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Antioxidative Effects of Ethanol Extracts from Rhus Verniciflua Stokes (RVS) on Mouse Whole Brain Cells (옻나무 에탄올 추출물의 쥐 뇌세포에 대한 항산화효과)

  • Lim, Kye-Taek;Shim, Jae-Han
    • 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.

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Purification of Human HtrA1 Expressed in E. coli and Characterization of Its Serine Protease Activity (E. coli에서 발현된 human HtrA1 단백질의 정제와 HtrA1의 serine protease 활성 조건에 관한 연구)

  • Kim, Kyung-Hee;Kim, Sang-Soo;Kim, Goo-Young;Rhim, Hyang-Shuk
    • 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.

Study for the Synthesis of $[^{123}I]$Idoxifene and Its Uptake in the Breast Cancer Cell ($[^{123}I]$Idoxifene 합성과 유방암의 세포섭취에 관한 연구)

  • Cho, Young-Sub;Yang, Seung-Dae;Suh, Yong-Sup;Chun, Kwon-Soo;Ahn, Soon-Hyuk;Lim, Soo-Jung;Lim, Sang-Moo;Yu, Kook-Hyun
    • The Korean Journal of Nuclear Medicine
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    • v.34 no.5
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    • pp.410-417
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    • 2000
  • Purpose: Idoxifene is currently entering phase II clinical trials for the treatment of advanced breast cancer. The radiolabeled idoxifene using $[^{123}I]$ provides an opportunity for clinical pharmacology with single photon emission computed tomography (SPECT). The purpose of this study was to prepare radiolabeled idoxifene using $[^{123}I]$ and to determine its cell uptake of breast cancer cell line. Materials and Methods: With a view to evaluating new anticancer drugs, we are investigating the novel antiestrogen pyrrolidino-4-iodotamoxifen (idoxifene). $[^{123}I]$Idoxifene has been prepared in no-carrier-added form using a tributyl stannylated precursor which has been synthesized by means of (2-chloroethoxy)benzene with (${\pm}$)-2-phenylbutanoic acid on the basis of previously reported standard methods. The biodistribution and dynamic behavior of the compound were investigated using the comparative breast cancer cell line, MCF-7 (estrogen receptor-positive) and MDA-MB-468 (non-estrogen receptor). Results and Conclusion: Acylation of (2-chloroethoxy)benzene with (${\pm}$)-2-phenylbutanoic acid gave the versatile ketone (81%) which reacted with 1,4-diiodobenzene to give triphenylethylene as a mixture of E and Z geometric isomers, which were separated by the recrystallization in ethanol. The E-isomer was treated with pyrrolidine to give idoxifene (67%). In order to incorporate radioactive iodine into the 4-position, the 4-stannylated precursor was prepared (30%). The yield of radioiodination was 90-92% with a high radiochemical purity greater than 98%. The ratio of tumor uptake of the breast cancer cell line between MCF-7 and MDA-MB-468 was about 1.7.

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Formation of Metastases in C57 BL/6 Mouse Lung (LL/2 Cells접종에 의한 C57 BL/6 Mouse 폐에서의 전이과정)

  • Kim, Ji-Hyun;Lee, Ji-Young;Kim, Woo-Kap;Hwang, Woo-Ik
    • Applied Microscopy
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    • v.29 no.2
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    • pp.137-147
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    • 1999
  • Comparative differences between the fine structure of cultured LL/2 cell in vitro and tumor cells in vivo which were induced in the lung by inoculation of LL/2 cells to C57 BL/6 mouse via tail vein during 21 days are not observed except for cell configuration which was changed spindle shape into oval shape. At first tumor cells appeared at lymphatic nodules and around capillary in the lung. Tumor cells divided actively by mitosis, so they became tumor nodules. The pulmonary aveoli around tumor nodules were observed somewhat flattened in shape but the cells in the aveoli appeared to be in normal condition. Furthermore the normal lung cells were observed in the tumor nodules and some apoptotic tumor cells appeared in the large tumor nodules. A lot of neutropiles were observed in the aveoli and tumor nodules of C57 BL/6 mouse lung after inoculation 22 days and 31days.

