• Title/Summary/Keyword: in-vitro

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Effect of Retrovirus Mediated TNF-$\alpha$ Gene Transfer to Tumor Necrosis Factor(TNF) Sensitive Tumor Cell Lines on Sensitivity to TNF (Retroviral Vector를 이용한 TNF-$\alpha$ 유전자의 이입이 암세포의 종양괴사인자(TNF) 감수성에 미치는 효과)

  • Oh, Yeon-Mok;Park, Kyeo-Yeong;Jung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Goo;Sim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.2
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    • pp.87-96
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    • 1994
  • Background : Since tumor necrosis factor was discovered in 1975, TNF has been well known about its cytotoxic effect on tumor cells in vivo and in vitro. According to the recent improvement of molecular biological techinques, it is possible that exogenous TNF gene is transferred to tumor cells and is expressed in theirs. By virtue of TNF gene transfer, we have expected that TNF expressed in TNF-gene-transferred tumor cells would kill tumor cells in vivo without systemic side effect. The expected mechanisms in which antitumor effects of TNF expressed in TNF-gene-transferred tumor cells are working would be as followings. In the first mechanism, TNF expressed in TNF-gene-transferred tumor cells would kill tumor cells around(like homicide). In the second mechanism, TNF expressed in TNF-gene-transferred tumor cells would kill themselves(like suicide). In the third mechanism, TNF expressed in TNF-gene-transferred tumor cells would recruit immune effector cells and kill tumor cells indirectly. In the last mechanism, TNF expressed in TNF-gene-transferred tumor cells would augment cytokine such as interferon-$\gamma$ to kill tumor cells. Among these four mechanisms of antitumor effect, only the second mechanism has not been established yet. Therefore, to elucidate the second mechanism, We performed this study. Method : We transferred TNF-$\alpha$ gene to NCI-H2058, a human mesothelioma cell line and WEHI164, a murine fibrosarcoma cell line by using retroviral vector(pLT12SNTNF). And, We determined by using MTT assay whether TNF expressed in TNF-gene-transferred tumor cell lines would kill themselves like suicide or not. Then, if TNF-gene-transferred tumor cell lines would not suicide themselves, I would know more about the TNF sensitivity of TNF-gene-transferred tumor cell lines to exogenous TNF also by MTT assay. Result : NCI-H2058 and WEHI164 which were sensitive to TNF, became far less sensitive to endogenous and exogenous TNF after being transferred TNF-$\alpha$ gene to. Conclusion : TNF-gene-transfer to NCI-H2058 and WEHI164 gave them resistance to TNF.

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Biological activity and analysis of α-glucosidase inhibitor from mulberry (Morus alba L.) wine (오디와인의 생리활성 및 α-glucosidase 저해제의 분석)

  • Son, Woo-Rim;Choi, Sang-Won
    • Food Science and Preservation
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    • v.20 no.6
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    • pp.877-885
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    • 2013
  • Wine extracts of four different berry fruits, such as mulberry, blueberry, strawberry, and raspberry, were investigated for antioxidant, anti-tyrosinase, and ${\alpha}$-glucosidase activities by using in vitro assays. Additionally, quantitative changes of ${\alpha}$-glucosidase inhibitor in mulberry wine were determined by HPLC according to mulberry cultivars and fermentation process. Among four berry wines examined, mulberry wine showed the most potent ${\alpha}$-glucosidase inhibitory activity with 69.37% at 0.23 mg/mL, while blueberry and strawberry wines exhibited the strongest inhibition against DPPH radical and tyrosinase activity, respectively. Four compounds were isolated and purified from mulberry wine by a series of isolation procedures, such as solvent fractionation, and Diaion HP-20, ODS-A, and Sephadex LH-20 column chromatographies. Among them, Comp. 4 exerted the strongest ${\alpha}$-glucosidase inhibitory activity ($I_C_{50}=31.57{\mu}M$), and its chemical structure was identified as quercetin by UV and NMR spectral analysis. Finally, the "Daeseongppong" (16.83 ppm) muberry wine had larger amount of quercetin than the "Iksuppong" (14.85 ppm) and "Cheongilppong" (8.92 ppm) mulberry wines, but their contents of three mulberry wines decreased considerably with aging process. These results suggest that mulberry wine containing quercetin acted as ${\alpha}$-glucosidase inhibitor may be useful as a potential functional wine for improving diabetic disorder.

