Effects of co-solvent polarity, citric acid, pressure, temperature, run time, and co-solvent ratio on extraction of major flavonoids from Lonicera Flos were investigated using supercritical fluid $CO_{2}(SF-CO_{2})$. HPLC analysis revealed addition of pure methanol resulted in low extraction yield of major flavonoids, luteoloin (Lu), Quercetin (Qu), Apigenin (Ap). Under same condition, as co-solvent polarity increased, yields of major flavonoids increased gradually, At optimum co-solvent extraction condirion of 60% aqueous methanol (10%, v/v), yields of Lu, Qu, and Ap were 42.09, 28.18, and 3.49 mg/100 g, respectively. Addition of citric acid to 60% aqueous methanol gave higher, with addition of 1% citrie acid resulting in highest yields of 63.2 (Lu), 39.35 (Qu), and 5.79 (Ap) mg/100 g. Optimum extraction conditions of major flavonoids were 200 bar, $50^{\circ}C$, 60 min, and $CO_{2}$-methanol-water(20: 1.8: 1.2).
Development of transgenic plant increasing crop yield or disease resistance is good way to solve the world food shortage. However, the persistence of marker genes in crops leads to serious public concerns about the safety of transgenic crops. In the present paper, we developed marker-free transgenic rice inserted high molecular-weight glutenin subunit (HMW-GS) gene ($D{\times}5$) from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation method. Two expression cassettes comprised of separate DNA fragments containing only the $D{\times}5$ and hygromycin resistance (HPTII) genes were introduced separately into Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring $D{\times}5$ or HPTII was infected into rice calli at a 3: 1 ratio of EHA105 with $D{\times}5$ gene and EHA105 with HPTII gene expressing cassette. Then, among 66 hygromycin-resistant transformants, we obtained two transgenic lines inserted with both the $D{\times}5$ and HPTII genes into the rice genome. We reconfirmed integration of the $D{\times}5$ and HPTII genes into the rice genome by Southern blot analysis. Wheat $D{\times}5$ transcripts in $T_1$ rice seeds were examined with semi-quantitative RT-PCR. Finally, the marker-free plants containing only the $D{\times}5$ gene were successfully screened at the $T_1$ generation. These results show that a co-infection system with two expression cassettes could be an efficient strategy to generate marker-free transgenic rice plants.
Koh, Eun Kyoung;Lee, Young Ju;Kim, Ji Eun;Kwak, Moon Hwa;Go, Jun;Son, Hong Joo;Lee, Hee Seob;Jung, Young Jin;Hwang, Dae Youn
Journal of Life Science
/
v.24
no.6
/
pp.595-602
/
2014
Styela Clava tunic (SCT) has found some applications in many areas of medical treatment including as an anti-inflammatory compound, a wound healing film, in guided bone regeneration, and as a food additive. The protective effect of SCT aqueous extract (AE-SCT) on cell death induced by $H_2O_2$ treatment was investigated by measuring the changes in cell viability in HepG2 cells after AE-SCT treatment. High concentrations of antioxidant compounds including flavonoids (3.3 mg/g) and phenolics (32.3 mg/g) were detected in AE-SCT but no significant cytotoxicity was observed in HepG2 cells treated with AE-SCT. The viability of HepG2 cells was also not changed by treatment with different concentrations of AE-SCT after $H_2O_2$ treatment. However, cell viability was significantly increased in cells treated with three different concentrations of AE-SCT before $H_2O_2$ treatment. The greatest increase in cell viability was observed in the group treated with $50{\mu}g/ml$ AE-SCT, when compared with vehicle-treated group. FACS and DAPI staining analysis indicated that the decrease in number of dead cells was dependent on the concentration of AE-SCT. Alterations in the Bax/Bcl-2 ratio after $H_2O_2$ treatment were significantly restored by treatment with different concentrations of AE-SCT. These results indicate that AE-SCT, which contains high levels of antioxidants, may protect cells against death induced by $H_2O_2$ treatment.
