• Title/Summary/Keyword: in vitro effect

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Practical Application of Dioscorea quinqueloba Extract for the Control of Citrus Green Mold (감귤 녹색곰팡이병 방제를 위한 천산용 추출물의 실용적 적용)

  • Lee, Ji Hyun;Kang, Sung Woo;Song, Jeong Young;Kim, Hong Gi
    • Research in Plant Disease
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    • v.18 no.4
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    • pp.354-360
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    • 2012
  • This study tested the antifungal compound obtained from a medicinal plant, Dioscorea quinqueloba Thunb., in order to search the possibility of practical application of this product in agriculture through evaluating its activity using the citrus fruits. The extract of D. quinqueloba Thunb., which has the strongest antifungal activity, was selected as a candidate among 101 plant extracts. Based on this examination concerning antifungal activity of the product on Penicillium digitatum in vitro, it was confirmed its effect of mycelial growth inhibition showed over 87% at 0.5 mg/ml concentration. This natural product showed the stability of the substance, as it was not significantly influenced by pH, temperature, or ultraviolet radiation. While citrus fruits were stored at room temperature, P. digitatum was inoculated into them in order to prepare a similar environmental conditions with epidemic occurrence of the mold. As the result of our investigation, the disease preventive effects of the active antifungal substance evidenced a 100% at 0.5 mg/ml. When the phytotoxicity of the selected natural product on citrus at 2 mg/ml was assessed, we noted no toxic effects. Based on the superior preventive effects from this natural product extracted from the plant, it is presumed to be very useful in agricultural applications for the control of green mold, P. digitatum, which has been occurred often the biggest problem in the storage of citrus fruits.

Role of Nuclear Factor (NF)-κB Activation in Tumor Growth and Metastasis (종양의 성장 및 전이에 있어서 NF-κB의 역할)

  • Ko, Hyun-Mi;Choi, Jung-Hwa;Ra, Myung-Suk;Im, Suhn-Young
    • IMMUNE NETWORK
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    • v.3 no.1
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    • pp.38-46
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    • 2003
  • Background: Platelet-activating factor (PAF) induces nuclear factor $(NF)-{\kappa}B$ activation and angiogenesis and increases tumor growth and pulmonary tumor metastasis in vivo. The role of $NF-{\kappa}B$ activation in PAF-induced angiogenesis in a mouse model of Matrigel implantation, and in PAF-mediated pulmonary tumor metastasis were investigated. Methods: Angiogenesis using Matrigel and experimental pulmonary tumor metastasis were tested in a mouse model. Electrophoretic mobility shift assay was done for the assessment of $NF-{\kappa}B$ translocation to the nucleus. Expression of angiogenic factors, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\alpha}$, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) were tested by RT-PCR and ELISA. Results: PAF induced a dose- and time-dependent angiogenic response. PAF-induced angiogenesis was significantly blocked by PAF antagonist, CV6209, and inhibitors of $NF-{\kappa}B$ expression or action, including antisense oligonucleotides to p65 subunit of $NF-{\kappa}B$ (p65 AS) and antioxidants such as ${\alpha}$-tocopherol and N-acetyl-L-cysteine. In vitro, PAF activated the transcription factor, $NF-{\kappa}B$ and induced mRNA expression of $TNF-{\alpha}$, $IL-1{\alpha}$, bFGF, VEGF, and its receptor, KDR. The PAF-induced expression of the above mentioned factors was inhibited by p65 AS or antioxidants. Also, protein synthesis of VEGF was increased by PAF and inhibited by p65 AS or antioxidants. The angiogenic effect of PAF was blocked when anti-VEGF antibodies was treated or antibodies against $TNF-{\alpha}$, $IL-1{\alpha}$, and bFGF was co-administrated, but not by antibodies against $TNF-{\alpha}$, $IL-1{\alpha}$, and bFGF each alone. PAF-augmented pulmonary tumor metastasis was inhibited by p65 AS or antioxidants. Conclusion: These data indicate that PAF increases angiogenesis and pulmonary tumor metastasis through $NF-{\kappa}B$ activation and expression of $NF-{\kappa}B$-dependent angiogenic factors.

