• Title/Summary/Keyword: Cysteine Oxidation

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Amino Acid-Based Material for the Complementary Therapy of Decubitus Ulcers

  • Nogueira, Frederico;Gouveia, Isabel C.
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.747-758
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    • 2017
  • Chronic wounds, pressure sores, lesions, and infections of microbial origin in bedridden, paralyzed, or malnutrition patients remain the object of study of many researchers. A variety of factors behind the development of these disorders are related to the patient's immune system, making it unable to respond effectively to the treatment of the wound. These factors can be properly controlled, giving particular importance to the ethiology and stage of the wound, as well as the time periods corresponding to the replacement of the dressings. The present research reports a novel foam/soft material, ${{\small}L}$-Cys-g-PCL, with an application for decubitus/pressure ulcers, especially for wounds with a difficult healing process due to infections and constant oxidation of the soft tissues. During this work, the interactions between S. aureus and ${{\small}L}$-Cys-g-PCL foam were studied under conditions that simulate decubitus ulcers; namely, pH and exudate. The effects of duration of grafting (1 or 8 h) and pH (7.0 and 8.9) on wettability, surface energy, swelling, and porosity were also evaluated. Results showed an effective microbicidal activity exhibiting an inhibition ratio of 99.73% against S. aureus. This new ${{\small}L}$-Cys-g-PCL soft material showed saftey to contact skin, ability to be shaped to fill in sunken holes (craters) - pressure ulcers stage III - and to act as a smart material responsive to pH, which can be tailored to develop better swelling properties at alkaline pH where exudates are normally higher, so as to address exudate self-cleaning and prevention of desiccation.

Purification and Reaction Mechanism of Rat Brain Succinic Semialdehyde Dehydrogenase

  • Kim, Kyu-Tae;Joo, Chung-No
    • BMB Reports
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    • v.28 no.2
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    • pp.162-169
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    • 1995
  • Rat brain succinic semialdehyde dehydrogenase (EC 1.2.1.24 SSADH) activity was detected in mitochondrial, cytosolic and microsomal fractions. Brain mitochondrial soluble SSADH was purified by ammonium sulfate precipitation, DEAE Sephacel, and 5'-AMP Sepharose 4B affinity chromatography. The purified enzyme was shown to consist of four identical subunits, and the molecular weight of a subunit was 55 kD. The $K_m$ for short chain aliphatic aldehydes and aromatic aldehydes were at the $10^{-3}M$ level but that for succinic semialdehyde was 2.2 ${\mu}M$. Either $NAD^+$ or $NADP^+$ can be used as a cofactor but the affinity for $NAD^+$ was 10 times higher than that for $NADP^+$. The brain cytosolic SSADH was also purified by ammonium sulfate precipitation, DEAE Sephacel, Blue Sepharose CL-6B and 5'-AMP Sepharose 4B affinity chromatography and its Km for short chain aliphatic aldehydes was at the $10^{-3}$ level but that for succinic semialdehyde was 3.3 ${\mu}M$. $NAD^+$ can be used as a cofactor for this enzyme. We suppose that both enzyme might participate in the oxidation of succinic semialdehyde, which is produced during GABA metabolism. The activity of both cytosolic and mitochondrial SSADH was markedly inhibited when the concentration of succinic semialdehyde was high. The reciprocal plot pattern of product inhibition and initial velocity indicated a sequential ordered mechanism for mitochondrial matrix SSADH. Chemical modification data suggested that amino acid residues such as cysteine, serine and lysine might participate in the SSADH reaction.

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Cloning of the Large Subunit of Replication Protein A (RPA) from Yeast Saccharomyces cerevisiae and Its DNA Binding Activity through Redox Potential

  • Jeong, Haeng-Soon;Jeong, In-Chel;Kim, Andre;Kang, Shin-Won;Kang, Ho-Sung;Kim, Yung-Jin;Lee, Suk-Hee;Park, Jang-Su
    • BMB Reports
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    • v.35 no.2
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    • pp.194-198
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    • 2002
  • Eukaryotic replication protein A (RPA) is a single-stranded(ss) DNA binding protein with multiple functions in DNA replication, repair, and genetic recombination. The 70-kDa subunit of eukaryotic RPA contains a conserved four cysteine-type zinc-finger motif that has been implicated in the regulation of DNA replication and repair. Recently, we described a novel function for the zinc-finger motif in the regulation of human RPA's ssDNA binding activity through reduction-oxidation (redox). Here, we show that yeast RPA's ssDNA binding activity is regulated by redox potential through its RPA32 and/or RPA14 subunits. Yeast RPA requires a reducing agent, such as dithiothreitol (DTT), for its ssDNA binding activity. Also, under non-reducing conditions, its DNA binding activity decreases 20 fold. In contrast, the RPA 70 subunit does not require DTT for its DNA binding activity and is not affected by the redox condition. These results suggest that all three subunits are required for the regulation of RPA's DNA binding activity through redox potential.

