• Environmental Engineering Research
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• 제17권4호
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• pp.205-210
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• 2012

Hydroperoxyl radical/superoxide anion radical ($HO_2{\cdot}/O_2^-{\cdot}$, $pK_a$=4.8) as an intermediate is of considerable importance in oxidation processes. Hence, the method of detecting $HO_2{\cdot}/O_2^-{\cdot}$ with high sensitivity is necessary to be developed. To achieve this objective, this study newly employed terephthalate (TA) as a probe for the measurement of $HO_2{\cdot}/O_2^-{\cdot}$ in the kinetic method presented in our previous study. This method was based on the hydroxylation of TA to produce mainly hydroxyterephthalic acid or hydroxyterephthalate (OHTA), which was analyzed by fluorescence detection (${\lambda}_{ex}$=315nm, ${\lambda}_{ex}$=425nm). The life-time of $HO_2{\cdot}/O_2^-{\cdot}$ and its concentration formed from the photolysis technique of $H_2O_2$ were reported in this study. At range of pH 2-10, the life-time of $HO_2{\cdot}/O_2^-{\cdot}$ was 51-422 sec. In particular, an increase in the life-time with pH was observed. The sensitivities of the kinetic method by using TA were always higher with 1.7-2.5 times at pH 8.0 than those by using benzoic acid. From these results, this study can contribute to understanding the basic functions of $HO_2{\cdot}/O_2^-{\cdot}$ in oxidation processes.

• Applied Biological Chemistry
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• 제47권2호
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• pp.279-282
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• 2004

An antioxidant was isolated from Korean mistletoe (Viscum album var. coloratum) by consecutive purification using silica gel, Sephadex LH-20, and RP-HPLC. The active principle was identified as homo-flavoyadorinin-B (3',7-dimethoxyluteolin-4'-O-[apiosyl $(1{\rightarrow}2)$ glucoside]) by spectral analyses. It inhibited 74.6% of hydroxyl radical and 30.6% of superoxide anion radical at 0.01 mM; however, th~compound did not show any scavenging activity against hydrogen peroxide radical. At 0.1 mM, above compound scavenged superoxide anion radical about twice as effective as positive controls, BHT and ${\alpha}-tocopherol$. Radical scavenging activities of homo-flavoyadorinin-B on DPPH, hydroxyl, and hydrogen peroxide radicals were almost same with those of positive controls.

• 방사선산업학회지
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• 제5권3호
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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.

• Preventive Nutrition and Food Science
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• 제14권1호
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• pp.86-89
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• 2009

In this study, we investigated the free radical [1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical (${\cdot}OH$) and superoxide anion (${O_2}^-$)] scavenging activity of MeOH extract and 3 fractions of Citrus junos. Of the tested fractions, the BuOH fraction showed the strongest DPPH scavenging activity, showing the $IC_{50}$ values of 63.4 mg/mL. Therefore, we continuously carried out DPPH, ${\cdot}OH$ and ${O_2}^-$ scavenging activity tests of BuOH fraction of Citrus junos. The BuOH fraction of Citrus junos inhibited DPPH radical to 97.5% at a concentration of 1000 mg/mL and the scavenging activities were increased concentration-dependently. In addition, BuOH fraction from Citrus junos also scavenged ${\cdot}OH$ in a concentration dependent manner from 5 to 1000 mg/mL. Furthermore, BuOH fraction showed about 56% ${O_2}^-$-scavengimg activity at 25 mg/mL concentration but, the scavenging activities were not enhanced in a dose dependent manner. The present results suggest that BuOH fraction of Citrus junos would have the protective potential from oxidative stress induced by free radicals.

