• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.747-748
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• 2003

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 1996.12a
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• pp.44-44
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• 1996

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 1999.04a
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• pp.57-57
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• 1999

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.2
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• pp.209-216
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• 1998

The present study was designed: (1) to determine whether or not hypoxia stimulates the release of endothelium-derived relaxing factors (EDRFs) from endothelial cells, and (2) to examine whether or not the hypoxia-induced EDRFs release is further augmented by previous hypoxia-reoxygenation, using bioassay system. In the bioassay experiment, rabbit aorta with endothelium was used as EDRFs donor vessel and rabbit carotid artery without endothelium as a bioassay test ring. The test ring was contracted by prostaglandin $F_{2{\alpha}}$ $(3{\times}10^{-6}\;M/L)$, which was added to the solution perfusing through the aortic segment. Hypoxia was evoked by switching the solution aerated with 95% $O_2/5%\;CO_2$ mixed gas to one aerated with 95% $N_2/5%\;CO_2$ mixed gas. When the contraction induced by prostaglandin $F_{2{\alpha}}$ reached a steady state, the solution was exchanged for hypoxic one. And then, hypoxia and reoxygenation were interchanged at intervals of 2 minutes (intermittent hypoxia). The endothelial cells were also exposed to single 10-minute hypoxia (continuous hypoxia). When the bioassay ring was superfused with the perfusate through intact aorta, hypoxia relaxed the precontracted bioassay test ring markedly. Whereas, when bioassay ring was superfused with the perfusate through denuded aorta or polyethylene tubing, hypoxia relaxed the precontracted ring slightly. The relaxation was not inhibited by indomethacin but by nitro-L-arginine or methylene blue. The hypoxia-induced relaxation was further augmented by previous hypoxia-reoxygenation and the magnitude of the relaxation by intermittent hypoxia was significantly greater than that of the relaxation by continuous hypoxia. The results suggest that hypoxia stimulates EDNO release from endothelial cells and that the hypoxia-induced EDNO release is further augmented by previous hypoxia-reoxygenation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.3
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• pp.186-192
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• 2004

Purpose: Human tooth proteins are highly heterogeneous, comprising diverse proteins derived from a number of genes. The attempts to identify protein for activity of tooth matrix proteins have been defied by several factors. First, the amount of proteins within teeth is very small relative to many extracellular matrix proteins of other tissues. Second, the bioassay system is tedious and needed for long time. Therefore we tried to find easy techniques, which increase the product rate, and an assay of small proteins, with which amino acid sequence is possible without additional procedures. Materials and Methods: Total protein were extracted from 300 g enamel removed teeth and 600 g teeth with 4 mol/L guanidine HCl and purified by gel chromatography. Aliquot of proteins was implanted into muscle pouches in Sprague-Dawley rats for bioassay. By SDS-PAGE and membrane blotting, molecular weight of each protein was estimated and a partial amino acid sequence was obtained. Each fraction blotted on the membrane was cut out and inserted in rat ectopic model. Results: In dissociative method, total tooth proteins were obtained 1mg/ml from enamel removed teeth and 3.5 mg/ml from teeth. In SDS-PAGE, four clear bands at the sites corresponding to 66, 40, 20 and 18 kD. Especially The 66 kD band was clearly exhibited. Amino acid sequencing from tooth could be possible using PVDF membrane blotting technique. In amino acid sequencing, 66 kD protein was identified as albumin. Conclusion: Compared with conventional method for extraction of teeth protein and bioassay of proteins, the methods in this study were easy, time-saving and more productive technique. The matured tooth proteins omitting additional procedure of mechanical removal of enamel were simply analyzed using blotted PVDF membrane. This method seems to make a contribution as a technique for bioassay and amino acid sequencing of protein.