EFFECTS OF FLUORIDE MOUTHRINSE ON CELL ACTIVITY OF GINGIVAL FIBROBLASTS OF CHILDREN (불소양치용액이 소아 치은 섬유아세포의 세포활성에 미치는 영향에 관한 연구)

  • Lee, Dong-Hyun;Lee, Kwang-Hee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.24 no.1
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    • pp.204-219
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    • 1997
  • The use of fluoride is one of the most effective methods for caries prevention. Fluoridation of public water supply has been recognized, for many years, as an effective way to reduce dental caries. The fluoride supplement has been recommended when the natural fluoride was unavailable or below the optimal range. However the mechanism of caries prevention by fluoride has not yet been clarified and it is well known that an overdose of fluoride results inacute and chronic toxicity, especially dental fluorosis. Fluoride mouthrinsing solution is widely used in dentistry due to its effectiveness in carrying anticariogenic action. Understanding the effects of fluoride mouthrinsing solution on human gingival fibroblasts will provide the safety rationale for its use during the caries preventive therapy. The purpose of this study was to evaluate the cytotoxic effect of fluoride mouthrinsing solution on the human gingival fibroblast in vitro. The human gingival fibroblasts were cultured from healthy gingiva on the extracted deciduous teeth of children. Cells were inoculated into a 24-well plate with $1{\times}10^4cells/well$ of medium at $37^{\circ}C$, 100% humidity, 5% $CO_2$ incubator for 24 hours. And the cells were counted by using the hemocytometer at each designed study. Human gingival fibroblasts were cultured in growth medium after one minute application range of 0.02%-0.2% NaF solution and 0.1% $SnF_2$ solution. The cells used in this study were between fifth to eighth passage number. The cell morphology was examined by inverted microscope and cell proliferation was measured by incorporating $[^3H]$-thymidine into DNA. DNA synthesis by human gingival fibroblasts was assessed by $[^3H]$-thymidine uptake assays while the cell activity was measured by MTT assay. Each concentrated fluoride mouthrinsing solution was estimated for its biocompatability with fibroblasts by the tissue culture technique. The results of this study were as follows : 1. It was observed that at 0.05%, 0.2% NaF mouthrinsing solution the cytoplasmic processes became globular. When 0.1% $SnF_2$ mouthrinsing solution was applied, the cytoplasmic process and cell morphology were disappeared. 2. DNA synthetic activity was reduced regardless of the concentration of the fluoride mouthrinsing solution. However, the result is statistically insignificant except 0.1% $SnF_2$ mouthrinsing solution(p<0.05). 3. Our results indicate that 0.02%, 0.05% concentrations of NaF mouthrinsing solution caused minimal cytotoxicity. But 0.2% NaF and 0.1% $SnF_2$ concentration were a significant difference between the cell activity in the experimental group and control group (p<0.05). 4. After appling 0.05% & 0.02% NaF fluoride mouthrinsing solution, cell activity was restored to the control groups level according to incubating time. The results suggest that direct exposure to fluoride solution inhibits gingival fibroblast activity. Therefore, for the most effective use of fluoride use, lowering the concentration of fluoride mouthrinsing is advisable because it maintains biocompatability and free ion in the oral fluid.

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Biologic Response of Human Deciduous Dental Pulp Cells on Newly Developed MTA-like Materials (새로 개발된 MTA 유사 재료에 대한 유치 치수세포의 생물학적 반응)

  • Lee, Haewon;Shin, Yooseok;Jung, Jaeeun;Kim, Seongoh;Lee, Jaeho;Song, Jeseon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.42 no.4
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    • pp.291-301
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    • 2015
  • This study compared the in vitro cell viability and differentiation potentials of human deciduous dental pulp cells (DPCs) on mineral trioxide aggregate (MTA)-like products (ProRoot MTA, RetroMTA and Endocem Zr). The experimental materials were prepared as circular discs, which were used to test the effects of the materials on the viability of human DPCs when placed in direct and indirect contact. Furthermore, the pH of the extracted materials was recorded, and their effect on cell differentiation potential was evaluated by evaluating the alkaline phosphatase (ALP) activity and Alizarin Red S staining of DPCs incubated with the test materials. In direct contact, the cell viability of human DPCs was higher with ProRoot MTA and RetroMTA than with Endocem Zr. However, when in indirect contact, the cell viability of human DPCs was generally higher in Endocem Zr than in ProRoot MTA and Retro MTA. With respect to pH, the alkalinity was lower for Endocem Zr than for the other test materials. The ALP activities of the cells were not enhanced by any of the experimental materials. Alizarin Red S staining of the tested human DPCs revealed that their differentiation potential was lower than for cells incubated with osteogenic induction medium. While there were differences in the responses of the human DPCs to the test materials, all displayed degrees of cytotoxicity and were unable to enhance either the viability or differentiation of human DPCs. However, Endocem Zr exhibited better cell viability and was less alkaline than the other test materials.

