• Journal of Microbiology
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• v.43 no.4
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• pp.375-380
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• 2005

Autonomous ultradian metabolic oscillation (T$\simeq$50 min) was detected in an aerobic chemostat culture of Saccharomyces cerevisiae. A pulse injection of GSH (a reduced form of glutathione) into the culture induced a perturbation in metabolic oscillation, with respiratory inhibition caused by $H_2S$ burst pro-duction. As the production of $H_2S$ in the culture was controlled by different amino acids, we attempted to characterize the effects of GSH on amino acid metabolism, particularly with regard to branched chain and sulfur-containing amino acids. During stable metabolic oscillation, concentrations of intra-cellular glutamate, aspartate, threonine, valine, leucine, isoleucine, and cysteine were observed to oscil-late with the same periods of dissolved $O_2$ oscillation, although the oscillation amplitudes and maximal phases were shown to differ. The methionine concentration was stably maintained at 0.05 mM. When GSH (100 $\mu$M) was injected into the culture, cellular levels of branched chain amino acids increased dramatically with continuous $H_2S$production, whereas the cysteine and methionine concentrations were noticeably reduced. These results indicate that GSH-dependent perturbation occurs as the result of the promotion of branched chain amino acid synthesis and an attenuation of cysteine and methionine synthesis, both of which activate the generation of $H_2S$. In a low sulfate medium containing 2.5 mM sulfate, the GSH injections did not result in perturbations of dissolved $O_2$ NAD(P)H redox oscillations without burst $H_2S$ production. This suggests that GSH-dependent perturbation is intimately linked with the metabolism of branched-chain amino acids and $H_2S$ generation, rather than with direct GSH-GSSG redox control.

• Molecules and Cells
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• v.28 no.5
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• pp.479-487
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• 2009

Glutathione reductase (GR) is an enzyme that recycles a key cellular antioxidant molecule glutathione (GSH) from its oxidized form (GSSG) thus maintaining cellular redox homeostasis. A recombinant plasmid to overexpress a GR of Brassica rapa subsp. pekinensis (BrGR) in E. coli BL21 (DE3) was constructed using an expression vector pKM260. Expression of the introduced gene was confirmed by semi-quantitative RT-PCR, immunoblotting and enzyme assays. Purification of the BrGR protein was performed by IMAC method and indicated that the BrGR was a dimmer. The BrGR required NADPH as a cofactor and specific activity was approximately 458 U. The BrGR-expressing E. coli cells showed increased GR activity and tolerance to $H_2O_2$, menadione, and heavy metal ($CdCl_2$, $ZnCl_2$ and $AlCl_2$)-mediated growth inhibition. The ectopic expression of BrGR provoked the co-regulation of a variety of antioxidant enzymes including catalase, superoxide dismutase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase. Consequently, the transformed cells showed decreased hydroperoxide levels when exposed to stressful conditions. A proteomic analysis demonstrated the higher level of induction of proteins involved in glycolysis, detoxification/oxidative stress response, protein folding, transport/binding proteins, cell envelope/porins, and protein translation and modification when exposed to $H_2O_2$ stress. Taken together, these results indicate that the plant GR protein is functional in a cooperative way in the E. coli system to protect cells against oxidative stress.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.81-81
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• 2019

Ginseng seeds are one of short-lived seeds species which loose their viability easily in the condition of conventional storage. Cryopreservation using liquid nitrogen (LN) has been recommended as a alternative storage for this kind of germplasm short lived or dessiccation-sensitive. This study was performed to find out whether cryopreservation could affect physiological change such as enzyme activity induced by reactive oxygen species. In this work, the redox ratio of ascorbate and glutathione were examined onto ginseng seedlings before and after LN storage of seeds for 1 day using spectrophotometer method. Reduced ascorbate (ASA) was increased while oxidized ascorbate (DHA) was decreased slightly for both after 1d-LN storage. And for glutathione also, reduced form (GSH) was increased while oxidized form (GSSG) was decreased slightly for both after 1d-LN storage. Consequently total phenol compound and ion leakage after LN storage showed no significant differences. Additionally root growth from the seeds after LN storage was not affected by ultra low temperature. From the above results, we may suggest that cryopreservation could be recommended for storage tool of ginseng seeds even with low viability also and expected to make slower seed aging process during preservation period through further study.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.1
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• pp.79-88
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• 1997

