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Potential Reproductive Toxicity Study of p53 Expressing Adenoviral Vector in Mice (아데노바이러스 유전자치료벡터의 생식독성 연구)

  • Rhee, Gyu-Seek;Kwack, Seung-Jun;Kim, Soon-Sun;Lee, Rhee-Da;Seok, Ji-Hyun;Chae, Soo-Young;Chung, Soo-Youn;Kim, Seung-Hee;Lee, Seung-Hoon;Park, Kui-Lea
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.151-158
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    • 2007
  • The possibility of inadvertent introduction of therapeutic gene expressing viral vectors has raised safety concerns about germ-line infection. Particularly, for indications such as prostate cancer and ovarian cancer, the proximity of the point of viral administration to organs of the reproductive system raises concerns regarding inadvertent germ-line transmission of genes carried by the virus vector. To evaluate the safety of in vivo adenovirus mediated gene transfer, we explored the biodistribution, persistance and potential germ-line transmission of p53-expressing adenovirus (Ad-CMV-p53). Both male and female Balb/c mice were injected with $1{\times}10^9$ PFU of Ad-CMV-p53. The PCR analysis showed that there were detectable vector sequences in liver, kidney, spleen, seminal vesicle, epididymis, prostate, ovary, and uterus. The RT-PCR analysis for detecting inserted gene, p53 showed that Ad-CMV-p53 viral RNA were present in spleen, prostate and ovary. Direct injected male and female mice of adenovirus vector into testis and ovary were mated and their of offspring were evaluated for germ-line transmission of the adenoviral vector. The PCR and RT-PCR analysis showed no evidence of germline transmission, although vector sequences were detected in DNA extracted from gonadal tissues. Real-time PCR result confirmed a significant decrease of adenovirus in gonad tissues 1 week after injection. We have also analysed the cell specific localization of viral DNA in gonad tissues by using in-situ PCR. Positive signals were detected in interstitial tissue but not in seminiferous tubule in sperm. In the case of ovary, adenovirus signal were localized to the stromal tissue, but no follicular signals were observed. Together, these data provide strong evidence that the risk of the Inadvertent germ-line transmission of vector sequences following intraperitoneal or direct injection into genito-urinary system of adenovirus is extremely low.

The Influence of 5-Fluorouracil Administration Mode on the Expression of Phospholipase C and Ras Oncoprotein Associated with Regeneration of Rat Intestinal Mucosa Following Radiation (방사선 조사후 백서 공장 점막의 재생과정에서 5-fluorouracil 투여가 phospholipsse C 와 ras 암유전자단백의 발현에 미치는 영향)

  • Park Kyung Ran;Lee Chung Sik;Kim Sung Sook;Lee Young Han;Ryu Sung Ho;Suh Pann-Ghill
    • Radiation Oncology Journal
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    • v.12 no.3
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    • pp.271-284
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    • 1994
  • Purpose : Phospholipase C(PLC) isozymes play significant roles in transmembrane signal transduction. PLC-${\gamma}1$ acts as the intracellular effector in signal transduction for cellular proliferation and differentiation. Ras oncoprotein is also involved in cell growth. We determined the biological significance of PLC and ras oncoprotein in regeneration following radiation and the effect of different modes of administration of 5-FU. Materials and Methods : To determine the effect of the administration mode of 5-FU on the regeneration of intestinal mucosa of rats following radiation, we compared the expression of PLC and ras oncoprotein in six groups. Group I had no treatment. Group II received radiation(8 Gy) only. Group III received radiation(8 Gy) and 5-FU(150mg/kg) continuous intravenous (iv) infusion for 12 hours. Group IV received radiation(8 Gy) and 5-FU(750mg/kg) iv bolus injection. Group V received only 5-FU(150mg/kg) continuous iv infusion for 12 hours, Group VI received only 5-FU (150mg/kg) iv bolus injection. Through immunoblotting and immunohistochemistry, we examined the expression of PLC and ras oncoprotein in rat jejunum at 96 hours after radiation or 5-FU administration and at 120 hours after radiation and 5-FU adminstration. We also investigated the histological findings using hematoxylin and eosin stain. Results : In the immunohistochemistry study, PLC-${\gamma}1$ expression was the highest in group III followed by groups II and VI in that order and was weakly positive in groups V and VI. PLC-${\gamma}1$ was hardly detected in the control group. The expression of ras oncoprotein was the same as the PLC-${\gamma}1$ expression for all groups. These results were confirmed by the histological findings regarding the mucosal regeneration. In the immunoblotting analysis, PLC-${\gamma}1$ expression was the highest in group III followed by group IV and II in that order. This difference between the immunoblotting and immunohistochemistry study was due to the high expression of PLC-${\gamma}1$ on the damaged surface epithelium rather than to its expression in the regeneration region as observed in the immunohistochemistry study for group IV. The expression of PLC-${\delta}1$ was positive only in group V and VI, which received both radiation and 5-FU, and the expression of PLC-${\beta}1$ was negligible for all groups. Conclusion : These results suggest that PLC-${\gamma}1$ mediated signal transduetion and ras oncoprotein may have a significant role in mucosal regeneration after radiation, and that continuous iv infusion of 5-FU may induce active regeneration in intestinal mucosa following radiation. In addition, the expression of PLC-${\delta}1$ in combined group of radiation and 5-FU implies that PLC-${\delta}1$ may be involved in signal transduction mediated by concerted action between radiation and 5-FU.

