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Production of Useful Proteins by Plant Cell Culture

  • Kwon, Tae-Ho;Kim, Dae-Hyun;Jang, Yong-Suk;Yang, Moon-Sik
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.07a
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    • pp.45-49
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    • 1999
  • Plant cell culture is emerging to express bioactive foreign proteins because it has several advantages in that it is safe, economical, genetically stable and eukaryotic expression system comparing with other expression systems. However several limitations such as slow growth rate, low expression level and lack of well established down stream process need to be answered. As a preliminary approach to produce the immunologically interested molecules through the plant cell culture, we tested if granulocyte-macrophage colony stimulating factors (GM-CSFs) from both murine (mGM-CSF) and human (hGM-CSF) are produced as a biologically active form through plant cell culture. The murine and human GM-CSF genes were cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring both recombinant GM-CSF (rGM-CSF) genes. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and cell suspension cultures. In addition, the biological activities of both murine and human GM-CSF from plant cell culture were confirmed by measuring the proliferation of the GM-CSF dependent FDC-PI and TF-1 cells, respectively.

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Antiviral activity of 20(R)-ginsenoside Rh2 against murine gammaherpesvirus

  • Kang, Soowon;Im, Kyungtaek;Kim, Geon;Min, Hyeyoung
    • Journal of Ginseng Research
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    • v.41 no.4
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    • pp.496-502
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    • 2017
  • Background: Ginsenosides are the major components of Panax ginseng Meyer, an herbal medicine used for the treatment of various diseases. Different ginsenosides contribute to the biological properties of ginseng, such as antimicrobial, anticancer, and immunomodulatory properties. In this study, we investigated the antiviral effects of 15 ginsenosides and compound K on gammaherpesvirus. Methods: The antiviral activity of ginsenosides was examined using the plaque-forming assay and by analyzing the expression of the lytic gene. Results: 20(R)-Ginsenoside Rh2 inhibited the replication and proliferation of murine gammaherpesvirus 68 (MHV-68), and its half-maximal inhibitory concentration ($IC_{50} $) against MHV-68 was estimated to be $2.77{\mu}M$. In addition, 20(R)-ginsenoside Rh2 inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced lytic replication of human gammaherpesvirus in the Kaposi's sarcoma-associated herpesvirus (KSHV)-positive cell line BC3. Conclusion: Our results indicate that 20(R)-ginsenoside Rh2 can inhibit the replication of mouse and human gammaherpesviruses, and thus, has the potential to treat gammaherpesvirus infection.

Development of Secretory Element in Murine Trachea and Larynx (백서 기관 및 후두에서 분비조직의 발생)

  • 조정일;박기현;김광문
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1993.05a
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    • pp.67-67
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    • 1993
  • Mucosal surface of the trachea and larynx is protected by several complex defense mechanism, namely mucociliary clearance, immunoglobulin, antibacterial secretory enzyme, which have also been demonstrated in the middle ear mucosa and eustachian tube. The morphologgy of secretory glands and cells of the trachea and larynx is well-known, but knowledge concerning their development related to secretory activity is still sketchy. The secretory element of the murine trachea and larynx, aging from gestational day 16 to postnatal day 21, was studied using hematoxylin & eosin and alcian blue/periodic acid-Schiff staining including lysozyme immunohistochemistry to investigate the development of secretory element of the murine trachea and larynx and to provide with basis of the future study for developmental morphology of the trachea and larynx. The results of this study suggest that the secretory activity starts to be established immediately after birth with the aeration of the lung.

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Effect of Myricetin on mRNA Expression of Different Antioxidant Enzymes in B16F10 Murine Melanoma Cells (B16F10 Murine Melanoma Cell에서 Myricetin이 항산화효소의 m-RNA 발현에 미치는 영향)

  • Yu Ji Sun;Kim An Keun
    • YAKHAK HOEJI
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    • v.49 no.1
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    • pp.86-91
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    • 2005
  • Flavonoids are class of polyphenolic compounds widely distributed in the plant kingdom, which display a variety of biological activities, including antiviral, antithrombotic, antiinflammatory, antihistaminic, antioxidant and free-radica 1 scavenging abilities. The antioxidant enzyme (AOE) system plays an important role in the defense against oxidative stress insults. To determine whether flavonoid, myricetin can exert antioxidative effects not only directly by modulating the AOE system but also scavenging free radical, we investigated the influence of the flavonoid myricetin on cell viability, different antioxidant enzyme activities, ROS level and the expression of different antioxidant emzyme in B16F10 murine melanoma cells. Myricetin in a concentration range from 6.25 to $50\;{\mu}M$ decreased superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzyme activities, but catalase (CAT) activity was increased. In the myricetin-treated group, ROS levels were decreased dose-dependently. Antioxidant enzyme expression was measured by RT-PCR. Myricetin treatment of B16F10 cells increased catalase expression. Expression levels of copper zinc superoxide dismutase (CuZn SOD) were not affected by exposure of myricetin. Manganese superoxide dismutase (Mn SOD) and GPx expression levels decreased slightly after myricetin treatment. In conclusion, the antioxidant capacity of myricetin was due to CAT and free-radical scavenging.