This experiment was conducted to investigate the protoporphyrin Ⅸ (PPIX)accumulation, activity of antioxidative enzymes and contents of antioxidant in tolerant-wheat and susceptible-barley to protoporphyrinogen oxidase (Protox) inhibiting-herbicides [oxyfluorfen(2-chloro-l-(3-ethoxy-nitrophenoxy-4-(trifluoromethyl) benzene, acifluorfen (5-[2-chloro-4-(trifl-uoromethyl) phenoxy]-2-nitrobenzoic acid), bifenox(methyl-5-(2, 4-dichlorophenoxy) 2-nitroben-zoate), and oxadiazon (5-tert-butyl-3-(2,4-dichloro-5-isopropoxyphenyl)-1,3,4-oxadiazol-2-one)]. The tolerant-wheat and susceptible-barley were soaked in these compounds at 10$^{-6}$ M for 2hrs and exposed to light for 2,4,6 or 8hrs to investigate change of the activity of antioxidative enzymes. The activities of monodehydroascorbate reductase(MDAR), catalase(CAL) and superoxide dismutase(SOD) were lower in the barley than in the wheat after the treatement of these compounds. The activity of peroxidase(POX) was lower in the barley than in the wheat at 8hrs after the treatment of oxyfluorfen but other compounds showed no difference in activity in wheat and barley. The activity of glutathione reductase(GR) was increased in wheat and barley according as hours of treatment of these compounds became increased but its activity was no difference between wheat and barley. In the case of the content of vitamin C due to the treatment of these compounds, the wheat decreased less than the barley. After the treatment of oxyfluorfen the content of vitamin E in the wheat was higher than in the barley but other compounds didn't have any difference between wheat and barley. And after the treatment of acifluorfen the content of carotenoid was greater in the wheat than in the barley but other compounds didn't have any difference between wheat and barley. The content of glutathione (GSH, GSSG) was greater in the barley than in the wheat. The content of protoporphyrin Ⅸ (PPIX) accumulation by the treatments of these compounds was more in the barley than in the wheat. Especially, the treatment of oxyfluorfen and acifluorfen were more accumulated 2.3 and 1.3 fold in the barley than in the wheat, respectively.

• Journal of Nutrition and Health
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• v.20 no.2
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• pp.111-121
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• 1987

Lipie peroxide formation, antiperoxidative s system and body adaptability for handling lipid p peroxide were examined in the first and second g generations of rats fed fish oil. Mackerel oil(MO) was used and four other dietary oils and fat, i.e. soybean oil(SO), perilla oil(PO), rapeseed oil(RO) and beef tallow(BT) were also employed to compare the effect of fish oil. Synthetic diets containing these five dietary fats at the level of 1O%(w/w), were given to the correspond­m ing groups of male and female rats weighing about 70 grams. After 34 days of feeding, male a and female rats were mated and their offsprings were raised throughout suckling (17, 26 days) and weanling (39 days) periods. Liver lipid pero­x xide level was highest in MO group of both first (mother rats after lactation) and second genera­t tions of 17 and 26 days old, but not of 39 days old. During suckling period, liver lipid peroxide level was well matched to total unsaturation of dietary fat. Brain lipid peroxide levels were not different among five groups. Liver $alpha$-tocopherol a and reduced glutathione (GSH) levels were lowest in MO fed first generation. In second generation, $alpha$-tocopherol level was also low in MO group, although the effect was less pronoun­c ced, but GSH level was not different from other groups. Oxidized glutathione (GSSG) level did not consistently vary by change in dietary fat. Glutathione peroxidase activity increased as young rats grew up to 39 days. Superoxide d dismutase activity change was insignificant by a age, but was shown as lowest in MO group. At the age of 26 and 39 days, liver glutatione peroxidase activity was increased as was level of lipid peroxide, suggesting that this is the one of the mechanisms responsible for body adapta­b bility for protection against the accumulation of lipid peroxide.

