• Applied Biological Chemistry
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• v.30 no.4
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• pp.364-370
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• 1987

The $O_2^-$ level of the extract from young rice leaves, which was cold treated for 2 days and then placed at room temperature for a period of time significantly higher than that from tissues untreated. $O_2^-$ level in leaves was practically unchanged during cold treatment for 48 hours. But it started to increase to arrive at maximum in 8 hours, once the plants were placed under room temperature. The abnormal production of $O_2^-$ in mitochondria during postchilling process was interpreted as a biochemical consequence of accumulation of glycolysis product(s) in cytosol and/or NADH in mitochondrial matrix due to disruption of catabolic balance at low temperature. Mitochondria isolated from the chilling injured tissue was found to have lost considerably their respiratory activity. This fact may imply the involvement of intramitochondrial accumulation of $O_2^-$ in the inactivation of electron transport chain system. The observation that mitochondria in the presence of the $O_2^--producing$ enzymatic system (Xanthine/Xanthine oxidase) lost their respiratory activity supports this inference. It was also found in this work that Superoxide dismutase (SOD) is a substrate inducible enzyme, and that SOD is a possible protective agent in plant cell against chilling injury.

• International Journal of Vascular Biomedical Engineering
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• v.2 no.2
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• pp.1-9
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• 2004

The history of studies in biology regarding reactive oxygen species (ROS) is approximately 40 years. During the initial 30 years, it appeared that these studies were mainly focused on the toxicity of ROS. However, recent studies have identified another action regarding oxidative signaling, other than toxicity of ROS. Basically, it is suggested that ROS are reactive, and degenerate to biomolecules such as DNA and proteins, leading to deterioration of cellular functions as an oxidative stress. On the other hand, recent studies have shown that ROS act as oxidative signaling in cells, resulting in various gene expressions. Recently ROS emerged as critical signaling molecules in cardiovascular research. Several studies over the past decade have shown that physiological effects of vasoactive factors are mediated by these reactive species and, conversely, that altered redox mechanisms are implicated in the occurrence of metabolic and cardiovascular diseases ROS is a collective term often used by scientist to include not only the oxygen radicals($O2^{-{\cdot}},\;{^{\cdot}}OH$), but also some non-radical derivatives of oxygen. These include hydrogen peroxide, hypochlorous acid (HOCl) and ozone (O3). The superoxide anion ($O2^{-{\cdot}}$) is formed by the univalent reduction of triplet-state molecular oxygen ($^3O_2$). Superoxide dismutase (SOD)s convert superoxide enzymically into hydrogen peroxide. In biological tissues superoxide can also be converted nonenzymically into the nonradical species hydrogen peroxide and singlet oxygen ($^1O_2$). In the presence of reduced transition metals (e.g., ferrous or cuprous ions), hydrogen peroxide can be converted into the highly reactive hydroxyl radical (${^{\cdot}}OH$). Alternatively, hydrogen peroxide may be converted into water by the enzymes catalase or glutathione peroxidase. In the glutathione peroxidase reaction glutathione is oxidized to glutathione disulfide, which can be converted back to glutathione by glutathione reductase in an NADPH-consuming process.

• BMB Reports
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• v.29 no.5
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• pp.423-428
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• 1996

A superoxide anion radical scavenger isolated from Capsella bursa-pastoris was characterized by infrared (IR) spectroscopy, sugar analysis, ultraviolet (UV) spectroscopy, $^{1}H$ and $^{13}C$ nuclear magnetic resonance (NMR) spectroscopies, and fast atom bombardment (FAB) mass analysis. The compound was assumed to be a flavonoid-O-glycoside from IR spectrum and UV absorption maxima. When the sugar composition of the compound was examined by thin layer chromatography (TLC) and gas chromatography (GC) of the acid hydrolysate, only glucose was detected. According to the results of UV spectrotroscopy by using shift reagents, the compound was supposed to be luteolin (5,7,3',4'-tetrahydroxy flavone) or chrysoeriol (5,7,4'-trihydroxy-3'-methoxy flavone) with glucose. Based on $^{1}H$- and $^{13}C-NMR$ spectroscopies, the compound was deduced as 7,4'-dihydroxy-5,3'-dimethoxy-${\alpha}$-6-c-glucosyl-${\beta}$-2"-o-glucosyl flavone. In FAB mass analysis the compound was finally characterized as 7,4'-dihydroxy-5,3'-dimethoxy-${\alpha}$-6-c-glucosyl-${\beta}$-2"-o-glucosyl flavone ($C_{29}H_{34}O_{16}$, M.W.=638).

