• Journal of Acupuncture Research
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• v.17 no.3
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• pp.176-187
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• 2000

Objectives : This study was purposed to investigate the antioxidative effects of Paeoniae radix aqua-acupuncture solution(PR) on culture liver cell system, lipid peroxidation and antioxidative enzyme activities in tert-butyl hydroperoxide(t-BHP) treatmented conditions. Methods : Cultured normal rat liver cell(Ac2F) were prepared and incubated with or without PR(at 2% volume in culture medium). After 16~18hr, cells placed in DMEM medium without serum, and then incubated with 1mM t-BHP for 2hr. Viable cells were detected by MTT assay, and the levels of lipid peroxide(LPO) were measured by TBA method. And catalase activity was measured as the decrease in hydrogen peroxide absorbance at 240nm on spectrophotometer using 30mM hydrogen peroxide. Superoxide dismutase(SOD) were assayed by recording the inhibition of nitro blue tetrazolium reduction with xanthine and xanthine oxidase. Glutathione peroxidase(GPX) activity was determined by the modified coupled assay developed by Paglia and Lawrence. The reaction was started by addition of 2.2mM hydrogen peroxide as substrate. The change in absorbance at 340nm was measured for 1min on spectrophotometer. Glutathione-S-transferase(GST) activity was assayed with CDNB as substrate and enzyme activity of GST towards the glutathione conjugation of CDNB. Results : Cell killing was significantly enhanced by addition of t-BHP compared to those of untreated group. PR pretreated cell resisted the toxic effects of t-BHP. LPO levels of t-BHP treatment group were significantly higher than other groups. This increased level was significandy reduced by PR pretreatment. The t-BHP treatment resulted in a decrease of catalase, GPX and GST activities. By contrast, PR pretreatment markedly increased compare to those of untreated groups. Conclusions : T-BHP which can produce intracellular free radical was used for inducer of the peroxidation of cellular lipids. PR protected the cell death induced by t-BHP and significantly increased cell viabiliry in the normal rat liver cell, and showed effective inhibition of lipid peroxidation, and elevations of catalase, GPX and GST activities. These results suggested that PR might play a protective role in lipid peroxidation by free radicals.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.4
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• pp.2145-2150
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• 2016

Background: Infection with hepatitis B virus (HBV) is a major global public health problem, with a wide spectrum of clinical manifestations. Human cytosolic glutathione-S-transferases (GSTs) include several classes such as alpha (A), mu (M), pi (P), sigma (S), zeta (Z), omega (O) and theta (T). The present study aimed to investigate the role of GST omega genes (GSTO1 and GSTO2) in different groups of patients infected with HBV. Materials and Methods: HBV groups were classified according to clinical history, serological tests and histological analysis into normal carriers (N), acute (A), chronic (CH), cirrhosis (CI) and hepatocellular carcinoma (HCC) cases. The study focused on determination of the genotypes of GST omega genes (GSTO1 and GSTO2) and GST activity and liver function tests. Results: The results showed that GSTO1 (A/A) was decreased in N, A, CH, CI and HCC groups compared to the C-group, while, GSTO1 (C/A) and GSTO1(C/C) genotypes were increased significantly in N, A, CH, CI and HCC groups. GSTO2 (A/A) was decreased in all studied groups as compared to the C-group but GSTO2(A/G) and GSTO2(G/G) genotypes were increased significantly. In addition, GST activities, albumin and TP levels were decreased in all studied groups compared to the C-group, while the activities of transaminases were increased to differing degrees. Conclusions: The results indicate that GSTO genetic polymorphisms may be considered as biomarkers for determining and predicting the progression of HBV infection.

• Proceedings of the Materials Research Society of Korea Conference
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• 2009.05a
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• pp.21.1-21.1
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• 2009

상변화 메모리는 비휘발성 메모리이면서 빠른 동작 속도, 낮은 동작 전압 등 다양한 장점을 지니고 있어 차세대 메모리로 주목 받고 있다. 최근 상변화 메모리의 동작 전류를 감소시키기 위해 상변화 물질 및 전극 물질에 대한 연구를 진행하고 있으며, 소자의 크기를 최소화 하기 위한 연구가 진행되고 있다. 본 연구에서는 나노 임프린트 리소그래피와 전도성 AFM을 이용하여 나노급 상변화 물질의 특성을 평가하였다. 나노급 상변화 물질을 형성하기 위해 열경화성 나노 임프린트 리소그래피를 이용하여 $Ge_2Sb_2Te_5$(GST)/Mo/SiO2 기판 위에 200nm급 홀 패턴을 형성하였다. 홀 패턴에 Cr을 증착하여 리프트 오프 한 뒤 Cr을 하드 마스크로 사용하여 GST를 식각하였다. 그 결과, Mo 하부 전극 위에200nm 지름과 100nm 높이를 가지는 GST 나노 기둥을 형성하였다. GST 나노 기둥의 전기적 특성 평가를 위해 저항 측정 장비 및 펄스 발생기와AFM을 사용하였다. AFM은 접촉 모드로 설정하였으며, Pt가 코팅된 AFM tip을 사용하여 Cr 하드 마스크와 함께 상부 전극으로 사용하였다. GST 나노 기둥을 초기화 시키기 위해 I-V sweep을 하였으며, 그 결과 $1M\Omega$에서 $10\;k\Omega$으로 저항이 변화함을 확인하였다. GST 나노 기둥은 2V, 5ns의 리셋 펄스에서 비정질로 변화하였으며, 1.3V, 150ns의 셋 펄스에서 결정질로 변화하였다. 이 동작 전압으로 5번의 스위칭 특성을 평가하였으며, 이 결과는 소자 형태의 200nm 급GST의 특성과 유사하여 나노급 상변화 물질을 테스트하는 새로운 방법으로 사용될 수 있을 것이다.