In vitro Antioxidant Effects of Sarijang (사리장의 항산화 효과)

  • Seo, Bo-Young;Choi, Mi-Joo;Choi, Eun-A;Park, Eunju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.4
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    • pp.618-623
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    • 2014
  • Sarijang, a soy sauce made from fermented black soybean (Rhynchosia nulubilis), sulfur fed duck, dried bark of Ulmus davidiana, Allium sativum, and bamboo salt, has been demonstrated to exert anti-inflammatory and anti-tumor activities. However, the antioxidant properties of Sarijang have not yet been elucidated. In this study, the antioxidant effects of Sarijang were investigated by determining total phenolic content (TPC), DPPH radical scavenging activity (DPPH RSA), total radical trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), and cellular antioxidant capacity (CAC). The inhibitory effects of Sarijang on oxidative stress-induced DNA damage (200 ${\mu}M$ $H_2O_2$, 250 ${\mu}M$ Fe-NTA, and 200 ${\mu}M$ HNE) in human leukocytes were evaluated by comet assay. The TPC of Sarijang was $1.04{\pm}0.01$ mg GAE/mL. DPPH RSA, TRAP, and ORAC values of Sarijang increased in a dose-dependent manner. The $IC_{50}$ for DPPH RSA of Sarijang was $11.2{\pm}0.3$ mg/mL, whereas $IC_{50}$ of TRAP was $209.5{\pm}2.0$ mg/mL. 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress and oxidative stress-induced DNA damage in HepG2 cells were effectively abrogated by all tested concentrations of Sarijang (1~100 ${\mu}g/mL$). These results suggest that Sarijang has antioxidative activity and protective effects against oxidative DNA damage.

Effect of Chlorella vulgaris on Immune-enhancement and Cytokine Production in vivo and in vitro

  • An, Hyo-Jin;Rim, Hong-Kun;Lee, Jong-Hyun;Seo, Min-Jun;Hong, Jin-Woo;Kim, Na-Hyung;Myung, Noh-Yil;Moon, Phil-Dong;Choi, In-Young;Na, Ho-Jeong;Kim, Su-Jin;Jeong, Hyun-Ja;Park, Hyeung-Suk;Han, Jae-Gab;Um, Jae-Young;Hong, Seung-Heon;Kim, Hyung-Min
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.953-958
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    • 2008
  • The object of this study was to investigate the immune-enhancing effects of Chlorella vulgaris (CV) on a deteriorated immune function by a protein-energy malnutrition (PEM) diet. Unicellular algae, CV were used as a biological response modifier. Male C57BL/6J mice were fed for 15 days with standard diet or a PEM diet, which is associated with decreased host immune defense. After 8 days, mice in the PEM diet group were orally administered by 0.05, 0.1, and 0.15 g/kg body weight of CV or distilled water. Nutritional parameters, and interferon (IFN)-$\gamma$ levels were significantly increased in the blood serum of the CV (0.15 g/kg)-treated group (29.6$\pm$2.8 pg/mL) compared to the non-treated PEM group (4.1$\pm$0.4 pg/mL, p<0.05). In addition, cell proliferation and production of cytokines were investigated via a CV (0.01, 0.1, and 1 mg/mL) treatment using a human T cell line MOLT-4 cell. The CV treatment (1 mg/mL) significantly increased the production of both IFN-$\gamma$ and interleukin (IL)-2 (51.3$\pm$3.4 and 285.9$\pm$18.8 pg/mL, respectively) compared to the control (51.3$\pm$3.4 and 442.6$\pm$14.3 pg/mL, respectively), but did not affect the production of IL-4. These results suggest that CV may be useful in improving the immune function.