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Analysis on the Components in Stem of the Lespedeza bicolor (싸리(Lespedeza bicolor) 줄기의 성분 분석)

  • Lee, Yang-Suk;Joo, Eun-Yong;Kim, Nam-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1246-1250
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    • 2005
  • This study was carried out to analyze the components of the stem of bush clover, Lespedeza bicolor to obtain basic data on the studies of the physiological function and a possibility of utilizing L. bicolor. General components of L. bicolor were 82.35$\%$ carbohydrate, 13.38$\%$ moisture, 2.80$\%$ crude protein, 0.86$\%$ crude ash and 0.61$\%$ crude fat. The content of reducing sugar was 147.58 mg/100 g. The total amount of free sugar was 52.4 mg/100 g that was composed of 31.0 mg/100 g fructose, 16.7 mg/100 g glucose and 4.7 mg/100 g sucrose. In the results of mineral analysis, the content of Ca was the highest (472.97 mg/100 g) and followed by 206.70mg/100 g of K. The total amount of hydrolyzed amino acid was 148.95 mg/100 g that consist of 31.74 mg/100g of essential amino acid and 117.24 mg/100 g of non-essential amino acid. Total free amino acids were contained 106.39 mg/100 g that was composed of 8.41 mg/100 g essential amino acids and 97.98 mg/100 g the non-essential, and proline (62.92 mg/100 g) was the highest that account for 59.8$\%$ of total free amino acids. Total content of amino acid derivatives was 30.01 mg/100g and that of $\gamma$ -aminoisobutyric acid was the highest as 12.57 mg/100 g among them.

Introduction of Integrated Coastal Management Program and Sustainable Development of Fishing Villages in Cheonsu Bay Region (연안통합관리계획의 도입과 천수만 어촌의 지속가능발전)

  • 김부성
    • Journal of the Korean Geographical Society
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    • v.38 no.2
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    • pp.184-205
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    • 2003
  • Sustainable Development(SD) is an important concept for the future of the coastal area, and for development of fishing villages. Since 1992 UN Conference on Environment and Development in Rio de Janeiro many governments and local authorities throughout the world have been engaged in preparing and implementing $\ulcorner$Agenda 21$\lrcorner$. Many projects which previously would have been identified as environmental protection are now presented under the banner of sustainable development. Integrated Coastal Management (ICM) is an extension of sustainable development. ICM was presented as a framework for resolution of coastal use conflicts. The aim of the present paper is to assess sustainable development potential of fishing villages in Cheonsu Bay Region according to implementation of ICM. Cheonsu Bay Region was known as one of the productive fishing grounds and Cheonsu Bay Region preserved unique characteristics of traditional fishing villages. But this region is now experiencing many changes through the massive reclamation projects like Seosan A B Project. After a brief overview of concepts and history of SD and ICM, the reclamation process and its impacts on both fishery and fishing communities in Cheonsu Bay Region are discussed. According to their changing environmental and socio-economic characteristics after the reclamation, ca 35 representative coastal villages in this region can be classified into 5 types. Many coastal villages shows diversity in their economic activities, as tourism and recreation function becomes more and more important in this region. In present-day Cheonsu Bay Region, it is possible to differentiate fishing village cooperatives(FVO) with high potential of sustainable fishery development, FVOs with medium potential, FVOs with low potential on the basis of 14 selected indicators.

Antioxidant and Anti-aging Effects of Extracts from Leaves of the Quercusaliena Blume on Human Dermal Fibroblast (피부 섬유아세포에서 갈참나무 잎 추출물의 항산화 및 항노화 효능)

