• Title/Summary/Keyword: Neutral red

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Expression of amino acid transport system L in the differentiation of periodontal ligament fibroblast cells (치주인대섬유모세포의 분화과정에서 아미노산 수송계 L의 발현)

  • Hwang, Kyu-Young;Kim, Do-Kyung;Kim, Chang-Hyun;Jang, Hyun-Seon;Park, Joo-Cheol;Choi, Seong-Mi;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.36 no.3
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    • pp.783-796
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    • 2006
  • The periodontium is a topographically complex organ consisting of epithelial tissue, soft and mineralized tissues. Structures comprising the periodontium include the gingiva, periodontal ligament (PDL) , cementum and the alveolar bone. The molecular mechanism of differentiation in PDL fibroblast cells remain unclear. Amino acid transporters play an important role in supplying nutrition to normal and cancer cells and for cell proliferation. Amino acid transport system L is a major nutrient transport system responsible for the Na+-independent transport of neutral amino acids including several essential amino acids. The system L is divided into two major subgroups, the L-type amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2). In this study, the expression pattern of amino acid transport system L was, therefore, investigated in the differentiation of PDL fibroblast cells. To determine the expression level of amino acid transport system L participating in intracellular transport of amino acids in the differentiation of PDL fibroblast cells, it was examined by RT-PCR, observation of cell morphology, Alizaline red-S staining and uptake analysis after inducing experimental differentiation in PDL fibroblast cells isolated from mouse molar teeth. The results are as follows. 1. The LAT1 mRNA was expressed in the early stage of PDL fibroblast cell differentiation. This expression level was gradually reduced by differentiation- inducing time and it was not observed after the late stage. 2. The expression level of LAT2 mRNA was increased in time-dependent manner during differentiation induction of PDL fibroblast cells. 3. There was no changes in. the expression level of 4F2hc mRNA, the cofactor of LAT1 and LAT2, during differentiation of PDL fibroblast cells. 4. The expression level of ALP mRNA was gradually increased and the expression level of Col I mRNA was decreased during differentiation of PDL fibroblast cells. 5. The L-leucine transport was reduced by time from the early stage to the late stage in PDL fibroblast cell differentiation. As the results, it is considered that among neutral ammo acid transport system L in differentiation of PDL fibroblast cells, the LATl has a key role in cell proliferation in the early stage of cell differentiation and the LAT2 has an important role in the late stage of cell differentiation for providing cells with neutral amino acids including several essential amino acids.

Comparison of Forage Yield and Quality, and Soil Improvement of Legumes (콩과작물의 사초생산성, 품질 및 토양개량 비교)

  • Kim Jong Duk;Kim Su Gon;Kwon Chan Ho;Abuel Sherwin J.;Chae Sang Heon;Kim Myoung Ki
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.25 no.3
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    • pp.151-158
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    • 2005
  • This experiment was conducted to evaluate forage production and quality, and soil improvement of three legumes at Cheonan Yonam College. The three legumes used in the experiment were crimson clover (Trifolium incarnatum L.) red clover(Trifolium pratense L.) and chinese milk vetch(Astragalus sinicus L.). Flowering stage was observed on the 20th of April for chinese milk vetch and on the 30th of April for crimson clover, but the red clover did not produce flower until harvest time. The dry matter(DM) content of crimson clover was the highest among the three legumes. In terms of DM yield, crimson clover has the highest yield, the highest yield in crude protein(CP) and total digestible nutrients(TDN) was red clover. In terms of forage quality, the highest CP content was red clover, while the chinese milk vetch was the lowest for both neutral detergent fiber(NDF) and acid detergent fiberr(ADF). Red clove. and chinese milk vetch were highest fur TDN content, and the chinese milk vetch has the highest for relative feed value(RFV). Assessment of the chemical properties of soil after harvest showed that the nitrogen content of soil increased in all legumes. The organic matter(OM) content of soil in both crimson and red clover increased, while the chinese milk vetch decreased. The available $P_2O_5$ decreased in all three legumes, but the biggest decrease was in the chinese milk vetch. The exchangeable potassium in the soil for both crimson and red clover increased, but in chinese milk vetch it decreased. Based on the results of this study, the chinese milk vetch was superior in terms of forage quality, but crimson and red clover are excellent in forage yield and in maintaining soil quality in upland.