Change of ROS Generation and Antioxidant Enzyme Activity of Flavonol Quercetin in the Presence of Vitamin E, L-Ascorbit acid, Reduced Glutathione on the B16F10 Murine Melanoma Cells (B16F10 세포에서 Quercetin과 Vitamin E, L-Ascorbic acid, 환원형 글루타치온과의 병용 투여에 의한 활성산소종 발생과 항산화 효소의 활성 변화)

  • 허정심;김안근
    • YAKHAK HOEJI
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    • v.47 no.6
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    • pp.432-437
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    • 2003
  • It has been known that quercetin, a bioflavonoid widely distributed in fruits and vegetables as dietary-derived flavonoid and exert significant multiple biological effects such as antioxidant and anti-inflammatory, anti-tumor effects. In addition, it has been shown to have a chemoprotective role in cancer, though complex effects on signal transduction involved in cell proliferation and angiogenesis. The present study investigated whether quercetin can enhance antioxidant enzyme activity (glutathione peroxidase: GPx, superoxide dismutase: SOD, catalase: CAT) and regulate the reactive oxygen species (ROS) generation in the presence of vitamin E, L-ascorbic acid, reduced glutathione (GSH) on B16F10 murine melanoma cells. After 48h treatment of cells with quercetin in the presence of vitamin E, L-ascorbic acid, GSH, we measured the cytotoxicities by MTT assay. The cells exhibited a dose-dependent inhibition in their proliferation in the presence of vitamin E, L-ascorbic acid, GSH respectively. We also investigated the effects of antioxidant enzyme activity and ROS generation. The antioxidant enzyme activity of quercetin in the presence of vitamin E was stronger than GSH, L-ascorbic acid, the same treatments decreased ROS generation in B16F10 murine melanoma cells. Taken together, these result demonstrate that the antioxidant effect of quercetin can enhanced in the presence of vitamin E and it might plays an important role in anti-oxidative effects.

Pycnogenol Supplementation Retards Immune Dysfunction in Murine AIDS (MAIDS) After LP-BM5 Leukemia Virus Infection by Modulating Cytokine Secretion

  • Lee, Jeong-Min;Park, Kun-Young;Hwang, Kwon-Tack;Watson, Ronald R.
    • Preventive Nutrition and Food Science
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    • v.10 no.2
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    • pp.161-166
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    • 2005
  • We investigated the effect of pycnogenol (PYC) supplementation on retarding the immune dysfunction of CS7BL/6 mice after murine AIDS (MAIDS) development. Dysfunction of T and B cell mitogenesis from primary cultured splenocytes has been observed with retrovirus infection and PYC supplementation partially recovered the dysfunction of T and B cells. There was an abnormal shift of cytokine pattern with retrovirns infection, which was designated by the decreased secretion of Th1 cytokines and increased secretion of Th2 cytokines. PYC supplementation increased IL-2 and $IFN-\gamma$ secretion and decreased IL-4, IL-6, and $TNF-\alpha$ secretion, but it was not sufficient enough to maintain the normal level of these cytokines. Hepatic vitamin E level was significantly decreased by retrovirns infection, in accordance with increased hepatic lipid peroxidation level, whereas PYC supplementation normalized the hepatic level of vitamin E and lipid peroxidation. This study suggests that PYC supplementation may partially help retard the incidence of symptoms during MAIDS.

Antifungal Effect of Brachyglottis repanda Ethanol Extract

  • Yook, Chan-Nam;Na, Young-Soon;Choi, Hwa-Jung;You, Il-Soo;Baek, Jong-Min;Baek, Seung-Hwa
    • Toxicological Research
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    • v.26 no.2
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    • pp.117-121
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    • 2010
  • The crude ethanol extract of B. repanda showed the cytotoxic activity against Polio virus (25% activity at $150{\mu}g$/disk) and the minor cytotoxic activity against BSC cells (African green monkey kidney). However, the crude ethanol extract of B. repanda was non-toxic to murine leukaemia cells CCL 46 P388D1 ($IC_{50}$, > 62,500 ng/ml). Cytotoxic and antifungal activities were strongly shown by Fr. 64-3 which was eluted with 90% $CH_3CN/H_2O$, 100% $CH_3CN$, and 50% $CH_3CN/H_2O$(SM 2 at $150{\mu}g$/disk). The fraction 64-3 also showed the most cytotoxic activity against murine leukaemia cells (128 mg, $IC_{50}$ 10,051 ng/ml at $75{\mu}g$/disk). These results suggest that this fraction has a potent antifungal activity against the dermatophytic fungus Trichophyton mentagrophytes ATCC 28185.

