• Title/Summary/Keyword: Sulfhydryl groups

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Preparation of IgG-$^{188}$Re Conjugate for Diagnosis of Abscess (농양진단을 위한 IgG-$^{188}$Re 표지화합물 제조)

  • 오옥두;최태현;임상무
    • Biomedical Science Letters
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    • v.3 no.2
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    • pp.131-138
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    • 1997
  • IgG-$^{188}$Re conjugate was prepared for the diagnosis of abscess. The IgG molecules reduced by 2-mercaptoethanol contained 1.5 free sulfhydryl groups per IgG molecule. The reduced IgG molecule was labelled with $^{188}$Re through chelate to 99% of labelling yield. The radiochemical purity of IgG-$^{188}$Re conjugate was maintained at 90% in the presence of human serum for 1 hour. The IgG-$^{188}$Re was intravenously administered into staphylococcal abscess-bearing rats and their biodistribution was monitored at 4 and 24 hours post injection. The IgG-$^{188}$Re conjugate was moderately localized in the abscess tissue. This result implies that the IgG-$^{188}$Re conjugate can be a tool for abscess diagnosis. This technique can be applied for the preparation of various monoclonal antibody labelled with $^{188}$Re.

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Antioxidant Effects of Serotonin and L-DOPA on Oxidative Damages of Brain Synaptosomes

  • Ham, Sang-Soo;Kim, Dong-Hyun;Lee, Suk-Ha;Kim, Yun-Sang;Lee, Chung-Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.2
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    • pp.147-155
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    • 1999
  • Antioxidant effects of serotonin and L-DOPA on neuronal tissues were examined by studying the oxidative damages of brain synaptosomal components. The study further explored the mechanism by which they exert protective actions. Serotonin and L-DOPA (1 ${\mu}M$ to 1 mM) significantly inhibited lipid peroxidation of brain tissues by either $Fe^{2+}$ and ascorbate or t-butyl hydroperoxide in a dose dependent fashion. Protective effect of serotonin on the peroxidative actions of both systems was greater than that of L-DOPA. Protein oxidation of synaptosomes caused by $Fe^{2+}$ and ascorbate was attenuated by serotonin and L-DOPA. Protein oxidation more sensitively responded to L-DOPA rather than serotonin. Serotonin and L-DOPA (100 ${\mu}M$) decreased effectively the oxidation of synaptosomal sulfhydryl groups caused by $Fe^{2+}$ and ascorbate. The production of hydroxyl radical caused by either $Fe^{3+},$ EDTA, H_2O_2$ and ascorbate or xanthine and xanthine oxidase was significantly decreased by serotonin and L-DOPA (1 mM). Equal concentrations of serotonin and L-DOPA restored synaptosomal $Ca^{2+}$ uptake decreased by $Fe^{2+}$ and ascorbate, which is responsible for SOD and catalase. Protective effects of serotonin and L-DOPA on brain synaptosomes may be attributed to their removing action on reactive oxidants, hydroxyl radicals and probably iron-oxygen complex, without chelating action on iron.

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Effect of Faba Bean Isolate and Microbial Transglutaminase on Rheological Properties of Pork Myofibrillar Protein Gel and Physicochemical and Textural Properties of Reduced-Salt, Low-Fat Pork Model Sausages

  • Geon Ho Kim;Koo Bok Chin
    • Food Science of Animal Resources
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    • v.44 no.3
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    • pp.586-606
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    • 2024
  • The study was performed to determine the effect of faba bean protein isolate (FBPI) alone or in combination with microbial transglutaminase (MTG) on the rheological properties of pork myofibrillar protein gel (MPG), and physiochemical and textural properties of reduced-salt, low-fat pork model sausages (LFMSs). The cooking yields of MPGs with MTG or FBPI alone decreased and increased, respectively. However, the combination of FBPI and MTG was similar to the control (CTL) without FBPI or MTG. Gel strength values of MPG added with both FBPI and MTG were higher than treatments with FBPI or MTG alone. The hydrophobicity values of CTL were lower than those of MPG with FBPI alone, whereas the addition of MTG decreased the hydrophobicity of MPGs. The incorporation of FBPI alone or in combination with MTG decreased sulfhydryl groups (p<0.05). Shear stress values of MPGs with MTG tended to be higher than those of non-MTG treatments at all shear rates, and the addition of FBPI into MPGs increased shear stress values. Reduced-salt (1.0%) LFMSs with FBPI alone or combined with MTG had both lower cooking loss and expressible moisture values than those of CTL and similar values to the reference sample (REF, 1.5% salt). Textural properties of reduced-salt LFMSs with FBPI or MTG were similar to those of REF. These results demonstrated that the combination of FBPI and MTG could improve the water binding capacity and textural properties of pork MPGs and LFMSs and might be suitable for application in the development of healthier meat products.