Antiviral and Antitumoral Activitivies of Domestic Medicinal Plants in Macrophages (대식세포에서 국산약용식물의 항암 및 항Virus에 대한 효과)

  • Um, Sung-Hee;Kim, Dae-Keun;Kwak, Jong-Whan;Lee, Kang-Ro;Rhee, Dong-Kwon;Pyo, Suhk-Neung;Zee, Ok-Pyo
    • Korean Journal of Pharmacognosy
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    • v.26 no.3
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    • pp.259-264
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    • 1995
  • In the present work, 70 extracts from 23 plants have been determined to induce cytotoxic and antiviral activities of macrophages using both MTT assay and neutral red dye uptake assay. We show that 13 extracts have induced cytotoxic activities and 5 extracts induced antiviral activity in mouse peritoneal macrophages. Among 13 extracts, macrophages treated with extracts from Salvia plebeia have demonstrated significant cytotoxicity but not antiviral activity. The present findings indicate that extracts from plants can stimulate macrophages to become resistant to virus and to kill tumor cells.

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In vitro cell recovery methoc as an altermative to human damaged skin recovery test

  • An, Su-Sun;Nam, Ki-Taek;Park, Jong-Ho;Koh, Jae-Sook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.23 no.3
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    • pp.97-100
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    • 1997
  • These days, the raw materials that have the cell recovering effect are used commonly in cosmetics. In this study, six materials were rested for the characteristics of recovering effect both on vivo and in vitro. Tested raw materials were Soypol, 3-APPA, Apple extract, Polygonatum japonicum extract, Scutellarkd baicalensis extract, Aloe extract. Among these materials, Soypol and 3-APPA were synthesized and others were made by extraction at the Pacific R&D Center. Human forearm skin and cultured skin cell were damaged by sodium lauryl sulfare and then raw materials were applied for open treatment on SLS damaged human skin or cells. The recovering effects of raw materials in vivo were evaluated by measuring transepidermal water loss, skin hydration and erythema and in vitro effects of proliferationg cells were assessed by neutral red uptake assay. In the in vivo study, only the evaluation by TEWL showed correlation with the visual score. Our of six materials, 3-APPA had the most positive effect in both in vivo and in vitro studies and the correlation was r=0.8286 (p=0.042).

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The Growth Inhibitory Effects of Epigallocatechin Gallate Against Human Skin Melanoma Cells and Human Oral Epitheloid Carcinoma Cells (Epigallocatechin gallate의 인체 피부흑색종세포와 인체 구강유상피암종세포에 대한 성장억제효과)

  • 한두석;박승택;백승화
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.98-103
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    • 1998
  • Epigallocatechin gallate (EGCG) was reported to exert weak cytotoxicity against normal healthy cells such as C3H10T1/2 cells, but profound inhibitory effects on the initiation or promotion stage of chemical carcinogenesis in mammary gland, blood and mouse skin. This study was carried out to develop antitumor agents with weak side effects and strong antitumor activity. Human skin melanoma cells (HBT 69) and human oral epitheloid carcinoma cells (OCL 17) were cultured in RPMI-1640 media containing 10% fetal bovine serum, antibiotic, and fungizone. After incubation for 24 hrs, the cells were treated with various amounts of (EGCG) for 48 hrs. The growth inhibitory effects of EGCG in human oral epitheloid carcinoma cells were evaluated by the 3- (4,5-djmethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR), and sulforhodamine B protein (SRB) assays of colorimetric methods. The light microscopic study was also carried out to observe morphological changes of the treated cells. These results obtained were as follows; 1. Significantly inhibitory effects of EGCG against cultured human oral epithelioid carcinoma cells. 2. Significantly inhibitory effects against cultured human skin melanoma cells treated with 50 $\mu$M EGCG, but decreased inhibitory effects in 100 $\mu$M EGCG. 3. Degenerative changes against cultured human oral epitheloid carcinoma cells. 4. Degenerative changes against human skin melanoma cells treated with 50 UM EGCG, but recovered degenerative changes in 100 $\mu$M EGCG.

