Purpose : FSGS do not respond well to any kind of therapy and gradually progress to end-stage renal disease. This study was conducted to investigate the difference of protein expression between MCNS and FSGS as a preliminary study for understanding the pathophysiology of FSGS. Methods : Renal biopsy samples of MCNS and FSGS were obtained, which was diagnosed by one pathologist. They were solubilized with a conventional extraction buffer for protein extraction. The solution was applied on immobilized linear gradient strip gel (pH 4-7) using IPGphor system. Silver staining was carried out according to standard method. Protein identification was done by searching NCBI database using MASCOT Peptide Mass Fingerprint software. Results : The differences in protein expressions between MCNS and FSGS were shown by increased or decreased protein spots. Most prominently expressed spot among several spots in FSGS was isolated and analyzed, one of which was glutathione S-transferase (GST) P1-1, whereas it was not found in MCNS. So GSTP1-1 was considered as the one of the key biomarkers in pathogenesis of FSGS. Conclusion : This result would be helpful in diagnosing FSGS and researching FSGS. Further studies for glutathione S-transferase P1-1 might be necessary to elucidate the mechanisms regarding FSGS.
Sohn, Eun Jeong;Shin, Min Jea;Kim, Dae Won;Son, Ora;Jo, Hyo Sang;Cho, Su Bin;Park, Jung Hwan;Lee, Chi Hern;Yeo, Eun Ji;Choi, Yeon Joo;Yu, Yeon Hee;Kim, Duk-Soo;Cho, Sung-Woo;Kwon, Oh Shin;Cho, Yong-Jun;Park, Jinseu;Eum, Won Sik;Choi, Soo Young
BMB Reports
/
v.49
no.7
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pp.382-387
/
2016
Reactive oxygen species generated under oxidative stress are involved in neuronal diseases, including ischemia. Glutathione S-transferase pi (GSTpi) is a member of the GST family and is known to play important roles in cell survival. We investigated the effect of GSTpi against oxidative stress-induced hippocampal HT-22 cell death, and its effects in an animal model of ischemic injury, using a cell-permeable PEP-1-GSTpi protein. PEP-1-GSTpi was transduced into HT-22 cells and significantly protected against H2O2-treated cell death by reducing the intracellular toxicity and regulating the signal pathways, including MAPK, Akt, Bax, and Bcl-2. PEP-1-GSTpi transduced into the hippocampus in animal brains, and markedly protected against neuronal cell death in an ischemic injury animal model. These results indicate that PEP-1-GSTpi acts as a regulator or an antioxidant to protect against oxidative stress-induced cell death. Our study suggests that PEP-1-GSTpi may have potential as a therapeutic agent for the treatment of ischemia and a variety of oxidative stress-related neuronal diseases.
Journal of The Korean Society of Grassland and Forage Science
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v.34
no.2
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pp.114-119
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2014
Salinity and drought stresses are probably the most significant abiotic factor limiting plant's growth, also negatively affect seed germination and early seedling development. To study on effect of NaCl and PEG stress on seed germination and gene expression pattern of tall fescue, the levels of NaCl and PEG-induced water stresses were determined in first experiment. Different concentration of NaCl (0 to 350 mM) and PEG (0 to 30%) were used for seed treatment. Seed Germination percentage reduced with increasing osmotic potential of growth medium either due to NaCl or PEG. Seeds were not germinate at 350 mM NaCl or 30% PEG treatment. On the basis of the results, Kentucky31(E-) had more resistant than Fawn in both stress conditions. Furthermore, we have used an annealing control primer-based differential display reverse transcription-polymerase chain reaction method to identify salt- and drought stress-induced differentially expressed genes (DEGs) in tall fescue leaves. Using 120 annealing control primers, a total of 4 genes were identified and sequenced. The possible roles of the identified DEGs are discussed in the context of their putative role during salinity and drought stresses.
