• KSBB Journal
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• v.18 no.4
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• pp.255-260
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• 2003

In vivo bioassays for biological medicines have been considered final resort to unequivocally assess the biological activities for them because there are some cases in which the biological activities obtained from in vivo bioassay and in vitro bioassay quite differ each other. The in vivo biological activity of EPO depends on its sialic acid contents which confer microheterogeneity-isoforms to this protein. We have devise a method which consists of a in vitro bioassay using BaF3 cell line and a capillary zone electrophoresis (CZE) for the measurement of the EPO isoform distribution. The biological activity of EPO obtained using in vitro bioassay with BaF3 cell line showed good correlation (C.V.(％) 7.34, 5.85, 8,16, 8.08, 8.8) to EPO content measured either spectrophotometric assay (A280 0.1 ％ ＝0.743) or radio immunoassay. The assay validation results of in vitro bioassay with 3 lot of in house EPO showed good results to EPO content measured either in vivo assay or radio immunoassay. and also showed good results the robustness of our method in terms of precision, accuracy, repeatability. The isoform distribution for EPO-BRP (1 : 1 mixture of epoetin-${\alpha}$ and epoetin-${\beta}$, European Pharmacopoeia) by CZE method resulted in isoform 2 through isoform 8. The major peaks in electrophoregram were composed of isoform 3 through 7. Our recombinant EPO (epoetin-${\alpha}$) having equivalent in vivo biological activity showed the isoform distribution of isoform 3 through 9. The major peaks consisted of isoform 4 through 8. The peak area of isoform 4 was always smaller than that of isoform 5. The preparations of recombinant epoetin-${\alpha}$ with lower in vivo biological activity than EPO-BRP showed the isoform 2 through 8 in their electrophoregrams whose major peaks consisted of the isoform 3 through 7. The peak area of isoform 4 was larger than that of isoform 5.

• Journal of the Korean Statistical Society
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• v.19 no.1
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• pp.71-79
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• 1990

In the linear model which consider both the multivariate parallel-line bioassay and the multivariate linear calibration, this paper presents a Bayesian procedure which is an extension of Hunter and Lamboy (1981) and has several advantages compared with the non Bayesian techniques. Based on the methods of this article we discuss the effect of multivariate calibration and give a numerical example.

• Archives of Pharmacal Research
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• v.21 no.1
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• pp.67-69
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• 1998

Three TNF-$\alpha$inhibitory lignans were isolated from the flower buds of Magnolia fargesii through bioassay-guided isolation. They were identified as eudesmin, magnolin and lirioresinol-B dimethylether on the basis of their spectroscopic data. All three lignans showed inhibitory effects on TNF-$\alpha$ production in LPS-stimulated murine macrophage cell line, RAW264.7 and eudesmin showed the strongest activity ($IC_{50}=51{\mu}M)$.

• 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.

• Korean Journal of Plant Resources
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• v.27 no.5
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• pp.401-414
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• 2014

In an attempt to evaluate licorice quality based on its biological activity, we grafted an on-line high-performance liquid chromatography (HPLC) bioassay method into the previously established HPLC analysis method. The common antioxidant peaks in licorices of various origin were observed through an on-line HPLC/DPPH system leading to a decrease in absorbance at 517 nm for 2,2-diphenyl-1-picrylhydrazyl (DPPH). Among them, the licorice from Youngju possessed the highest activity. Therefore, three active standard compounds from the dehydroglyasperin C, dehydroglyasperin D, and isoangustone A, were isolated and elucidated by medium pressure liquid chromatography (MPLC) and instrumental analysis such as nuclear magnetic resonance (NMR), respectively. On-line HPLC/ABTS analysis method with the simultaneous determination of three standard compounds and their radical scavenging activity was established for the quality evaluation of licorices. 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid radicals (ABTS) which is stable and effective was used in replace of DPPH. The radical scavenging activity of three standards is compared with that of Trolox, known as antioxidant, showing a negative peak with a decrease in absorbance at 734 nm for ABTS. This on-line HPLC/ABTS analysis method was validated for specificity, linearity, precision and accuracy in compliance with international conference on harmonization (ICH) guideline.

• Journal of Life Science
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• v.16 no.6
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• pp.919-924
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• 2006

Toxicity evaluation systems for various chemicals and their metabolites have been developed during last decades. In this study, the acute toxicity of endocrine disrupting chemicals, such as endosulfan, bisphenol A, vinclozolin, and 3,5-dichloroaniline, was evaluated using HepG2 cell line, Lumbricus rubellus and Saccharomyces cerevisiae, respectively. The extents of toxicity of the chemicals in different bioassay systems varied substantially, such as endosulfan>3,5-dichloroaniline> bisphenol A in HepG2 cell line system, endosulfan>bisphenol A>3,5-dichloro aniline in L. rubellus system, and 3,5-dichloroaniline>endosulfan>bisphenol A in S. cerevisiae system. Meanwhile, no cytotoxicity was observed by treatment of vinclozolin in the evaluation systems. Our results suggest that earthworm and yeast are useful to evaluate acute toxicity of endocrine disrupting chemicals, and direct comparison of toxicity data from different bioassay systems is unattainable. Based on our results, we propose that the bioassay system with earthworm or yeast, a rapid, simple and economic system, could be applied as pre-test for the toxicity evaluation using human cell line or animals.