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Isolation and Evaluation of an Antiviral Producing Serratia spp. Strain Gsm01 against Cucumber mosaic virus in Korea (한국에서 CMV에 항바이러스 효과를 나타내는 Serratia spp. Gsm01 균주의 분리 동정 및 효과 검정)

  • Ipper, Nagesh S.;Lee, Seon-Hwa;Suk, Jung-Ki;Shrestha, Anupama;Seo, Dong-Uk;Park, Duck-Hwan;Cho, Jun-Mo;Park, Dong-Sik;Hur, Jang-Hyun;Lim, Chun-Keun
    • The Korean Journal of Pesticide Science
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    • v.10 no.4
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    • pp.344-350
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    • 2006
  • An Antiviral producing bacterial strain was isolated from ginseng root environment in Hongcheon, Kangwon province of Republic of Korea. Identification of this bacterial strain was performed by physiological and biochemical tests along with 16S rRNA analyses. The results revealed that the bacterium was closer to genus Serratia, which was named as Gsm01. The strain was grown in Mannitol-Glutamate-Yeast (MGY) broth for 48 h. The culture was centrifuged and the filtrate obtained was tested for its ability to control Cucumber mosaic virus strain Y (CMV-Y) in greenhouse and field experiments. In the green house experiments, CF was evaluated for its ability to protect local host, Chenopodium amaranticolor and systemic host of CMV, Nicotiana tabacum cv. Xanthi-nc. It was found that, CF treatment reduced viral infection by 98% in local host; C. amaranticolor. The N. tabacum cv. Xanthi-nc plants treated with CF did not show visible viral symptoms 15 days post inoculation (dpi) and remained symptomless throughout the periods of the study. To evaluate effectiveness of CF under field conditions, experiment was carried out in a polyvinyl house. It was observed that, 52% plants were protected from viral diseases compared to non-treated plants, increasing the crop yield. This is the first report showing antiviral activity of a Serratia spp. against CMV.

Studies on the Enzyme of Rhizopus oryzae - Part I. Production of Acid Protease and Enzymatic Characteristics - (Rhizopus oryzae의 효소(酵素)에 관(關)한 연구(硏究) - 제(第)1보(報), 산성(酸性) Protease 생산(生産) 및 효소(酵素)의 특성(特性) -)

  • Hou, Won-Nyong;Chung, Man-Jae
    • Applied Biological Chemistry
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    • v.22 no.3
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    • pp.135-141
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    • 1979
  • These experiments were conducted to investigate the conditions of the acid protease production by Rhizopus oryzae and the characteristics of crude enzyme. The results obtained were as follows: 1. The optimum culture time and the optimum amount of added water to the wheat bran medium were about 48 hrs and $80{\sim}120%$, respectively. 2. The addition of $(NH_4)_6Mo_7O_{24},\;(NH_4)_2SO_4,\;NH_4NO_3$, casein, and albumin, respectively, as nitrogen sources to the wheat bran medium was effective. Of these, the optimum concentrations of addition of $(NH_4)_6Mo_7O_{24}$ and casein which were the most effective were 0.1% and 1.0%, respectively. 3. The addition of glucose, galactose, maltose, lactose, and soluble starch, respectively, as carbon sources to the wheat bran medium was effective. Of these, the optimum concentration of addition of glucose which was the most effective was 3.0%. 4. The addition of $KH_2PO_4$ as a phosphate salt to the wheat bran medium was effective. The optimum concentration of addition of $KH_2PO_4$ was 0.3%. 5. The optimum pH for the enzyme action was 2.4, the optimum temperature about $40^{\circ}C$, and the stable pH range $2.0{\sim}5.0$. The enzyme was stab1e below $40^{\circ}C$. 6. The enzyme activity increased rapidly for 10 minutes after addition, thereafter it increased slowly. 7. The enzyme activity increased rapidly to 2 ml of addition, but nearly did not increase at the amounts greater than 2ml.