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산삼의 배양 및 그 응용에 관한 연구

  • Sin, Mi-Hui
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.151-162
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    • 2001
  • Korea mountain ginseng known as oriental miracle drug is an important medicinal plant. The effect of mountain ginseng adventitious roots extract has been described. The valuable root of mountain ginseng contained several kinds of ginsenosides that have been confirmed to have many active functions for the human body. However, the study of mountain ginseng has a limit because the price of wild ginseng is very expensive and rare. The mountain ginseng adventitious roots were derived from mountain ginseng callus that were induced from mountain ginseng roots. Adventitious roots were separated from callus and grown in solid media(Murachige and stoog media). It was cultured in a 20L bioreactor. After culturing for 40days, adventitious roots were harvested. Afterwards the harvested mountain ginseng adventitious roots were dryed and extracted. We examined the effect on melanogenesis of mountain ginseng adventitious roots extrac. Here, we report the inhibitory effect of melanin biosynthesis on the adventitious roots extract of In vitro test. Also, we assessed the safety of adventitious roots extract. In vitro, cytotoxicity of adventitious roots extract was assessed in mouse fibroblast using two method: The neutral red uptake assay and the MTT assay. In vivo, the allergic and irritant were patch tested in 30 patients. Consequently, extract of mountain ginseng adventitious roots have inhibitory effect on melanin biosynnthesis in B-16 melanoma cell test, tyrosinase inhibitory test and DOPA auto-oxidation test. There were decreased 86%(0.5% concentration), 45%(1% concentration) and 61%(1% concentration), respectively

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Effects of Airborne Samples Collected in Yeochun on Gap Junctional Inter cellular Communication in WBF-344 Rat Liver Epithelial Cells (여천공단 일부지역의 대기오염물질이 WBF-344간 상피세포의 Gap Junctional Intercellular Communication에 미치는 영향)

  • 양재만;박재학;김윤신;이영순
    • Journal of Korean Society for Atmospheric Environment
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    • v.13 no.3
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    • pp.207-214
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    • 1997
  • We collected airborne complex mixtures in a industrial area of Yeochun, and examined whether these complex mixtures could affect gap junctional intercellular communication (GJIC) in a cultured WBF-344 rat liver epithelial cells (LEC). Since the reduction of GJIC plays an important role in chemical carcinogenesis, measurement of changes of GJIC is a meaningful method to screen carcinogenicity of these mixtures. High and low volume samples were dissolved in dimethyl sulfoxide (DMSO) and tested. Blank filter extractions were also examined for exclud-ing possible toxicity of filter itself, and TPA (12-O-tetradecanoylphorbol-13-acetate) and DMSO were used as positive and negative control, respectively. When the cells were exposed to samples at concentration below that required to maintain rather than 85% cell viability based on the result of neutral red uptake assay, maximal inhibition of GJIC was observed at 1hr after treatment with both high and low volume samples by scrape-loading dye transfer assay. In fluorescence recovery after photobleaching assay, recovery rates via gap junctions were 33%/min in high volume sample and 62%/min in low volume sample. In together, airborne samples collected in Yeochun inhibited GJIC in a cultured WBF-344 rat LEC. These results suggest airborne samples tested in this experiment may attribute to cause a certain type and degree of cancers in in vivo when exposured for some periods.