• 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.

• YAKHAK HOEJI
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• v.44 no.3
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• pp.213-223
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• 2000

SD-994 was prepared from methanol extract of Artemisia argyi by stepwise purification of solvent partioning and silica gel chromatography. In the course of this purification, fractions obtained at each step were investigated for their cytotoxicities against L1210 cells. Fractions A~G prepared from chloroform fraction showed considerable cytotoxicities raging 40~90% against L1210 cells. Subfractions I~IX obtained from fraction A exhibited various cytotoxicities and subfraction I (SD-994) was found to be the most effective compound. $IC_{50}$ values of SD-994 were measured to be $0.5{\;}{\mu\textrm{g}}/ml and less than $0.05{\;}{\mu\textrm{g}}/ml against L1210 cells and normal lymphocytes, respectively: When SD-994 was added to L1210 cell as cytotoxic agent, significantly increased amount of superoxide ($O_2^-$) and dramatically augmented activities of superoxide dismutase (SOD), specially MnSOD and glutathione peroxidase (GPx) were observed according to the concentration and incubation time. Whereas, in case of normal lymphocytes under the same condition, cytotoxicities were not apparent and the generation of superoxide ($O_2^-$) or the activity changes of SOD and GPx were insignificant. These results together indicate that the cytotoxic action of SD-994 against L1210 cell may be achieved via necrosis and/or apoptosis induced by reaction oxygen species which could not probably be completely abolished even by drastically increased antioxidant enzymes, SOD and GPx activities.

• Archives of Pharmacal Research
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• v.27 no.6
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• pp.610-614
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• 2004

The in vitro antioxidant properties of some flavone-6(4)-carboxaldehyde oxime ether deriva-tives (Ia-f, lIa-f） were determined by their effects on the rat liver microsomal NADPH-dependent lipid peroxidation (LP） levels by measuring the formation of 2-thiobarbituric acid reactive substances. The free radical scavenging properties of the compounds were also examined in vitro by determining their capacity to scavenge superoxide anions and interact with the stable free radical 2, 2-diphenyl-1-picrylhydrazyl (DPPH). The most active compounds, lib (Flavone-4＇-carboxaldehyde-O-ethyl oxime） and Id (Flavone-6-carboxaldehyde-O-［2-(1-pyrolidino） ethyl］ oxime）, caused 98 and 79％ inhibition of superoxide anion production and DPPH stable free radical at $10^{-3}{\;}M$, respectively.

• Parasites, Hosts and Diseases
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• v.51 no.4
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• pp.479-484
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• 2013

Neutrophils are the predominant inflammatory cells found in vaginal discharges of patients infected with Trichomonas vaginalis. In this study, we examined superoxide anion ($O^{\cdot}_{2^-}$) production by neutrophils activated by T. vaginalis. Human neutrophils produced superoxide anions when stimulated with either a lysate of T. vaginalis, its membrane component (MC), or excretory-secretory product (ESP). To assess the role of trichomonad protease in production of superoxide anions by neutrophils, T. vaginalis lysate, ESP, and MC were each pretreated with a protease inhibitor cocktail before incubation with neutrophils. Superoxide anion production was significantly decreased by this treatment. Trichomonad growth was inhibited by preincubation with supernatants of neutrophils incubated for 3 hr with T. vaginalis lysate. Furthermore, myeloperoxidase (MPO) production by neutrophils was stimulated by live trichomonads. These results indicate that the production of superoxide anions and MPO by neutrophils stimulated with T. vaginalis may be a part of defense mechanisms of neutrophils in trichomoniasis.