Bioequivalence of Bucilin Tablet to Rimatil Tablet (Bucillamine 100 mg) (리마틸 정(부시라민 100 mg)에 대한 부시린 정의 생물학적 동등성)

  • Cho, Hea-Young;Lee, Moon-Seok;Oh, In-Joon;Kim, Dong-Hyun;Moon, Jai-Dong;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.31 no.2
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    • pp.125-130
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    • 2001
  • Bucillamine is a novel cysteine derivative with two free intramolecular sulfhydryl groups, and has a preventive and therapeutic effect on adjuvant arthritis, suggesting its antirheumatic action. With respect to the effect on the immune system, bucillamine-exerted such immunoregulating actions are to nomalize an excessive reduction or acceleration in immune reaction. It is useful not only in patients with early stage of rheumatoid arthritis (RA) but also in those with active RA retained for more than 10 years. The purpose of the present study was to evaluate the bioequivalence of two bucillamine tablets, $Rimatil^{TM}$ (Chong Kun Dang Pharmaceutical Co., Ltd.) and $Bucilin^{TM}$ (Kuhn Il Pharmaceutical Co., Ltd.), according to the guidelines of Korea Food and Drug Administration (KFDA). Eighteen normal male volunteers, $23.67{\pm}2.09$ years in age and $65.03{\pm}6.73\;kg$ in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After three tablets containing 100 mg of bucillamine per tablet were orally administered, blood was taken at predetermined time intervals and the concentrations of bucillamine in serum were determined using GC/MS with mass selective detector. Pharmacokinetic parameters such as $AUC_t$, $C_{max}\;and\;T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters. The results showed that the differences in $AUC_t$, $C_{max}\;and\;T_{max}$ between two tablets were -0.29%, -3.20% and 8.22%, respectively, when calculated against the $Rimatil^{TM}$ tablet. The powers $(1-{\beta})$ for $AUC_t\;and\;C_{max}$ were 84.31 % and 91.16%, respectively. Minimum detectable differences $({\Delta})$ at ${\alpha}=0.10$ and $1-{\beta}=0.8$ were less than 20% (e.g., 18.58% and 16.51% for $AUC_t\;and\;C_{max}$, respectively). The 90% confidence intervals were within ${\pm}20%$ (e.g.,$-12.77{\sim}12.20$ for $AUC_t$ and $-14.30{\sim}7.90$ for $C_{max}$). Two parameters met the criteria of KFDA for bioequivalence, indicating that $Bucilin^{TM}$ tablet is bioequivalent to $Rimatil^{TM}$ tablet.

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Nucleotide Activation of Catabolic Threonine Dehydratase from Serratia marcescens (뉴클레오타이드에 의한 Serratia marcescens Catabolic Threonine Dehydratase의 활성화)

  • Choi, Byung-Bum
    • The Korean Journal of Food And Nutrition
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    • v.23 no.2
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    • pp.171-177
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    • 2010
  • The catabolic threonine dehydratase from Serratia marcescens ATCC 25419 was purified to homogeniety using Sephadex G-200 gel filtration and AMP-Sepharose 4B affinity chromatography. The molecular weight of the native enzyme was 120,000 by native pore gradient PAGE. The enzyme was composed of four identical subunits with subunit molecular weights of 30,000 by SDS-PAGE. The Km values of the enzyme for L-threonine with and without AMP were 7.3 and 92 mM, respectively. There were 2 moles of pyridoxal phosphate and 16 moles of free -SH groups per 1 mole of enzyme. The enzyme was inhibited by $\alpha$-ketobutyrate, pyruvate, glyoxylate, and phosphoenol pyruvate(PEP) in the presence of AMP, yet stimulated by cAMP and ADP. For enzyme properties in comparison with S. marcescens, E. coli, and S. typhimurium enzyme, such as the PLP content, number of free sulfhydryl groups, and existence of ADP binding site, the S. marcescens enzyme was more similar to the S. typhimurium enzyme than the E. coli enzyme. Of the three enteric bacteria, the E. coli and S. typhimurium enzyme was increased the activity by ADP and cAMP, respectively, but only the S. marcescens enzyme was increased the activity by both ADP and cAMP. Therefore, the subtle differences in the properties between enzymes from the three enteric bacteria may represent minor structural differences among these enzymes and warrants further study.