• Journal of the Korean Society of Food Science and Nutrition
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• v.16 no.2
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• pp.147-155
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• 1987

In order to examine the effect of dietary fish oil on lipid peroxide formation and antiperoxidative efficiency in liver and brain, a group of male and female rats weighing about 70 grams were fed for three months, diet containing mackerel oil(MO) at the level of 10% (w/w). Results were compared, according to sex and source of dietary fat, i.e., in addition to MO, perilla oil(PO), soybean oil(SO), rapeseed oil(RO) or beef tallow(BT). Liver lipid peroxide level was significantly higher and levels of ${\alpha}-tocopherol$ and reduced glutathione(GSH) were lower in MO group than in other groups. This phenomenon was less clear in male than in female. Liver GSH level was lower in male, compared to female, but oxidized glutathione (GSSG) level did not vary, depending on either sex or dietary fat source. Brain lipid peroxide and ${\alpha}-tocopherol$ levels were not different among five experimental groups. Activities of liver and brain glutathione peroxidase and superoxide dismutase were not changed by dietary fat source, but glutathione peroxidase activity was higher in female than in male. The present study shows (a) that there is sex-related difference in antiperoxidatiye activity and (b) that fish oil containing $C_{20-22}({\omega}3)$ fatty acids, increases body lipid peroxide level and consumes more of cellular antioxidant, although it has lower total PUFA content than perilla or soybean oils.

• Proceedings of the Microbiological Society of Korea Conference
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• 2008.05a
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• pp.39-41
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• 2008

The yeast Saccharomyces cerevisiae has been shown to contain three isoforms of citrate synthase (CS). The mitochondrial CS, Cit1, catalyzes the first reaction of the TCA cycle, i.e., condensation of acetyl-CoA and oxaloacetate to form citrate [1]. The peroxisomal CS, Cit2, participates in the glyoxylate cycle [2]. The third CS is a minor mitochondrial isofunctional enzyme, Cit3, and related to glycerol metabolism. However, the level of its intracellular activity is low and insufficient for metabolic needs of cells [3]. It has been reported that ${\Delta}cit1$ strain is not able to grow with acetate as a sole carbon source on either rich or minimal medium and that it shows a lag in attaining parental growth rates on nonfermentable carbon sources [2, 4, 5]. Cells of ${\Delta}cit2$, on the other hand, have similar growth phenotype as wild-type on various carbon sources. Thus, the biochemical basis of carbon metabolism in the yeast cells with deletion of CIT1 or CIT2 gene has not been clearly addressed yet. In the present study, we focused our efforts on understanding the function of Cit2 in utilizing $C_2$ carbon sources and then found that ${\Delta}cit1$ cells can grow on minimal medium containing $C_2$ carbon sources, such as acetate. We also analyzed that the characteristics of mutant strains defective in each of the genes encoding the enzymes involved in TCA and glyoxylate cycles and membrane carriers for metabolite transport. Our results suggest that citrate produced by peroxisomal CS can be utilized via glyoxylate cycle, and moreover that the glyoxylate cycle by itself functions as a fully competent metabolic pathway for acetate utilization in S. cerevisiae. We also studied the relationship between Cit1 and apoptosis in S. cerevisiae [6]. In multicellular organisms, apoptosis is a highly regulated process of cell death that allows a cell to self-degrade in order for the body to eliminate potentially threatening or undesired cells, and thus is a crucial event for common defense mechanisms and in development [7]. The process of cellular suicide is also present in unicellular organisms such as yeast Saccharomyces cerevisiae [8]. When unicellular organisms are exposed to harsh conditions, apoptosis may serve as a defense mechanism for the preservation of cell populations through the sacrifice of some members of a population to promote the survival of others [9]. Apoptosis in S. cerevisiae shows some typical features of mammalian apoptosis such as flipping of phosphatidylserine, membrane blebbing, chromatin condensation and margination, and DNA cleavage [10]. Yeast cells with ${\Delta}cit1$ deletion showed a temperature-sensitive growth phenotype, and displayed a rapid loss in viability associated with typical apoptotic hallmarks, i.e., ROS accumulation, nuclear fragmentation, DNA breakage, and phosphatidylserine translocation, when exposed to heat stress. Upon long-term cultivation, ${\Delta}cit1$ cells showed increased potentials for both aging-induced apoptosis and adaptive regrowth. Activation of the metacaspase Yca1 was detected during heat- or aging-induced apoptosis in ${\Delta}cit1$ cells, and accordingly, deletion of YCA1 suppressed the apoptotic phenotype caused by ${\Delta}cit1$ mutation. Cells with ${\Delta}cit1$ deletion showed higher tendency toward glutathione (GSH) depletion and subsequent ROS accumulation than the wild-type, which was rescued by exogenous GSH, glutamate, or glutathione disulfide (GSSG). Beside Cit1, other enzymes of TCA cycle and glutamate dehydrogenases (GDHs) were found to be involved in stress-induced apoptosis. Deletion of the genes encoding the TCA cycle enzymes and one of the three GDHs, Gdh3, caused increased sensitivity to heat stress. These results lead us to conclude that GSH deficiency in ${\Delta}cit1$ cells is caused by an insufficient supply of glutamate necessary for biosynthesis of GSH rather than the depletion of reducing power required for reduction of GSSG to GSH.