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Development of Anticancer Agents from Korean Medicinal Plants. Part 7. Cytotoxic Activity of the Chloroform soluble Fraction of Perrila frutescens Against Human Oral Epitheloid Carcinoma Cells (한국산 생약으로 부터 항암물질의 개발 (제7보), 소엽의 Chloroform 가용성 분획이 인체 구강유상피암종세포에 미치는 세포독성작용)

  • Han, Du-Seok;Kim, Young-Il;Choi, Kyw-Eun;Kwag, Jung-Suk;Baek, Seung-Hwa
    • Environmental Mutagens and Carcinogens
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    • v.18 no.1
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    • pp.37-42
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    • 1998
  • In the present study, we have evaluated cytotoxic effects of the chloroform soluble fraction of the methanolic extract of Perilla frutescens in human oral epitheloid carcinoma cells. The light microscopic study showed morphological changes of the treated cells. Cell membrane damaging activity was measured by the lactate dehydrogenase (LDH) assay and disruptions in cell organelles were determined by 3-(4,5-dime-thylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR) and sulforhodamine protein B (SRB) of colorimetric assay. These results suggest that Perilla frutescens retains a potential antitumor activity.

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Electrochemical Activation of Nitrate Reduction to Nitrogen by Ochrobactrum sp. G3-1 Using a Noncompartmented Electrochemical Bioreactor

  • Lee, Woo-Jin;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.836-844
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    • 2009
  • A denitrification bacterium was isolated from riverbed soil and identified as Ochrobactrum sp., whose specific enzymes for denitrification metabolism were biochemically assayed or confirmed with specific coding genes. The denitrification activity of strain G3-1 was proportional to glucose/nitrate balance, which was consistent with the theoretical balance (0.5). The modified graphite felt cathode with neutral red, which functions as a solid electron mediator, enhanced the electron transfer from electrode to bacterial cell. The porous carbon anode was coated with a ceramic membrane and cellulose acetate film in order to permit the penetration of water molecules from the catholyte to the outside through anode, which functions as an air anode. A non-compartmented electrochemical bioreactor (NCEB) comprised of a solid electron mediator and an air anode was employed for cultivation of G3-1 cells. The intact G3-1 cells were immobilized in the solid electron mediator, by which denitrification activity was greatly increased at the lower glucose/nitrate balance than the theoretical balance (0.5). Metabolic stability of the intact G3-1 cells immobilized in the solid electron mediator was extended to 20 days, even at a glucose/nitrate balance of 0.1.

Enrichment of Hydrogenotrophic Methanogens in Coupling with Methane Production Using Electrochemical Bioreactor

  • Jeon, Bo-Young;Kim, Sung-Yong;Park, Yong-Keun;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1665-1671
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    • 2009
  • Anaerobic digestion sludge was cultivated in an electrochemical bioreactor (ECB) to enrich the hydrogenotrophic methanogens. A modified graphite felt cathode with neutral red (NR-cathode) was charged with electrochemical reducing power generated from a solar cell. The methane and carbon dioxide collected in a Teflon bag from the ECB were more than 80 ml/l of reactant/day and less than 20 ml/l of reactant/day, respectively, whereas the methane and carbon dioxide collected from a conventional bioreactor (CB) was around 40 ml/l of reactant/day, respectively. Moreover, the maximal volume ratios of methane to carbon dioxide (M/C ratio) collected in the Teflon bag from the ECB and CB were 7 and 1, respectively. The most predominant methanogens isolated from the CB on the $20^{th}$, $80^{th}$, and $150^{th}$ days of incubation were hydrogenotrophs. The methanogenic diversity analyzed by temperature gradient gel electrophoresis (TGGE) of the 16S rDNA variable region was higher in the ECB than in the CB. The DNA extracted from the TGGE bands was more than 95% homologous with hydrogenotrophic methanogens in the ECB, but was an aceticlastic methanogen in the CB. In conclusion, the ECB was demonstrated as a useful system for enriching hydrogenotrophic methanogens and increasing the M/C ratio of the gas product.