• BMB Reports
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• v.40 no.4
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• pp.511-516
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• 2007

A glutathione S-transferase (GST) related to the phi (F) class of enzymes only found in plants has been cloned from the Oryza sativa. The GST cDNA was cloned by PCR using oligonucleotide primers based on the OsGSTF5 (GenBank Accession No. $\underline{AF309382}$) sequences. The cDNA was composed of a 669-bp open reading frame encoding for 223 amino acids. The deduced peptide of this gene shared on overall identity of 75% with other known phi class GST sequences. On the other hands, the OsGSTF5 sequence showed only 34% identity with the sequence of the OsGSTF3 cloned by our previous study (Cho et al., 2005). This gene was expressed in Escherichia coli with the pET vector system and the gene product was purified to homogeneity by GSH-Sepharose affinity column chromatography. The expressed OsGSTF5 formed a homo-dimer composed of 28 kDa subunit and its pI value was approximately 7.8. The expressed OsGSTF5 displayed glutathione conjugation activity toward 1-chloro-2,4-dinitrobenzene and 1,2-epoxy-3-(p-nitrophenoxy)propane and glutathione peroxidase activity toward cumene hydroperoxide. The OsGSTF5 also had high activities towards the herbicides alachlor, atrazine and metolachlor. The OsGSTF5 was highly sensitive to inhibition by S-hexylGSH, benastatin A and hematin. We propose from these results that the expressed OsGSTF5 is a phi class GST and appears to play a role in the conjugation of herbicide and GPOX activity.

• Bulletin of the Korean Chemical Society
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• v.26 no.3
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• pp.433-439
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• 2005

To gain further insight on the relationship between structure and functions of glutathione S-transferase (GST), the three tyrosine 108 mutants, Y108A, Y108F, and Y108W, of human GST P1-1 were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. The substitution of Tyr 108 with alanine resulted in significant decrease of the GSH-conjugation activity and the GSH peroxidase activity, but approximately 63% increase of steroid isomerase activity toward ${\Delta}^5$–[androstene 3,17-dione. On the other hand, the substitution of Tyr 108 with phenylalanine resulted in decreases of $k_{cat}\;and\;k_{cat}/K_m{^{EPNP}}$ by 2 orders of magnitude, suggesting that Tyr 108 residue of hGSTP1-1 are considered to be important for the catalysis and the binding of the epoxide substrates. The substitution of Tyr 108 with tryptophan resulted in significant decreases of the specific activities toward EPNP, cumene hydroperoxide and ${\Delta}^5$–ndrostene 3,17-dione, but approximately 2-fold increase on the enzyme-catalyzed addition of GSH to DCNB. We conclude from these results that Tyr 108 in hGST P1-1 plays very different roles depending upon the nature of the electrophilic substrates.

• Journal of Environmental Health Sciences
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• v.30 no.3
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• pp.230-238
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• 2004

The adverse health effects of a number of environment pollutions are related to the formation of free radicals. Induction of antioxidant defensive system in the response to an oxidative attack is an essential element of the cell to survive. CYP2E1 is easily induced by organic solvents and induces continuous formation of reactive oxygen species (ROS). ${\gamma}$-Glutamyltranspeptidase (${\gamma}$GT) plays an important role in glutathione metabolism and xenobiotic detoxification. To evaluate the characteristic of oxidative stress which induces GGT expression and to understand human antioxidant defensive response against oxidative stress induced by CYP2E1, we studied regulation of ${\gamma}$GT enzyme expression in response to various oxidative stresses in human HepG2 cells. The ${\gamma}$GT activity was not modified after exposure of acute oxidative stress inducing agents (ferric nitrilotriacetate, cumene hydroperoxide, ADP-Fe, O-tetradecanoylphorbol-13-acetate, tumor necrosis factor-alpha). To induce continuous exposure of cells to ROS, HepG2 cells were transfected by human CYP2E1 gene transiently. The CYP2E1 activity was verified with chlorzoxazone hydroxylation. Transfection of CYP2E1 showed continuous 60% increase in intracellular ROS and 240 % increase in microsomal ROS. CYP2E1 overexpressing cells showed increased ${\gamma}$GT activity (2.5-fold). The observed enhancement of ${\gamma}$GT activity correlated with a significant increase of ${\gamma}$GT mRNA (2.1-fold). Treatment with antioxidant strongly prevented the increase in ${\gamma}$GT activity. The CYP2E1 overexpression did not modify toxicity index and increased glutathione levels. These results show that continuous exposure of cells to ROS produced by CYP2E1 up-regulates ${\gamma}$GT; This may be one of the adaptive antioxidant responses of cells to oxidative insult. Present study also suggests that the induction of ${\gamma}$GT could be used as a marker of oxidative stress induced by exposure to organic solvents.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.27 no.1
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• pp.23-37
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• 2017