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Radioprotective Effect of Red Ginseng in Irradiated Mice with ${\gamma}$-ray (생쥐에서 홍삼의 감마선조사에 의한 방어효과)

  • Seung, Ka-Yeon;Lee, Heung-Man;Kim, Jong-Sang;Lee, Jun-Haeng
    • Journal of the Korean Society of Radiology
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    • v.4 no.1
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    • pp.31-35
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    • 2010
  • Recently, the incidents of direct or indirect radiation exposure due to increase of use of radiation or radioisotope are on the increase in medical and industrial circles. If cells are irradiated, free radicals are created through biological process, and cells are directly or indirectly damaged. This research intends to explore into the effect of saponin at the level of cell (in vitro) and entity (in vivo), using red ginseng extract "saponin", as radioprotective agent. In the experiment implemented at the level of cell (in vitro), degree of cell activity was measures by adding mouse mesenchymal stem cells "C3H/10T1/2 cells" into red ginseng extract "saponin(0, 0.05, 0.2, and 0.4 g/L)", and then the optimal concentration of saponin influencing cells was calculated, in 24, 48, 72, and 96 hours after gamma irradiation at the optimal concentration of saponin, each cell survival rate was observed through XTT assay. The best time period of cultivation for the optimal activity of C3H/10T1/2 cells was as 48 hours, and the degree of optimal activity was shown at 0.05 g/L. In 48 hours after irradiation of 5 Gy to C3H/10T1/2 cells at 0.05 g/L, the degree of activity of cells increased by 10%. In the experiment implemented at the level of entity (in vivo), red ginseng extract "saponin" at a dose of 100 mg/kg/day was injected into the abdominal cavity of six-week immature mouse for two weeks. Right after the last abdominal injection, total body irradiation of gamma rays was carried out at a dose of 5 Gy and 10 Gy. And after irradiation, the blood sample was taken, and then the number of red corpuscles was counted. In result, the decrement of experimental group treated with red ginseng extract "saponin" was 2.3 times larger than that of control group. In view of the results so far achieved, it was revealed that red ginseng extract "saponin" has a radiation exposure protection effect in the experiment implemented at the level of cell (in vitro). In case of animal experiment, the decrement of number of red corpuscles decreased. Finally, it is necessary to carry out more various researches continuously.

Protective Effects of a Herbal Composition (HemoHIM) Against Apoptosis Induced by Oxidative Stress of Hydrogen Peroxide (과산화수소의 산화적 스트레스로 유도된 Apoptosis에 대한 생약복합조성물(HemoHIM)의 방호효과 평가)

  • Shin, Sung-Hae;Kim, Do-Soon;Kim, Mi-Jung;Kim, Sung-Ho;Jo, Sung-Kee;Byun, Mung-Woo;Yee, Sung-Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.9
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    • pp.1127-1132
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    • 2006
  • In our previous study, a novel herb mixture (HIM-I) of Angelica gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix was developed to protect the intestinal and immune systems and promote its recovery against radiation damage. A new herbal composition (HemoHIM) with the high immune modulating activity was developed from HIM-I. HIM-I was fractionated into ethanol fraction (HIM-I-E) and polysaccharide fraction (HIM-I-P). And HemoHIM was prepared by adding HIM-I-P to HIM-I. HemoHIM showed more effective than HIM-I in immune modulation as well as radioprotection. The present study is designed to investigate the protective effects of HIM-I, HIM-I-P, and HemoHIM on hydrogen peroxide $(H_2O_2)$ induced apoptosis of human promyelocytic leukemia (HL-60) cells. It was shown that $H_2O_2$ treatment reduced the viability of cells, and increased appearance of DNA ladders, hypodiploid (subG1) cells, and phosphatidylserine translocation level. Pretreatment of HemoHIM significantly reduced the cytotoxic effect induced by $H_2O_2$, associated with reducing the translocation of phosphatidylserine, hypodiploid cells and DNA ladders. HemoHIM appeared to be more protective than HIM-I against $H_2O_2$ induced apoptosis whereas, it exhibited similar activity to HIM-I-P. These results indicated that HemoHIM might be an useful agent for protection against oxidative stress $(H_2O_2)-induced$ apoptosis as well as immune modulation, especially since it is a relatively nontoxic natural product.