Effect of Different Storage-Temperature Combinations on Longissimus dorsi Quality upon Sous-vide Processing of Frozen/Thawed Pork

  • Ji, Da-Som;Kim, Ji-Han;Yoon, Dong-Kyu;Kim, Jung-Ho;Lee, Ha-jung;Cho, Won-Young;Lee, Chi-Ho
    • Food Science of Animal Resources
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    • v.39 no.2
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    • pp.240-254
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    • 2019
  • This study investigated the effect of storage state (chilled state on sous-vide, CS; frozen state without thawing on sous-vide, FS; and frozen/thawed states on sous-vide, TS) and sous-vide cooking temperature ($65^{\circ}C$ and $72^{\circ}C$) on the longissimus dorsi muscle quality of pork. FS showed a higher moisture content than that of CS and TS (p<0.001), whereas both FS and CS showed higher expressible moisture loss than that of TS (p<0.001). FS showed a lower cooking loss (p<0.001) than that of CS and TS. FS and TS exhibited significantly higher lipid oxidation than that of CS. Carbonyl and sulfhydryl content were not significantly affected by the storage treatment. FS and TS exhibited lower shear force than that of CS (p<0.001). FS and TS showed higher springiness than that of CS (p<0.001), FS exhibited lower gumminess than that of CS and TS (p<0.01). Sous-vide treatment at $65^{\circ}C$ exhibited significantly higher moisture content and lower expressible moisture loss, cooking loss, and total and sarcoplasmic protein than those at $72^{\circ}C$. Shear force and springiness of $65^{\circ}C$-treated groups were lower than those of $72^{\circ}C$-treated groups (p<0.01). Cooking temperature significantly influenced overall acceptability, whereas the storage state did not affect the overall acceptability. These results indicated that meat quality might be improved upon cooking from the frozen or frozen/thawed state using sous-vide when compared with traditional processing.

Growth Inhibitory Activity of Sulfur Compounds of Garlic against Pathogenic Microorganisms (마늘 황화합물의 병원성미생물 번식억제작용)

  • Kyung Kyu-Hang
    • Journal of Food Hygiene and Safety
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    • v.21 no.3
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    • pp.145-152
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    • 2006
  • Efforts have been made to explore the possibility of using garlic as an antimicrobial therapeutic agent since garlic extract and its individual sulfur compounds show antimicrobial activities against all kinds of microorganisms including bacteria, molds, yeasts and protozoa. Staphylococcus aureus has been the most studied bacteria along with many other Gram positive and negative pathogenic bacteria, including species of the genera Clostridium, Mycobacterium, Escherichia, Klebsiella, Bacillus, Salmonella and Shigella. Candida albicans has been the most studied among the eukaryotic microorganisms. A pathogenic protozoa, Giardia intestinalis, was also tested. All the microorganisms tested was inhibited by garlic extract or its sulfur components. Garlic has been known to be growth inhibitory only when fresh garlic is crushed, since allicin-generating reaction is enzyme-catalyzed. Allicin is known to be growth inhibitory through a non-specific reaction with sulfhydryl groups of enzyme proteins that are crucial to the metabolism of microorganisms. Another plausible hypothesis is that allicin inhibits specific enzymes in certain biological processes, e.g. acetyl CoA synthetase in fatty acid synthesis in microorganisms. Allicin transforms into other compounds like ajoene and various sulfides which are also inhibitory to microorganisms, but not as potent as their mother compound. It is reported recently that garlic heated at cooking temperatures is growth inhibitory especially against yeasts, and that the growth inhibitory compound is allyl alcohol thermally generated from alliin in garlic.

Reduced Protein Denaturation in Thermotolerant Cells by Elevated Levels of HSP70 (열내성이 유도된 세포에서 HSP70 단백질 증가에 의한 단백질 변성 감소)

  • Han, Mi-Young;Park, Young-Mee
    • The Korean Journal of Pharmacology
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    • v.32 no.3
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    • pp.433-444
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    • 1996
  • We describe a novel approach to evaluate quantitatively the amounts of denatured proteins in cells upon heat exposure. A thiol compound, diamide [azodicarboxylic acid bis (dimethylamide)] causes protein cross-linking with exposed sulfyhydryl residues of denatured proteins. Since denatured proteins expose normally well-hidden sulfhydryl groups, these will be preferentially cross-linked by diamide. Thus diamide acts to 'trap' denatured proteins. We observed that protein aggregates (high molecular weight protein aggregates, HMA) appeared on SDS-polyacrylamide gels run under non-reducing conditions and that the amount of HMA can be quantified by scanning the gels using a gas flow counter. Heating cells followed by a fixed dose of diamide exposure resulted in HMA increases in a heat-dose dependent manner, demonstrating that the quantitation of HMA could serve as a measure of heat-denatured proteins. We compared thermotolerant and nontolerant cells and found decreased HMA in tolerant cells upon heat treatment. As an attempt to examine the kinetics of protein renaturation (or 'repair'), we measured the amounts of aggregates formed by the addition of diamide at various times after heat shock. Such experiments demonstrate an equally rapid disappearance of HMA in previously unheated and in thermotolerant cells. Levels of HMA in tolerant cells increased significantly after electroporation of HSP70 specific mAbs, suggesting an involvement of HSP70 in reducing HMA levels in thermotolerant cells upon heat exposure. Immunoprecipitation studies using anti-HSP70 antibody indicated an association of HSP70 with heat-denatured proteins. Our results suggest that heat induces protein denaturation, and that elevated level of HSP70 present in thermotolerant cells protects them by reducing the level of protein denaturation rather than by facilitating the 'repair' (or degradation) process.