• Journal of Korean Academy of Fundamentals of Nursing
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• v.5 no.1
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• pp.125-141
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• 1998

The purpose of this research is to analyze the effects of aerobic exercise on body composition, cardiopulmonary function, serum lipid level and antioxidants of obese and normal college female students. The subject group was made up of 13 normal students (below 30% body fat ratio) and 12 obese students (above 30% body fat ratio). After a pretest, the subjects were given an 8-week aerobic program. Then the subjects were given a posttest and analyzed of body composition, serum lipid level, antioxidants and cardiopulmonary function after the 6th and the 8th week of the program. The program schedule was made up of 4 days per week, 60 minutes per day. Test includes B.W., subscapular and triceps subcutaneous fat thickness, change of respiratory gas, and two blood sampling before treadmill exercise and post all out state, which analyzed serum lipid and antioxidants. The subjects performed treadmill exercise starting with 4km/hr of walking and then gradually increase the speed of 1km/hr per minute until all out state. The obtained data were analyzed using SAS program. The statistical methods employed here were one-way ANOVA with repeated measure, Duncan Multiple range test, paired-t test and t-test. The test results and conclusion of this research were as follows. 1. The effects of aerobic exercise on body composition were as follows ; Percent body fat was significantly reduced 6 weeks after the program and lean body mass was significantly increased 8 weeks after the program in both groups(obese group: F=3.44 P=.044, normal group: F=3.30 P=.048). subscapular skinfold of the obese group showed a remarkable decrease after the 6th week(F=4.33 P=.021) triceps skinfold of the normal group showed a remarkable decrease after the 6th and the 8th week(F=4.55 P=.017) compared with readings before the aerobic program, the aerobic program made a bigger difference concerning body fat, lean body mass, subscapular skinfold in the obese group than in the normal group(t=2.41 P=.024, t=2.40 p=.025, t=2.43 p=.028). 2. The effects of aerobic exercise on cardiopulmonary function were as follows ; Maximal $O_2$ uptake/kg was significantly increased 6 weeks after the program in the obese group(F=3.20 P=.054), but not much difference was observed in the normal group. Maximal pulse rate was significantly reduced in both groups after 6 weeks of the program(obese group: F=2.77 P=.087, normal group: F=7.17 P=.001). 3. The effects of aerobic exercise on serum lipid level were as follows ; In a resting period, total cholesterol, Triglyceride, and LDL-cholesterol were slightly higher in the obese group than in the normal group, but HDL-cholesterol was higher in the normal group. But, with the aerobic program, total-cholesterol, Triglyceride, LDL-cholesterol were reduced gradually and HDL-choleterol got increased in both groups, but not much change was noticed in the normal group. However, in the obese group, serum HDL-cholesterol level got increased significantly(F=5.12 P=.012). 4. The effects of aerobic exercise in serum antioxidants were as follows ; In a resting period, the obese group's serum Free Radical and GSSG content were higher than the normal group's and the normal group's serum GSH content was higher than the obese group's. After 6 weeks of the aerobic program, Free Radical was reduced significantly in both groups(obese group: F=13.87 P=.000, normal group: F=18.60 P=.000) In the obese group, 8 weeks after the program, GSH was increased significantly(F=13.78, P=.000). In the normal group, 6 weeks after the program, GSH was reduced but increased again after 8 weeks(F=6.07 P=.005). Plasma GSSG was significantly increased after 8 weeks of exercise in both groups(obese group: F=19.75 P=.000, normal group: F=22.42 P=.000,) Compared with readings before the aerobic program, the aerobic program made a bigger difference serum GSH in the normal group than in the obese group(t=3.37 p=.003). As this result shows, it is known that the regular aerobic exercise improves cardiopulmonary function, body composition, serum lipid effectively and through the serum Free Radical reduction and antioxidant system activation, oxidant stress was suppressed. This effect was higher in the obese group than in the normal one. At least 6weeks exercise period need for improvement of body composition, cardiopulmonary function and activation of antioxidant system. This result suggest that improvement of serum lipid profile was needed longer than 8weeks exercise period.

