• Proceedings of the Ginseng society Conference
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• 1988.08a
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• pp.92-98
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• 1988

The present study was performed to find the effects of ginseng and its saponins. which is written in Chung Yao Ta Tsu Tien as anti-amnesia in its chief indication. on experimental amnesia in mice. In the step through test. ginsenoside $Rb_1\;(GRb_1)\;and\;GRg_1$ facilitated the registration of memory and antagonized the electroconvulsive shock (ECS)-induced inhibition of the retention of memory. Moreover. $GRg_1$ antagonized the EtOH-induced inhibition of the retrieval of memory. In the step down test. $GRb_1\;GRb_2\;and\;GRg_1$ antagonized the ECS-induced inhibition of the retention of memory. Moreover. $GRg_1$ antagonized the EtOH-induced inhibition of the retrieval of memory and facilitated the acquisition of short term memory. In the shuttle hox and lever press tests. they have no effects on acquisition and retrieval of memory. except $GRb_1\;GRb_1$ depressed the retrieval of conditioned avoidance response in the shuttle box test. After the end of four tests. the effects of these orally administered drugs on sedative. analgesic. antipyretic and anticonvulsant actions. and on spontaneous and exploratory movements were tested in doses of less than 500mg/kg. but they had none of these effects. Present study may indicate that $GRg_1$ had effects on the retrieval of memory and on the acquisition process of learning response. The recent research on the role of NGF for the survival. regeneration and regulation of brain in adult animals. indicated the importance of NGF on dementia and amnesia. During our research on the specificity of the neurite out growth induced by NGF. we found that the effect of NGF was potentiated by $GRb_1$ in organ cultures of chick embryonic dorsal root ganglia. Then. the effect of $GRb_1$ on neuronal cell survivalin cell culture system was studied. $GRb_1$ potentiated the NGF-mediated increase of neurofilaments in cell cultures of chick embryonic sensory and sympathetic neurons. NGF with $GRb_1$ also showed a tendency to increase the number of surviving neurons of rat embryonic cerebral cortex. NGF increased choline acetyl transferase activity in cell cultures of rat embryonic septum area neurons. but $GRb_1$ did not potentiate NGF activity in cell cultures of rat embryonic septum area neurons. Present study may indicate that $GRb_1$ plays an important role for the survival or regeneration of neurons in the brain.

• Korean Journal of Plant Resources
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• v.15 no.3
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• pp.237-242
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• 2002

Loaves, stems, cotyledons, and roots of Hovenia dulcis Thunb grown in test tube were cultured on media containing different concentrations of single or combined growth regulators. In MS media containing 2mg/ι BA, the shoot formation rate was 95.5% and it was the highest frequency of shoot formation. MS media showed most efficiency in the shoot formation at 0.01mg/ι TDZ for the callus formation, but the color of callus changed to brown at a higher concentration of TDZ. Callus formation was 89.% at 0.5mg/ 2.4-D, but IAA, IBA, and NAA were not effective on the formation of callus. Calli were formed only on wound area when IAA, IBA, and NAA were added into MS media. Combined growth regulators (BA + auxin) were more effective in roots and nodes than leaves and cotyledons on the formation of shoot. More than 97％ of shoot formation was obtained on MS media containing BA and auxin. For the production of multiple shoot, nodes of Hovenia dulcis were used and effect of growth regulators on the formation of multiple shoot was evaluated on MS media. Highest shoots (5.3) of Hovenia dulcis were induced on MS media supplied with 0.1mg/ι BA and 0.1mg/ι NAA, and an average of 6.4 shoots per explant were obtained in 1/2 MS media containing same concentration and growth regulators. An average of 7 shoots per explant after 4 weeks of culture from nodes of Hovenia dulcis was produced on a woody plant medium(WPM) containing 0.1mg/ι BA and 0.1mg/ι NAA. Shoot length was 6.0 cm in average.

• Korean Journal of Plant Resources
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• v.25 no.5
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• pp.568-577
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• 2012