Background and Objectives: Individual genetic susceptibilities to chemical carcinogens have been recognized as a major important host factors in human cancers. The cytochrome P450 family (CYPs) and glutathione S-transferase(GST) have been reported to be associated with risks to the smoking-related human cancers. Inactivation of tumor suppressor genes like p53 playa key role in tumor progression. The purpose of this study is to demonstrate an association between p53 overexpression and the prevalence of the genetic polymorphisms of CYP1A1 and GSTs in Korean head and neck squamous cell carcinoma (HNSCC). Materials and Methods: The polymorphisms of CYPIA1 and GSTs were analyzed by PCR and PCR-RFLP in 98 Korean head and neck squamous cell carcinoma patients. The expression of p53 was analyzed by immunohistochemistry with anti-p53 Ab (DO7). Results: Overexpression of p53 detected in 45.9% of HNSCC. The odds ratio for p53 overexpression in GSTM1(-), GSTT1(-), GSTP1(val/val) and CYP1A1(val/val) were 1.53, 1.83, 1.17 and 1.47, respectively. Among the combined genotypes, the odds ratio of the CYP1A1 val/val, GSTM1 (-), CYP1A1 val/val, GSTT1(-), and CYP1A1 val/val, GSTT1(-) were 2.0, 2.34 and 4.68, respectively. Conclusion: Based on our results, it might be suggested that p53 overexpression is slightly increased in GSTM1(-), GSTT1(-), GSTP1 val/val, CYP1A1 val/val genotypes. The further study is needed to evaluate the relationship and mechanism between the p53 overexpression and the specific CYP1A1 and GSTs genotypes.
Journal of the Korean Society of Food Science and Nutrition
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v.31
no.1
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pp.81-86
/
2002
The present study was conducted to investigate the effect of dietary supplementation of vitamin A or $\beta$-carotene on oxidative damage induced by acute ethanol administration. Sprague-Dawley rats were fed on the experimental diets supplemented with retinyl acetate (2.86 mg/kg diet) or $\beta$-carotene (15.2 mg/kg diet) for 5 weeks. After fed the diet, rats were administered 20% ethanol solution (3g/kg B.W.) acutely. Lipid peroxide values in hepatic tissue, hepatic antioxidative enzyme activities and contents of antioxidative nutrient such as vitamins A and E in serum and hepatic tissue were measured. Hepatic level of malondialdehyde decreased in $\beta$-carotene group compared to the control group. However, there was no significant difference between retinal acetate and $\beta$-carotene groups. Superoxide dismutase activity was higher in retinal acetate group than in the control group. Hepatic glutathione-S-transferase activity of retinal acetate and $\beta$-carotene groups significantly decreased as compared with that of control group. The hepatic content of retinol increased in retinal acetate and $\beta$-carotene groups, especially, in retinyl acetate group. But there was no significant difference in serum content of retinol among the groups. Hepatic content of $\alpha$-tocopherol was significantly increased in retinyl acetate and $\beta$-carotene groups. In conclusion, acute ethanol administration might induce lipid peroxidation, and the dietary supplementation of retinyl acetate or $\beta$-carotene improve partly the antioxidative system through activation of superoxide dismutase and retention of hepatic $\alpha$-tocopherol in ethanol-treated rats.
Objective : The purpose of this study was to investigate the effects of fidget spinner training on the hand function and handwriting legibility of lower grade elementary school studens. Methods : This study randomly assigned a study group of 12 children and control group of 12 children from 24 children in grade 1 and 2 (ages 7 through 8), whose are dominantly right handed. The study used was a pre-post process. The intervention was conducted only on the study group twice a week for 5 weeks and for 40 minutes per session, for a total of ten sessions. The measuring instruments used to compare the hand functions and handwriting legibility were the Jebsen-Taylor Hand Function Test, Grip Strength Test, and Legibility Test. The data analysis used a Wilcoxon signed rank, Mann-Whitney U and Chi-Square cross analysis. Results : The fidget spinner training showed significant improvement in the study group's hand function(grip strength and handwriting legibility) and a significant difference was shown between the control and study groups. Conclusion : This study confirmed the value and utility of a fidget spinner as a tool for improving the hand function and handwriting legibility of elementary school students in lower grades. Future studies are expected to verify the effectiveness of the fidget spinner training based on the present study.