Objectives : This study was conducted to evaluate the level of exposure to volatile organic compounds (VOCs) among workers handling rust preventive oils. Methods : A total of 30 bulk samples and 54 personal air samples were collected using diffusive samplers at 22 workplaces handling rust preventive oils in Daegu and Gyongsangbuk-do Province from March to October 2013. We also investigated detailed information on the related work conditions, such as kinds of products, handling methods, local exhaustive ventilation systems, and the status of the wearing personal protective equipment. All bulk samples and air samples were analyzed using gas chromatography mass spectrometry (GC-MS) to identify components to which workers potentially were exposed. Quantitative airborne concentrations of VOCs were confirmed using gas chromatography with flame ionized detectors. Results : In terms of qualitative analyses for the 30 bulk samples, we found carcinogenic, mutagenic and reproductive toxic(CMR) substances such as butane(carcinogenic Group 1A, mutagenic Group 1B), butoxy ethanol(carcinogenic Group 2), cumene (carcinogenic Group 2), ethyl benzene(carcinogenic Group 2), methyl isobutyl ketone(carcinogenic Group 2) and toluene (reproductive toxic, Group 2). As a result of full-shift based personal air samples, eight substances such as n-hexane, n-heptane, octane, nonane, decane, toluene, ethyl benzene and xylene were detected. Among them, n-hexane and n-heptane were detected in all of 54 air samples with $13.13mg/m^3$ and $8.61mg/m^3$ of maximum concentration, respectively. The level of airborne concentration from all of samples were bellow the occupational exposure limit in Korea. Conclusions : Based on the results of this study, workers handling rust preventive oils could be exposed to CMR substances contained in rust preventive oils and n-hexane and n-heptane were found as the most frequent sources of VOC exposure.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.6 no.2
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• pp.187-201
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• 1996

This study was to evaluate the efficiency of diffusive monitor using activated carbon fiber(ACF, KF-1500) in measuring airborne organic solvents. The following characteristics were identified and studied as critical to the performance of diffusive monitor; recovery, sampling rate, face velocity, reverse diffusion and storage stability. For the evaluation of the performance of this monitor, MIBK, PCE, toluene were used as organic solvents. In the sampling rate experiments, eight kinds of solvents (n-hexane, MEK, DIBK, MCF, TCE, CB, xylene, cumene) as well as the above solvents were used. The results were as follows: 1. The desorption efficiencies(DE's) of ACF diffusive monitor ranged from 83 % to 101 %. In contrast, those of coconut shell charcoal ranged from 78 % to 102 %. Especially, the DE's of ACF for the polar solvents such as MEK were superior to those of charcoal. 2. Experimental sampling rates on ACF were average 42ml/min(37-46ml/min) for 11 organic solvents at $24{\pm}2^{\circ}C$, $50{\pm}5%RH$. However ideal sampling rates(DA/L) were 33 % higher than experimental sampling rates. 3. The initial response(15~16 min) of the testing monitor was 2 times higher than the actual concentration determined by the reference methods at $24{\pm}2^{\circ}C$, $8{\pm}5%RH$ and $80{\pm}5%RH$. Within 1 hours, the curve reached a linear horizontal line at low humidity condition. But sampling efficiencies decreased with respect to time at high humidity condition. And sampling efficiencies were higher at high humidity condition than low humidity condition for MIBK. 4. At very low velocity (less than 0.02 m/sec), the concentration of ACF diffusive monitor were poorly estimated. But ACF diffusive monitor were not affected at higher velocity(0.2 m/sec-0.6 m/sec). 5. There was no significant reverse diffusion when the ACF monitors were exposed to clean air for 2 hours after being exposed for 2 hours at the level of 1 TLV. 6. There was no significant sample loss during 3 weeks of storage at room temperature and 5 weeks of storage at refrigeration.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.16 no.4
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• pp.314-323
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• 2006

This study was conducted to identify ingredients of thinners and to confirm reliability of material safety data sheets (MSDS) of thinners for public and workers' health. The 41 thinner products were collected from paint shops located in Busan and Gyeongnam province. The 12 thinner products among them were identified using product MSDS. GC-MSD was used to analyze 41 kinds of thinners qualitatively and quantitatively. The 12 products MSDS were compared with thinner's component through qualitative analysis to confirm MSDS. Chemical ingredients, such as Benzene, Toluene, and Xylene etc., of thinners were analysed in quantity. The 41 thinner products contained 17 disclosed specific, trade name, or generically described chemical solvent ingredients. These 17 ingredients came under 6 classes: alcohols, aromatic hydrocarbons, esters, glycol ethers, ketones, and mixtures. These 17 ingredients were important in the view of industrial hygiene and had occupational exposure limit in the ambient, such as toluene, xylene, acetone, nonane, EGEE, heptane, cumene, MIBK, indene, tri-methyl benzene, etc, were found in 41 kinds of thinners. Aromatic hydrocarbons were the most identified ingredient in thinners. Especially, the benzene, which induces leukemia, was found in 4 kinds of thinners. The content rates of benzene in thinners were 0.25~1.18%. The benzene in enamel thinner, which were 0.39~0.72%, was highest from chemical classification. The contents of toluene, which was found from 27 kinds of thinners, were 5.35~64.16%, which were highest in sobu thinner as 58.80%. Xylene was found from 22 kinds of thinners and contents of xylene were 4.61~72.42%. Acrylic thinner's contents of xylene were 12.06~51.05%, which was most high. It was found that contents of benzene were increased and frequency of detection was decreased through comparison with other study. The MSDS possession rate of paint shops was low as 29.27%. So it did not provide information with public or workers. Mean of agreement rate between MSDS and components of thinners through qualitative analysis was 42.01% and it has wide range from 8.3% to 75%. There are many deficiencies in MSDS about component of thinners. In some case of sample, expecially, despite containing benzene, information was not written it on MSDS.