• Korean Journal of Fisheries and Aquatic Sciences
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• v.36 no.4
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• pp.346-351
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• 2003

Single-cell gel electrophoresis (comet assay) was used to detect DNA single strand break in blood cells from several marine fish species. Three fish species were collected from Georgia coastal area. Mummichog, Fundulus heteroclitus showed higher DNA damage than sea bass, Lateolabrax japonicus and trout, Oncorhynchus masou masou under the same experimental conditions. Mummichogs had more alkaline-labile sites on their DNA than other fish species. The comet assay with mummichog blood cells at pH 12.5 showed a dose-response curve with the increasing concentrations of hydrogen peroxide. While the isolated leucocytes showed no increase of DNA damage after in vitro exposure to 2-methyl-1,4-naphthoquinone (MNQ), erythrocytes showed dose-dependent DNA damage. These results indicate that the comet assay can be applied successfully as a bioassay using erythrocyte for environmental monitoring.

• BMB Reports
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• v.30 no.4
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• pp.258-261
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• 1997

A simple quantitative assay method for ribonuclease activity has been developed. This method is based on the decrease of fluorescence intensity emitted by the ethidium bromide bound to RNA due to the degradation of RNA by ribonuclease. The substrate RNA was reacted with ribonuclease A and the fluorescence intensity was measured after the addition of ethidium bromide. The intensity difference was calculated using a blank reaction mixture containing no RNase. Whole cellular RNA substrate produced a significant error and was not suitable for this assay method possibly because of local microheterogeniety caused by high molecular weight rRNA. but satisfying results were obtained with tRNA substrate. The intensity difference increased linearly by raising enzyme concentration up to $2{\times}10^{-4}$ Kunitz Units of ribonuclease A. More refined and reliable results were obtained by use of initial reaction velocities which were calculated from the plots of intensity difference vs time. A linear relationship between initial velocities and enzyme concentrations was observed up to 0.01 Kunitz Units of enzyme.

• The Journal of Korean Medicine
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• v.23 no.2
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• pp.57-69
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• 2002

Objective: The aim of this study is to investigate the effect of Injin butanol fractions with Thin Layer Chromatography on Fas-mediated Apoptosis. Method: Injin-butanol fraction separated by TLC. MIT assay, cell cycle analysis, Caspase-3 protease assay, DNA fragmentation assay and quantitative RT-PCR were performed to evaluate the effects of TLC extraction of lnjin-butanol fraction on cell viability, cell cycle progression and apoptosis. Results: Scopoletin, luteolin, apigenin and unknown powder was isolated by TLC. Fas-mediated apoptosis analysis shows that scopoletin has inhibiting function on apoptosis. Caspase- 3 protease assay analysis shows that scopoletin inhibits activity of caspase-3. Quantitative RT-PCR analysis shows that no activity on caspase-3, but apoptosis inhibition cytokine -Bcl-2- is activated, and apoptosis activating cytokine -Bax- is unactivated. Conclusion: These results show that each fraction of Injin-butanol TLC extraction, especially scopoletin, acts as a protective function on liver cell viability, and inhibitory function on apoptosis. (J Korean Oriental Moo 2002;23(2):57-69)

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.2
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• pp.316-321
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• 2000

This study carried out to eluciate the cancer chemoprevention of Korean native fermented food, baechu kimchi using Comet assay (in other words, single cell microgel electrophoresis). For this purpose, baechu kimchi was fractionated by water, n-hexane, chloroform and ethyl acetate. 5 strains of dominant fermented bacteria were isolated from baechu kimchi. The water fraction, n-hexane fraction, chloroform fraction, ethyl acetate fraction and water insoluble fractions showed no antigenotoxicitie in non-tumoral normal 3T3 cells. Among 5 bacteria isolates from baechu kimchi, two isolates bacteria 1 and 2 strongly inhibited genotoxicity of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in non-tumoral normal 3T3 cells (p<0.05). Bacteria 3, 4 and 5 were also not antigenotoxic.

• Journal of the Korean Applied Science and Technology
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• v.32 no.2
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• pp.312-319
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• 2015

In this study, n-butanol and ethyl acetate extracts were prepared from raw yam (Dioscorea batatas DECNE.). Their antioxidative potencies were investigated employing various in vitro methods, such as ferrous ion chelating, ${\beta}$-carotene bleaching assay, lipid peroxidation inhibition and nitric oxide (NO) radical scavenging and nitrite scavenging activity. The n-butanol fraction was assayed to possess stronger antioxidant activity by ${\beta}$-carotene bleaching assay, lipid peroxidation inhibition and NO radical scavenging activity. However, ethyl acetate extract was more effective in chelating ferrous ion and scavenging nitrite. Based on the results obtained, yam is a potential active ingredient that could be applied in antioxidation as well as bio-health functional food to take a good part in prevention of human diseases and aging.