Effect of Solvent Fractions from Methanol Extract of Doenjang on Inhibition of Growth and DNA Synthesis of Human Cancer Cells. (인체 암세포 성장 및 DNA 합성 억제에 미치는 된장 분획물의 영향)

  • LIM Sun-Young;Rhee Sook-Hee;Park Kun-Young
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.685-691
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    • 2005
  • Growth and DNA synthesis inhibitory effects of doenjang methanol extract and its solvent fractions on AGS human gastric adenocarcinoma cells, Hep 3B human hepatocellular carcinoma cells, HT-29 human colon cancer cells and MG-63 human osteosarcoma cells were studied. The treatment of doenjang methanol extract ($ 200{\mu}g/ml $) with the AGS, Hep 3B, HT-29 and MG-63 cancer cells after 6 days of incubation inhibited the growth of cancer cells by $32\%$, $51\%$, $84\%$ and $33\%$, respectively. To separate active compounds of doenjang, doenjang methanol extract was fractionated with dichloromethane, ethylacetate, and buthanol. Among the solvent fractions, the dichloromethane and ethylacetate fractions showed the highest growth inhibitory effects on various cancer cells. For example, the dichloromethane and ethylacetate fractions ($200a{\mu}g/ml$) sig-nificantly inhibited the growth of various cancer cells by $89\∼96\%$ and$62\∼86\%$, respectively. DNA synthesis of AGS and Hep 3B cancer cells was significantly inhibited by adding dichloromethane fraction ($200{\mu}g/ml$) up to $94\%$ and $80\%$, respectively. Similarly, the ethylacetate fraction ($ 200\mug/ml $) showed a $ 95\% $ inhibition rate of DNA synthesis in AGS cells. These results suggest that the dichloromethane and ethylacetate fractions have specific active compounds, which will explain this anticancer effect of doenjang.

Influence of Sodium Hypochorite & EDTA on the Microtensile Bond Strength of Ethanol Wet Bonding (Ethanol Wet Bonding에서 NaOCl과 EDTA가 결합강도에 미치는 영향)

  • Kim, Deok-Joong;Song, Yong-Beom;Park, Sang-Hee;Kim, Hyoung-Sun;Lee, Hye-Yoon;Yu, Mi-Kyung;Lee, Kwang-Won
    • Journal of Dental Rehabilitation and Applied Science
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    • v.29 no.1
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    • pp.37-44
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    • 2013
  • Sodium hypochlorite and ethylene diamine tetra acetic acid are substances usually used during endodontic treatment. Several studies found that the bonding was negated with certain irrigants and some of the used irrigants have demineralizing and chealating effects, so it was advocated to omit the etching step in etch and rinse adhesive systems. The purpose of this in vitro study was to evaluate the influence of NaOCl & EDTA on the bonding strength of ethanol wet bonding. Thirty human molars were selected and mesiodistally sectioned into halves, thus providing sixty specimens. The specimens were randomly assigned to 4 groups(n=15) according to the irrigant regimen used : (1) irrigated with distilled water for 10min (control); (2) irrigated with 5.25% NaOCl(10min), flushed with 5.25% NaOCl(1min) (3) irrigated with 5.25% NaOCl, flushed with 17% EDTA (4) irrigated with 5.25% NaOCl, flushed with 17% EDTA. Each group was acid-etched with 37% phosphoric acid(except group 4) and had their dentin surfaces dehydrated with ethanol solutions : 50%, 70%, 80%, 95%, 3x100%, 30s for each application. After dehydration, a primer( 50% all bond 3 resin + 50% ethanol) was used, followed by the adhesive(ALL-BOND 3 RESIN) application. Resin composite build-ups were then prepared using an incremental technique. Specimens were sectioned into beams and submitted to a tensile load using a Micro Tensile Tester(Bisco Inc.). The data were statistically analyzed using one-way ANOVA and Tukey HSD at p<0.5 level. There was no significant difference on G1(control) and G2(irrigated with NaOCl only ). (p>0.05). G3(flushed with EDTA) showed significantly high tensile bonding strength compared to the G2 (p<0.05). G4( treated with EDTA but no acid-etching) was significantly lower value than G3. (p<0.05) Although there was no significant difference, 5.25% NaOCl seemed to have an adverse effect on the bonding strength of ethanol wet bonding. The flushing with EDTA after NaOCl irrigation prevents the decrease of bonding strength. The use of 17% EDTA as a final flush can enhance the bonding strength but EDTA flushing can't substitute for a acid-etching.

