• International Journal of Stem Cells
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• v.16 no.3
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• pp.356-362
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• 2023

Glutathione (GSH) is a chief cellular antioxidant, affecting stem cell functions. The cellular GSH level is dynamically altered by the redox buffering system and transcription factors, including NRF2. Additionally, GSH is differentially regulated in each organelle. We previously reported a protocol for monitoring the real-time GSH levels in live stem cells using the reversible GSH sensor FreSHtracer. However, GSH-based stem cell analysis needs be comprehensive and organelle-specific. Hence, in this study, we demonstrate a detailed protocol to measure the GSH regeneration capacity (GRC) in living stem cells by measuring the intensities of the FreSHtracer and the mitochondrial GSH sensor MitoFreSHtracer using a high-content screening confocal microscope. This protocol typically analyses the GRC in approximately 4 h following the seeding of the cells onto plates. This protocol is simple and quantitative. With some minor modifications, it can be employed flexibly to measure the GRC for the whole-cell area or just the mitochondria in all adherent mammalian stem cells.

• Asian-Australasian Journal of Animal Sciences
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• v.8 no.4
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• pp.379-385
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• 1995

Ninety Wistar male rats were used to study the effects of vitamin E and Se supplementation to diets containing aflatoxin $B_1$ on the contents of liver lipids and various blood parameters. Two levels of dietary aflatoxin (0 and 1 ppm), 3 levels of vitamin E (30, 60 and 120 IU/kg), and 3 levels of Se (0.1, 1 and 2 ppm) were used to design a $2{\times}3{\times}3$ factorial experiment. Rats, weighing about 200 g, were randomly allotted to 18 cages, 5 rats per cage. The aflatoxin significantly (p < .05) decreased growth rate, feed intake and feed efficiency. Aflatoxin increased the glucose level and decreased the cholesterol level in blood significantly. Levels of blood triglyceride, total protein, and albumin were not affected by aflatoxin, vitamin E or Se. Activities of blood alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were significantly increased by aflatoxin; however, the glutathione peroxidase (GSH-Px) activity in the blood was decreased by aflatoxin even in the presence of Se. The vitamin E supplementation decreased the AST activity significantly, while GSH-Px activity increased significantly as the levels of dietary Se increased. The levels of total cholesterol and free cholesterol in the liver were significantly lower in rats receiving aflatoxin, while the extra vitamin E supplementation increased these hepatic cholesterol levels. It was concluded that the extra dietary vitamin E or Se supplementation might partially alleviate some of the harmful effects of aflatoxin in rats.

• Journal of agricultural medicine and community health
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• v.19 no.2
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• pp.107-118
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• 1994

Health effects of pesticide among vinylhouse workers in Sangjoo County, Kyungpook Province were assessed by measuring cholinesterase, glutathion reductase, and methemoglobin. Activities of cholinesterases among vinylhouse workers and general farmers were 3.89U and 3.98U in serum and 5.29U and 5.50U in red blood cells, respectively. While levels of methemoglobin between vinylhouse workers(0.16%) and general farmers(0.17%) were very similar, glutathione reductase among vinylhouse workers were 8% lower than that of general farmers. Inhibition of RBC cholinesterase among vinylhouse workers was two times greater than general farmers, suggesting a greater exposure of vinylhouse workers to the organophosphate pesticides. Methemoglobin level among vinylhouse workers with more than or equal to 10 vinylhouse units was 13% higher than that of the workers with less than 10 units. Vinylhouse workers using protective gears during pesticide spray showed higher level of reduced glutathione and lower level of methemoglobin, as compared to the non-users. This indicates that protective gears play an important role against pesticide exposure. Vinylhouse workers practicing good personal hygiene showed a higher level of reduced glutathione, as compared to those with poor personal hygiene, indicating that personal hygiene is also an important factor in reducing pesticide exposure. The present study represents a first attempt to analyze cholinesterase, methemoglobin and glutathione reductase activities among the vinylhouse workers. While neither vinylhouse workers nor general farmers showed abnormal findings among biochemical parameters used in the present study, increased level of methemoglobin and decrease of glutathione reductase activity among vinylhouse workers suggest that these workers have been subject to a higher exposure of pesticide than general farmers.

• Korean Journal of Acupuncture
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• v.18 no.1
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• pp.1-9
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• 2001

Artemisia asiatica Nakai aqua-acupuncture solution (ANAS) was administered once daily for 10 days before the tumor implantation ($1{\times}10^6\;cells$). Body weight, spleen weight and the number of ascitic tumor cells were measured at 6 days after tumor implantation. The change of body weight and the survival rate of mice were observed for 21 days. It was used three biomarkers (quinone reductase, glutathione, glutathione S-transferase) to test chemopreventive potentials of ANAS. ANAS exerted antitumor activity by inhibiting the growth of Ehrlich ascites tumor cells in vivo. Mice given Ehrlich cells and ANAS at $CV_{12}$ and $BL_{18}$ had 57.1% to 49.2% survival after 21 days. Quinone reductase activity and glutathione levels were increased with ANAS. However, glutathione S-transferase level was 1.1-fold with ANAS. These results suggest that ANAS has chemopreventive potential by inducing QR activity and increasing GSH level.