  • Choi, Sun-Il;Lee, Jong Seok;Lee, Sarah;Yeo, Joohong;Jung, Tae-Dong;Cho, Bong-Yeon;Choi, Seung-Hyun;Sim, Wan-Sup;Han, Xionggao;Lee, Jin-Ha;Kim, Jong Dai;Lee, Ok-Hwan
    • Journal of Food Hygiene and Safety
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    • v.33 no.2
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    • pp.140-145
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    • 2018
  • The skin of the human body occupies the largest surface area of the body and acts as a protection for the person's internal organs. As such, the skin is a major target of oxidative stressors, and these oxidative stressors are known to contribute to skin aging over the course of time. For the most part, an antioxidant is an effective approach to utilize to prevent symptoms related to the reactive oxygen species (ROS)-induced aging of the skin. Therefore, we investigated the antioxidant and anti-aging activity of the leaves of the Quercusaliena Blume extract (QBE). In our study, we confirmed that the cell viability tested with XTT {2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide innersalt} assay was not affected up to a concentration of $100{\mu}g/mL$. In addition, the cell viability of HDF cells induced by hydrogen peroxide was recovered from 81% to 104% after treatment with QBE, which showed the greater protective effect than that of ascorbic acid. Treatments of QBE dose-dependently inhibited reactive oxygen species (ROS) production in HDF cells induced by hydrogen peroxide, which correlated with their protective effects on cell viability. Since QBE treatment exhibited the suppression effect of skin aging by decreasing the ROS production, QBE could be used as a not only natural anti-aging but also antioxidant resource.

Effect of Ethanolic Extract of Schizandra chinensis for the Delayed Ripening Kimchi Preparation (오미자(Schizandra chinensis) 추출물이 김치의 과숙억제에 미치는 영향)

  • Moon, Young-Ja;Park, Sun;Sung, Chang-Keun
    • The Korean Journal of Food And Nutrition
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    • v.16 no.1
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    • pp.7-14
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    • 2003
  • This study mainly focused on to investigate the effects of Schizandra chinensis on the growth of a bacterium, CS6 which was isolated from kimchi. CS6 was final]y identified to lactobacillus plantarum that caused acidification of kimchi. The ethanolic extract of Schizandra chinensis(EES) inhibited the growth of L. plantarum. Minimum inhibition concentration of crude EES on L. plantarum was 62.5mg/$m\ell$. In broth culture, 5$\mu\textrm{g}$/$m\ell$ of EES completely inhibited the growth of L. plantarum during fermentation. The addition of 0.4% of EES has no apparent effect on quality including the taste and color on kimchi. It was expected that EES-containing kimchi could extend the period of preservation. Analysis of organic acids in water fractions of EES was carried out by HPLC. It is apparent that antimicrobial active fractions contained the highest concentration of succinic acid, a little tartaric acid and malic acid. Among these organic acids, succinic acid showed the strong inhibitory effect against L. plantarum CS6 in vitro. Succinic acid-containing kimchi with a concentration of 0.4 and 0.5% had the inhibitory effect on growth of L. plantarum. Inhibitory effect of EES on amylase, cellulase and pectinase was also tested. In conclusion, the present experiment demonstrated that EES inhibited the growth of L. plantarum, and various enzyme activity. EES-containing kimchi was sustained the hardness, and initial acidity during fermentation. EES was considered as the possible additive of kimchi process and EES added in kimchi increase the quality, and storage period of kimchi.

Effects of the Glycoprotein Isolated from Pteridium aquilinum on the Immune Function of Mice (고사리 단백다당(Pteridium aquilinum Glycoprotein, PAG)이 마우스 면역활성에 미치는 영향)

  • Park, Hyeon-Ae;Kweon, Mee-Hyang;Han, Hyung-Mee;Sung, Ha-Chin;Yang, Han-Chul
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.976-982
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    • 1998
  • The effects of the glycoprotein (PAG) isolated from Pteridium aquilinum on the immune function was examined in mice. PAG was intraperitoneally administered into BALB/C mice for 14 days and the antibody forming ability to hen egg lysozyme (HEL) and the blastogenic responses of splenocytes were measured. PAG treatment significantly increased antibody formation to HEL in a dose-dependent manner. Blatogenesis of splenocytes in response to lipopolysaccharide (LPS, B-cell specific mitogen) or phytohemagglutinin (PHA, T-cell specific mitogen) was also increased after treatment with PAG, indicating that the PAG increases both humoral and cellular immunities. To examine whether the immune function of PAG was via a direct effect on the lymphocytes, splenocytes were isolated from BALB/C mice, exposed to various concentrations of PAG in vitro and the blastogenic responses were measured. In vitro exposure to PAG significantly increased blastogenesis of splenocytes to LPS up to $500{\;}{\mu}g/kg$, whereas the blastogenic response to PHA was not altered by PAG treatment. To identify the fraction responsible for the increase in the immune function, the effect of periodate digest, pronase digest or purified polysaccharide on the antibody production to HEL was examined. Crude protein fraction of PAG significantly increased the antibody formation to HEL. On the other hand, both crude and purified polysaccharide fractions did not have any effects on the antibody production ability. These data indicated that 1) PAG increased both humoral and cellular immune functions, 2) the increase in humoral immunity was probably via a direct action of PAG on lymphocytes and 3) the protein portion of PAG was responsible for the increase in humoral immunity.

