• The Korean Journal of Physiology and Pharmacology
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• 제7권1호
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• pp.15-23
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• 2003

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.

• Journal of Microbiology and Biotechnology
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• 제23권5호
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• pp.699-706
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• 2013

We have previously reported that N-acetylcysteine (NAC) not only delayed apoptosis but also enhanced the production of recombinant erythropoietin (EPO) in Chinese hamster ovary (CHO) cell culture. To investigate the production enhancement mechanism, the effects of similar thiol-reducing agents were studied. Intriguingly, all mild reducing agents examined including mercaptoethanesulfonic acid (MESNA), thiolactic acid (TLA), and thioglycolate (TG) were shown to block apoptosis and increase EPO production. A pulse-chase study of EPO secretion revealed that all four thiol-reducing agents increased the EPO secretion rate; among them TLA showed the highest rate. In terms of product quality, the sialic acid content of the glycoprotein is one of the most important factors. It was reported that a number of glycoproteins produced by CHO cells often have incomplete sialylation, particularly under high-producing conditions. Human ${\alpha}2,3$-sialyltransferase (${\alpha}2,3$-ST) was introduced into EPO-producing CHO cells in order to compensate for the reduced sialylation during supplementation with NAC. When ${\alpha}2,3$-ST was expressed in the presence of NAC, reduced sialylation was restored and an even more sialylated EPO was produced. Thus, our study is significant in that it offers increased EPO production while still allowing the prevention of decreased sialylation of EPO.

• Food Science and Biotechnology
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• 제16권6호
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• pp.1029-1034
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• 2007

Curcumin (diferuloyl methane), originated rhizomes of Curcuma longa L. has been suggested as an anti-inflammatory and anti-carcinogenic agent. In the present study, modulation of cytotoxic effects of curcumin on HeLa cells by different types of antioxidants was investigated. Cytotoxic effects of curcumin were significantly enhanced in the presence of superoxide dismutase (SOD) by decreasing $IC_{50}$ to 15.4 from $26.0\;{\mu}M$ after 24 hr incubation; the activity was not altered by catalase. The effect of curcumin was significantly less pronounced in the presence of 4 mM N-acetylcysteine (NAC). Low concentration (<1 mM) of NAC, however, increased the efficacy of curcumin. Cysteine and ${\beta}$-mercaptoethanol that have a thiol group, showed the similar biphasic patterns as NAC for modulating curcumin cytotoxicity, which was, however, constantly enhanced by ascorbic acid, a non-thiol antioxidant. In the presence of SOD, ascorbic acid, and 0.5 mM NAC, cellular levels of curcumin were significantly increased by 31-66%, whereas 4 mM NAC decreased the level. The present results indicate that thiol reducing agents showed a biphasic effect in modulating cytotoxicity of curcumin; it is likely that their thiol group is reactive with curcumin especially at high concentrations.

• Toxicological Research
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• 제32권2호
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• pp.133-140
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• 2016

Sulfasalzine is a widely administered drug against inflammatory-based disorders in human. However several cases of liver injury are associated with its administration. There is no stabilized safe protective agent against sulfasalazine-induced liver injury. Current investigation was designed to evaluate if N-acetylcysteine (NAC) and dithioteritol (DTT) as thiol reducing agents and/or vitamins C and E as antioxidants have any protective effects against sulfasalazine-induced hepatic injury in an ex vivo model of isolated rat liver. Rat liver was canulated and perfused via portal vein in a closed recirculating system. Different concentrations of sulfasalazine and/or thiol reductants and antioxidants were administered and markers of organ injury were monitored at different time intervals. It was found that 5 mM of sulfasalazine caused marked liver injury as judged by rise in liver perfusate level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) (p < 0.05). A significant amount of lipid peroxidation and hepatic glutathione depletion were detected in drug-treated livers, accompanied with significant histopathological changes of the organ. Administration of NAC ($500{\mu}M$), DTT (${400\mu}M$), Vitamin C ($200{\mu}M$), or vitamin E ($200{\mu}M$) significantly alleviated sulfasalazine-induced hepatic injury in isolated perfused rat liver. The data obtained from current investigation indicate potential therapeutic properties of thiol reductants and antioxidants against sulfasalazine-induced liver injury.