Cyclopamine, an Antagonist of Hedgehog (Hh) Signaling Pathway, Reduces the Hatching Rate of Parthenogenetic Murine Embryos

  • Park, Jaehyun;Moon, Jeonghyeon;Min, Sol;Chae, Stephan;Roh, Sangho
    • Journal of Embryo Transfer
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    • v.33 no.4
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    • pp.237-243
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    • 2018
  • Hedgehog (Hh) pathway plays a key role in development from invertebrate to vertebrate. It is known to be involved in cell differentiation, polarity, proliferation, including the development of vertebrate limb and the establishment of flies' body plan. To investigate how the regulation of Hh pathway affects the development of parthenogenetic murine embryos, the parthenogenetically activated murine embryos were treated with either cyclopamine (Cyc), an antagonist of Hh pathway, or purmorphamine, an agonist of Hh pathway. While Cyc did not affect the blastocyst formation and its total cell number, the chemical reduced the hatching rate of embryos and the expression levels of Fn1 mRNA. The results of the present study show the possibility that Cyc may affect the development of embryos at blastocyst stage by blocking Hh pathway and this may cause detrimental effect to the embryos at peri-, and post-implantation stages.

Effecs of Soyangin Indongdeungjikolpitang on the Complication of Diabetes (소양인(小陽人) 인동등지골피탕(忍冬藤地骨皮湯)이 중소합병증(中消合倂症)에 미치는 영향(影響))

  • Seo, Yong-bae;Song, Jeong-mo
    • Journal of Sasang Constitutional Medicine
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    • v.12 no.1
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    • pp.186-200
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    • 2000
  • The purpose of this research was to investigate the effects of Indongdeungjikolpitang water extract(IJTE) on the complication of diabetes. IJTE did not affect the level of blood glucose in alloxan- or streptozotocin-induced hyperglycemic mice, but inhibited the motility of gastrointestine. IJTE inhibited the writhing syndrome induced by acetic acid, the permeability of evans blue into peritoneal cavity induced by acetic acid, the paw edema induced by histamine, and the formation of cotton pellet granuloma. IJTE increased the cell viability of thymocytes and splenocytes. IJTE decreased the release of ${\gamma}-interferone$(${\gamma}-IFN$) and interleukin-2(IL-2), but did not affect the release of interleukin-4(IL-4) from murine thymocytes. IJTE increased the release of IL-4 and decreased the release of tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) and $interleukin-1{\beta}$($IL-1{\beta}$), but did not affect of ${\gamma}-IFN$ and IL-2 from murine splenocytes. IJTE decreased the release of $TNF-{\alpha}$ and $IL-1{\beta}$ from murine peritoneal macrophages. IJTE decreased the production of niric oxide(NO) from murine peritioneal macrophages and increased the phagocytic activity of murine peritoneal macrophages. These results suggest that IJTE has an anti-inflammatory action via the inhibition of $TNF-{\alpha}$, $IL-1{\beta}$ and NO production from immune cells.

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The Role of nerve Growth Factor on Corneal Wound Healing in Dogs

  • 우흥명
    • Journal of Veterinary Clinics
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    • v.18 no.4
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    • pp.418-423
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    • 2001
  • To investigate the modulation of nerve growth factor (NGF) during corneal epithelial wound healing and the effect of topical NGF on corneal epithelial wound healing in dogs. An axial epithelial defect was created in the right eye using 6mm axial corneal mechanical debridement while the left served as an unwounded control. The tears were collected from both eyes during 1 week and the corneal epithelium was processed for the measurement of NGF at day 0 and 7. The NGF content of tears and corneal epithelium was determined by enzyme-linked immunosorbent assay. In another experiment, the animals were divided into 3 groups. The right eyes in each group were treated every six hours with 200 ug/ml of recombinant human (rh) NGF, murine NGF, or 600 ug/ml of anti-NGF blocking antibody. The left eye of each animal was treated with bovine serum albumin (BSA) to serve as controls. Wound healing was analyzed using NIH image software. Tear NGF was markedly increased in the wounded eyes, relative to tears from control eyes during the early healing period. The NGF content of the corneal epithelium was elevated in the wounded eye (p=0.024). Time to wound closure and rate of epithelial migration were not significantly different between the NGF treated or the NGF antibody treated, and the control BSA treated eyes. Corneal epithelial wounding increased NGF content only on the wounded side during the early healing period. Neither topical recombinant human or murine NGF affected corneal epithelial wound healing in the normal dog.

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