• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2003.04a
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• pp.65-71
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• 2003

Bax, a mammalian pro-apoptotic member of the Bcl-2 family, induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorho-damine 123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MBP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to play a novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

• Journal of Life Science
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• v.21 no.5
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• pp.746-752
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• 2011

The contents of bioactive materials (e.g. polyphenolics compounds, flavonoids, minerals, and fatty acids) and antioxidative activities (DPPH (${\alpha}$,${\alpha}$'-diphenyl-${\beta}$-picrylhydrazyl) free radical scavenging activity, peroxidation of linoleic acid and rat hepatocyte microsome, and Fe/Cu reducing power) were tested by in vitro experimental models using water, ethanol and methanol extracts of leaves (TOL) and fruits (TOF) from Thuja orientalis. Methanol extract from TOL showed the highest extraction yield (12.90%) as well as contents of polyphenolic compounds (16.02%) and flavonoids (0.25%). Major minerals were Ca, K, and Mg. Major fatty acids were palmitic and lauric acids in TOL and palmitic and decanoic acids in TOF. In oxidation of in vitro models using DPPH free radical scavenging activity, Fe/Cu reducing power, $Fe^{2+}$/ascorbate-induced linolenic acid peroxidation by ferric thiocyanate and thiobarbituric acid (TBA) methods, and autooxidation of rat hepatic microsomes membrane, anti-oxidative activities were stronger in all extracts of TOL than in those of TOF in a dose-dependent manner. From these results, methanol extract of TOL was shown to have the most potent anti-oxidative properties and the highes content of antioxidative compounds such as polyphenolic compounds and flavonoids.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.1
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• pp.14-19
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• 2010

This study was to investigate antioxidant effects of mistletoe extracts by extraction conditions. Electron donating ability of 50% ethanol extract had shown 92.27% at 50 mL/g, which was higher than 1.0% and 0.1% L-ascorbate solutions (p<0.05). SOD-like activities were the most effective in all samples at 50 mL/g, which were barely detectable. The total polyphenol contents of water extracts including all extracts were the highest at 50 mL/g extracts (p<0.05). The nitrite scavenging effects were different depending on pH value; they were decreased overall as pH value was increased. Especially, nitrite scavenging effects were the most effective in pH 1.2, which showed more than 80% (p<0.05). Tyrosinase-inhibitory activities ranged at fewer than 10%. These results indicate that mistletoe extracts are potential sources of natural antioxidant.

• Journal of Korean Neurosurgical Society
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• v.53 no.4
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• pp.207-212
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• 2013

Objective : Recent studies have shown encouraging progress toward the use of autogenic and allogenic mesenchymal stem cells (MSCs) to arrest, or even lead to partial regeneration in, intervertebral disc (IVD) degeneration. However, this technology is still in its infancy, and further development is required. The aim of this study was to analyze whether rat adipose-derived mesenchymal stem cells (ADMSC) can differentiate towards IVD-like cells after treatment with transforming growth factor ${\beta}3$ (TGF-${\beta}3$) in vitro. We also performed quantitative analysis of gene expression for ADMSC only, ADMSCs treated with TGF-${\beta}3$, and co-cultured ADMSCs treated with TGF-${\beta}3$. Methods : ADMSCs were sub-cultured to homogeneity and used in fluorocytometry assays for CD11, CD45, and CD90/Thy1. ADMSCs were differentiated in spheroid culture towards the chondrogenic lineage by the presence of TGF-${\beta}3$, dexamethasone, and ascorbate. We also co-cultured pure ADMSCs and nucleus pulposus cells in 24-well plates, and performed immunohistochemical staining, western blotting, and RT-PCR for quantitative analysis of gene expression. Results : Results of fluorocytometry were positive for CD90/Thy1 and negative for CD11 and CD45. TGF-${\beta}3$-mediated induction of ADMSCs led to the expression of the differentiation markers of intervertebral disc-like cells, such as aggrecan, collagen II, and sox-9. Co-cultured ADMSCs treated with TGF-${\beta}3$ showed higher expression of differentiation markers and greater extracellular matrix production compared with ADMSCs treated with TGF-${\beta}3$ alone. Conclusion : ADMSC treated with TGF-${\beta}3$ may be an attractive source for regeneration therapy in degenerative IVD. These findings may also help elucidate the pathologic mechanism of MSC therapy in the degeneration of IVD in vivo.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.7
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• pp.942-948
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• 2011

