• Korean Journal of Food Science and Technology
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• v.54 no.1
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• pp.1-7
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• 2022

Riboflavin (Rb), in the presence of methionine (Met) under light, generates superoxide radicals that can reduce nitro blue tetrazolium (NBT) to its corresponding formazan. The Rb-Met/NBT system has been used to measure the superoxide dismutase (SOD)-like activities of various antioxidants. However, the reaction mechanisms have not been clearly defined, and the assay conditions are not consistent. In this study, the effects of different photosensitizers and amino acids on NBT reduction in different solvents were investigated. NBT reduction in the Rb-Met/NBT system was more pronounced in phosphate-buffered saline, compared to distilled water or Tris (pH 7.5); histidine (His) instead of Met also led to considerable Rb-induced NBT reduction. Among the photosensitizers, methylene blue with His caused potent NBT reduction in Tris. Rb-induced NBT reduction combined with Met or His was quantitatively inhibited by SOD or gallic acid, but did not affect MB-induced reduction sensitively.

• Applied Biological Chemistry
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• v.36 no.5
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• pp.364-369
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• 1993

Triton X-100 enhances to a marked extent the analytical sensitivity of the nitrobluetetrazolium (NBT) reduction method for the assay of superoxide$(O^-_{\.{^.2}})$ production. In the present work, it was attempted to elucidate the physicochemical nature of this Triton X-100 effect, focusing on not only the surfactant-caused stabilization of the water-insoluble formazan colloid but also the kinetic competition between the $NBT-O^-_{\.{^.2}}$ reaction and the autodisproportionation of concommitantly occuring in aqueous media. The measurements of formazan and $H_2O_2$ formed in a number of reaction systems, as prepared by vortex-mixing potassium superoxide dissolved in an aprotic solvent with aqueous solutions of NBT, revealed that Triton X-100 exerts its effect both through preventing formazan colloid from aggregation and thereby increasing the formazan absorbance and through suppressing the autodisproportionation reaction of $O^-_{\.{^.2}}$. It also turned out that the relative shares of the colloid stabilization effect and the kinetic effect in the contribution to the sensitivity amplification of the NBT method are dependent upon the reaction conditions, particulary the molar concentration ratio of NBT to $O^-_{\.{^.2}}$ in the reaction systems.

• Journal of fish pathology
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• v.14 no.1
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• pp.3-10
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• 2001

The present study was to obtain a basic research data about medicinal herbs by screening in vitro antimicrobial activity and the production of superoxide anion($O_2^-$) from the head kidney phagocytes of olive flounder, Paralichthys olivaceus. The following fourteen kinds of medicinal herbs extracted by boiling water were used : Gosam, Gwijeonu, Gujeolcho, Bagha, Bangpung, Yeongyo, Yagssug, Jiyu, Sambaegcho, Samjiguyeobcho, Sangbaegpi, Sohwehyang, Pyeonchug, Palgag. Antimicrobial activity against fish pathogenic bacteria, Listonella anguillarum, Vibrio sp., Vibrio alginolyticus, Edwardsiella tarda, Streptococcus sp. and Lactococcus garvieae, and the production of superoxide in kidney macrophage of olive flounder were examined by disk method and nitroblue tetrazolium(NBT) reaction, respectively. Among the tested herbs, Yagssug showed the highest antimicrobial activity against those fish pathogenic bacteria and stimulation of $O_2^-$ production.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.37 no.4
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• pp.275-280
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• 2004

In this study, the effcts of kelp (Ascophyllum nodosum) meal on growth and immune responses of parrot fish (Oplegnathus fasciatus) were studied. Fish were fed an experimental diet supplemented with $2{\%}\;and\;5{\%}$ kelp meal in a controled diet. Several factors such as weight gain, hematological parameters and nonspecific immune responses were evaluated far 0, 2, 4 and 8 weeks after the administration of the kelp meal supplemented diet. Weight gain in the fish fed the diet supplemented with $2{\%}\;and\;5{\%}$ kelp meal was not significant among the tested groups. The NBT reaction of the phagocytes in the head kidney and the phagocytic rate/index of phagocytes in the peripheral blood were significantly higher than the control group. But, there was no significant difference both in complement bactericidal activity, mucus Iysozyme activity and hematology among each group.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.4
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• pp.940-945
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• 2007