Effects of Saccharomyces cerevisiae and phytase co-fermentation of wheat bran on growth, antioxidation, immunity and intestinal morphology in broilers

  • Chuang, Wen-Yang;Lin, Li-Jen;Hsieh, Yun-Chen;Chang, Shen-Chang;Lee, Tzu-Tai
    • Animal Bioscience
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    • v.34 no.7
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    • pp.1157-1168
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    • 2021
  • Objective: The aim of this study was to investigate the effects of different amounts of wheat bran (WB) inclusion and postbiotics form by Saccharomyces cerevisiae and phytase co-fermented wheat bran (FWB) on the growth performance and health status of broilers. Methods: Study randomly allocated a total of 300 male broilers to a control and 4 treatment groups (5% WB, 5% FWB, 10% WB, and 10% FWB inclusion, respectively) with each pen having 20 broilers and 3 pens per treatment. Results: The WB does not contain enzymes, but there are 152.8, 549.2, 289.5, and 147.1 U/g dry matter xylanase, protease, cellulase and β-glucanase in FWB, respectively. Furthermore, FWB can decrease nitric oxide release of lipopolysaccharide stimulated chicken peripheral blood mononuclear cells by about two times. Results show that 10% FWB inclusion had significantly the highest weight gain (WG) at 1 to 21 d; 5% FWB had the lowest feed conversion rate at 22 to 35 d; 10% WB and 10% FWB inclusion have the highest villus height and Lactobacillus spp. number in caecum; and both 5% and 10% FWB can increase ash content in femurs. Compared to control group, all treatments increase mucin 2, and tight junction (TJ), such as occludin, claudin-1, zonula occludens-1, and mRNA expression in ileum by at least 5 folds. In chicken peripheral blood mononuclear cells, nicotinamide adenine dinucleotide phosphate-oxidase-1 mRNA expression decreases from 2 to 5 times, and glutamate-cysteine ligase catalytic subunit mRNA expression also increases in all treatment groups compared to control group. The mRNA expression of pro-inflammatory cytokines, including interleukin-6 (IL-6), nuclear factor-κB, and IL-1β, decreases in 5% and 10% FWB groups compared to control group. Conclusion: To summarize, both WB and FWB inclusion in broilers diets increase TJ mRNA expression and anti-oxidation and anti-inflammation, but up to 10% FWB groups have better WG in different stages of broiler development.

The impaired redox status and activated nuclear factor-erythroid 2-related factor 2/antioxidant response element pathway in wooden breast myopathy in broiler chickens

  • Pan, Xiaona;Zhang, Lin;Xing, Tong;Li, Jiaolong;Gao, Feng
    • Animal Bioscience
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    • v.34 no.4
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    • pp.652-661
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    • 2021
  • Objective: Wooden breast (WB) is a novel myopathy affecting modern broiler chickens, which causes substantial economic losses in the poultry industry. The objective of this study was to evaluate the effect of WB abnormality on meat quality, redox status, as well as the expression of genes of the nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway. Methods: A total of 80 broilers (Ross 308, 42 days of age, about 2.6 kg body weight) raised at Jiujin farm (Suqian, Jiangsu, China) were used. Twelve unaffected (no detectable hardness of the breast area) and twelve WB-affected (diffuse remarkable hardness in the breast muscle) birds were selected from the commercial broiler farm according to the criteria proposed by previous studies. Results: The results indicated that WB showed histological lesions characterized by fiber degeneration and fibrosis, along with an increase of muscle fiber diameter (p<0.05). Moreover, higher pH value, lightness, yellowness, drip loss and cooking loss were observed in the WB group (p<0.05). Compared with the normal breast (NOR) group, the WB group showed higher formation of reactive oxygen species (p<0.05), increased level of oxidation products and antioxidant activities (p<0.05), accompanied with mitochondrial damages and lower mitochondrial membrane potential (p<0.05). Meanwhile, the relative mRNA expressions of Nrf2 and its downstream antioxidant genes including heme oxygenase-1, NAD(P)H qui none dehydrogenase 1, glutathione peroxidase, superoxide dismutase, and glutamate-cysteine ligase were higher than those of the NOR group (p<0.05). Conclusion: In conclusion, WB myopathy impairs meat quality by causing oxidative damages and mitochondrial dysfunction in broilers, even though the activated Nrf2/antioxidant response element pathway provides protection for the birds.