This study was performed to evaluate the anti-oxidative, anti-inflammatory, anti-allergy, and whitening effects of Zizania latifolia ethanol extracts prepared from 5 different ethanol concentrations (10, 30, 50, 70, and 90%). As the ethanol concentration in the extraction solvent was increased, the radical scavenging activities also increased. The inhibitory activity of Z. latifolia ethanol extracts on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells tended to increase as the content of ethanol increased. The highest inhibitory activity was obtained with 70% ethanol extract. The antiallergy effects of Z. latifolia ethanol extracts were tested by measuring the release of ${\beta}-hexosaminidase$ in IgE-sensitized RBL-2H3 cells. The suppressive effect of Z. latifolia ethanol extracts increased in a dose-dependent manner as the proportion of ethanol increased, except for the 10% ethanol extract. Furthermore, the inhibitory effects of Z. latifolia ethanol extracts against melanin production in ${\alpha}-melanocyte$ stimulated hormone (MSH)-stimulated B16F0 cells increased as the ethanol ratio increased, and 70 and 90% ethanol extracts showed similar inhibitory activities to arbutin, a positive control, at $250{\mu}m$. The present study confirmed the efficacy of Z. latifolia ethanol extracts in various areas, demonstrating antioxidative, anti-inflammation, antiallergy, skin protective, and skin whitening effects, with no cytotoxicity. It could be used as a raw material in functional foods, as well as in cosmetics.
The optimization of supercritical water oxidation (SCWO) process for decomposing nitromethane was studied by means of a design of experiments. The optimum operating region for the SCWO process to minimize COD and T-N of treated water was obtained in a lab scale unit. The authors had compared the results from a SCWO pilot plant with those from a lab scale system to explore the problems of scale-up of SCWO process. The COD and T-N in treated waters were selected as key process output variables (KPOV) for optimization, and the reaction temperature (Temp) and the mole ratio of nitromethane to ammonium hydroxide (NAR) were selected as key process input variables (KPIV) through the preliminary tests. The central composite design as a statistical design of experiments was applied to the optimization, and the experimental results were analyzed by means of the response surface method. From the main effects analysis, it was declared that COD of treated water steeply decreased with increasing Temp but slightly decreased with an increase in NAR, and T-N decreased with increasing both Temp and NAR. At lower Temp as $420{\sim}430^{\circ}C$, the T-N steeply decreased with an increase in NAR, however its variation was negligible at higher Temp above $450^{\circ}C$. The regression equations for COD and T-N were obtained as quadratic models with coded Temp and NAR, and they were confirmed with coefficient of determination ($r^2$) and normality of standardized residuals. The optimum operating region was defined as Temp $450-460^{\circ}C$ and NAR 1.03-1.08 by the intersection area of COD < 2 mg/L and T-N < 40 mg/L with regression equations and considering corrosion prevention. To confirm the optimization results and investigate the scale-up problems of SCWO process, the nitromethane was decomposed in a pilot plant. The experimental results from a SCWO pilot plant were compared with regression equations of COD and T-N, respectively. The results of COD and T-N from a pilot plant could be predicted well with regression equations which were derived in a lab scale SCWO system, although the errors of pilot plant data were larger than lab ones. The predictabilities were confirmed by the parity plots and the normality analyses of standardized residuals.
Journal of the Korean Institute of Traditional Landscape Architecture
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v.28
no.3
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pp.114-121
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2010
Traditional temples in Korea are the important cultural heritage because of artistic traditonal buildings and structures, paintings, sculptures, and big forest areas which are most ecological and beautiful in Korea. Since traditional temples still function as religious places for very long time, the sense of places intrinsic to the temples are very strong and vivid. The sense of place is very closely related to the conservation of the original landscape type. Recently however, there is a strong tendency to use solar energy in traditional temples because of the low energy efficiency of the old traditional architecture which may have negative impact on landscape which again in turn may lead to the destruction of the sense of place. The purpose of this study was to suggest some landscape design guidelines to protect the sense of place of traditional temple by investigating the impact of solar energy facility on the landscape experience of traditional temple. In order to do perform this purpose, Tongdosa was selected as a study site and four kinds of measurement tools(landscape image, temple identity, landscape satisfaction, degree of landscape improvement) were used as questionnaire items. 180 college students participated in the questionnaire survey. The analysis showed that the solar energy facility had very negative impact on landscape experience such as three landscape image factors(scenic beauty, openness, complexity), landscape satisfaction, temple identity, and landscape improvement. Based on the results, three landscape improvement plans were suggested. First, solar energy facility should be built in the forest in order not to be exposed to visitors, if possible. Second, the landscape management of traditional temple should emphasize on sustaining scenic beauty and temple identity along with the provision of openness. Lastly, detailed landscape guideline should be prepared to regulate the scale, ratio, and the form of the artificial buildings and structures to protect the sense of place of traditional temple.