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Use of a Xanthine-Xanthine Oxidase System on In Vitro Maturation and Fertilization in Pig

  • Sa, S.J.;Park, C.K.;Cheong, H.T.;Yang, B.K.;Kim, C.I.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.13-13
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    • 2001
  • This study was undertaken to evaluate the effects of catalase using xanthine (X) - xanthine oxidase (XO) system on in vitro maturation and fertilization in pig. When follicular oocytes were cultured in maturation medium with X and/or XO, the maturation rates were not significantly different between in medium with and without catalase despite of different culture periods. However, significantly (P<0.05) higher maturation rates were obrained in culture with X-XO system. The rates of degenerated oocytes were increased with culture periods prolonged, and were significantly (P<0.05) higher in medium without than with catalase at 120 h of culture. On the other hand, the parthenogenetic oocytes were observed with high proportions at 72 h of culture, hut were not different in medium with and without catalase at various times of culture. In another experiment, the frozen-thawed boar spermatozoa treated with X-XO system for in vitro fertilization. The penetration rates were higher in medium with that than without catalase during the in vitro fertilization with, none (P<0.05), XO and X+XO. On the other hand, when sperm were treated with none, X, XO and X+XO, lipid peroxidation were higher in medium without that than with catalase. However, the changes in sperm penetration and lipid peroxidation showed opposite patterns. The sperm suspensions were also treated with X and/or XO for assay of sulfhydryl (-SH) group content. Under the above all conditions, sperm-SH group were higher detected In medium with that than without catalase. The activity of sperm binding to zona pellucida was also evaluated through binding to salt-stored porcine oocytes. In control group, sperm binding to zona pellucida were higher than in medium with X, XO and X+XO groups. No significant differences, however, were observed between medium with and without catalase. In conclusion, the exposure of follicular oocytes and spermatozoa to X-XO system may be caused stimulating in vitro maturation and fertilization in pig. This work was supported by grant No. 2000-1-22200-001-3 from the Basic Research Program of the Korea Science & Engineering Foundation.

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Characterization of Antioxident Enzymes in the Lung of Rat Exposed to Cigarette Smoke (흡연한 흰쥐 폐조직 항산화효소들의 특성)

  • 이영구;손형옥;임흥빈;이동욱;박준영
    • Journal of the Korean Society of Tobacco Science
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    • v.15 no.1
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    • pp.3-14
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    • 1993
  • Oxidants in environment or cigarette smoke are known to be implicated in the oxidative damages of pulmonary system. Such cellular damages are prevented by the presence of adequate levels of antioxidants in the tissue. In the present study, we investigated the influences of smoking duration and concentration of smoke on lung antioxidant defense in rats. Subchronic exposure of rats to smoke generated from 6 cigarettes per day for 90 days caused the activities of catalase and superoxide dismutase (SOD) to increase. However, glutathione peroxidase (GP-Xase) was not significantly changed. Total sulfhydryl compounds (Total-SH) in the lung homogenates from the rats inhaled with cigarette smoke for 15 days was decreased by 44% , thereafter it was returned to the level of normal rats. On the contrary, when rats were daily exposed to a different concentration of smoke generated from 1 to 20 cigarettes per day for 15 days, the activity of catalase was increased gradually with dose, but total SOD activity was increased only in the rats of low dose groups less than 5 cigarettes. Three types of SOD (one Cu, Zn-SOD with pI 4.9, and two Zn-SOD with pI 4.7 and 7.9)were detected in the lung homogenates and Zn-SOD with pI 4.7 was the major and cigarette-smoke inducible form. These results indicate that the protection of lung against oxidants from cigarette smoke seems to be accomplished by the induction of catalase and SOD, especially a cyanide resistant Zn-SOD with pI 4.f, following the consumption of antioxidants such as GSH in the beginning of inhalation period.

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