Oxidative Inactivation of Peroxiredoxin Isoforms by H2O2 in Pulmonary Epithelial, Macrophage, and other Cell Lines with their Subsequent Regeneration (폐포상피세포, 대식세포를 비롯한 각종 세포주에서 H2O2에 의한 Peroxiredoxin 동위효소들의 산화에 따른 불활성화와 재생)

  • Oh, Yoon Jung;Kim, Young Sun;Choi, Young In;Shin, Seung Soo;Park, Joo Hun;Choi, Young Hwa;Park, Kwang Joo;Park, Rae Woong;Hwang, Sung Chul
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.1
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    • pp.31-42
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    • 2005
  • Background : Peroxiredoxins (Prxs) are a relatively newly recognized, novel family of peroxidases that reduce $H_2O_2$ and alkylhydroperoxide into water and alcohol, respectively. There are 6 known isoforms of Prxs present in human cells. Normally, Prxs exist in a head-to-tail homodimeric state in a reduced form. However, in the presence of excess $H_2O_2$, it can be oxidized on its catalytically active cysteine site into inactive oxidized forms. This study surveyed the types of the Prx isoforms present in the pulmonary epithelial, macrophage, endothelial, and other cell lines and observed their response to oxidative stress. Methods : This study examined the effect of exogenous, excess $H_2O_2$ on the Prxs of established cell lines originating from the pulmonary epithelium, macrophages, and other cell lines, which are known to be exposed to high oxygen partial pressures or are believed to be subject to frequent oxidative stress, using non-reducing SDS polyacrylamide electrophoresis (PAGE) and 2 dimensional electrophoresis. Result : The addition of excess $H_2O_2$ to the culture media of the various cell-lines caused the immediate inactivation of Prxs, as evidenced by their inability to form dimers by a disulfide cross linkage. This was detected as a subsequent shift to its monomeric forms on the non-reducing SDS PAGE. These findings were further confirmed by 2 dimensional electrophoresis and immunoblot analysis by a shift toward a more acidic isoelectric point (pI). However, the subsequent reappearance of the dimeric Prxs with a comparable, corresponding decrease in the monomeric bands was noted on the non-reducing SDS PAGE as early as 30 minutes after the $H_2O_2$ treatment suggesting regeneration after oxidation. The regenerated dimers can again be converted to the inactivated form by a repeated $H_2O_2$ treatment, indicating that the protein is still catalytically active. The recovery of Prxs to the original dimeric state was not inhibited by a pre-treatment with cycloheximide, nor by a pretreatment with inhibitors of protein synthesis, which suggests that the reappearance of dimers occurs via a regeneration process rather than via the de novo synthesis of the active protein. Conclusion : The cells, in general, appeared to be equipped with an established system for regenerating inactivated Prxs, and this system may function as a molecular "on-off switch" in various oxidative signal transduction processes. The same mechanisms might applicable other proteins associated with signal transduction where the active catalytic site cysteines exist.

Effect of L-ascorbic acid on the degradation of aflatoxin $B_1$ (아스콜빈산에 의한 Aflatoxin $B_1$의 파괴에 관한 연구)

