• KOREAN JOURNAL OF CROP SCIENCE
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• v.59 no.3
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• pp.341-349
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• 2014

Field emergence of Azuki bean is poor due to hard seed coat as compared to other legumes. In this study, an attempt was made to develop prediction method with regression analysis based on various seed vigor tests in laboratory for field emergence of azuki bean. Azuki bean seeds artificially aged to provide various levels of seed quality were evaluated by the standard germination test (SGT), cold germination test (CT), cool germination test (CGT), complex stressing vigor test (CSVT), tetrazolium(TZ) vigor test and electroconductivity test. The SGT was suitable for predicting the field emergence in the unaged high vigor seeds. The abnormal seedling percentage and shoot length in the CGT were highly correlated with field emergence of moderate vigor seeds artificially aged for 2 days. Electroconductivity, seed viability in the CSVT, and vigor and predicted germinability in the tetrazolium vigor test were also useful for predicting field emergence. Percent of ungerminated seed in the CSVT was correlated with field emergence in the low vigor seeds artificially aged for 4 days. In a stepwise multiple regression analysis, seed viability in the SGT, normal seedling percentage and dry matter weight in the CGT accounted for 86.9% of the predicted value of field emergence in azuki bean.

• Korean Journal of Food Science and Technology
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• v.43 no.6
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• pp.754-759
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• 2011

Zinc protoporphyrin (ZnPP) is produced endogenously during heme metabolism and treated in cells as a heme oxygenase inhibitor. In the present study, the effects of ZnPP on the color response of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, a commonly-used method for analyzing cell viability, were investigated. ZnPP induced rapid decolorizaion of MTT formazan under light; the degradation rates were 10- and 20- folds faster in the presence of 5 and $10{\mu}M$ ZnPP, respectively. Methylene blue (MB), another type of photosensitizer, also accelerated degradation of formazan under light. Butylated hyroxytoluene did not inhibit ZnPP- or MB-induced formazan degradation. The color degradation of formazan dye was signficantly delayed in the presence of N-acetylcysteine or ${\beta}$-carotene. The present results suggest that certain photosensitizing compounds may affect the color and stability of MTT formazan, which should be carefully considered when conducting the MTT assay.

• Journal of Pharmacopuncture
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• v.20 no.1
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• pp.52-56
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• 2017

Objectives: Neutrophils represent the front line of human defense against infections. Immediately after stimulation, neutrophilic enzymes are activated and produce toxic mediators such as pro-inflammatory cytokines, nitric oxide (NO) and myeloperoxidase (MPO). These mediators can be toxic not only to infectious agents but also to host tissues. Because flavonoids exhibit antioxidant and anti-inflammatory effects, they are subjects of interest for pharmacological modulation of inflammation. In the present study, the effects of rutin on stimulus-induced NO and tumor necrosis factor $(TNF)-{\alpha}$ productions and MPO activity in human neutrophils were investigated. Methods: Human peripheral blood neutrophils were isolated using Ficoll-Hypaque density gradient centrifugation coupled with dextran T500 sedimentation. The cell preparations containing > 98% granulocytes were determined by morphological examination through Giemsa staining. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without rutin ($25{\mu}M$) for 45 minutes, and stimulated with phorbol 12-myristate 13-acetate (PMA). Then, the $TNF-{\alpha}$, NO and MPO productions were analyzed using enzyme-linked immunosorbent assay (ELISA), Griess Reagent, and MPO assay kits, respectively. Also, the viability of human neutrophils was assessed using tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and neutrophils were treated with various concentrations of rutin ($1-100{\mu}M$), after which MTT was appended and incubated at $37^{\circ}C$ for 4 hour. Results: Rutin at concentrations up to $100{\mu}M$ did not affect neutrophil viability during the 4-hour incubation period. Rutin significantly decreased the NO and $TNF-{\alpha}$ productions in human peripheral blood neutrophils compared to PMA-control cells (P < 0.001). Also, MPO activity was significantly reduced by rutin (P < 0.001). Conclusion: In this in vitro study, rutin had an anti-inflammatory effect due to its inhibiting NO and $TNF-{\alpha}$ productions, as well as MPO activity, in activated human neutrophils. Treatment with rutin may be considered as a therapeutic strategy for neutrophil-mediated inflammatory/autoimmune diseases.