The physiological activities of extracts from the leaf, stem, and fruit of Cudrania tricuspidata were investigated. The electron-donating ability (EDA) of the 70% (v/v) ethanolic extract of stem was 90.20%; this was the highest value of all the extracts tested and higher than the L-ascorbate solutions. The total polyphenol contents were the highest in the leaf extracts under all extraction conditions. Especially, 70% (v/v) methanolic extract of leaf contained the highest total polyphenol content of 224.48 mg%. SOD-like activity showed the highest activity in water extract of leaf at 64.53%. Tyrosinase-inhibitory activities were the most effective in all extracts of fruit. ACE inhibitory activities were the highest in water extract of fruit. Nitrite-scavenging abilities under acidic conditions (pH 1.2 and pH 3.0) were the most effective in all the extracts. The results of this study will be useful for understanding the physiological activities of Cudrania tricuspidata extracts.

• Journal of Korean Society of Forest Science
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• v.99 no.6
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• pp.878-884
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• 2010

Ozone sensitivity of physiological indicators and the difference of ozone tolerance on 4 families of Quercus aliena seedlings were investigated on the basis of the standardized physiological indicators. Photosynthetic parameters, photosynthetic pigment and malondialdehyde (MDA) content, and antioxidative enzyme activities were measured or analyzed from the leaves of Q. aliena seedlings at the end of ozone fumigation, and ozone tolerance indices among 4 families were calculated with the standardized physiological parameters. After ozone treatment, the reduction of carboxylation efficiency was observed in the leaves of four families, and their reduction were ranged from -24.1% to -56.9% of control seedlings. Photosynthetic pigment content differed significantly among 4 families and treatments. The reduction of total chlorophyll content showed the highest in family SU4 (-40.6%) and the lowest family US2 (-18.8%). Ascorbate-peroxidase (APX) activity showed significant difference among families and treatments, and increased as compared with control in three families, except for family US2. On the basis of the physiological indices, ozone tolerance of four families was ranked in the order of US1 > SU4 > US2 > SU1. In conclusion, photosynthetic parameters, pigment content and APX activity were recommended as appropriate indicators to assess the tolerance against ozone stress of Q. aliena.

• Journal of Life Science
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• v.13 no.5
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• pp.658-667
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• 2003

Antioxidative enzymes (superoxide dismutase; SOD, ascorbate peroxidase; APX, glutathione reductase; GR) play major roles in scavenging mechanism of reactive oxygen species which were involved in various stress conditions including salt. In order to investigate the relation between their growth responses (dry weight) and the changes of antioxidative enzymes activity, salt-tolerant spinach beet having 15cm of shoot length were treated with various salt levels (0, 50, 200, 1000 mM NaCl) for 24 hours. Spinach beet exhibited an increase in the activity of antioxidative enzymes by salt, the maximal activity at 200 mM NaCl and the lowest activity at 50 mM NaCl in 2 hrs. after treatments. As a result of PAGE, it has been confirmed that spinach beet contained 3 isoforms (Fe-SOD, CuZn-SOD and Mn-SOD) of SOD and main isoform was CuZn- SOD form. In case of APX, isoforms of the low molecular weight(No. 7, 8) were showed strong expression especially at 200 and 400 mM NaCl treatment. Meanwhile, GR did not show specific pattern of isoforms among the salt treatments. Especially, in case of 50 mM treatment, plant showed the lowest activity of SOD with the best growth, a low enzyme activity was induced by inactivation of the Mn-SOD. Therefore, we suggested that the decrease of SOD activity at a low salt level (50 mM NaCl) or the increase of enzyme activity at a high salt level (200 mM NaCl) may be related to expression of the Mn-SOD isoform. These antioxidative enzymes showed the increase of activity in a short time by salt addition. So, it is considered that spinach beet copes effectively with a stressful condition such as salt by operating effective antioxidative defense mechanism rapidly under high salt level.

• Food Science of Animal Resources
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• v.31 no.4
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• pp.588-595
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• 2011

In this study, the effects of red ginseng extracts (RGE), which has been used as an antioxidant and antimicrobial agent, on pork sausage were evaluated. The treatments were as follows; addition of 0.01% sodium ascorbate (V), 0.5% RGE (T1), 1.0% RGE (T2) and 1.5% RGE (T3) to the basal formula (C). T3 had a significantly higher pH, cooking loss and yellowness ($CIEb^*$) and lower lightness (CIE $L^*$) and redness (CIE $a^*$) than the other samples. The hardness and surface hardness values of 1.5% RGE treated sample were significantly lower (p<0.01) than those of C. However, the cohesiveness values of the RGE samples were higher than the others (p<0.05). In the sensory evaluation, no significant differences in color, taste, texture, juiciness and acceptability were observed among the tested samples, while, the aroma scores of T2 and T3 were higher than those of the C and V samples (p<0.05). The TBARS values of RGE treated groups were higher (p<0.05) than the C sample after 1, 2 and 3 weeks of storage; however, this value did not increase with storage time (p>0.05). When the RGE concentration was high, the reduction in total plate counts and VBN value at week 3 and 4 of storage (p<0.01) decreased. In conclusion, red ginseng extracts seemed to have a positive impact on lipid oxidation, aroma and the microbial characteristics of pork sausage.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.203-203
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• 2017