  • Park, Kun-Young;Kweon, Mee-Hyang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.16 no.1
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    • pp.1-9
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    • 1987
  • Large amount of aflatoxin $B_1(AFB_1)$ is disappeared in the presence of L-ascorbic acid(AA) in buffer solution at pH values from 1 to 7 during 5 days of Incubation at $37^{\circ}C$. $AFB_1$ was quite stable at pH's between 5 and 7 when AA was absent(control), however, $50{\sim}60%$ of APB, was degraded in its presence after 5 days. The rate of disappearance of $AFB_1$ increased with a decreasing of pH in the presence of AA, even though $AFB_1$ in the control degraded increasingly with the decrease in $pH(pH{\leq}4)$. The level of $AFB_1$, decreased as the reaction temperature increased when $AFB_1$ reacted with AA. The aflatoxin could not be detected at all after 3 days when the reaction occurred at $60^{\circ}C$, while the aflatoxin was stable at $5^{\circ}C$ thoughout the reaction period. $90{\sim}96%$ of $AFB_1$ was found to be degraded in a far when $AFB_1$ reacted with AA plus different concentrations of $CuSO_4{\cdot}5H_{2}O$, showing remarkably faster rate than the control; however, different concentrations of L-cysteine instead of $CuSO_4\;5H_{2}O$ protected the degradation of aflatoxin and no $AFB_1$ was degraded for a day and resulted in less $AFB_1$ disappeared than the control. The degradation of $AFB_1$ was dependent on AA concentration and the rate of disappearance as the concentration of AA decrease, but $AFB_1$ concentration did not influence the rate. The product formed when $AFB_1$ reacted with AA was identified to $AFB_{2a}$ by using HPLC chromatographic examinations, and by UV spectrum of $AFB_1$ reacted with AA. The disappearance of $AFB_1$ was correlated well in the appearance of $AFB_2a$. From the results, the degradation of $AFB_1$ in the presence of AA is probably due to one or more of the oxidative products of AA which was produced during the AA oxidation.

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Expression and Characterization of Thiol-Specific Antioxidant Protein, DirA of Corynebacterium diphtheriae (코리네박테리움 디프테리아 티올 특이성 항산화단백 DirA의 발현 및 특성)

  • Myung-Jai Choi;Kanghwa Kim;Won-Ki Choi
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.1-9
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    • 1998
  • A Corynebacterium diphtheriae iron-repressible gene dirA, that was homologous to TSA of Saccharomyces cerevisiae and AhpC subunit of Salmonella typhimurium alkyl hydroperoxide reductase, was amplified with PCR and expressed in E. coli. The DirA purified from the transformed E. coli crude extracts prevented the inactivation of enzyme caused by metal-catalyzed oxidation (MCO) system containing thiols but not by ascorbate/Fe$^{3+}$/$O_2$ MCO system. The DirA concentration, which inhibited the inactivation of glutamine synthetase by 50% (IC$_{50}$) against MCO system, was 0.12 mg/ml. The multimeric forms of DirA were converted to the monomeric form in SDS-PAGE under the thioredoxin system comprised of NADPH, Saccharomyces cerevisiae thioredoxin reductase, and thioredoxin. Also, DirA showed thioredoxin dependent peroxidase activity. All of these results were consistent with the characteristics of a thiol specific antioxidant (TSA) protein having two conserved cysteine residues.

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Effect of Onion Consumption on Cardiovascular Disease in Human Intervention Studies: A Literature Review (국내.외 인체중재연구를 통해 살펴 본 양파의 심혈관계 질환 개선효과에 관한 고찰)

  • Kim, Jung-Mi;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.10
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    • pp.1565-1572
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    • 2010
  • Onion (Allium cepa L.) production in Korea has increased gradually over the past 15 years, placing second in food consumption survey with 20.6 g daily intake in 2006. Onions, used as an ingredient in many dishes and accepted by almost all traditions and cultures, have been reported to have a range of health benefits which include anticarcinogenic, antiasthmatic, antibiotic, and antioxidative effects. These effects may be attributable to a powerful flavonoid pigment-containing compounds, such as quercetin and alk(en)yl cysteine sulphoxides (ACSOs). Although antiplatelet and antithrombotic activities of onion have been confirmed by many of in vitro or animal studies, only a few human intervention studies have been examined. The majority of human studies identified that onion improves some cardiovascular markers such as lipid profile and platelet coagulant. With regard to antioxidative effects, somewhat positive effects are confirmed through strengthening the resistance of oxidative DNA damage in lymphocyte and urine, while most studies failed to find inhibitory effects on LDL oxidation. The discrepancies among studies might be ascribed to producing area, processing methods of onion, dosage, subject characteristics, study duration, and measurement methods. In this review, we focused on the preventive effect of cardiovascular disease through onion consumption in human intervention studies.