• Korean Journal of Environmental Biology
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• v.32 no.3
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• pp.234-242
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• 2014

In this study, gametotoxicity and embryotoxicity experiments using Hemicentrotus pulcherrimus were carried out to investigate the ecotoxicological effects of bisphenol A (BPA). We examined the effects of BPA on fertilization and normal embryogenesis at various concentrations (0, 300, 500, 800, 1000, and 1500 ppb). The results demonstrated that the fertilization rates were not changed. The normal embryogenesis rates were gradually decreased in a dose-dependent manner, and were significantly lowered following 800 ppb BPA treatment ($EC_{50}$=1056.1 ppb, 95% Cl=981.8~1163.9 ppb). The observed effective concentration and the lowest observed effective concentration of the normal embryogenesis rate were 500 ppb and 800 ppb, respectively. The embryos showed retarded development at each tested concentration, indicating the fact the embryonic development was delayed due to the increasing concentrations of BPA. Furthermore, we examined the expression of glutathione S-transferase (GST) mRNA at various concentrations of BPA in H. pulcherrimus. Interestingly, it was found that the expression level of GST mRNA was significantly increased in the experimental group exposed to BPA. Based on these results, we suggested that BPA at greater than 800 ppb has a toxic effect during the early embryonic stages of H. pulcherrimus, and GST mRNA may be used as a biomarker for risk assessment of BPA contamination.

• Korean Journal Plant Pathology
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• v.13 no.4
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• pp.248-254
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• 1997

Expression vector (pGEX-Tu) for the coat protein (CP) gene of turnip mosaic virus Ca strain (TuMV-Ca) was constructed by incorporation of TuMV CP gene into pGEX-KG vector which had ${\beta}$-galactosidase gene and IPTG (isopropylthio-${\beta}$-D-galactoside) induction site. The results of ELISA and western hybridization indicated that optimal condition of the expression were when IPTG and western hybridization indicated that optimal condition of the expression were when IPTG induction was carried out on YTA medium with ampicillin in 2 hours after the E. coli seed inoculation ($A_{595}$=0.1/ml). TuMV CP gene was expressed with GST (Glutathion S-Transferase) gene fusion system, and the size of fusion protein was estimated to be 59kDa, for TuMV CP was 33 kDa and GST was 26 kDa.

• Korean Journal of Environmental Agriculture
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• v.16 no.1
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• pp.37-43
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• 1997

Tolerance mechanism to simazine (6-chloro-N,N'-diethyl-1,3,5-triazine-2,4-diamine) in Coix lacryma-jobi was investigated with respect to herbicide detoxification via glutathione conjugation. Simazine was initially absorbed by seedlings of C. lacryma-jobi and corn, but after 12 hours of treatment, no significant difference in simazine absorption was found in both species. Simazine absorbed was rapidly metabolized to glutathione-simazine conjugate. One to six hours after treatment, metabolism was approximately 2-fold faster in C. lacryma-jobi than in corn. Glutathione content was found 1.5- and 2.3-fold higher in coleoptile and root of C. lacryma-jobi, respectively, compared with corn. In both species, the highest concentration of glutathione was found in coleoptile tissue. Glutathione S-transferase that exhibits activity with 1-chloro-2,4-dinitrobenzene was not significantly different between two species. However, glutathione S-transferase activity with simazine was approximately 2-fold greater in C. lacryma-jobi than in corn. The glutathione S-transferase activity was 20 to 30% greater in shoot of either species than in root. Fast protein liquid chromatography-anion exchange column was used to separate glutathione S-transferase isozymes in coleoptiles of C. lacryma-jobi and corn. A peak of glutathione S-transferase activity with 1-chloro-2,4-dinitrobenzene and two peaks of glutathione S-transferase activity with simazine from C. lacryma-jobi were coeluted with those from corn, but showed greater activity than in the case of corn. Another glutathione S-transferase isozyme that exhibits activity with simazine was detected in the elution of C. lacryma-jobi extract, but not in corn. Electron transport in chloroplast thylakoids isolated from leaves of both species was equally sensitive to simazine applied at 1 to 100 nM. These results indicate that the simazine tolerance in C. lacryma-jobi is due to its capacity to detoxify the herbicide via glutathione conjugation, which is positively correlated with the level of glutathione content and glutathione S-transferase activity.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.211-218
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• 2001