Biological Activities of Oenothera laciniata Extracts (Onagraceae, Myrtales) (애기달맞이꽃(Oenothera laciniata Hill) 추출물의 생리활성 탐색)

  • Lee, Jung-A;Kim, Ji-Young;Yoon, Weon-Jong;Oh, Dae-Ju;Jung, Yong-Hwan;Lee, Wook-Jae;Park, Soo-Yeong
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.810-815
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    • 2006
  • The biological activities of Oenothera laciniata extracts were measured, including antioxidant activity and cytotoxic effects. O. laciniata is an endemic species of Jeju Island, Korea with a seaside habitat. The concentration of total polyphenolic compounds from ethanol (EtOH), n-hexane, dichloromethane ($CH_2Cl_2$), ethylacetate (EtOAc), butanol (BuOH), and water fractions of O. laciniata was 63.96, 8.49, 28.11, 172.64, 114.56, and 34.91 mg/g, respectively. The EtOAc fraction contained the highest antioxidative activities ($IC_{50}$), measured as follows: 16.19 ${\mu}g/mL$ in DPPH radical scavenging capacity, 220.37 ${\mu}g/mL$ in xanthine oxidase inhibitory activity, 42.07${\mu}g/mL$ in superoxide radical scavenging capacity, and 421.33 ${\mu}g/mL$ in nitric oxide scavenging capacity. The cytotoxicity of O. laciniata extracts was examined through their effect on the growth of HL-60 cells. Incubation of HL-60 cells with the EtOAc fraction resulted in the greatest inhibition of cell growth; high DNA fragmentation and numerous sub-G1 hypodiploid cells were observed in HL-60 cell cultures treated with the EtOAc fraction. These results suggest that the EtOAc fraction of O. laciniata has potent apoptotic and antioxidative activities in vitro.

Study on Summer Forage Crop Cultivation Using SCB (Slurry Composting-Biofilteration) Liquid Fertilizer on Reclaimed Land (간척지에서 SCB 액비를 이용한 여름철 사료작물 재배에 관한 연구)

  • Jo, Nam-Chul;Shin, Jae-Soon;Kim, Sun-Ho;Yoon, Sei-Hyung;HwangBo, Soon;Jung, Min-Woong;Lee, Kyung-Dong;Kim, Won-Ho;Seo, Sung;Kim, Jong-Geun;Song, Chae-Eun;Choi, Ki-Choon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.30 no.2
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    • pp.121-126
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    • 2010
  • Until now, The experiment about the forage crop have been almost not conducted on the reclaimed land. Therefore, this experiment was carried out in order to know productivity of summer forage crop using slurry composting-biofilteration (SCB) liquid fertilizer on reclaimed land of Hwaong and Sukmoon in korea from 2008 to 2009. The forage crops used in this experiment were corn and sorghum${\times}$sorghum hybrid which are used as summer forage crops in South Korea. The experiment was treated with chemical fertilizer (CF), swine slurry (SS) and SCB liquid fertilizer. Dry matter (DM) yield of corn was higher than those of sorghum${\times}$sorghum hybrid in both reclaimed lands but the effect of SCB liquid fertilizer was not appeared. The neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents of corn were lower than those of sorghum${\times}$sorghum hybrid. The crude protein (CP) content and in vitro dry matter digestibility (IVDMD) of corn were higher than those of sorghum${\times}$sorghum hybrid. In generally feed values of corn were higher than those of sorghum${\times}$sorghum hybrid. The results of this study showed that summer forage crop cultivation using uses SCB liquid fertilizer on reclaimed land are possible.

Comparative Analysis about the Effect of Isolated Phosphatidylcholine and Sodium Deoxycholate for the Viability of Adipocyte (Phosphatidylcholine과 Sodium Deoxycholate가 지방세포 생존에 미치는 영향의 비교 분석)