• Molecules and Cells
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• 제20권3호
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• pp.364-370
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• 2005

We show that sulforaphane inhibits osteoclastogenesis in the presence of macrophage colony-stimulating factor (M-CSF) and receptor for activation of nuclear factor-${\kappa}B$ ligand (RANKL) in osteoclast (OC) precursors. Sulforaphane, an aliphatic isothiocyanate, is a known cancer chemo-preventative agent with anti-oxidative properties. Nuclear factor-${\kappa}B$ (NF-${\kappa}B$) is a critical transcription factor in RANKL-induced osteoclastogenesis, and electrophoretic mobility shift assays (EMSAs) and assay of NF-${\kappa}B$-mediated secreted alkaline phosphatase (SEAP) revealed that sulforaphane selectively inhibited NF-${\kappa}B$ activation induced by RANKL. Inhibition may involve interaction of sulforaphane with thiol groups, since it was prevented by reducing agents.

• The Korean Journal of Physiology and Pharmacology
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• 제25권3호
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• pp.227-237
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• 2021

Carbon monoxide (CO) is a cardioprotectant and potential cardiovascular therapeutic agent. Human cardiac fibroblasts (HCFs) are important determinants of myocardial structure and function. Large-conductance Ca2+-activated K+ (BK) channel is a potential therapeutic target for cardiovascular disease. We investigated whether CO modulates BK channels and the signaling pathways in HCFs using whole-cell mode patch-clamp recordings. CO-releasing molecules (CORMs; CORM-2 and CORM-3) significantly increased the amplitudes of BK currents (IBK). The CO-induced stimulating effects on IBK were blocked by pre-treatment with specific nitric oxide synthase (NOS) blockers (L-NG-monomethyl arginine citrate and L-NG-nitroarginine methyl ester). 8-bromo-cyclic GMP increased IBK. KT5823 (inhibits PKG) or ODQ (inhibits soluble guanylate cyclase) blocked the CO-stimulating effect on IBK. Moreover, 8-bromo-cyclic AMP also increased IBK, and pre-treatment with KT5720 (inhibits PKA) or SQ22536 (inhibits adenylate cyclase) blocked the CO effect. Pre-treatment with N-ethylmaleimide (a thiol-alkylating reagent) also blocked the CO effect on IBK, and DL-dithiothreitol (a reducing agent) reversed the CO effect. These data suggest that CO activates IBK through NO via the NOS and through the PKG, PKA, and S-nitrosylation pathways.

• 한국식품조리과학회지
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• 제16권2호
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• pp.128-134
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• 2000

분자 내에 반응성이 큰 thiol(-SH)기를 갖고 환원제로서 밀가루 반죽에 작용하여 반죽의 탄성을 감소시키고 점성은 증가시키는 cysteine을 첨가하여 밀가루 가공품의 산화적 갈변을 억제함으로써 색을 개선시키는 방법에 대하여 시험하였다. 한국산 밀가루와 수입 밀가루를 이용한 국수 반죽에 cysteine을 첨가하여 국수의 물성과 색 및 조리 특성에 미치는 영향을 조사하고 최적의 첨가량과 최적의 조리시간을 찾고자 하였다. Cysteine을 첨가하였을 때 건면과 생면은 오히려 명도가 감소하고 황색도가 증가되었으나 조리된 국수의 색은 cysteine첨가량이 증가될수록 국수의 명도가 증가되며 특히 우리밀 국수의 명도가 높아졌다. Cysteine 첨가에 따른 국수의 수분활성도는 뚜렷한 차이를 나타내지 않았으나 cysteine의 화학작용에 의한 반죽의 물성 변화에 따라 cysteine을 넣지 않은 국수에 맞추어 동일한 시간 조리된 cysteine이 첨가된 국수는 첨가량이 증가됨에 따라서 유기물의 용출로 인한 조리손실 율이 증가하고 국물의 탁도가 증가되었다. 또한 관능검사 결과 국수의 색은 옅게 평가되지만 씹을 때의 단단한 정도가 급격히 감소하였다. 반면에 각각의 국수를 가장 적당한 조리시간에 맞추어 최적 시간 조리하였을 때 국수의 색(P<0.001)과 점착성(P<0.01)에 대한 차이는 식별하였으나 국수의 단단한 정도와 씹는 횟수 및 맛과 품질의 전반적인 선호도에 대하여 cysteine 첨가에 따른 차이는 나타나지 않았다. 따라서 1% cysteine의 첨가량은 조리된 국수의 명도를 가장 증가시키며 조리에 필요한 시간을 단축함으로써 물성 변화에 따른 제품의 문제점을 해결하였다.

