• Korean Journal of Plant Resources
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• v.26 no.6
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• pp.686-694
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• 2013

Shade treatment of Synurus deltoides (Aiton) Nakai was carried out with 0, 35, and 55% shading net, and samples were marked as no shade, 35% shade, and 55% shade, respectively. We examined in vitro antioxidant and anti-inflammatory capacities using a 1,1-diphenyl-2-2-pricylhydrazyl (DPPH) radical scavenging assay, a reducing power assay, a total antioxidant assay, a metal chelating assay, a superoxide radical scavenging assay, and a nitric oxide inhibition assay. As a result, no shade and 35% shade possessed higher DPPH radical scavenging activity and reducing power ability than that of 55% shade. Notably, no shade had significantly higher total phenolic and flavonoid contents than those in the other samples. No shade exhibited significantly higher total antioxidant activity than that of 35% shade and 55% shade. However, the chelating ability of 55% shade was significantly greater than that of no shade and 35% shade; 55% shade also showed significantly higher anti-inflammatory capacity than that of no shade or 35% shade.

• Journal of Applied Biological Chemistry
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• v.57 no.4
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• pp.301-305
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• 2014

The Antioxidant activity screening identification of five kind compounds in Artemisiae annuae herba with the on-line screening high performance liquid chromatography (HPLC) $ABTS^+$ assay. The various experimental variables such as the extraction time (h) and extraction solvent composition (%) of dipping method were investigated efficiently extraction at the room temperature $25^{\circ}C$. The results, the highest yield of total extract amount (0.458 g, 15.250%) was obtained by dipping method with 100% water and extraction time to 3 h. And the on-line screening HPLC-$ABTS^+$ assay method was rapid and efficient to search for bioactivity from natural products.

• BMB Reports
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• v.30 no.6
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• pp.443-447
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• 1997

We have developed a novel method for assays of spermidine and spermine synthase (aminopropyltransferase) activities using antibody against 5'-deoxy-5'-methylthioadenosine (MTA). A new assay is reported here which is based on the observation that MTA is formed as a stoichiometric by-product of the spermidine and spermine synthases reactions. In order to determine MTA, a radioimmunoassay method with sensitivity and rapidity was used. (Lee and Cho, 1997). In this assay, adenine must be added in the reaction mixture, since it effectively inhibits the action of MTA phosphorylase by which MTA is metabolized. This assay is a improvement in term of sensitivity and time saving, compared to the currently used methods. It has a level of sensitivity (100 fmol) sufficient to monitor aminopropyltransferase activities in incubations containing as little as $10{\mu}g$ protein prepared from rat tissue homogenate. The results obtained showed that this method is particularly useful for cultured cells with low enzyme concentration. Moreover, this assay has the advantage which allows studies using alternative substrates (other amines). Spermidine synthase activity was high in rat liver, but low in rat kidney. The activity of spermine synthase was in most rat tissues very low as compared to that of spermidine synthase, but was high in brain.

• Korean Journal of Environmental Biology
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• v.29 no.1
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• pp.23-30
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• 2011

In this study, the Comet assay (evaluation of DNA damage) used the fish hepatocellular carinoma cell, PLHC-1, was tried to the sediment extract obtained from freshwater to understand its applicability as a tool for monitoring sediment toxicity. In parallel, induced EROD (7-ethoxyresorufin- O-deethylase) activity and DNA damage (TEM values) in PLHC-1 cells were measured for establishing the tandem endpoints of the PLHC-1cell test to test the ecotoxicity of sediment. Among several study sites in a small river passed through downtown and industrial park area, one of them, site B, showed a higher level of EROD activity and DNA damage than other sites. It indicates that a tandem endpoints of PLHC-1 cells could be useful tools for assessing the toxicity of sediment. The sensitivity of Comet assay with PLHC-1 cells was a little higher than that with a blood cell of frog tadpoles to the solvent extract of sediment. According to the results, a PLHC-1 cell-Comet assay could be used as a useful tool for evaluating ecotoxicity of the freshwater sediment. In addition, more detailed studies are needed to the contaminated site.