Assays of Maturity and Antifungal Activity against Plant Pathogen during the Animal Manure Composting Process (가축분 퇴비화 과정에서 부숙도 및 퇴비의 항균활성 검정)

  • Seo, Myung-Chul;So, Kyu-Ho;Park, Won-Mok
    • Korean Journal of Soil Science and Fertilizer
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    • v.32 no.3
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    • pp.285-294
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    • 1999
  • Changes of chemico-physical properties and mturitiy during pig manure composting were analysed using three kinds of bulking agents with rice hull(T1), rice hull and extruding hull mixture (T2, 1:1, v/v), and extruding hull(T3). During composting process, temperature of T1, T2 and T3 were maintained over $50^{\circ}C$ for 31, 21, and 35 days respectively. Organic matter content of each treatment was decreased from 82.2%, 82.0%, and 82.8% to 70.5%, 68.9% and 69.7% and pH increased to 8.85, 9.91, and 8.80, respectively. Total nitrogen content of all treatments gradually decreased, but C/N ratio, phosphorous, and potassium content did not, show any changes during composting process. Both germination rate and early growth were tested using radish seeds for composting maturity. From those results, it was concluded that all treatments were stabilized after 45th day and extruding hull(T3) added compost was superior to others. The test of suppressive effect showed that all treatment have no effect against Fusarium oxysporum, Alternaria altemata, Botrytis cinerea. Compost supplemented with rice hull showed an inhibitory effect after 30th days, while compost supplemented with rice hull and extruding hull(T2) had an inhibitory effect during all period against Rhizoctonia solani. But treatment with extruding hull(T3) added compost did not have any inhibitory effect against Rhizoctonia solani. Only 63th samples in T1 and T2 treatment showed inhibitory effect against Colletoerichum gloeosporioides. However, T3 did not. Suppressive effect of extracts from 67 kinds of composts was investigated in vitro against plant pathogens, such as Fusauum oxysporum. Alternaria alternata, Colletotrichum gloeospoioides, Rhizoctonia solani, and Botrytis cinerea. Thirty two of them showed inhibitory effect against more than one phytopathogen, nine against one pathogen, four against two, six against three, six against four, and seven against five.

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Immuno-stimulating and Antitumor Effects of Crude Polysaccharides Extracted from Fruiting Body of Grifola frondosa (잎새버섯(Grifola frondosa)의 자실체에서 추출한 조다당류의 면역증강 및 항암효과)