The purpose of this study was to investigate the effect of vitamin E and selenium on the antioxidative defense mechanism in the liver of streptozotocin(STZ)-induced diabetic rats. Sprague-Dawley male rats(120$\pm$10gm) were randomly assigned to one control and five STZ-diabetic groups. Diabetic groups were classified to STZ-0E (vitamin E free diet), STZ-40E(40mg vitamin E/kg of diet), STZ-400E(400mg vitamin E/kg of diet), STZ-S(0.5ppm Se/kg of diet) and STZ-400ES(400mg vitamin E and 0.5ppm Se/kg of diet) according to the level of vitamin E and selenium supplementation. Diabetes was experimentally induced by intravenous adminstration of 55mg/kg of STZ in citrate buffer(pH 4.3) after 4-weeks feedng of six experimental diets. Animals were sacrificed at the 4th day of diabetic states. Activities of the serum glutamic oxaloacetate transaminase(GOT) and the glutaminc pyruvate transaminase(GPT) in STZ-0E, STZ-40E and STZ-S rats were higher than those of control. Liver xanthine oxidase activities were similar to serum GOT and GPT. Liver superoxide dismutase(SOD) activities were higher in STZ-0E and STZ-40E groups by 33%, 22%, respectively than that of control. Glutathione S-transferase(GST) activities of liver were similar to GSH-Px activities. The contents of vitamin E in liver tissue were significantly lower STZ-0E, STZ-40E and STZ-S groups by 50%, 36%, 45% than that of control. Reduced glutathione(GSH) contents of liver were lower STZ-0E, STZ-40E, STZ-400E, STZ-S and STZ-400ES groups by 57%, 51%, 19%, 18%, 12% than that of control. Lipid peroxide values (LPO) in liver were higher 5.6, 2.3 and 2.3 times in STZ-0E, STZ-40E and STZ-S group than that of control. The present results indicate that STZ-induced diabetic rats are more sensitive to oxidative stress, leading to the acceleration of lipid peroxidation process, which can be more accelerated by feeding the low level of dietary vitamin E. In the coincident supplementation of high dietary vitamin E and selenium antioxidative enzymes activities and physiolosical antioxidants were increased more than those of the separate supplementation of vitamin E or selenium. Therefore, dietary vitamin E and selenium reduced peroxidative damage of tissue, promoting antioxidative defense mechanism against lipid peroxidation by diabetes.
Journal of the Korean Society of Food Science and Nutrition
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v.33
no.4
/
pp.653-658
/
2004
Oxidative stress is currently suggested as a mechanism underyling diabetes. Accordingly, the present study was designed to evaluate the effect of Aralia elate water extracts (AEW) on activities of hepatic oxygen free radical generating and scavenging enzymes in streptozotocin (STZ)-induced diabetic rats. Male Wistar rats divided into nondiabetic group, diabetic group, and diabetic-AEW supplemented group. The extract was supplemented in 1.14% of raw Aralia elata/kg diet for 7 weeks. Diabetes was induced by injecting STZ (55 mg/kg BW, ip) once 2 weeks before sacrifying. The hepatic cytochrome P-450 content, xanthine oxidase and aminopyrine N-demethylase activities were significantly lowered in the diabetic group compared to the nondiabetic group. Whereas, the activities of aniline hydroxylase and oxygen free radical scavenging enzymes, superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glucose-6-phosphate dehydrogenase and glutathione S-transferase, were significantly higher in the diabetic group than in the nondiabetic group. However, the supplementation of AEW normalized these enzyme activities in STZ-induced diabetic rats. When the AEW was supplemented with the diabetic rats, hepatic glutathione content was markedly elevated as well as lipid peroxide level was significantly lowered compared to those of the diabetic group. Thus, these results suggested that AEW supplement enhanced the activities of oxygen species metabolizing enzymes in STZ-induced diabetic rats.