• Journal of Life Science
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• v.23 no.3
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• pp.341-346
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• 2013

The disappearance of seaweed flora in some rocky areas, which is known as algal whitening, barren ground, coralline flats, or deforested areas, is associated with some species of coralline algae. To determine the biological characteristics of a representative species of crustose coralline alga, the 18S rDNA gene was sequenced to identify the genus Lithophyllum. According to its morphological and distributional characteristics, it was deduced to be L. yessoense. Viability was measured using triphenyl tetrazolium chloride and showed high viability from December to February. Culture conditions of $16^{\circ}C$, a 16 hr light, 8 hr dark cycle, and 30 ${\mu}E/m^2/s$ light intensity were optimal for maintaining the viability of the alga for up to five days. Included in the fatty acids was 9.7% ${\omega}$-3 eicosapentaenoic acid. An electron microscopy scan of the surface structure revealed round craters about 3.6 ${\mu}m$ in diameter, which were covered with rough, irregular, and angular polygon-shaped structures about 1.0 to 3.7 ${\mu}m$ in size. Based on the composition and structure found in our study, biomimetic coralline alga might become an environmentally friendly antifouling material against the attachment of soft foulants.

• Journal of Digital Convergence
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• v.13 no.5
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• pp.337-344
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• 2015

Medicinal plants containing wellness-fusion-complex compound are increasingly being pursued as suitable alternative sources of various biological properties. In this study, inhibitory effect of Quintinia acutifolia, which is a New Zealand plant, on P388 murine lymphocytic leukemia cells using MTT [3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazolium bromide] assay. Based on $^1H-NMR$, $^{13}C-NMR$ spectral data and other spectral analysis, 2,3,2'',3''-tetrahydroochanaflavone (1) and 2'',3''-dihydroochana-flavone (3) inhibited the leukemia cells were purified from the plants. 2,3,2'',3''-tetrahydroochanaflavone (1) and 2'',3''-dihydroochana-flavone (3) are biflavonoids possessing two basic flavonoids and actively inhibited growth of P388 murine lymphocytic leukemia cells with a 50% inhibitory concentration ($IC_{50}$) of $8.2{\mu}g/mL$ and $3.1{\mu}g/mL$, respectively. Specially, 2'',3''-dihydroochana-flavone (3) possessed unconjugated flavonone system, which isn't consist of a pair with B ring of 2,3,2'',3''-tetrahydroochanaflavone (1). Therefore, the two compounds could be considered as a candidate for development of anticancer drugs and need to much studies in the future.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.34 no.2
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• pp.134-141
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• 1989

In order to estimate field seedling emergence and grain yield by the information collected from various laboratory seed vigor tests, each two malting and naked barley cultivar seeds were artificially aged according to the standard aging treatment suggested by the Association of Official Seed Analysts. The seed vigor tests adopted were warm- and cold-germination test, tetrazolium vigor test and electrocon. ductivity test. Field emergence of malting barley (Y) was estimated by Y=-2.962+0.229X$_1$ (% warm germination) -0.001X$_2$ (vigor of warm germination test: WGT) +0.354X$_3$ (vigor of cold germination test: CT) -0.558X$_4$ (% cold germination). The multiple correlation coefficient indicated that % warm germination. was contributed 64 % of the variation in seedling emergence rate of malting barley. The vigor of warm and cold germination tests, and % cold germination contributed additional 4, 7, and 9%, respectively, upon addition of the variables into regression. For naked barley, the regression equation of emergence rate was less efficient(R$^2$=54%) than that of malting barley(R$^2$=84%). A model to predict grain yield by the results of various seed vigor tests was not evaluated for both malting and naked barlev.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.1
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• pp.47-54
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• 2000

Transition metal ions including $Se^{2+},\;Cd^{2+},\;Hg^{2+}\;or\;Mn^{2+}$ have been thought to disturb the bone metabolism directly. However, the mechanism for the bone lesion is unknown. In this study, we demonstrated that MC3T3E1 osteoblasts, exposed to various transition metal ions; selenium, cadmium, mercury or manganese, generated massive amounts of reactive oxygen species (ROS). The released ROS were completely quenched by free radical scavengers-N-acetyl cysteine (NAC), reduced glutathione (GSH), or superoxide dismutase (SOD). First, we have observed that selenium $(10\;{\mu}M),$ cadmium $(100\;{\mu}M),$ mercury $(100\;{\mu}M)$ or manganese (1 mM) treatment induced apoptotic phenomena like DNA fragmentation, chromatin condensation and caspase-3-like cysteine protease activation in MC3T3E1 osteoblasts. Concomitant treatment of antioxidant; N-acetyl-L-cysteine (NAC), reduced-form glutathione (GSH), or superoxide dismutase (SOD), prevented apoptosis induced by each of the transition metal ions. Catalase or dimethylsulfoxide (DMSO) has less potent inhibitory effect on the apoptosis, compared with NAC, GSH or SOD. In line with the results, nitroblue tetrazolium (NBT) stain shows that each of the transition metals is a potent source of free radicals in MC3T3E1 osteoblast. Our data show that oxidative damage is associated with the induction of apoptosis in MC3T3E1 osteoblasts following $Se^{2+},\;Cd^{2+},\;Hg^{2+}\;or\;Mn^{2+}$ treatment.