  • Kim, Jeong-Hwa;Cha, Youn-Jeong;Shim, Mi-Ja;Lee, Min-Woong;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.68-77
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    • 2011
  • 80% methanol and 0.9% neutral saline soluble and hot water substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were extracted from fruiting bodies of Grifola frondosa. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180 and RAW 264.7 at the concentration of 10~2000 ${\mu}g/ml$, but crude polysaccharides from Fr. NaCl was slightly toxic to HT-29 and NIH3T3 at the concentration of 2000 ${\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited life prolongation effect of 25.0~52.9% in mice previously inoculated with Sarcoma 180. Fr. HW increased the numbers of spleen cells by 1.3 fold at the concentration of 200 ${\mu}g/ml$ compared with control. Fr. NaCl improved the immuno-stimulating activity of B lymphocyte by increasing the alkaline phosphatase activity by 1.5 fold compared with control at the concentration of 200 ${\mu}g/ml$. 10~14 ${\mu}M$ of nitric acid were generated when Fr. NaCl was added to RAW 264.7 at the concentration of 50~500 ${\mu}g/ml$, while the control group produced 4.3 ${\mu}M$ of nitric oxide. The Fr. NaCl, Fr. HW and Fr. MeOH increased the production of TNF-${\alpha}$, IL-$1{\beta}$, Il-2 and IL-6 by more than 1.4 times compared with the control group. The Fr. of MeOH increased the numbers of peritoneal exudate cells and circulating leukocytes by 3.0 and 2.0 folds compared with the control at the concentration of 50 ${\mu}g/ml$, respectively. Therefore, the crude polysaccharides extracted from fruiting bodies of Grifola frondosa could improve antitumor activity of mice.

Human Impact on Sedimentary Environment of Estuarine Coastal Salt Marches, Southern Coastal Region of Korea Peninsula (인위적 환경변화에 따른 해안지역 퇴적환경의 변화)

  • 박의준
    • Journal of the Korean Geographical Society
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    • v.36 no.2
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    • pp.111-125
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    • 2001
  • An estuary is semi-inclosed inlets, located between terrestrial and marine environment. Since many estuaries along south-western coasts of Korean peninsula were affected by human settlements and activities, significant changes in sedimentation environments have been observed. The research area is divided into three distinct morpho-stratigraphic units: fluvial dominated area(Area1), mixed area(Area 2), tide-dominated area(Area3). The landform of this area has been changed by reclamation and river channel change. Temporal variations affected by dam construction, periodic freshet was iterrupted. Sediments began to continuously accmulate on estuary banks by tide. Meanwhile, because of the continuous but reduced discharge of fresh water, the salinity of estuarine sediments was declined. That processes made vegetated area( Phregmites lonivalvis and Suaeda japonica) to be expanded. It indicates that the magnitude and frequency of geomorphic processes has been significantly changed.

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Analysis of Chemical Composition and in vitro Anti-oxidant Properties of Extracts from Sea Buckthorn (Hippophae rhamnoides) (비타민나무(Sea Buckthorn, Hippophae rhamnoides) 추출물의 이화학적 성분 분석과 항산화 활성효과)

  • Kim, Kyung-Min;Park, Min-Hee;Kim, Kyung-Hee;Im, Sang-Hyun;Park, Yoo-Hwa;Kim, Young-Nam
    • Journal of Applied Biological Chemistry
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    • v.52 no.2
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    • pp.58-64
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    • 2009
  • Total polyphenol contents and antioxidative activity of water and ethanol extracts from Sea Buckthorn (Hippophae rhamnoides) leaves, branches (bough and twig) and roots were analyzed. The level of crude ash and crude protein in water extract of leaves, branches (bough and twig), and roots were shown to be a bit higher than ethanol extracts. Especially crude protein contents from water extract of leaves, bough, twig, and roots were 14.90, 18.60, 18.03, and 16.61% respectively. Total polyphenol content of ethanol extracts of all parts of Sea buckthorn was ranged from 106.33${\pm}$2.32 ${\mu}g/g$ to 147.78${\pm}$3.06 ${\mu}g/g$ showing higher amount than water extracts. To investigate antioxidative activity of Sea buckthorn, DPPH free radical scavenging activity, hydroxy radical scavenging activity, and SOD-like activity were analyzed. The results showed that the antioxidative activity of ethanol extracts was relatively higher than water extracts. The $IC_{50}$ values for DPPH free radical scavenging activity was ranged from 23.58${\pm}$0.84 ${\mu}g/mL$ to 59.35${\pm}$1.69 ${\mu}g/mL$. Compared to 68.85${\pm}$1.44% of SOD-like activity from L-ascorbic acid used as a control, the ethanol extract of Sea buckthorn branches showed relatively strong activity of 35.03${\pm}$2.33%. The highest hydroxy radical scavenging activity was shown as 66.12${\pm}$8.73% from ethanol extract of Sea buckthorn roots which was similar value to 72.47${\pm}$2.83% of L-ascorbic acid.