• The Korean Journal of Malacology
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• v.22 no.2
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• pp.167-173
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• 2006

The embryos of marine bivalves have been commonly used in bioassays for quality assessments of marine environments. Although several standard protocols for the developmental bioassay of bivalves have been proposed, only a few trials for application of these protocols in environmental assessments or for the development of a new protocol with Korean species have been conducted. As such, there is a strong need to establish standard bioassay protocols with bivalves commonly found in Korean waters. To determine the sensitivity of Mytilus galloprovincialis to establish a standard bioassay, their fertilized eggs were exposed to six metals (Ag, Cd, Cr, Cu, Ni, and Zn). The order of biological impact was Ag > Cu > Ni > Zn > Cr > Cd and their lowest observed effective concentration were 5, 16.4, 25.4, 142, 187 and 1,500${\mu}g/l$, respectively. The proportion of normal larvae appeared to decrease linearly with the logarithm of each toxicant concentration within the tested range. The average values of median effective concentrations $(EC_{50})$ from the triplicate experiments for Ag, Cd, Cr, Cu, Ni, and Zn were 6.8, 1,797, 786, 16.6, 68.1, and 139.2${\mu}g/l$, respectively. There was a more than 100-fold difference in $EC_{50}$ values of Cu and Cd. The value of $EC_{50}$ or median lethal concentration of Cu was within the range observed for other bivalve developmental bioassays. The overall sensitivity of M. galloprovincialis in the present developmental bioassay was also similar to that of other marine organisms commonly used in aquatic bioassays (e.g. sea urchins, oysters). Hence, the bioassay using the embryo of M. galloprovincialis is considered to be a useful tool to monitor and evaluate the quality of marine aquatic environments.

• The Korean Journal of Physiology and Pharmacology
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• v.10 no.6
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• pp.349-354
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• 2006

The aromatic hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates many of the biological and toxicological effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related chemicals. The application of recombinant reporter plasmid such as the firefly luciferase gene has proven to be a very effective method to detect these chemicals. The bioassay system, CALUX, is sensitive in directly detecting AhR-agonists from a variety of environmental and biologic materials. However, responses of the AhR-dependent bioassays are dependent on the cell types used. Thus, we developed a sensitive bioassay using the recombinant mouse hepatoma cell (Hepa1c1c7) for the determination of dioxins. The recombinant cell line was stably transfected with firefly luciferase reporter gene (pGudLuc1.1). The transfected cells showed the highest induction of luciferase activity at 4.5 hr and a decrease beyond this time point. The system showed the highest sensitivity of detection ever reported. Upon TCDD exposure cells showed 2 fold increase at 10 pM and 7 fold increase at 100 pM, respectively. The passage number after the transfection played an important role in the sensitivity. The increase of passage number tended to increase the sensitivity of the cells up to 15. The media without phenol red showed a higher induction rate than with phenol red, suggesting the preferable use of phenol red-free media for the bioassay. Since each of the assays has unique characteristics that make them suitable for some screening applications and not others, development of sensitive bioanalytical methods based on a variety of cellular systems in a key to the successful determination of dioxins. The bioassay system developed in this study will contribute to further development of successful screening the AhR agonists among the environmental mixture. In addition, the rapid and sensitive nature of this cellular system can be applied as a valuable tool to screen the dioxin-like moieties among the prodrugs at the initial stage, thereby expediting the new drug discovery.

• Applied Biological Chemistry
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• v.37 no.3
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• pp.182-188
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• 1994

Similar to the $^{42}K$ uptake, $^{86}Rb$ uptake by the roots of Hordeum distichum grown in the hydroponic culture was negatively correlated with the concentration of K supplied previously, showing that $^{86}Rb$ can be used for the K-bioassay. $^{86}Rb$ having longer half life (18.86 day) than $^{42}K$ (12.36 hr) allowed the use of larger number of root samples. $^{86}Rb$ uptake of 3 years old Citrus unshiu Marc. grown in water culture decreased drastically with the increase of K concentration of the culture solution, thus demonstrating that the nutrition status of K for citrus trees can be diagnosed by K-bioassay using $^{86}Rb$ tracer. $^{86}Rb$ uptake by the excised roots of Hordeum distichum grown in the pot with different K fertilizations was well correlated with the exchangeable K in soil. The amount of exchangeable K in soil for the optimal plant growth can be determined by its relationship. $^{42}K$ and $^{86}Rb-uptake$ by the Hordeum distichum roots were markedly inhibited by $5{\times}10^{-3}\; M$ KCN in the bioassay solution, indicating that uptake is energy-dependent. There was no significant relationship between K content in citrus leaves and K concentration in the water-culture medium. It is concluded that K-bioassay is a potentially useful tool for determining of K requirement in citrus trees.

• 대한방사선방어학회:학술대회논문집
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• 2009.11a
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• pp.108-109
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• 2009

• Journal of Korean Society of Water and Wastewater
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• v.17 no.4
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• pp.471-478
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• 2003