We have examined the induction of HL-60 cell differentiation by treatment of 2-chloromethyl-1-dihydroxyphosphinyl pyrrolidine(2C-1DPP), which is derivative of piperidine and pyrrolidine by ${\alpha}-phosphoramidoakylation$ reaction. It was observed that HL-60 cell proliferation was dose- and time-dependently inhibited by treatment with 2C-1DPP. 2C-1DPP treatment caused a significant change in NBT reduction and enhanced ATRA-induced NBT reduction. Treatment of 2C-1DPP to HL-60 cells increased only CD11b expression in the cells, and also increased markedly G0/G1 stage arrest of HL-60 cells. These results can suggest that 2C-1DPP induced the differentiation of HL-60 cells to granulocytes lineage and enhanced ATRA-induced differentiation. Moreover, DNA expression levels of p27 were up-regulated during 2C-1DPP-dependent HL-60 cell differentiation. Our results suggest that 2C-1DPP have potential as a therapeutic agent in human leukemia.

• Journal of Radiation Industry
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• v.5 no.3
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• pp.221-225
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• 2011

To detect superoxide anion ($O_2{\cdot}^-$) or singlet oxygen ($^1O_2$) in biological systems during gamma irradiation, specific chemical probes, 4,5-dihydroxy-1,3-benzene disulfonic acid (Tiron) or 2,2,6,6-tetramethyl-piperidine (TEMP), were evaluated. Tiron or TEMP spin adducts was structurally stable in aqueous solution during gamma irradiation up to 500 or 1,000 Gy, respectively. The signal of Tiron semiquinone radical, a spin adduct of Tiron upon reaction with $O_2{\cdot}^-$, was slightly increased by gamma irradiation. This trend was dose-dependently manifested in $O_2$-saturated aqueous solution using nitro blue tetrazolium (NBT), a common probe for both hydrated electron ($e{^-}_{aq}$) and $O_2{\cdot}^-$. In contrast, a spin adduct of TEMP, was never inducible by gamma irradiation, while its signal was substantially enhanced by photosensitization of riboflavin. These results suggest that Tiron and NBT or TEMP could be utilized to detect $O_2{\cdot}^-$ or $^1O_2$ in biological systems during gamma irradiation, although $O_2{\cdot}^-$ or $^1O_2$ are not the main reactive oxygen species produced by water radiolysis.

• The Korean Journal of Pharmacology
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• v.21 no.1
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• pp.12-26
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• 1985

The generation of $O^{-}_{2}{\cdot}$ and its toxic effects were studied with rat brain mitochondria. The production of $O^{-}_{2}{\cdot}$ from mitochondria in the presence of succinate and antimycin was demonstrated by SOD-inhibitable reduction of NBT. Although succinate can support the $O^{-}_{2}{\cdot}$ formation, the highest rate needs antimycin indicating that blockade of electron flow in the respiratory chain augments the univalent reduction of molecular oxygen. Under this condition, $H_2O_2$ was also observed to be produced. But its formation appears to be derived from the dismutation of the primary product, $O^{-}_{2}{\cdot}$ since the rate of $H_2O_2$ production was markedly decreased by NBT and ferricytochrome c. The $O^{-}_{2}{\cdot}$ and $H_2O_2$ produced were able to cause toxic actions to mitochondrial and extra-mitochondrial components as shown by lipid peroxidation of mitochondrial membrane, and inactivation and lysis of isocitrate dehydrogenase and erythrocytes added to the medium, respectively. In all the toxic actions observed, $Fe^{++}$ was required. It appears that in the toxic actions $OH{\cdot}$ generated from the iron-catalyzed Haber-Weiss reaction acts as a mediator. This was supported by the finding that mitochondria in the presence of succinate and antimycin produced ethylene from methional, and $Fe^{++}$ added increased the ethylene production. The observed toxic actions of mitochondrial $O^{-}_{2}{\cdot}$ may provide evidence supporting a potential role of mitochondria as a source of oxygen radicals to cause tissue damage.