An in-vitro wear study of human enamel opposing heat-pressed ceramics (2종의 열가압 도재와 법랑질 간의 마모에 관한 연구)

  • Park, Chan-Yong;Jeon, Young-Chan;Jeong, Chang-Mo;Yun, Mi-Jung
    • The Journal of Korean Academy of Prosthodontics
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    • v.47 no.1
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    • pp.21-28
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    • 2009
  • Purpose: The purpose of this study was to compare the wear characteristics of human enamel opposing 2 heat-pressed ceramics (e.max Press and Empress Esthetic), conventional feldspathic porcelain (Ceramco 3) and type III gold alloy. Material and methods: Intact cusps of extracted premolars were used for enamel specimens. Five disk samples were made for each of two heat-pressed ceramics groups, conventional feldspathic porcelain group and type III gold alloy group. Wear tests were conducted in distilled water using a pin-on-disk tribometer. The amount of enamel wear was determined by weighing the enamel specimens before and after wear tests, and the weight was converted to volumes by average density. The wear tracks were analyzed by scanning electron microscopy and surface profilometer to elucidate the wear characteristics. Results: 1. Ceramco 3 led to the greatest amount of enamel wear followed by Empress Esthetic, e.max Press and type III gold alloy. However, there was no significant difference between Ceramco 3 and Empress Esthetic (P>.05), and there were also no significant differences among Empress Esthetic, e.max Press and type III gold alloy (P>.05). 2. The average surface roughness of e.max Press after wear test was smallest followed by Empress Esthetic and Ceramco 3, but there was no significant difference between Empress Esthetic and Ceramco 3 (P>.05). 3. There were no significant differences among the depth of wear tracks of all the groups (P>.05). The group that showed the largest width of wear track was Ceramco 3 followed by Empress Esthetic, e.max Press and type III gold alloy. However, there was no significant difference between e.max Press and Empress Esthetic (P>.05), and there was also no significant difference between Empress Esthetic and Ceramco 3 (P>.05). Conclusion: Within the limits of this study, heat-pressed ceramics were not more abrasive than conventional feldspathic porcelain.

Metabolic comparison between standard medicinal parts and their adventitious roots of Cynanchum wilfordii (Maxim.) Hemsl. using FT-IR spectroscopy after IBA and elicitor treatment (IBA 및 elicitor 처리에 따른 백수오 기내 생산 부정근 및 표준품의 FT-IR 스펙트럼 기반 대사체 비교 분석)

  • Ahn, Myung Suk;So, Eun Jin;Jie, Eun Yee;Choi, So Yeon;Park, Sang Un;Moon, Byeong Cheol;Kang, Young Min;Min, Sung Ran;Kim, Suk Weon
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.250-256
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    • 2018
  • To determine whether metabolite fingerprinting for whole cell extracts based on Fourier transform infrared spectroscopy (FT-IR) can be used to discriminate and compare metabolic equivalence, standard medicinal parts of Cynanchum wilfordii (Maxim.) Hemsl. and their adventitious roots were subjected to FT-IR. The principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) from FT-IR spectral data showed that whole metabolic pattern from the adventitious root of Cynanchum wilfordii was highly similar to its standard medicinal parts. These results clearly showed that mass proliferation of adventitious roots could be applied for the novel supply of standard medicinal parts of medicinal plants. Furthermore, FT-IR spectroscopy combined with multivariate analysis established in this study could be applied as an alternative tool for discriminating of whole metabolic equivalence from standard medicinal parts. Thus, it is proposed that these metabolic discrimination systems from the adventitious root of Cynanchum wilfordii could be applied for metabolic standardization of in vitro grown Cynanchum wilfordii.