO3 layer by field emission treatment, the catalytic activity of the tip has been lost. 3-mercapto isobutyric acid is a chiral bi-functional molecule which has two functional groups, carboxylic acid group and thiol group, at each end. 3-Mercapto Isobutyric Acid adsorbs at Ge(100) surface only through carboxylic acid group at room temperature and this adsorption was enhanced by the tunneling electrons between a STM tip and the surface. Using this enhancement, it is possible to make thiol group-terminated surface where we desire. On the other hand, surprisingly, the WO3 covered W tip of STM was found to act as a chemical catalyst to catalyze the adsorption of 3-mercapto isobutyric acid through thiol group at Ge(100) surface. Using this catalysis, it is possible to make carboxylic acid group-terminated surface where we want. This functional group-selective adsorption of bi-functional molecule using the catalysis may be used in positive lithographic methods to produce semiconductor substrate which is terminated by desired functional groups.
Purpose: Due to the increasing prevalence of obesity worldwide, non-alcoholic fatty liver disease (NAFLD) has reached epidemic dimensions over time. NAFLD is the most common cause of childhood chronic liver disease. There is a relationship between NAFLD and oxidative stress. This study aims to investigate the changes in thiol/disulfide homeostasis parameters to determine the oxidant/antioxidant balance in obese rats with diet-induced NAFLD and healthy rats. Methods: Twelve Wistar albino rats were used in this study. Experimentally produced NAFLD obese rats (n=6) and healthy rats were compared. Experimental NAFLD model was created with a special fatty liver diet (Altromin® C1063, Fatty Liver Diet, Exclusivet, Lage, Germany). The biochemical and histopathological features of the groups, as well as serum thiol/disulfide homeostasis parameters, were analyzed and compared. Results: In the experimentally induced NAFLD rat model, they gained more weight than the control group. Steatosis (at least grade 2) occurred in all rats fed with special fatty liver diet for 12 weeks. Histopathologically, no high-grade inflammation was observed in rats with experimental NAFLD after feeding a diet for 12 weeks. Results revealed that aspartate transaminase and alanine transaminase levels were high, albumin levels were low, oxidant stress parameters increased, and antioxidant thiol groups decreased. Conclusion: Experimental NAFLD is characterized by increased oxidant stress accompanying fatty tissue in the liver. Analysis of thiol/disulfide homeostasis parameters in NAFLD can be used in further studies to develop effective treatment options.
In order to evaluate the usefulness of O. javanica for the phytoremediation, it was grown for 1, 3, 7, 14, 21 days and was exposed to $50\;{\mu}M\;of\;CdCl_2$ in hydroponic medium after 3 weeks. Its biomass and contents of chlorophylls were analyzed. The growth of O. javanica showed little difference between cadmium treated and non-treated groups, while its contents of chlorophylls of Cd-treated group decreased up to 50% compared to the case of non-treated group. Its accumulated cadmium concentrations were 2.1, 7.3 and $113\;{\mu}moles\;Cd/g$ dry weight in the leaf, stem and root, respectively. The total contents of thiol increased 0.5, 1 and 7 times in the leaf, stem and root, respectively, while the contents of glutathione tended to decrease by 43%, 70% and 47% in the leaf, stem and root, respectively. Using HPLC analysis, the reasonable peaks of thiol compounds in shoot and root of Cd-treated sample were compared to those of non-treated sample in O. javanica, and found to be phytochelatins. In case of Nicotiana tabacum cv. Xanthi tested as control plant, the cadmium treatment for 3 weeks resulted in the decrease of both biomass and chlorophyll up to 70% and 75%, respectively. The roots of tobacco became rotten and eventually died. These results suggested that Oenanthe javanica is cadmium-tolerant hyperaccumulator.(Received December 20, 1996; accepted March 17, 1997)