Journal of the Korean Institute of Traditional Landscape Architecture
/
v.34
no.3
/
pp.28-37
/
2016
Through the research and analysis on the hydrological environment and soil environment of habitats through documentary and field studies over 14 habitats of Euryale ferox Salisb. within Jeollabukdo, with the objective of acquiring the basic data for forming an environment based on plantation of reservoirs that are composed with Euryale ferox, the following results were obtained. As a result of analyzing the construction period of the habitats of Euryale ferox from a total of 14 places, the average period of duration after construction of all subject reservoirs appeared to be 71.8 years. Moreover, when examining the relationship between the age of reservoirs and eutrophication, it could be judged that at least the eutrophication of subsoil and water environment is not an obstacle to the growth of Euryale ferox grows in habitats that have a reservoir age of approximately 70 years or more. As a result of analyzing the gardening of soil sediment of the Euryale ferox habitats, the component ingredients appeared to be composed of 80.2% of clay, 16.7% of silt and 3.1% of sand, and the soil class pursuant to such was classified as 'heavy clay'. The organic matter contents of soil sediment appeared to be an average of 36g/kg, and there appeared to be no noticeable difference between the habitats and non-habitats of Euryale ferox. The water quality environment of Euryale ferox habitat appeared to be pH 6.5~7.9, concentration of dissolved oxygen to be $1.8{\sim}8.8mg/{\ell}$, concentration of COD to be $6.8{\sim}74mg/{\ell}$, floating materials to be $2.0{\sim}213mg/{\ell}$, total nitrogen to be $0.422{\sim}10.723mg/{\ell}$, and phosphate to be $0.003{\sim}0.126mg/{\ell}$. The average DO concentration of Aedang Reservoir at Jeongeup, Daejeong Reservoir at Imsil, and Myeongdeokji at Gimje with high vitality and green coverage ratio of Euryale ferox appeared to be $3.5mg/{\ell}$, total nitrogen to be $1.33mg/{\ell}$, and concentration of phosphorus-phosphate to be $0.061mg/{\ell}$. When comparing such with the entire average value, the DO and total nitrogen concentration appeared to be rather low, and the phosphorus-phosphate concentration appeared to be higher by two times or more, thus, an in-depth study on the correlation of the vitality of Euryale ferox Salisb. and concentration of phosphorate-phosphorus will be needed in the future.
$TiO_{2}$/Se : Te heterojunction for color sensor has been fabricated by RF reactive sputtering and thermal evaporation methods onto glass substrate. The optimum deposition condition of $TiO_{2}$ films was such that RF power was 120 W, substrate temperature was $100^{\circ}C$, oxygen concentration was 50%, working pressure was 50 mTorr for the $TiO_{2}$ film thickness of $1000{\AA}$. In this case, the optical transmittance of $TiO_{2}$ film at 550 nm-wavelength was 85%, resistivity was $2{\times}10^9{\Omega}{\cdot}cm$, refractive index was 2.3, and optical bandgap was 3.58 eV. The composition ratio of 0 to Ti by AES analysis was 1.7. When $TiO_{2}$ films were annealed at $400^{\circ}C$ for 30 min. in $O_{2}$ ambient, the optical transmittance of $TiO_{2}$ films at the wavelength range of $300{\sim}580$ nm was improved from 0 to 25%. When Se : Te films were annealed at $190^{\circ}C$ for 1 min., photosensitivity under illumination of 1000 lux was 0.75. The optical bandgap of Se : Te films was 1.7 eV. The structures of Se : Te films were the hexagonal with (100) and (110) orientation. The spectral response of a-Se was improved by the addition of Te, especially in the long wavelength region. The $TiO_{2}$/Se : Te heterojunction showed wide spectral response, and more improved one than that of a-Si film in the blue light region.