• Food Science of Animal Resources
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• v.24 no.3
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• pp.293-297
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• 2004

This study was carried out to investigate the antitumor activity from sulfur fed duck. The antitumor substances were crude purified by solvent extraction, silica gel column chromatography, and HPLC using C18 column. In MTT assay, the active compounds exhibited more cytotoxic activity on tumor cell lines than normal cell line. In addition of 100 $\mu\textrm{g}$/mL concentrations of crude purified active compounds, the growth inhibition rate of tumor cell lines was 56% (Hep-2j human larynx), 58% (KB; human epidermoid of mouth carcinoma), and 28% (MDBK; bovine normal kidney), respectively. The survival rate of clonogenic assay was 26% in Hep-2 and 28% in KB at 200 $\mu\textrm{g}$/mL.

• CELLMED
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• v.3 no.3
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• pp.22.1-22.10
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• 2013

Traditional Chinese medicine (TCM) in single herb or formula prescription has been used for thousands of years. Many of them possess antioxidant activity and the activity may contribute the therapeutic effect. This paper would review the relationship of traditional herbal medicine and antioxidant with particular reference to ginseng. This medicinal herb has been used worldwide with extensive tonic effect. The comet assay, a technique for DNA protecting and damaging investigation would be introduced and the application of comet assay on TCM would be discussed.

• 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 Aquaculture
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• v.19 no.1
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• pp.33-39
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• 2006

Anticoagulant activities of a fermented edible brown alga, Laminaria ochotensis was investigated. L. ochotensis was fermented with 15% sugar (w/v) at $25^{\circ}C$ for 10 weeks. Anticoagulant activity was measured from the supernatant of algal mixture at biweekly intervals up to $10^{th}$ week by activated partial thromboplastin (APTT), prothrombin time (PT) and thrombin time (TT) assay using citrated human plasma. Sample having high APTT activity $(6^{th}\;week)$ was filtered, ethanol precipitated and freeze-dried. The polysaccharide compound having anticoagulant activity was purified by DEAE ion exchange chromatography followed by Sepharose-4B gel filtration chromatography. Anticoagulant activity, polysaccharide concentration, and heparin like activity were determined for the collected fractions by APTT, $phenol-H_2SO_4$, and glycosaminoglycan assay, respectively. The anticoagulant activity assay showed that the activity was increased up to $6^{th}$ week, and decreased thereafter. The concentration of our purified compound was $31.0{\mu}g/ml$ and showed higher APTT activity than commercial heparin. At the same concentration of $31.0{\mu}g/ml$, the heparin showed 186.5 sec activity while our purified compound showed an activity of 386 sec. Single spot on agarose gel electrophoresis showed that the compound was purified and polyacrylamide gel electrophoresis (PAGE) results revealed that the molecular mass of the purified polysaccharide compound was between 60 and 500 kDa. Therapeutic interest of the algal polysaccharide as an anticoagulant has recently been in highlighted. This purified anticoagulant compound from fermented L. ochotensis can be used as a model for anticoagulant agent or could be developed as an anticoagulant agent. This study can be extended to identify the structure and chemical composition of the purified polysaccharide, and to establish a relationship between structure and the function of the identified anticoagulant compounds.

• Environmental Analysis Health and Toxicology
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• v.25 no.1
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• pp.85-94
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• 2010

Platycodon grandiflorum, Doraji as Korean name, is one of the most widely used traditional oriental medicine for bronchial diseases and also used as a folk remedy for geriatric diseases and inflammatory diseases. In recent studies, it has been reported that some effect of P. grandiflorum is derived from its antioxidant activity, although there is still a lack of evidence to establish its oxy-radical scavenging activity. In this study, total oxy-radical scavenging capacity (TOSC) assay was used to evaluate antioxidant activity of total extracts (T-PG), polysaccharide fraction (Po-PG), and saponin fraction (Sa-PG) isolated from P. grandiflorum against peroxyl radicals and peroxynitrites. And MTT assay was taken to assess cyto-protective effects of T-PG, Po-PG and Sa-PG in H4IIE cells treated with hydrogen peroxide and tert-butylhydroperoxide. In the TOSC assay, Sa-PG showed strong oxy-radical scavenging capacity compared with T-PG and Po-PG. In cell-based assay, T-PG and Po-PG protected cells from oxidative stress, but Sa-PG did not protect cells because of cytotoxicity of Sa-PG. These results suggest that the saponin components of P. grandiflorum have relatively strong antioxidant capacity and cytotoxicity in rat hepatoma cells.