The effect of thiol compounds on development and intracellular glutathione(GSH) concentrations of bovine embryos produced by in vitro maturation and in vitro fertilization(IVM/IVF) was examined in CRlaa medium with or without $\beta$-mercaptoethanol(0, 10, 25 and 50$\mu$MME) and cysteamine(0, 25, 50 and 75 $\mu$M). Numbers of cells comprising blastocysts were also counted using double fluorescence stain and the total glutathione levels(oxidized and reduced form) of morula and blastocyst embryos were than measured by an enzymatic method. Following routine IVM/IVF procedures oocytes and zygotes were cultured for 40 to 44h in CRlaa medium. Then 2 to 8-cell embyos had cumulus cell removed and were allotted randomly to the experimental medium. In Experiment 1, the proportion of embryos developing to and beyond morulae stages in 0, 10, 25 and 50 $\mu$M $\beta$-ME was 42.9%, 50.0%, 53.7% and 65.6%, respectively. Fifty $\mu$M $\beta$-ME group was significantly higher than those of any other groups (P<0.05). In Experiment 2, the percentages of embryos developed beyond morulae stages in 0, 25, 50 and 75 $\mu$M cysteamine was 42.9%, 40.4%, 60.0% and 59.2%, respectively. Fifty and 75$\mu$M cysteamine groups were significantly higher than in 0 and 25 $\mu$M cysteamine groups, but all of culture medium containing cysteamine(52.6%) was not significantly difference in control group(42.9%). In Experiment 3, the intracellular GSH concentrations of morulae and blastocyst embryos in 0 and 50 $\mu$M $\beta$-ME was 42.4 pM and 44.9 pM, 49.5 pM and 67.8 pM, respectively. Morulae embryos were not difference, but blastocyst embryos were significantly difference between treatments(P<0.05). In Experiment 4, the intracellular GSH concentrations of morulae in CRlaa with or without cysteamine were 39.8 pM and 45.6 pM, and blastocysts were 59.3 pM and 66.8 pM, respectively. Cell numbers of blastocysts were similar to in all experimental groups. These experiments indicate that thiol compounds can increase the proportion of embryos that developing to and beyond morulae stage and the intracellular GSH concentrations.
The effects of protective colloids on the colloid stability of poly(vinyl acetate) (PVAc) latex was investigated. The stability of PVAc latex in reactive poly(vinyl alcohol) mono thiol (PVALT) (DP=1080) having 78.4% saponification value was better than poly (vinyl alcohol)(PVA) (DP=1100) having 81.6% saponification value. The colloidal stability of PVAc latex particles improved drastically with increase of the reactive PVALT. The particle surface morphology of PVAc latex was examined by transmission electron microscopy (TEM). It was shown that particle size of 1ha latexes decreased with increasing reactive PVALT concentration. Therefore, the stabilities of latex for reactive PVALT protective colloid was superior to that of PVA ones. This result is due to the introduction of many thiol groups that induce chemical bonds at PVAc latexes surface, so that the formation of PVALT-b-PVAc block copolymer via the reaction of PVAc with reactive PVALT. In addition, zeta potential of the PVAc latexes decreased with increasing sodium carbonate concentration.
Changes of thiol methyltransferase (TMT) activity in cholestatic rat liver were studied. Hepatic subcellular and serum TMT activities were determined in cholestatic rat induced by common bile duct (CBD) ligation over a period 28 days. The mitochondrial and microsomal TMT activities in cholestatic rat liver were found to be significantly increased between the 1st and the 28th day after CBD ligation. The TMT activity in serum was significantly increased throughout the experiments. The Vmax values of the above hepatic TMT in cholestatic rat were significantly increased at the 7th day after CBD ligation. However, the Km values of the above hepatic enzymes did not vary in all the experimental groups. Therefore, the results indicate that the biosynthesis of TMT was increased in cholestatic rat liver. The elevated serum TMT activity is most likely caused by increased hepatocytes membrane permeability due to cholestasis mediated liver cell necrosis.
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