The $M_r$ 63,000 glycoprotein (GP63) and lipophosphoglycan (LPG) of Leishmania donovani were evaluated as vaccine candidates against visceral leishmaniasis. Mice were immunized with liposomeencapsulated GP63 and/or LPG that were purified from the soluble extract of L. donovani promastigotes, and were challenged with virulent amastigotes. Mice immunized with GP63/LPG in liposomes plus BCG resulted in a 27.4% reduction of amastigotes in the liver compared to the control group (liposomes plus BCG), and mice immunized with liposome-GP63 plus BCG failed to induce a protective immune response against the challenge infection. Immunization of mice with GP63 fused to the Schistosoma japonicum glutathione S-transferase (GP63-GST) plus BCG also failed to elicit protective immunity. To analyze the cause of failure to induce protection, cytokine release from the spleen cells of immunized mice and Leishmania-specific serum antibodies were analyzed. Spleen cells from mice immunized with GP63-GST plus BCG that were exposed to soluble extract of L. donovani in vitro produced 10-fold greater quantities of IFN-gamma and 3-fold greater quantities of IL-5 than cells from mice receiving BCG only or saline. Western blot analysis revealed that sera from these mice had Leishmania-specific antibodies recognizing 1 to 3 antigens of L. donovani with M. W. of 60-65 kDa. Although immunization of mice with GP63-GST induced a strong Th1 response, this study indicated that GP63-GST simultaneously elicited the Th2 response of the CD4+ T-cell, which was known to abrogate the protective immune response conferred by the Th1 effector function.

• Journal of Life Science
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• v.9 no.3
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• pp.253-261
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• 1999

Leptin gene, an obesity gene, has been known to involve in the regulation of food intake and body weight. It is also thought to be related to the glucose metabolism, insulin secretion and type II diabetes mellitus. Recently, the production of recombinant leptin protein has been attempted for the application in the treatment of obesity and the correction of hereditary obesity and type II diabetes. In the present study, leptin cDNA was cloned from mouse fat cells by RT-PCR and prokaryotic expression of leptin was attempted in order ot prepare a leptin-specific antigen. Immunization of a rabbit with the leptin-specific antigen into a rabbit resulted in the generation of leptin-specific antiserum that could be useful in the detection of leption expressed in various tissues. The sequence of leptin cDNA prepared in the present study wa identical to the previously reported one. Transformation of E. coli(DH5a) cells with the leptin cDNA-inserted translation vector, pGEX-4T-3-leptin followed by treatment with IPTG (0.1mM) resulted in the expression of a large amount of GST-leptin fusion protein with a molecular weight of 44 KDa as an inclusion body. Denaturation of the insoluble fusion protein by 8M urea, 6M guanidium-HCI or 0.1% 2-mercaptoethanol followed by a slow oxidation could not solubilize the inclusion body. The cell extract was subjected to SDS-PAGE and GST-leptin protein electroeluted from the gel was then injected into a rabbit subcutaneously for the immunization. Anti-GST-leptin rabbit antiserum which had a cross reactivity to the GST-leptin protein was generated. Leptin protein expressed in mouse brain and fat tissues was detected by Western blot immunodetection system using the antiserum generated in the present study.

• 대한예방의학회:학술대회논문집
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• 1999.10a
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• pp.132-133
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• 1999

• Journal of Environmental Health Sciences
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• v.29 no.2
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• pp.80-86
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• 2003

To evaluate the effect of cyclohexane(CH) treatment on the serum levels of glutathion S-transferase(GST) activity in liver damaged animals, damaged liver was induced with pretreatment of 50% $CCl_4$ dissolved in olive oil (0.1 m1/100g body weight) intraperitoneally 17 times every other day. To $CCl_4$-treated rats, CH (1.56 g/kg body weight, i.p) was injected once and then the animals were sacrificed at 4 hours after injection of CH. The $CCl_4$-treated animals were identified as severe liver damage on the basis of liver functional findings, 1,e, increased serum levels of alanine aminotransferase(ALT), alkaline phosphate(ALP) and xanthine oxidase(XO) activities. On the other hand, $CCl_4$-treated animals injected with CH once($CCl_4$-pretreated animals) showed more decreased serum levels of ALT and XO, and more increased those of ALP rather than $CCl_4$-treated animals. In case of comparing the GST with ALT activity in liver, both $CCl_4$-treated and pretreated animals showed similar changing pattern of enzyme actvity. Especially $CCl_4$-pretreated animals showed significantly increased serum level of GST actvity compared with the $CCl_4$-treated those, whereas those of ALT showed reversed tendency. In aspects of GST enzyme kinetics, $CCl_4$-pretreated animals showed higher Vmax of liver GST enzyme than $CCl_4$-treated animals. In conclusion, injection of CH to the liver damaged rats led to enhanced liver damage and more increased activity of serum GST which may be chiefly caused by the enzyme induction.