A Medium-Maturing and Good Quality Japonica Rice Variety, "Cheongan" (벼 중생 고품질 신품종 "청안")

  • Yang, Sae-Jun;Kim, Yeon-Gyu;Choi, Im-Soo;Cho, Young-Chan;Hwang, Hung-Goo;Hong, Ha-Cheol;Kim, Myeong-Ki;Oh, Myung-Kyu;Shin, Young-Seop;Lee, Jeom-Ho;Choi, Yong-Hwan;Choi, In-Bea;Kang, Kyung-Ho;Yea, Jong-Doo;Lee, Jeong-Heui
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.649-653
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    • 2009
  • "Cheongan" is a new japonica rice variety developed from a cross between SR15225-B-22-1-2-1 and Iksan431 in summer season, 1997 by National Institute of Crop Science, RDA. The line SR15225-B-22-1-2-1 has good canopy architecture and multi-disease and insect resistance, and Iksan431 has translucent milled rice and good eating-quality. Heading date of Cheongan is August 13 in central lowland and mid-mountainous areas. "Cheongan" having culm length of 84 cm shows relatively semi-erect pubescent leaf blade and rigid culm, tolerance to lodging with and good canopy architecture. This variety has 14 tillers per hill and 126 spikelets per panicle. It shows tolerance to heading delay and spikelet sterility comparable to Hwaseongbyeo when exposed to cold stress. Leaf senescence of Cheongan progresses slowly during the ripening stage and the viviparous germination ratio was 59 %, similar to that of Hwaseongbyo. "Cheongan" shows moderately resistance to blast disease, but susceptible to stripe virus and brown planthopper. The milled rice of "Cheongan" exhibits translucent, clear non-glutinous endosperm and medium short grain. It shows similar amylose content of 18.7%, gelatinization temperature, and similar palatability of cooked rice compared to Hwaseongbyeo. The milled rice yield of this cultivar is about 5.54 MT/ha at ordinary season culture in local adaptability test for three years. Especially, "Cheongan" has better milling properties of higher 98.4% and 73.9% in the percentage of head rice in milled rice and milling recovery of head rice, respectively, than those of Hwaseongbyeo. "Cheongan" could be adaptable to the central and mid-southern plain area, and mid-western coastal area of Korea.

Estimation of Precipitable Water from the GMS-5 Split Window Data (GMS-5 Split Window 자료를 이용한 가강수량 산출)

  • 손승희;정효상;김금란;이정환
    • Korean Journal of Remote Sensing
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    • v.14 no.1
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    • pp.53-68
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    • 1998
  • Observation of hydrometeors' behavior in the atmosphere is important to understand weather and climate. By conventional observations, we can get the distribution of water vapor at limited number of points on the earth. In this study, the precipitable water has been estimated from the split window channel data on GMS-5 based upon the technique developed by Chesters et al.(1983). To retrieve the precipitable water, water vapor absorption parameter depending on filter function of sensor has been derived using the regression analysis between the split window channel data and the radiosonde data observed at Osan, Pohang, Kwangiu and Cheju staions for 4 months. The air temperature of 700 hPa from the Global Spectral Model of Korea Meteorological Administration (GSM/KMA) has been used as mean air temperature for single layer radiation model. The retrieved precipitable water for the period from August 1996 through December 1996 are compared to radiosonde data. It is shown that the root mean square differences between radiosonde observations and the GMS-5 retrievals range from 0.65 g/$cm^2$ to 1.09 g/$cm^2$ with correlation coefficient of 0.46 on hourly basis. The monthly distribution of precipitable water from GMS-5 shows almost good representation in large scale. Precipitable water is produced 4 times a day at Korea Meteorological Administration in the form of grid point data with 0.5 degree lat./lon. resolution. The data can be used in the objective analysis for numerical weather prediction and to increase the accuracy of humidity analysis especially under clear sky condition. And also, the data is a useful complement to existing data set for climatological research. But it is necessary to get higher correlation between radiosonde observations and the GMS-5 retrievals for operational applications.