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
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pp.36-36
/
2017
Out of twenty common protein amino acids, there are many kinds of non protein amino acids (NPAAs) that exist as secondary metabolites and exert ecological functions in plants. Mimosine (Mim), one of those NPAAs derived from L. leucocephala acts as an iron chelator and reversely block mammalian cell cycle at G1/S phases. Cysteine (Cys) is decisive for protein and glutathione that acts as an indispensable sulfur grantor for methionine and many other sulfur-containing secondary products. Cys biosynthesis includes consecutive two steps using two enzymes-serine acetyl transferase (SAT) and O-acetylserine (thiol)lyase (OASTL) and appeared in plant cytosol, chloroplast, and mitochondria. In the first step, the acetylation of the ${\beta}$-hydroxyl of L-serine by acetyl-CoA in the existence of SAT and finally, OASTL triggers ${\alpha}$, ${\beta}$-elimination of acetate from OAS and bind $H_2S$ to catalyze the synthesis of Cys. Mimosine synthase, one of the isozymes of the OASTLs, is able to synthesize Mim with 3-hydroxy-4-pyridone (3H4P) instead of $H_2S$ for Cys in the last step. Thus, the aim of this study was to clone and characterize the cytosolic (Cy) OASTL gene from L. leucocephala, express the recombinant OASTL in Escherichia coli, purify it, do enzyme kinetic analysis, perform docking dynamics simulation analysis between the receptor and the ligands and compare its performance between Cys and Mim synthesis. Cy-OASTL was obtained through both directional degenerate primers corresponding to conserved amino acid region among plant Cys synthase family and the purified protein was 34.3KDa. After cleaving the GST-tag, Cy-OASTL was observed to form mimosine with 3H4P and OAS. The optimum Cys and Mim reaction pH and temperature were 7.5 and $40^{\circ}C$, and 8.0 and $35^{\circ}C$ respectively. Michaelis constant (Km) values of OAS from Cys were higher than the OAS from Mim. Inter fragment interaction energy (IFIE) of substrate OAS-Cy-OASTL complex model showed that Lys, Thr81, Thr77 and Gln150 demonstrated higher attraction force for Cys but 3H4P-mimosine synthase-OAS intermediate complex showed that Gly230, Tyr227, Ala231, Gly228 and Gly232 might provide higher attraction energy for the Mim. It may be concluded that Cy-OASTL demonstrates a dual role in biosynthesis both Cys and Mim and extending the knowledge on the biochemical regulatory mechanism of mimosine and cysteine.
Kim, In-Hye;Cho, Kang-Jin;Ko, Jeong-Sook;Kim, Jae-Hyun;Om, Ae-Son
The Korean Journal of Food And Nutrition
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v.25
no.1
/
pp.123-131
/
2012
The objective of the present study was to evaluate the potential antidiabetic and antioxidant effect of the ethanol extract from Chrysanthemum cornarium L. var. spatiosum(CSE) against alloxan-induced oxidative stress in pancreatic ${\beta}$-cells, HIT-T15. In this study, the antidiabetic effect of CSE was examined using the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliu bromide(MTT) cell proliferation assay, lactate dehydrogenase(LDH) release assay, $NAD^+$/NADH ratio and insulin secretion. To further investigate whether CSE is involved in the antioxidant activity of alloxan-damaged HIT-T15 cells, its antioxidant effect against alloxan-induced oxidative stress was measured in HIT-T15 cells by determining the levels of antioxidant enzymes including superoxide dismutase(SOD), glutathione S-transferase(GST), glutathione reductase(GR) and glutathione peroxidase(GPx). The results of this analysis showed that alloxan significantly decreased cell viability, increased LDH leakage, and lowered $NAD^+$/NADH ratio and insulin secretion in HIT-T15 cells. However, CSE significantly increased the viability of alloxan-treated cells and lowered LDH leakage. The intracellular NAD+/NADH ratio and insulin secretion were also significantly increased by 1.7-fold and 1.3-fold, respectively, after treatment with 100 ${\mu}g/m{\ell}$ CSE. The HIT-T15 cells treated with alloxan showed significant decreases in the activities of antioxidant enzymes, while CSE significantly elevated the levels of antioxidant enzymes. These findings suggest that CSE could have a protective effect against cytotoxicity and dysfunction of pancreatic cells in the presence of alloxan-induced oxidative stress.
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