• Journal of Applied Biological Chemistry
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• v.48 no.4
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• pp.202-206
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• 2005

Boiled-water extract (BWE) of Hizikia fusiformis has antioxidant effects. To investigate antioxidant effects of BWE, 1,1-dephenyl-2-picrylhydrazyl (DPPH) and thiobarbituric acid (TBA) methods were used. BWE exhibited strong radical-scavenging activity (87.3% at final conc. $2000\;{\mu}g$ BWE/ml in an assay mixture) on DPPH and good inhibitory activity (64% at final conc. $1000\;{\mu}g$ BWE/ml in an assay mixture) on lipid peroxidation. Raw 264.7 cells treated with lipopolysaccharide (LPS) showed 26.7-fold increase in nitric oxide (NO) ($48.9\;{\mu}M$) compared with control group ($1.83\;{\mu}M$). When treated with BWE and LPS, NO production was inhibited by BWE dose-dependently.

• Journal of Microbiology
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• v.34 no.1
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• pp.40-48
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• 1996

We have examined the chromatin structure of the HMRE/HSP82 and HMRa/HSP82 allels using three complementary approaches : DNase I chromating footprinting, micrococcal nuclease (MNase) nucleosome-protected ladder assay, and an in vivo E. coli dam methylase accessibility assay. The footprinting results indicate that the promoter and silencer sequences are assembled into nucleoprotein complexes which exhibit no detectable change in structure, despite a 70-fold range in expression levels. In addition, the promoter region of the HMRa/HSP82 allele is cleaved randomly by MNase in all cases, indicating the absence of anonical nucleosomes over this region irrespective of SIR4 or heat-shock. Finally, no discernible difference in the accessibility of the HMRE/HSP82 locus to dam methylase in SIR4 vs. sir4 cells was seenm which again suggests that the chromatin structure of HMRE/HSP82 allele is identical regardless of SIR4. Altogether, our results indicate that in contrast to other observations of the silent mating-type loci, no discernible structural alteration is detected at either HMR/HSP82 allele regardless of SIR genetic background or transcriptional state of the gene.

• Journal of Forest and Environmental Science
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• v.23 no.2
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• pp.87-91
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• 2007

The needles of L. kaempferi was extracted with 95% ethanol and successively partitioned with n-hexane, $CH_2Cl_2$ and EtOAc. Repeated column chromatography on the EtOAc and $H_2O$ soluble fractions gave three flavan-3-ols, one flavone glycoside, six flavonol glycosides and one lignan derivative. Their structures were elucidated on the basis of chemical and spectroscopic evidences. The antioxidant activities of the isolated compounds were evaluated by DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging method. Flavan compounds indicated good antioxidative potentials compared with BHT (butylated hydroxytoluene) and ${\alpha}$-tocopherol as controls. In the anti-inflammatory test on most of the isolated compounds, NO (nitric oxide) assay against the RAW 264.7 (Mouse Macrophage) showed similar inhibitory potentials to NO production of the control. The cytotoxicity was determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and most of the isolated compounds indicated no toxicity in various concentration.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.13 no.1
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• pp.100-115
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• 2000

Taklysodoksan(TSS) was a drug used in the treatment of carbuncle and cellulitis in oriental medicine. The purpose of this Study is to investigate the anti-cancer effect of TSS, the proliferation of immunocytes and nitric oxide(NO) production from peritoneal macrophages of mice. This Study estimated the proliferation of Ll210 cell lines, mouse thymocytes and splenocytes and NO production from peritoneal macrophages in vitro and vivo. The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). From the in vitro and vivo Study of TSS treatment, it did not effect the proliferation of L1210 cells. It also did not have any positive prodution of NO in peritoneal macrophages. This results suggest that TSS treatment in WanBingHuiChun(萬病回春) did not have significant anti-cancer effect and immuno-action comparing with TSS treatment of WaiKeZhengZhong(外科正宗).

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.15 no.2
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• pp.132-144
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• 2002

Shinsuwuisaeng-Tang was a drug that treated carbuncle and cellulitis. So, the purpose of this Study was to investigate effect of Shinsuwuisaeng-Tang on the anti-cancer and nitric oxide(NO) production of peritoneal macrophages. We used Shinsuwuisaeng-Tang extract(SWT) with freeze-dried, 8wks-old male mice. and cancer cell lines(L1210, sarcoma-180) for this Study. The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). The results of this Study were obtained as follow; SWT was showed cytotoxicity on the L1210 and sacoma-180(S-180) cell lines, SWT inhibited significantly proliferation of L1210 cells in L1210 cells transplanted mice, SWT accelerated NO production of peritoneal macrophages in L1210 cells transplanted mice. And SWT inhibited significantly tumor weight, increased significantly body weight and mean survival days in S-180 cells transplanted mice. This results suggest that SWT has anti-cancer by producing NO of peritoneal macrophages.