• 한국수산과학회지
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• 제20권3호
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• pp.213-218
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• 1987

고도불포화지방산함량이 많은 적색육어류지질의 산화에 의한 DNA 손상작용기구를 조사하기 위하여 고등어를 시료로 하여 추출한 지질을 plasmid pBR 322DNA와 $37^{\circ}C$에서 반응시켜 경시적인 DNA 손상정도를 조사하였으며 이와 함께 지질과 DNA의 반응계에 각종 활성산소소거제를 일정농도씩 첨가하고 지질산화로 일어나는 DNA 손상에 대한 활성산소종의 영향을 비교${\cdot}$ 검토하였다. 고등어 지질의 농도가 클수록 DNA 손상작용이 컸으며 이러한 작용은 고등어 지질의 POV가$100millieq{\cdot}/kg$이하에서 진행된 것으로 나타났다. 또한 활성산소종중 일중항산소$(^1O_2)$와 superoxide anion$({\cdot}O_2^-)$ DNA 손상작용이 가장 컸으며 수산 radical과 과산화 수소는 반응1일이후 거의 영향을 미치지 않아 반응초기의 DNA 손상작용에 관여할 것으로 생각된다. 또한, 고등어 지질의 산화에 대한 활성산소종의 영향을 살펴 본 결과, DNA 손상에 대한 결과와 마찬가지로 일중항산소와 superoxide anion의 영향이 큰 것으로 보아 고등어 지질의 산화반응이나 DNA 손상작용에 있어서 활성산소종이 밀접한 관계가 있는 것으로 나타났다.

• Journal of Acupuncture Research
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• 제15권2호
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• pp.135-145
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• 1998

Bupleury radix has been used for the treatment of fever, liver disease, inflammation in traditional medicine. The present study was carried out to evaluate the antioxidant effects of Bupleury radix aqua-acupuncture solution (BRAS) in vitro. Oxygen derived free radicals produced in the course of normal aerobic life, such as superoxide anion radical($O_2^-$ ), hydroxyl radicaI( OH), hydrogen peroxide($H_2O_2$) and singlet oxygen($^1O_2$) can attack polyunsaturated fatty acid in cell membranes, enzymes, other cell compounds, and give rise to lipid peroxidation, DNA damage, lipofuscin accumulation, structure alteration of cell membrane and cell death. In this study, antioxidant effects of BRAS on lipid peroxidation were determined according to the method of TBA. BRAS inhibited markedly peroxidation of linoleic acid during the autoxidation, and also inhibited lipid peroxidation induced by hydroxyl radical derived from $H_2O_2-Fe^{2+}$ in rat liver homogenate. And BRAS showed 30% scavenging effect on DPPH radical, also exhibited a 30% inhibitory effect on superoxide generation from xanthine-xanthine oxidase system. In addition, BRAS protected the cell death induced by tert-butyl hydroperoxide(t-BHP) and significantly increased cell viability in the normal rat liver cell(Ac2F).

• Preventive Nutrition and Food Science
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• 제9권2호
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• pp.150-155
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• 2004

The free radical scavenging activities and the protective effects of Rhus javanica extracts against oxidative damage induced by reactive oxygen species (ROS) were investigated. n-Hexane, ethyl acetate and water fractions were prepared from a methanol extract. DPPH radical, superoxide anion and hydroxyl radical scavenging activities were estimated. Intracellular ROS formation was quantified using fluorescent probes, 2', 7'-dichlorofluorescin diacetate (DCFH-DA) for hydroxyl radical and dihydroethidium (DHE) for superoxide anion. The oxidative DNA damage was investigated by the comet assay in HepG$_2$ cells exposed either to $H_2O$$_2$ or to menadione. The highest $IC_{50}$/ values for DPPH radical scavenging activity was found in the ethyl acetate fraction with a value of 5.38 $\mu\textrm{g}$/mL. Cells pretreated with $\geq$ 1 $\mu\textrm{g}$/mL of the ethyl acetate extract had significantly increased cell viability compared to control cells, which were not pretreated with the extract. Intracellular ROS formation and DNA damage in HepG$_2$ cells, which were pretreated with the various concentrations of Rhus javanica ethyl acetate extract and then incubated either with $H_2O$$_2$ or with menadione, reduced in a dose-dependent manner. These findings suggest that Rhus javanica might have biologically active components which have strong protective effects against ROS induced oxidative damages to the biomolecules, such as cell membranes and DNA.