The objective of this study was to investigate the antioxidative capacity of ethanol extracts from Rumex crispus L. The concentration of R. crispus L. extract at which DPPH radical scavenging activity was inhibited by 50% was 2.15 mg/mL, which was lower than that of ${\alpha}$-tocopherol (0.43 mg/mL), as compared to 100% by pyrogallol as a reference. Total antioxidant status was examined by total antioxidant capacity against ABTS radical reactions. Total antioxidant capacities of R. crispus L. extract at concentrations of 0.1 and 1 mg/mL were 0.47 and 2.33 mM Trolox equivalents, respectively, which were higher than those of ${\alpha}$-tocopherol. Superoxide scavenging activities of R. crispus L. extract at concentrations of 0.1 and 1 mg/mL were 21.5 and 78.9%, respectively, which were not significantly (p>0.05) different from those of catechin. Oxygen radical absorbance capacities of R. crispus L. extract at concentrations of 20 and 100 ${\mu}g/mL$ were 62.5 and 156.4 ${\mu}M$ Trolox equivalents, respectively, which were lower than those of ascorbic acid. Cupric reducing antioxidant capacities of R. crispus L. extract at concentrations of 0.1 and 1 mg/mL were 0.28 and 1.88 mM Trolox equivalents, which were similar or significantly (p<0.05) higher than those of ${\alpha}$-tocopherol, respectively. R. crispus L. extract prevented supercoiled DNA strand breakage induced by hydroxyl radical and peroxyl radical. Total phenolic contents of R. crispus L. extract at concentrations of 0.5 and 5 mg/mL were 0.58 and 3.85 mM gallic acid equivalents, respectively. R. crispus L. extract at concentration of 0.1 and 0.5 mg/mL inhibited 0.2 mM tert-butyl hydroperoxide-induced cytotoxicity by 38.5 and 63.5%, respectively, in HepG2 cell culture system. Thus, strong antioxidant and cytotoxicity-inhibiting effects of R. crispus L. extract seem to be due to, at least in part, the prevention from free radicals-induced oxidation as well as high levels in total phenolic contents.

• Korean Journal of Plant Resources
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• v.21 no.5
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• pp.368-373
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• 2008

In order to investigate the effects of low temperature pretreatment of floral bud and plant growth regulators on anther-derived callus and shoot differentiation, anthers were cultured on 1/2 MS medium supplemented with 2,4-D, NAA, BA and TDZ. This plant depends on the plant growth regulators, for these anthers couldn't respond on 1/2 MS medium without plant growth regulators. 2,4-D was a prerequisite substance in this experiment, especially 52.6% of callus formation on MS medium with 2.0mg/L 2,4-D alone. However, the optimum medium was on 1/2 MS medium with 0.1 mg/L 2,4-D and 1.0mg/L BA for continuous growth and shoot differentiation from the anther. Calli derived from on MS medium with 2.0mg/L 2,4-D transferred to the 1/2MS medium with TDZ and BA. TDZ were less superior to BA, only one anther could produce shoot on MS media with 1.0mg/L TDZ. On the other hand, when the calli transferred to the medium with 3.0mg/L BA, adventitious shoots were proliferated, subsequently, regenerated shoots elongated from the embryogenic calli. After floral buds of one week before anthesis were incubated at $5^{\circ}C$ refrigerator for eight or fifteen days, anthers seperated from floral buds were cultured on 1/2MS medium supplemented with 0.1mg/L 2,4-D and 1.0mg/L BA. Callusing and shoot differentiation on anthers from treated at $5^{\circ}C$ for eight days were more effective than those of fifteen days or control.

• Korean Journal of Food Science and Technology
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• v.29 no.6
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• pp.1248-1254
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• 1997

To measure antioxidative activities, the various extracts from RVS (Rhus Verniciflua Stokes) were tried out with either DPPH or thiocyanate method. Also we used the GO (Glucose Oxidase) 20 mU/mL hydroxyl radical system in mouse whole brain cell culture. Chloroform, n-hexane or ethanol were used as extract solutions which had different polarity respectively. In DPPH and thiocyanate method, the antioxidative activities of the crude ethanol extracts were stronger than other extracts. The crude ethanol extracts were fractionated 5 peaks by glass column. Among of them, antioxidative activity of peak II $(P_{II})$ was shown stronger than other fractions, a little for peak III $(P_{III})$ and peak IV $(P_{IV})$, and none for peak I $(P_I)$ and Peak V $(P_V)$. In the antioxidative effects of crude ethanol extracts (30 mg/mL), cell viabilities were evaluated $1\;{\mu}L\;(297\;{\mu}g/mL)$, $2\;{\mu}L\;(588\;{\mu}g/mL)$ of crude ethanol extracts 59%, 68% respectively. $10\;{\mu}L\;(2,727\;{\mu}g/mL)$ addition of crude ethanol extracts had 95% cell viabilities, 0.01% significant, comparing control. In addition, the compounds related to antioxidative effect of crude ethanol extract might be glycoproteins by means of SDS-PAGE. Comparison to antioxidative effects between several antioxidants (ascorbic acid, ${\alpha}-tocopherol$, catalase) $273\;{\mu}L/mL$ addition of crude ethanol extracts corresponds to $1\;{\mu}g/mL$ catalase in antioxidative effects.