• Bulletin of the Korean Chemical Society
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• 제8권4호
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• pp.285-291
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• 1987

The approximate rates, stoichiometries and products of the reaction of potassium triethylborohydride $(KEt_3BH)$ with selected organic compounds containing representative functional groups under the standard condition $(0^{\circ}C,$ THF) were examined in order to explore the reducing characteristics of this reagent as a selective reducing agent. Primary alcohols, phenols and thiols evolve hydrogen rapidly whereas secondary and tertiary alcohols evolve very slowly. n-Hexylamine is inert to this reagent. Aldehydes and ketones are reduced rapidly and quantitatively to the corresponding alcohols. Reduction of noncamphor gives 3% exo- and 97% endo-norboneol. Anthraquinone is cleanly reduced to 9,10-dihydro-9,10-dihydroxyanthracene stage. Carboxylic acids liberate hydrogen rapidly and quantitatively but further reduction does not occur. Anhydrides utilize 2 equiv of hydride to give an equimolar mixture of acid and alcohol. Acid chlorides, esters and lactones are rapidly and quantitatively reduced to the corresponding alcohols. Epoxides are reduced at moderate rates with Markovnikov ring opening to give the more substituted alcohols. Primary amides liberate 1 equiv of hydrogen rapidly. Further reduction of caproamide is slow whereas benzamide is not reduced. Tertiary amides are reduced slowly. Benzonitrile utilizes 2 equiv of hydride in 3 h to go to the amine stage whereas capronitrile takes only 1 equiv. The reaction of nitro compounds undergo rapidly whereas azobenzene and azoxybenzene are reduced slowly. Cyclohexanone oxime rapidly evolves hydrogen without reduction. Phenyl isocyanate utilizes 1 equiv of hydride to proceed to formanilide stage. Pyridine N-oxide and pyridine is reduced rapidly. Disulfides are rapidly reduced to the thiol stage whereas sulfoxide, sulfonic acid are practically inert to this reagent. Sulfones and cyclohexyl tosylate are slowly reduced. Octyl bromide is reduced rapidly but octyl chloride and cyclohexyl bromide are reduced slowly.

• Bulletin of the Korean Chemical Society
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• 제33권2호
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• pp.483-488
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• 2012

We successfully decorated multi-wall carbon nanotubes (MWNTs) with silver by reacting Ag-NPs with thiolfunctionalized MWNT-SH. Ag alkylcarbamate complex was used as an Ag precursor. Uniform Ag-NPs (5-10 nm) were effectively prepared by microwaving within 60 s using 1-amino-4-methylpiperazine (AMP), which acts as a reaction medium, reducing agent, and stabilizer. The MWNTs were functionalized with 2-aminoethanethiol. Exploiting the chemical affinity between thiol and Ag-NPs, Ag-MWNT nanohybrids were obtained by spontaneous chemical adsorption of MWNT-SH to Ag through Ag-S bonds. The Ag-S-MWNTs were characterized by TGA, XRD, and TEM to confirm that Ag-NPs were uniformly decorated onto the MWNTs. The Ag-S-MWNTs were then employed as conducting filler in epoxy resin to fabricate electrically conducting polymer composites. The electrical properties of the composites were measured and compared with that containing MWNT-SH. The electrical conductivity of composites containing 0.4 wt % Ag-S-MWNT was four orders of magnitude higher than those containing same content of MWNT-SH, confirming Ag-S-MWNT as an effective conducting filler.

• 한국음향학회지
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• 제35권6호
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• pp.473-479
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• 2016

본 연구는 센서 표면에 고정된 티오에스터 분자와 금 나노입자를 이용하여 수용액 중의 시안화이온(cyanide)을 선택적이고, 고민감도로 검출할 수 있는 200 MHz 표면음향파(Surface Acoustic Wave, SAW) 센서의 개발에 관한 것이다. SAW 센서표면에 형성된 티오에스터 단분자막은 시안화이온의 친핵성 첨가반응에 의해 가수분해되어 티올(thiol)이 만들어지고, 티올 분자는 다시 금 나노입자와 반응에 의해 티올-금 나노입자 복합체를 형성한다. 이후 신호증폭을 위해, gold(III) chloride trihydrate와 hydroxylamine hydrochloride 조합에 의한 금 나노입자의 사이즈 확대반응을 수행하였다. SAW 센서는 수용액 중에서 시안화이온에 대한 검출 능력이 17.7 uM이었으며, 공진주파수 변화량은 시안화이온의 농도가 커지면서 포화되는 현상을 보여주었다. 한편, 제작된 SAW 센서는 시안화이온 이외의 플루오라이드(fluoride), 아세테이트(acetate), 그리고 설페이트(sulfate) 이온 등의 다른 음이온에는 전혀 반응성이 없었으며, 다른 음이온에 의한 간섭현상도 나타나지 않았다. 끝으로 모든 실험은 재현성 있는 실험 결과를 얻기 위해서 자체 제작한 유체제어 모듈과 센서를 이용하여 진행하였다.