• 약학회지
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• 제34권2호
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• pp.112-116
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• 1990

8-Acyl-2-hydroxyphenzaine -5, 10-dioxides and 8-acyl-2-aminophenazine-5, 10-dioxides bearing n-butanoyl, n-hexanoyl and n-octanoyl groups were synthesized. It was attempted to observe the effect of phenazine dioxide derivatives on the hepatic xanthine oxidase activity in this study. As the activity of xanthine oxidase, the key enzyme in the generation of superoxide anion radical ($O_2$), was measured in the presence of phenazine dioxide derivatives, the action of 8-acyl-2-hydroxyphenazine-5,10-dioxide derivatives inhibited with increase of numbers of carbon atom bearing 8-acyl group. Moreover, when plotted on double reciprocal form, the Vmax value decrease as increase of numbers of carbon atom bearing acyl groups without affecting the Km value. However, the hepatic xanthine oxidase activity was not changed by 8-acyl-2-aminophenazine-5,10-dioxide derivatives.

• 대한한방내과학회지
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• 제29권2호
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• pp.421-431
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• 2008

Objectives : This study was to investigate the inhibitory effects of Ulmus davidiana on the generation of peroxynitrite $(ONOO^{-})$, nitric oxide (NO) and superoxide anion radicals $(O_{2}^{-})$ in the endothelial cells of rat vessels. The effects of Ulmus davidiana on the expression of inflammation-related proteins, $NF-{\kappa}B$ (p50, p65), COX-2, and iNOS, were examined by western blotting. Methods : For this study, fluorescent probes, namely dihydrorhodamine 123 (DHR 123), 4,5-diaminofluorescein (DAF-2) and 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) were used. Western blotting was performed via using anti-$NF-{\kappa}B$ (p50, p65), anti-COX-2, and anti-iNOS, respectively. Results : Ulmus davidiana inhibited the generation of $ONOO^{-}$, NO and $(O_{2}^{-})$ in the lipopolysaccharide (LPS)-treated endothelial cells of rat vessels in vitro. Ulmus davidiana inhibited the expression of COX-2 and iNOS genes by means of decreasing the $NF-{\kappa}B$ activation. Conclusions : These results suggest Ulmus davidiana is effective on inhibiting the generation of $ONOO^{-}$, NO and $O_{2}^{-}$, and that therefore it might have a potential role as a treatment for the inflammatory process and inflammation-related diseases.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.503-511
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• 2023

본 연구는 동백나무(Camellia japonica L.) 유박 추출물 및 분획물의 in vitro 항산화 활성에 대해 조사하였다. 동백나무 유박은 에탄올을 사용하여 추출하였으며, 이 후 부탄올(BuOH), 에틸아세테이트(EtOAc), 클로로포름 및 헥산을 이용하여 분획물을 조제하였다. 동백 유박 추출물 및 분획물의 항산화 활성을 평가하기 위해 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS⁺), hydroxyl radical, superoxide anion (O₂^-) radical 소거능을 측정하였다. 동백 유박 추출물 및 분획물은 농도의존적으로 DPPH, ABTS⁺ 및 O₂^- radical 소거능을 증가시켜 항산화 활성을 나타내었다. 특히 동백 유박 BuOH 분획물은 다른 추출물 및 분획물에 비해 가장 높은 radical 소거능을 나타내었다. 동백 유박 BuOH 분획물의 총 폴리페놀 및 플라보노이드 함량은 각각 23.26 mg GAE/g 및 32.39 mg QE/g이었으며, 이는 다른 추출물 및 분획물보다 높은 수치임을 알 수 있었다. 동백 유박 BuOH 분획물 및 EtOAc 분획물의 cameliaside B 함량은 각각 102.37, 165.05 ㎍/g임을 확인하여 다른 추출물 및 분획물에 비해 높은 수치를 나타냄을 알 수 있었다. 따라서 동백 유박 추출 및 분획물 중에서 BuOH 분획물 및 EtOA c 분획물은 다른 추출 및 분획물에 비해 radical 소거능이 우수할 뿐 아니라 항산화 물질 함유량이 높아, 항산화 기능성 소재로 활용될 수 있을 것으로 사료된다.