• Korean Journal of Medicinal Crop Science
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• v.17 no.4
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• pp.243-250
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• 2009

Fifty percent ethanol extract of Lythrum salicaria Linne root (LSR) was tested in vitro on antioxidant activity, and furthermore was investigated on antioxidative and fibrosis protecting activities in $CCL_4$-induced liver fibrosis rat model. Ratio of hepatic GSH/GSSG (reduced glutathione/oxidized glutathione) as bio-parameter of antioxidant level in $CCL_4$ plus LSR-treated rats for 6 weeks significantly increased from 2.8- to 5.7-fold than that of $CCL_4$-treated rats at p < 0.05. Thiobarbituric acid reactive substances (TBARS) contents in $CCL_4$ plus LSR-treated rats ranged from 1.57- to 2.19-fold of normal rats and were lower than those in $CCL_4$ plus silymarin-treated rats ($1.78{\sim}2.46$-fold of normal rats) (p < 0.05). Amounts of hydroxyproline of liver tissue showing the content of total collagen, a parameter of fibrosis, in $CCL_4$ plus LSR-administrated rat livers were $4.9{\sim}8.8{\mu}g$/mg ($-47{\sim}-71%$, compared with that in $CCL_4$-treated rat livers ($16.6{\mu}g$/mg tissue), which were significantly lower than those in $CCL_4$ plus silymarin-administrated rats being $8.4{\sim}11.7{\mu}g$/mg ($-30{\sim}-50%$). This collagen reducing effect of liver tissue in $CCL_4$ plus LSR-treated rats was supported by histological observation using microscopy assay. From the results, we conclude that the root of L. salicaria have efficient antioxidant potential and effective antifibrotic activities.

• Korean Journal of Environmental Agriculture
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• v.19 no.4
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• pp.309-313
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• 2000

To investigate the effects of different UV-B levels on growth and biochemical defense response in plants, cucumber plants were subjected to three levels of biologically effective ultraviolet-B $(UV-B_{BE})$ radiation [daily dose: 0.03 (No), 6.40 (Low) and $11.30\;(High)\;kJ{\cdot}m^{-2}$, $UV-B_{BE}$] in the growth chambers for 3 weeks during the early growth period. Enhanced UV-B radiation drastically decreased both dry weight and leaf area of cucumber. With increasing UV-B intensity, chlorophyll content was decreased, however the level of malondialdehyde was highly increased linearly. Total contents of ascorbic acid and glutathione were tended to increase by UV-B, while the ratios of dehydroascorbate/ascorbate and oxidized glutathione/reduced glutathione were significantly increased with increasing UV-B intensity in cucumber. All the enzyme activities investigated (superoxide dismutase, ascorbate peroxidase, dehydroascorbate reductase, guaiacol peroxidase etc.) in cucumber were increased by the UV-B enhancement. These results suggested that enhanced UV-B irradiation caused photooxidative stress in cucumber plant and resulted in significant reduction in plant growth. Biochemical protection responses might be activated to prevent the leaves from damaging effects of oxidative stress generated by UV-B irradiation.