• Korean Journal of Food Science and Technology
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• v.33 no.3
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• pp.378-383
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• 2001

Formation of superoxide anion $O_2\bar{{\bullet}}$ in the autoxidation of L-ascorbic acid (AsA) in the presence of heavy metal ions were determined. The generation of $O_2\bar{{\bullet}}$ was studied by using superoxide dismutase (SOD) in aqueous and buffer solution, and using nitro bule tetrazolium (NBT) in methanol solution. The remaining amount of AsA was significantly higher in the presence of SOD than in its absence. It suggested that SOD stabilizes AsA in aqueous and buffer solution because of scavenging $O_2\bar{{\bullet}}$ formed during the autoxidation reaction of AsA in the presence of heavy metal ions. NBT has an absorption maximum at about 560 nm in methanol solution. The absorbance at 560 nm increased during the oxidation of AsA, suggested the formation of $O_2\bar{{\bullet}}$in methanol solution. Thus, the formation of $O_2\bar{{\bullet}}$ was confirmed during the autoxidation of AsA not only in aqueous solution but also in methanol solution in the presence heavy metal ions.

• Journal of Plant Biology
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• v.33 no.2
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• pp.111-118
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• 1990

In order to characterize the chloroplasts of Korean ginseng as a semi-shade plant and radish as a sun plant, effects of growth light intensity on photosynthetic electron transport (PS) activity in chloroplasts and superoxide (O2.-) production in thylakoid membrane by irradiation were investigated. High-light chloroplasts of both plants showed higher PS activities than those grown under ow growth light intensity. High PS II and low PS I activities in ginseng chloroplasts (ratio of PS II/PS I : 1.1) were observed, but radish chloroplasts showed low PS II and high PS I activities (ratio of PS II/PS I : 0.3). PS II activity of both plants was little affected by temperature in range of 15-35$^{\circ}C$. Activities of whole -chain (PS II+I) in ginseng and PS I in radish were increased at high temperature (4$0^{\circ}C$). Preincubation of chloroplasts at 4$0^{\circ}C$ during 30 min, as a mild heat stress, caused rapid decrease in PS II and PS II+I activities of both plants. However PS I activity was not decreased in ginseng and rather increased in radish. O2.- production (NBT reduction) in Mehler reaction in the thylakoid membrane was inhibited by DCMU in both plants. DMBIB inhibited O2.- production in ginseng, but radish was insensitive to DMBIB. Electron flow system in ginseng thylakoid membrane was more susceptible to damage of photooxidation than that of radish.

• Korean Journal of Acupuncture
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• v.28 no.2
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• pp.23-36
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• 2011

목적 : 이 연구는 관절 및 심혈관계 질환 치료에 사용되는 송절(松節)(Pinus densiflora Gnarl)을 약침용 시료로 조제하여 본 약물의 항산화 효능을 규명하고자 하였으며 이를 다양한 시스템에서 검토하였다. 방법 : $FeCl_2$-ascorbic acid system에서 흰쥐 간조직의 지질과산화 반응을 관찰하였고, Fenton reaction system에서 자유기에 의한 plasmid DNA 분절을 유도하였다. 또한 deoxyribose assay를 통해 hydroxyl radical 소거능을 관찰하였고, NBT reduction assay로 superoxide radical 소거능을 검토하였다. 또한 human low-density lipoprotein(LDL)의 산화를 유도하기 위해 $CuSO_4$와 AAPH를 사용하였으며 relative electrophoretic mobility (REM) assay로 LDL 산화 억제 효능을 대조 항산화물질과 비교 검토하였다. 결과 : 송절 약침액은 자유기에 의한 간조직의 지질과산화(p < 0.01)및 DNA 분절을 현저하게 억제하였으며, hydroxyl radical, superoxide radical (p < 0.01), nitric oxide 및 peroxynitrite를 강하게 소거하였다. 또한 $CuSO_4$ ($IC_{50}=9.2{\pm}0.2\;{\mu}g/ml$)와 AAPH ($IC_{50}=34.8{\pm}5.1\;{\mu}g/ml$)에 의해 유도된 human LDL의 산화를 억제하였고, REM assay에서도 산화 억제 효능을 재확인할 수 있었다. 결론 : 송절 약침액은 활성산소종 및 활성질소종를 소거하였고, 지질과산화를 억제하였으며, 특히 human LDL의 산화적 손상을 방어하였다. 이에 본 약물은 자유기에 의한 심혈관의 산화적 손상을 효과적으로 보호할 것으로 판단된다.