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Components and Their Antioxidative Activities of Methanol Extracts from Sarcocarp and Seed of Zizyphus jujuba var. inermis Rehder (생대추(풋대추) 과육 및 씨 추출물의 항산화 효과 및 항산화 성분)

  • Yu, Mi-Hee;Im, Hyo-Gwon;Lee, Hyo-Jung;Ji, Young-Ju;Lee, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.38 no.1
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    • pp.128-134
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    • 2006
  • Antioxidant activities of methanol extracts from sarcocarp and seed of Zizyphus jujuba var. inermis Rehder were investigated in vitro. Contents of total polyphenols in methanol extracts from sarcocarp and seed were 98.83, $138.99\;{\mu}g/mg$, respectively. Radical-scavenging activities of methanol extracts were examined using ${\alpha},{\alpha}-diphenyl-{\beta}-picrylhydrazyl$ and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radicals, and hydrogen peroxide assay. Inhibition effects of methanol extracts on peroxidation of linoleic acid were examined by ferric thiocyanate and thiobarbituric acid methods. Both sarcocarp and seed of Zizyphus jujuba var. inermis Rehder showed relatively high antioxidant activities in various systems.

Effect of the extracts from Schisandra chinensis Fruit and Morus alba Leaf on Insulin Secretion in Glucose-induced HIT-T15 Cells (오미자와 뽕잎 추출물이 glucose에 의해 유도된 HIT-T15세포의 인슐린 분비능에 미치는 영향)

  • Jeong, Yoo-Seok;Hong, Joo-Heon;Jung, Hee-Kyoung
    • Food Science and Preservation
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    • v.18 no.6
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    • pp.1002-1008
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    • 2011
  • This study aimed to examine the effect of the Schizandra chinensis fruit and Morus alba leaf on insulin expression in HIT-T15 cells, which is exposed by glucose. The total polyphenol contents of the S. chinensis fruit ethanol extract and the M. alba leaf hot-water extract were $20.11{\pm}0.35$ mg/g and $50.02{\pm}0.62$ mg/mL, respectively. The S. chinensis fruit ethanol extract and the M. alba leaf hot-water extract contained $2.85{\pm}0.15$ and $8.76{\pm}0.43$ mg/g flavonoids, respectively. The antioxidant ability of the M. alba leaf hot-water extract was found to be superior to that of the S. chinensis fruit ethanol extract. Compared to the HIT-T15-treated 10 mM 2-deoxy-D-glucose, the $100{\mu}g/mL$ S. chinensis ethanol extract was found to have a two fold increase in insulin productivity. Moreover, the $100{\mu}g/mL$ M. alba leaf hot-water extract promoted the insulin secretion of high-glucose-damaged HIT-T15 almost ten fold. The above results showed that the S. chinensis fruit ethanol extract and M. alba leaf hot-water extract may improve the insulin productivity of the beta cell with glucose-induced oxidative damage. These data suggest that the S. chinensis fruit ethanol extract and the M. alba leaf hot-water extract can be used as food materials for the regulation of insulin secretion.

Effect of 17β-estradiol on Ecdysteroid Pathway Related Genes in the Brackish Water Flea Diaphanosoma celebensis (17β-estradiol이 기수산 물벼룩의 Ecdysteroid 경로에 미치는 영향)

  • In, Soyeon;Yoo, Jewon;Cho, Hayoung;Lee, Young-Mi
    • Journal of Marine Life Science
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    • v.5 no.2
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    • pp.35-42
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    • 2020
  • 17β-estradiol (E2) is a natural hormone secreted by ovary, and continuously discharged from household and livestock wastewater into aquatic environment. Due to its strong estrogenic activity, it has adverse effects on development and reproduction in crustacean as an endocrine disrupting chemical. Although ecdysteroid signaling pathway play a key role in development in crustacean, little information on transcriptional modulation of ecdysteroid-related genes in response to E2 is available in small crustacean. Here, we investigated the acute toxicity of E2 to obtain 24-h LCx values in the brackish water flea Diaphanosoma celebensis. Time-dependent expression patterns of seven ecdysteroid pathway - related genes (CYP314a1, EcRA, EcRB, USP, ERR, Vtg, VtgR) were further examined using quantitative real time reverse transcriptase polymerase chain reaction (qRT-PCR). As results, 24-h LC50 and LC10 values were 9.581 mg/l and 4.842 mg/l, respectively. The mRNA expression of CYP314a1, EcRA, USP, VtgR was significantly up-regulated at 12 or 24 h after exposure to E2. These findings indicate that E2 can affect their molting and reproduction by modulating the expression of ecdysteroid pathway - related in D. celebensis. This study will be useful for better understanding of molecular mode of action of endocrine disrupting chemicals on molting process in small crustacean.