The Earth's climate is rapidly changing because of increasing carbon dioxide content in atmosphere so, climate prediction models anticipate that earth surface temperature will rise by 3 to $5^{\circ}C$ in next 50 to 100 years. Therefore, frequency of un-expected weather events such as drought, salinity, low or high temperature and flooding etc. will be increasing worldwide. Furthermore, increased atmosphere temperature can influence pests and pathogens spread as well. Therefore, to protect enormous grain loss from unexpected weather conditions, studies related with combine stress conditions like abiotic plus biotic stress condition are really required. Thus, our research focused on physiological responses under combined abiotic and biotic stress condition in rice plant. To induce uniform stress condition, we used NaCl (100 mM) and salicylic acid (0.5 and 1.0 mM SA) as each stress a stimulator. Each artificial abiotic and biotic stress inducer was applied to hydroponically grown rice seedlings alone or together for four day. The data were collected in a time-dependent manner [1, 2, 3 and 4 day(s) after treatment (DAT)] and were matched with our anticipation that shoot length and shoot fresh weight was decreased in solo and combined abiotic and biotic stress condition. The lipid peroxidation content was significantly increased ($1.5{\pm}0.2$ to $2.7{\pm}0.1mg$ mg of $MDA\;g^{-1}FW$) in the first two days in both stress exposed plants, and showed the opposite trend ($0.5{\pm}0.01$ to $0.1{\pm}0.001mg$ of $MDA\;g^{-1}FW$) in last two days under multi stress condition. Superoxide dismutase (SOD) activity did not showed difference in only biotic stress condition (alone 0.5 and 1.0 mM SA) as compared to control however, it was significantly increased in multi stress condition or solo abiotic stress condition whereas, catalase (CAT), and ascorbate peroxidase (APX) activities were significantly decreased in solo biotic and combined abiotic and biotic condition. In particular, both enzymes activities were more decreased in multi stress condition as compared to solo biotic stress condition. The results for relative mRNA expression level of CAT and APX enzymes were in agreement with results of spectrophotometric values. Correlation value between each stress condition and phenotypic data showed that biotic stress condition showed high correlation with activity of CAT and APX whilst, abiotic stress condition revealed significant correlation with SOD activity.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.127-127
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• 2017

Aluminum is the most abundant metallic element in the Earth's crust and considered as the most limiting factor for plant productivity in acidic soils. The inhibition of root growth is recognized as the primary effect of Al toxicity. Seeds of wheat cv. Keumkang (Korean cultivar) were germinated on petridish for 5 days and then transferred hydroponic apparatus which was treated with $0{\mu}M$ $AlCl_3$ (control), $100{\mu}M$ $AlCl_3$ and $150{\mu}M$ $AlCl_3$ for 5 days. The length of roots, shoots and fresh weight of wheat seedlings were decreased under aluminum stress. The concentrations of $K^+$, $Mg^{2+}$ and $Ac^{2+}$ were decreased whereas $Al^{3+}$ and $P_2O_5{^-}$ concentration was increased under aluminum stress. Using confocal microscopy, the fluorescence intensity of aluminum was increased with morin staining. In this study, a proteome analysis was performed to identify proteins, which is responsible to aluminum stress in wheat roots. In 10-day-old seedlings, proteins were extracted from roots and separated by 2-DE, stained by CBB. Using image analysis, a total of 47 differentially expressed protein spots were selected, whereas 19 protein spots were significantly up-regulated such as s-adenosylmethionine, oxalate oxidase, malate dehydrogenase, cysteine synthase, ascorbate peroxidase and 28 protein spots were significantly down-regulated such as heat shock protein 70, o-methytransferase 4, enolase, amylogenin by aluminum stress following protein spots analyzed by LTQ-FTICR mass spectrometry. The results provide the global picture of Al toxicity-induced alterations of protein profiles in wheat roots, and identify the Al toxicity-responsive proteins related to various biological processes that may provide some novel clues about plant Al tolerance.