• Plant Biotechnology Reports
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• v.5 no.4
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• pp.353-365
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• 2011

The present study investigates the possible regulatory role of exogenous nitric oxide (NO) in antioxidant defense and methylglyoxal (MG) detoxification systems of wheat seedlings exposed to salt stress (150 and 300 mM NaCl, 4 days). Seedlings were pre-treated for 24 h with 1 mM sodium nitroprusside, a NO donor, and then subjected to salt stress. The ascorbate (AsA) content decreased significantly with increased salt stress. The amount of reduced glutathione (GSH) and glutathione disulfide (GSSG) and the GSH/GSSG ratio increased with an increase in the level of salt stress. The glutathione S-transferase (GST) activity increased significantly with severe salt stress (300 mM). The ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), catalase (CAT) and glutathione peroxidase (GPX) activities did not show significant changes in response to salt stress. The glutathione reductase (GR), glyoxalase I (Gly I), and glyoxalase II (Gly II) activities decreased upon the imposition of salt stress, especially at 300 mM NaCl, with a concomitant increase in the $H_2O_2$ and lipid peroxidation levels. Exogenous NO pretreatment of the seedlings had little influence on the nonenzymatic and enzymatic components compared to the seedlings of the untreated control. Further investigation revealed that NO pre-treatment had a synergistic effect; that is, the pre-treatment increased the AsA and GSH content and the GSH/GSSG ratio, as well as the activities of MDHAR, DHAR, GR, GST, GPX, Gly I, and Gly II in most of the seedlings subjected to salt stress. These results suggest that the exogenous application of NO rendered the plants more tolerant to salinity-induced oxidative damage by enhancing their antioxidant defense and MG detoxification systems.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.79-91
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• 2021

γ-Glutamylcysteine synthetase (Gcs1) and glutathione reductase (Glr1) activity maintains minimal levels of cellular methylglyoxal in Candida albicans. In glutathione-depleted Δgcs1, we previously saw that NAD(H)-linked methylglyoxal oxidoreductase (Mgd1) and alcohol dehydrogenase (Adh1) are the most active methylglyoxal scavengers. With methylglyoxal accumulation, disruptants lacking MGD1 or ADH1 exhibit a poor redox state. However, there is little convincing evidence for a reciprocal relationship between methylglyoxal scavenger genes-disrupted mutants and changes in glutathione-(in)dependent redox regulation. Herein, we attempt to demonstrate a functional role for methylglyoxal scavengers, modeled on a triple disruptant (Δmgd1/Δadh1/Δgcs1), to link between antioxidative enzyme activities and their metabolites in glutathione-depleted conditions. Despite seeing elevated methylglyoxal in all of the disruptants, the result saw a decrease in pyruvate content in Δmgd1/Δadh1/Δgcs1 which was not observed in double gene-disrupted strains such as Δmgd1/Δgcs1 and Δadh1/Δgcs1. Interestingly, Δmgd1/Δadh1/Δgcs1 exhibited a significantly decrease in H2O2 and superoxide which was also unobserved in Δmgd1/Δgcs1 and Δadh1/Δgcs1. The activities of the antioxidative enzymes erythroascorbate peroxidase and cytochrome c peroxidase were noticeably higher in Δmgd1/Δadh1/Δgcs1 than in the other disruptants. Meanwhile, Glr1 activity severely diminished in Δmgd1/Δadh1/Δgcs1. Monitoring complementary gene transcripts between double gene-disrupted Δmgd1/Δgcs1 and Δadh1/Δgcs1 supported the concept of an unbalanced redox state independent of the Glr1 activity for Δmgd1/Δadh1/Δgcs1. Our data demonstrate the reciprocal use of Eapx1 and Ccp1 in the absence of both methylglyoxal scavengers; that being pivotal for viability in non-filamentous budding yeast.