• Journal of Life Science
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• v.17 no.5 s.85
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• pp.671-677
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• 2007

This study was performed to investigate the effect of the water-extract from non-fermented or fermented Chaga mushrooms (Inonotus obliquus) on the proliferation and apoptosis of the NIH3T3 mouse normal fibroblast cells and various human cancer cell lines including HCT-15 human colon carcinoma, AGS human gastric carcinoma, MCF-7 human breast adenocarcinoma, Hep3B human hepatocellular carcinoma and HeLa human cervical carcinoma using MTT(3-[4,5-dimethylthiazol-2-yl] -2,5-diphenyl tetrazolium bromide) assay and DNA fragmentation. In an anti-cancer test using various human cancer cells, fermented Chaga mushroom extract showed higher antiproliferating effect than that of non-fermented Chaga mushroom extract. Mouse normal NIH3T3 cells were exhibited 80% above survival under fermented or non-fermented Chngn mushroom extract of various concentrations(0, 0.5 and 1 mg/ml). Fermented Chaga mushroom extract significantly inhibited cell growth on HCT-15 cells in a dose-dependent manner. HCT-15 cells treated with non-fermented or fermented Chaga mushrooms extract produced a distinct oligonucleosomal ladder with different sizes of DNA fragments, a typical characteristic of cells undergoing apoptosis. These results suggest that fermented Chaga mushroom extract suppresses growth of HCT-15 human colon carcinoma cells through apoptosis.

• Polymer(Korea)
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• v.32 no.5
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• pp.403-408
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• 2008

Keratin is the major structural fibrous protein providing outer covering such as wool, hair, and nail. Keratin is useful as natural protein. We developed the keratin loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds (keratin/PLGA) for the possibility of the application of the tissue engineering using bone marrow mesenchymal (BMSCs). Keratin/PLGA (contents 0%, 10%, 20% and 50% of PLGA weight) scaffolds were prepared by solvent casting/salt leaching method. We characterized porosity, wettability, and water uptake ability, DSC of keratin/PLGA scaffold. We seeded BMSCs isolated from the femurs of rat into the inner core of the hybrid scaffold. Celluar viability were assayed by 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT) test. We confirmed that keratin/PLGA scaffold is hydrophilic by wettability, and water uptake ability measurement results. In MTT assay results, cell viability in scaffolds impregnated 10 and 20 wt% of keratin were higher than other scaffolds. In conclusion, we suggest that keratin/PLGA scaffold may be useful to tissue engineering using BMSCs.

• The Journal of the Korea Contents Association
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• v.19 no.7
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• pp.186-191
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• 2019

Introduction: In aged people, stroke incidence is increased. But standardized experimental animal protocol study for the research of stroke therapy is rare. There is little report on the success rate of cerebral artery occlusion model using standardized Nylon thread length of precise thread end-size controlled. Method: In this study, the operator intended the occlusion of middle cerebral artery (MCA) using $0.18{\pm}0.02mm$ end 5-0 Nylon thread. Middle cerebral artery occlusion was induced for 60min under isoflurane anesthesia. After 60min, the operator removed the Nylon thread and reperfusion was induced for 23hrs. The mice was killed 23hrs after reperfusion and infarction area of brain was confirmed by 1.5% TTC (2,3,5-tryphenyl tetrazolium chloride) staining. Results: According to end size and insert length of Nylon thread, Middle cerebral artery occlusion (n=50), internal carotid artery occlusion (n= 14), distal middle cerebral artery occlusion (n= 36), anterior cerebral artery (n= 1) were induced. And no infarction (n= 50) was observed. Conclusion: According to weight of mice, the operator induced reversible cerebral artery occlusion model by different insert length (30.0~36.9g : 9.0mm, 37.0~40.0g : 9.5mm) of Nylon thread. Success of cerebral artery occlusion model was confirmed by checking infarction area using TTC staining. The success rate (66.9%, 101/151) of reversible cerebral artery occlusion model in the mouse and the operational conditions are shown.