  • Rha, Eun-Young;Kang, Jo-A;Lee, Jung-Ho;Oh, Deuk-Young;Seo, Je-Won;Moon, Suk-Ho;Ahn, Sang-Tae;Rhie, Jong-Won
    • Archives of Plastic Surgery
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    • v.37 no.5
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    • pp.531-534
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    • 2010
  • Purpose: Lipobean$^{(R)}$s, widely used in lipodissolving techniques, contain phosphatidylcholine and sodium deoxycholate as its main substances. They have been approved only as medication for liver disease by the FDA. However, they have been used under various clinical settings without exact knowledge of its action mechanism. The authors designed an in vitro study to analyze the effects of different concentrations of phosphatidylcholine and sodium deoxycholate on adipocytes and other types of cells. Methods: Human adipose-derived stem cell were cultured and induced to differentiate into adipocytes. Fibroblasts extracted from human inferior turbinate tissue, and MC3T3-E1 osteoblast lines were cultured. Phosphatidylcholine solution dissolved with ethanol was applied to the culture medium at differing concentrations (1, 4, 7, 10 mg/mL). The sodium deoxycholate solution dissolved in DMSO applied to the medium at differing concentrations (0.07, 0.1. 0.4. 0.7 mg/mL). Cells were dispersed at a concentration of $5{\times}10^3$ cells/well in 24 well plates, and surviving cells were calculated 1 day after the application using a CCK-8 kit. Results: The number of surviving cells of adipocytes, fibroblasts and osteoblasts decreased as the concentration of sodium deoxycholate increased. However, all types of cells that had been processed in a phosphatidylcholine showed a cell survival rate of over 70% at all concentrations. Conclusion: This study shows that sodium deoxycholate is the more major factor in destroying adipocytes, and it is also toxic to the other cells. Therefore, we conclude that care must be taken when using Lipobean$^{(R)}$s as a method of reducing adipose tissue, for its toxicity may destroy other nontarget cells existing in the subcutaneous tissue layer.

Derivation of MSC Like-Cell Population from Feeder Free Cultured hESC and Their Proteomic Analysis for Comparison Study with BM-MSC (Feeder Free 상태에서 배양된 인간 배아 줄기세포를 이용한 중간엽 줄기세포 분화 및 단백체학을 이용한 골수 유래 중간엽 줄기세포와의 비교)

  • Park, Soon-Jung;Jeon, Young-Joo;Kim, Ju-Mi;Shin, Jeong-Min;Chae, Jung-Il;Chung, Hyung-Min
    • Reproductive and Developmental Biology
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    • v.34 no.3
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    • pp.143-151
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    • 2010
  • Pluripotency of human embryonic stem cell (hESC) is one of the most valuable ability of hESCs for applying cell therapy field, but also showing side effect, for example teratoma formation. When transplant multipotent stem cell, such as mesnchymal stem cell (MSC) which retains similar differentiation ability, they do not form teratoma in vivo, but there exist limitation of cellular source supply. Accordingly, differentiation of hESC into MSC will be promising cellular source with strong points of both hESC and MSC line. In this study, we described the derivation of MSC like cell population from feeder free cultured hESC (hESC-MSC) using direct differentiation system. Cells population, hESC-MSC and bone marrow derived MSC (BM-MSC) retained similar characteristics in vitro, such as morphology, MSC specific marker expression and differentiation capacity. At the point of differentiation of both cell populations, differentiation rate was slower in hESC-MSC than BM-MSC. As these reason, to verify differentially expressed molecular condition of both cell population which bring out different differentiation rate, we compare the molecular condition of hESC-MSC and BM-MSC using 2-D proteomic analysis tool. In the proteomic analysis, we identified 49 differentially expressed proteins in hESC-MSC and BM-MSC, and they involved in different biological process such as positive regulation of molecular function, biological process, cellular metabolic process, nitrogen compound metabolic process, macromolecule metabolic process, metabolic process, molecular function, and positive regulation of molecular function and regulation of ubiquitin protein ligase activity during mitotic cell cycle, cellular response to stress, and RNA localization. As the related function of differentially expressed proteins, we sought to these proteins were key regulators which contribute to their differentiation rate, developmental process and cell proliferation. Our results suggest that the expressions of these proteins between the hESC-MSC and BM-MSC, could give to us further evidence for hESC differentiation into the mesenchymal stem cell is associated with a differentiation factor. As the initial step to understand fundamental difference of hESC-MSC and BM-MSC, we sought to investigate different protein expression profile. And the grafting of hESC differentiation into MSC and their comparative proteomic analysis will be positively contribute to cell therapy without cellular source limitation, also with exact background of their molecular condition.