Soybean [$Glycine$$max$ (L.) Merr.] protein is a high quality source for food and feed. But, antinutritional factors in the raw mature soybean are exist. Kunitz trypsin inhibitor (KTI) protein is a main antinutritional factor in soybean seed. Also, P34 protein, referred as $Gly$$m$ Bd 30K, has been identified as a predominant immunodominant allergen. Genetic relationship between KTI protein and P34 protein could be useful in soybean breeding program for the genetic elimination or reduction of these factors. The objective of this study was to determine the independent inheritance or linkage between KTI protein and P34 protein in soybean seed. A total of 479 $F_2$ seeds were obtained from the cross of 07B1 and PI567476 parents. KTI protein and relative amount of P34 protein were analysed from $F_2$ seeds harvested from the F1 plants by using SDS-PAGE and Western blot analysis. The segregation ratios of 3 : 1 for KTI protein (353 KTI protein present : 126 KTI protein absent) and relative amount of P34 protein (363 normal amount of P34 protein : 116 low amount of P34 protein). The segregation ratio of 3 : 1 suggested that KTI protein and relative amount of P34 protein in mature soybean seed were controlled by a single major gene. The segregation ratios of 9 : 3 : 3 : 1 (266 KTI protein present, normal amount of P34 protein: 88 KTI protein present, low amount of P34 protein: 102 KTI protein absent, normal amount of P34 protein: 23 KTI protein absent, low amount of P34 protein) and Chi-square value (${\chi}^2$=3.31, P=0.346) were observed in $F_2$ seeds. This data showed that KTI protein was inherited independently with relative amount of P34 protein in soybean. These results will be helpful in breeding program for selecting the line with lacking KTI protein and reduced amount of P34 protein in soybean.
Antifungal compounds from Bacillus polyfermenticus CJ6 were purified using SPE, preparative HPLC, and reverse phase-HPLC. Antifungal compounds from B. polyfermenticus CJ6 were separated into three fractions (8, B, C) using preparative HPLC. LC/MS analysis of antifungal peaks suggested that B. polyfermenticus CJ6 produces lipopeptides; two kinds of iturin A ($C_{14}$, $C_{15}$), three kinds of surfactins ($C_{13}$, $C_{14}$, $C_{15}$), four kinds of fengycin A ($C_{14}$, $C_{15}$, $C_{16}$, $C_{17}$) and two kinds fengycin B ($C_{16}$, $C_{17}$). The antifungal activity of fraction 8, which was presumed as inturin A, was found to be stable after the pH, heat or proteolytic enzyme treatment, but it was unstable at 50-$70^{\circ}C$ for 24 hr. The antifungal activity of fraction B, which presumed as surfactins and fengycin A, was found to be stable after the heat treatment, but it was unstable in the pH 3.0 and after the protease (type I) or ${\alpha}$-chymotrypsin treatment. The antifungal activity of fraction C, which was presumed as fengycin A and B, was found to be stable in the pH 3.0-9.0 range and the heat treatment, but it was unstable with the treatment of protease (type I). The amino acid composition of the purified peaks 8-1 and 8-2 were Asx, Tyr, Gln, Pro, and Ser in a molar ratio of 3:1:1:1:1, which showed the same amino acid composition as iturin. From these results, we confirmed that antifungal compounds from B. polyfermenticus CJ6 most likely belonged to iturin A as well as surfactins and fengycins. As lipopeptides are known to act in a synergistic manner, the antifungal compounds from B. polyfermenticus CJ6 might have potential uses in biotechnology and biopharmaceutical applications.
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