• Korean Journal of Food Science and Technology
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• v.17 no.5
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• pp.389-398
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• 1985

To shorten the processing of cheese slurry, four different slurries, ie, Control, Cheddar 1 and 2, and Italian-type that were made of Na-caseinates, cream, trace elements, lactic culture, and enzymes were fermented at $30^{\circ}C$ for 7days with daily stirring. PH, titratable acidity, soluble nitrogen, viable cell count, active SH groups, total volatile fatty acid, free fatty acid, electrophoretic patterns of degraded caseins, and viscosity were analyzed to investigate physicochemical properties of fermented slurries. Acid production was accelerated in the cheese slurries with protease than that without the enzyme and PH of the former was decreased after three days of fermentation to 4.90. The Change of titratable acidity agreed to PH patterns. Soluble nitrogen of the Control slurry was increased slowly for four days and then rapidly to 40% of total nitrogen while those containing protease to 70%. The protease of lactic cultures used (Streptococcus lactis and Streptococcus cremoris) broke down as-casein more rapidly than $\beta$-casein and most proteins were degraded to peptides and amino acids after three days of fermentation. Total volatile fatty acids were increased by added lipase and free fatty acids composition analyzed by GLC in cheddar slurry with 0.00001% lipase was similar to that of commercial cheddar cheese, while that in Italian-type slurry was a half of that in commercial Italian cheese. Active SH groups were increased in the cheese slurries with glutathione from fourth day of fermentation. The viscosity of slurries decreased very rapidly by addition of protease.

• Journal of Life Science
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• v.13 no.3
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• pp.324-332
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• 2003

The objective of this study was to examine if transient transfection of CAR can transactivate CYP2B1 PBRU reporter gene in COS cells in which the endogenous CYP2B1 gene is not induced by PB. In non-transfeced cells of both Hep G2 and COS, the endogeneous expression of CAR was not detected by antibody against CAR. When cultured cells were transfected with CAR expression plasmid, mCAR1-GFP, both cell types expressed high levels of CAR protein and could allow to examine the effect of CAR in PBRU transactivation. Both cell types expressed endogenous RXR and transfection of RXR expression plasmid dramatically increased its protein expression. Whereas CAR transactivated PBRU2C1Luciferase about 12 fold as compared to 2C1Luciferase in Hep G2 cells, it did not stimulate the luciferase activity of the PBRU reporter gene in COS cells. These results indicate that Hep G2 cells can respond to CAR differently from COS cells, and suggest that factors other than CAR and RXR may be required in inducing PBRU activation and the expression of these factors may be different between liver and kidney.

• Journal of Life Science
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• v.16 no.7 s.80
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• pp.1133-1140
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• 2006

Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.

• Journal of Life Science
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• v.21 no.1
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• pp.119-126
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• 2011

There is a growing recognition of the significance of $H_2S$ as a biological signaling molecule involved in vascular and nervous system functions. In mammals, two enzymes in the transsulfuration pathway, cystathionine ${\beta}$-synthase (CBS) and cystathionine ${\gamma}$-lyase (CGL), are believed to be chiefly responsible for $H_2S$ biogenesis. Genetic inborn error of CGL leads to human genetic disease, cystathioninuria, by accumulating cystathionine in the body. This disease is secondarily associated with a wide range of diseases including diabetes insipidus and Down's syndrome. Although the human CGL (hCGL) overexpression is essential for the investigation of its function, structure, reaction specificity, substrate specificity, and protein-protein interactions, there is no clear report concerning optimum overexpression conditions. In this study, we report a detailed analysis of the overexpression conditions of the hCGL using a bacterial system. Maximum overexpression was obtained in conditions of low culture temperature after inducer addition, performing low aeration during overexpression, and using a low concentration inducer (0.1 mM, IPTG) for induction. Expressed hCGL was purified by His-tag affinity column chromatography and confirmed by Western blot using hCGL antibody and enzyme activity analysis. We also report that the His tag with TEV site attached protein exhibits 76% activity for ${\alpha}-{\gamma}$ elimination reaction with L-cystathionine and 88% for ${\alpha}-{\beta}$ elimination reaction with L-cysteine compared to those of wild type hCGL, respectively. His tag with TEV site attached protein also exhibits a 420 nm absorption maximum, which is attributed to the binding cofactor, pyridoxal 5'-phosphate (PLP).

• Journal of Life Science
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• v.21 no.12
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• pp.1752-1760
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• 2011

In this study, the effect of the Opuntiahumifusa extracts on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and ROS level of a cell was investigated using an osteoblast. Opuntiahumifusawas separated intoOpuntiahumifusapeel (OH-P), seed (OH-Se) and stem (OH-St).These were subjected to extraction by using hot water and ethanol. The proliferation of the MC3T3-E1 osteoblastic cells that were treated with OH-Se water extract were increased by approximately 120%. Regarding the effects of OH-Se on ALP activity, the $50{\mu}g/ml$ ethanol extract group showed the highest activity. The synthesis of collagen increased significantly in response to treatment with OH-Se water extract. The ROS scavenging effects of Opuntiahumifusawere investigated for involvement of oxidativedamage, cell culture and staining. Also, when OH-Se water extract $100{\mu}g/ml$ was added, the ROS level decreased by 54%. These results indicate that Opuntiahumifusa extracts have an anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.