• Journal of Applied Biological Chemistry
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• v.44 no.4
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• pp.159-162
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• 2001

Plant stress incurred by flooding was studied in terms of oxidative stress, using greened rice seedlings subjected to a complete submergence followed by re-exposure to air under illumination ($30W/m^2$). It appeared that shoot tissues of the seedlings suffered oxygen deficiency during the flooding treatment, pertinent to the general concept. Interestingly enough, however, membrane peroxidation in shoots was enhanced by the submergence, as assessed by the content of 2-thiobarbituric acid-reactive substances (TBARS), and the re-aeration resulted in a rapid reduction of TBARS content. Such pattern of response was also seen in the change in the steady state level of $H_2O_2$. In contrast, superoxide dismutase and glutathione reductase that are involved in the detoxifying processes of superoxide in plant cells were significantly activated only during the re-aeration. These results allowed us to suggest the followings as a working hypothesis. Photorespiration-linked production of $H_2O_2$ may largely contribute to the increase in $H_2O_2$ level as well as TBARS production in shoots during the submergence. An abrupt re-supply of $CO_2$ by the re-aeration brings the photosynthetic apparatus back to full operation, suppressing photorespiration and probably causing a momentary, excess formation of superoxide and its dismutation product through side reaction, which gives rise to activating substrate-inducible antioxidative enzymes.

• Applied Biological Chemistry
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• v.37 no.6
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• pp.433-440
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• 1994

The exposure of Clamydomonas reinhardtii to low temperatures resulted in an accumulation of cellular pyruvate that dissipated when the chilled cells returned to ambient temperature. The dissipation of pyruvate accumulation was accompanied by an increase in the production level of superoxide radicals $(O_2^-)$ in cells. The formation of $O_2^-$ at an excessive level during the post-chilling period was apparently countered by a substantial activation of superoxide dismutase (SOD). All these results are similar to those observed previously in rice seedlings subjected to the cold-treatment, implicating that a common mechanism is probably underlying for the primary processes of chilling injury both in higher plants and in algae. It was also observed that the activation of Mn-containing SOD contributes the major share in the increase of SOD activity of whole algal cells. Because Mn-SOD is present only in mitochondria, the observation corroborates the concept that the $O_2^-$ scavenging enzyme would be induced to cope with the cold treatment-caused adverse situation in mitochondria where the toxic active oxygen is produced at rates far exceeding the normal rate.

• Applied Biological Chemistry
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• v.32 no.1
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• pp.23-29
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• 1989

Biochemical consequence of the accumulation in cells of superoxide $(O^{-}_{2})$ which was proposed to be probably a common chemical factor in the secondary process of the mechanism of chilling injury as well as in the visible light photodamage in cells of higher plants, has been investigated in the present work. Especially focused was the destructive effect of $O^{-}_{2}$ on the biochemical activity of mitochondria, as informations which support the suggestion that mitochondrial inner membrane is the major site of $O^{-}_{2}$ production have been collected. Mitochondria and submitochondrial particles (SMP) were prepared from soybean hypocotyls for this case study. When SMP were treated with the electrolytically produced $O^{-}_{2}$ they suffered not only inhibition of the membrane-bound enzymes as demonstrated by cytochrome c oxidase, but also lipid peroxidation of membrane as proved by malondialdehyde production. Malate dehydrogenase present in the protein extract from mitochondrial matrix was also inhibited by the $O^{-}_{2}$ treatment. These results exhibited the chaotic effect of the overproduction and accumulation of $O^{-}_{2}$ in cells under a certain abnormal circumstance such as environmental stress on the physiological function of mitochondrial; disruption of the cellular metabolic pathways and the structural integrity of membrane.