• Korean Journal of Acupuncture
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• v.25 no.2
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• pp.143-158
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• 2008

Objectives : The purpose of this study is to observe the anti-oxidative effects of electroacupuncture at GB34(Yangreungcheon) in rats. Methods : The authors performed several experimental items including measurements of body weight, relative liver weight, levels of albumin, total bilirubin, LDL-cholesterol, LDH, GOT and GPT in blood serum, levels of SOD, glutathione, catalase, NO and MDA in liver, and histological analysis of liver. The conclusions are as follows. Results : 1. In the GB34-EA group, liver index was lower than those of the control group and the holder group. 2. In the GB34-EA group, the levels of albumin and total bilirubin in serum were increased significantly compared to those of the control and the holder group. LDL-cholesterol level was decreased significantly compared to those of the control group and the holder group. 3. In the GB34-EA group, liver SOD, glutathione, catalase activity were increased, and NO concentration in liver was decreased significantly compared to those of control and the holder group. 4. In the GB34-EA group, the density of liver tissue was more similarly maintained to that of the normal group compared to those of the control group, holder group and sham-EA group. Conclusions : These results suggest that electroacupuncture at GB34 may have an antioxidant effect in rat.

• The Korean Journal of Physiology and Pharmacology
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• v.20 no.3
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• pp.269-277
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• 2016

Ellagic acid (EA), an antioxidant polyphenolic constituent of plant origin, has been reported to possess diverse pharmacological properties, including anti-inflammatory, anti-tumor and immunomodulatory activities. This work aimed to clarify the skin anti-photoaging properties of EA in human dermal fibroblasts. The skin anti-photoaging activity was evaluated by analyzing the reactive oxygen species (ROS), matrix metalloproteinase-2 (MMP-2), total glutathione (GSH) and superoxide dismutase (SOD) activity levels as well as cell viability in dermal fibroblasts under UV-B irradiation. When fibroblasts were exposed to EA prior to UV-B irradiation, EA suppressed UV-B-induced ROS and proMMP-2 elevation. However, EA restored total GSH and SOD activity levels diminished in fibroblasts under UV-B irradiation. EA had an up-regulating activity on the UV-B-reduced Nrf2 levels in fibroblasts. EA, at the concentrations used, was unable to interfere with cell viabilities in both non-irradiated and irradiated fibroblasts. In human dermal fibroblasts, EA plays a defensive role against UV-B-induced oxidative stress possibly through an Nrf2-dependent pathway, indicating that this compound has potential skin antiphotoaging properties.

• Journal of Environmental Health Sciences
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• v.19 no.2
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• pp.78-87
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• 1993

The accumulations of mercury, lactate dehydrogenase and antioxidant enzymes activities of which are glutathione peroxidase, catalase and superoxide dismutase, and pathological changes were investigated in liver and kidney of mice which were fed the water supplemented with two levels (0.5 mM and 1.0 mM) of mercury chloride (HgCl$_2$). During the mercury feeding, the weight gain of mice in experimental groups was less than that of control group mice, while no overt signs related to mercury toxicity were noted in any experimental groups. Mercury concentrations in liver and kidney increased significantly in the early period (1~2 weeks) after mercury administration, which were measured as high as 100 times in liver and kidney in comparison to those of the control groups, but there were relatively stable for the levels of accumulation in following periods. The lactate dehydrogenase activities in liver and kidney were relatively increased in the period of 2~3 weeks of mercury administration in the experimental groups, there were normal levels in other periods of administration without the dose-dependencies. The glutathione peroxidase activities were not affected by the dosages of mercury chloride and the duration of ingestion. But the catalase activities significantly increased in 2~3 weeks after ingestion, and the superoxide dismutase activities of kidney also showed a peak in 3 weeks of ingestion while this peak was not found in the results measured in liver tissues.

• Journal of Nutrition and Health
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• v.30 no.8
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• pp.920-929
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• 1997

This study examined the effect of dietary salt levels on the incidence and cure of gastric ulcer in rats. Two sets of experiment were conducted . In the first experiment, the rats were divided into 3 groups. The 3 groups were fed 0%, 4%, and 8% NaCl diets respectively for 20days. The rats were given water -immersion restraint stress at the end of the dietary period , and sacrificed. The ulcer index by histological test was higher in rats fed the 8% NaCl diet than those in the other groups. The hexosamine and glutathione levels were significantly lower in the rats fed the 8% NaCl diet. Hematocrit and total iron binding capacity(TIBC) showed lower values caused by bleeding of gastric mucosa. The second experiment was designed to determine the effect of soldium concentration on the cure of gastric ulcer . As the gastric ulcer was recovered, ulcer length was gradually deceased in the control group but not changed in the 8% NaCl diet group. The gastric hexosamine and hepatic glutathione were increased in the control group but decreased in the 8% NaCl diet group. The hematologic indices of stressed rats showed the same tendency. As a result, dietary salt per se did not cause gastric ulcer . Once an ulcer is formed by stress or any other factor, higher levels of dietary salt may be detrimental to gastric mucosa, thereby delaying the healing of the ulcer. It is recommended that dietary salt intake be reduced in stress-prone people.