• Korean Journal of Medicinal Crop Science
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• v.27 no.3
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• pp.208-217
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• 2019

Background: In the present study, we identified two cytotoxic compounds from the root of Rumex crispus L. using a bioassay-based method. Methods and Results: Compared with the other fractions, the diethyl ether ($Et_2O$) fraction of R. crispus root extract exhibited the strongest of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging effect [scavenging concentration 50% $(SC_{50})=63.8{\pm}1.47{\mu}g/m{\ell}$], nitric oxide (NO) production inhibitory effect on the mouse macrophage cell line RAW264.7 [inhibitory concentration 50% $(IC_{50})=60.9{\pm}7.52{\mu}g/m{\ell}$] and cytotoxicity effect on the human hepatoma cell line, HepG2 [lethal concentration 50% $(LC_{50})=115.4{\pm}1.86{\mu}g/m{\ell}$]. According to the bioassay-based method, two cytotoxic compounds were purified from the $Et_2O$ fraction by using column chromatography and preparative high performance liquid chromatography (prep-HPLC). These two compounds were identified as parietin and chrysophanol by using nuclear magnetic resonance (NMR) and liquid chromatography quadruple time of flight mass spectrometry (LC-QTOF-MS). In addition, both parietin and chrysophanol exhibited a cytotoxicity effect on HepG2 cells, their $LC_{50}$ values were $169.1{\pm}17.67{\mu}M$ and $111.5{\pm}6.62{\mu}M$, respectively. Conclusions: Parietin and chrysophanol isolated from the $Et_2O$ fraction of the R. crispus root extract showed cytotoxicity in HepG2 cell.

• Journal of Ginseng Research
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• v.48 no.2
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• pp.171-180
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• 2024

Background: Epimers of ginsenoside Rg3 (Rg3) have a low bioavailability and are prone to deglycosylation, which produces epimers of ginsenoside Rh2 (S-Rh2 and R-Rh2) and protopanaxadiol (S-PPD and R-PPD). The aim of this study was to compare the efficacy and potency of these molecules as anti-cancer agents. Methods: Crystal violet staining was used to study the anti-proliferatory action of the molecules on a human epithelial breast cancer cell line, MDA-MB-231, and human umbilical vein endothelial cells (HUVEC) and compare their potency. Cell death and cell cycle were studied using flow cytometry and mode of cell death was studied using live cell imaging. Anti-angiogenic effects of the drug were studied using loop formation assay. Molecular docking showed the interaction of these molecules with vascular endothelial growth factor receptor-2 (VEGFR2) and aquaporin (AQP) water channels. VEGF bioassay was used to study the interaction of Rh2 with VEGFR2, in vitro. Results: HUVEC was the more sensitive cell line to the anti-proliferative effects of S-Rh2, S-PPD and R-PPD. The molecules induced necroptosis/necrosis in MDA-MB-231 and apoptosis in HUVEC. S-Rh2 was the most potent inhibitor of loop formation. In silico molecular docking predicted a good binding score between Rh2 or PPD and the ATP-binding pocket of VEGFR2. VEGF bioassay showed that Rh2 was an allosteric modulator of VEGFR2. In addition, SRh2 and PPD had good binding scores with AQP1 and AQP5, both of which play roles in cell migration and proliferation. Conclusion: The combination of these molecules might be responsible for the anti-cancer effects observed by Rg3.

• Korean Journal of Organic Agriculture
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• v.18 no.4
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• pp.573-586
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• 2010

This study was carried out to evaluate characteristics of Chrysanthemum sp. and to bioassay Tetranychus urticae, Nilaparvata lugens, and Aphis gossypii by different extracts part and method. Flower type of Chrysanthemum sp. line was anemone except 0706 line which was pompon. The flowering date was from April to June but 0721 line kept up November. They could overwinter except 0706 line. The Ethanol extracts of 0718 line of flower was high mortality as 63.8% on Tetranychus urticae. Ethanol mixed emulsifier extracts were the highest mortality as 69.4% at 5 days after. Mortality in water extract was lower than in ethanol and ethanol mixed emulsifier extracts. Ethanol mixed emulsifier extracts were higher mortality as 69.4% at 5 days after in 0718 line on Nilaparvata lugens. Mortality in water extract was lower than in ethanol and ethanol mixed emulsifier extracts. On Aphis gossypii, Chrysanthemum flower extracts could decrease the density of aphids in early days but the density of that was increase as time goes by because aphids produce offspring.

• Korean Journal of Pharmacognosy
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• v.39 no.4
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• pp.347-351
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• 2008

The methanol (MeOH) extract of the rhizome of Alpinia officinarum Hance (Zingiberaceae) demonstrated a potent inhibition on the proliferation of cultured human tumor cell lines such as MES-SA (human uterine carcinoma cell line), MESSA/DX5 (multidrug resistant subline of MES-SA), HCT-15 (human colorectal adenocarcinoma cell line), HCT15/CL02 (multidrug resistant subline of HCT15). The MeOH extract was fractionated into four portions by serial solvent partition, ie., methylene chloride (CH2Cl2) soluble part, ethylacetate (EtOAc) soluble part, n-butanol (BuOH) soluble part and remaining water layer. Among them, the $CH_2Cl_2$ soluble part of the extract exhibited a most potent inhibition on the proliferation of tested tumor cell lines. Bioassay-guided fractionation of the $CH_2Cl_2$ soluble part led to the isolation of five diarylheptanoid and two flavonoid constituents, i. e., galangin (1), 7-(4"-hydroxy-3"-methoxyphenyl)-1-phenylhept-4-en-3-one (2), 1,7-diphenyl-5-hydroxy-3-heptanone (3), trans,trans-1-(3'-methoxy-4'-hydroxyphenyl)-7-phenyl-5-ol-4,6-dien-3-heptanone (4), 5-methoxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone (5), kaempferide (6), 5-hydroxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone (7). Structures of the isolated active components (1 - 7) were established by chemical and spectroscopic means.