• Title/Summary/Keyword: H9c2 cells

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Isolation of the Efficacy Constituent for Neuronal Regeneration from Cibotium barometz (구척으로부터 신경재생 효능 성분 분리)

  • 김상태;한용남;손연경;장형석;김수장;신준식
    • YAKHAK HOEJI
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    • v.46 no.6
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    • pp.398-404
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    • 2002
  • A phytochemical study on the root of Cibotium barometz J. Smith led to the isolation of onitin (I), daucosterol (II) and a new compound (III). Compound III was characterized as 2-Ο-(9Z,12Z-octadecadienoyl)-3-Ο-[$\alpha$-D-galactopyranosyl-(1"-6")-Ο-$\beta$-D-galactopyranosyl] glycerol, named shinbarometin by $^1$H-, $^{13}$ C-NMR and LC/MS data. Compound III exerted an induced neuronal regeneration on nogo-A induced neuroblastoma cells.

Comparison of Environmental Stress Tolerance Between Lactobacillus fermentum Strains with High and Low Cell Surface Hydrophobicity

  • Li, Shao-Ji;Jeon, Jeong-Min;Hong, Sang-Won;So, Jae-Seong
    • Food Science and Biotechnology
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    • v.17 no.2
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    • pp.257-261
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    • 2008
  • Previous studies have suggested a possible correlation between cell surface hydrophobicity (CSH) and stress tolerance in Bifidobacterium. In this study, the relationship was examined between CSH and environmental stress tolerance in Lactobacillus spp. By measuring the adhesion to hexadecane, 2 Lactobacillus fermentum strains- KLB 261 and KLB 231 were found to have high and low CSH, respectively. To measure their tolerance to various stresses, cells were subjected to salt (2 M NaCl), acid (pH 2), $H_2O_2$ (0.01 %, v/v), ethanol (20%, v/v), heat ($60^{\circ}C$), and cold ($-20^{\circ}C$). Compared with KLB 231, the hydrophobic KLB 261 was found to be much more resistant to the various stresses examined. After being subjected to different stresses for a period of time, KLB 261 and KLB 231 showed 50 and 0% survivability in 2 M NaCl, 108.2 and 0.6% in 0.01 %(v/v) $H_2O_2$, 40.2%(v/v), and 3.7% at $60^{\circ}C$ incubation, 4 and 0.6% at $-20^{\circ}C$, 12.9 and 0.1 % in pH 2, 33.8 and 0.2% in 20%(v/v) ethanol, respectively. Autoaggregation test and morphological observation were also conducted in an attempt to explain these differences. These results suggested that high CSH could strengthen the stress tolerance of lactobacilli.

Synthesis of Electrolyte Films for Low-Temperature Solid Oxide Fuel Cells by Sol-Gel Coating and Their Characteristics (졸-겔 코팅에 의한 저온형 고체산화물 연료저지용 전해질막의 합성 및 특성)

  • 현상훈;김승구;장운석
    • Journal of the Korean Ceramic Society
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    • v.36 no.4
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    • pp.391-402
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    • 1999
  • Characteristics of composite electrolytes which were prepared by coating a thin film of YSZ (yttria sta-bilized zirconia : (ZrO2)0.92 (Y2O3)0.08) on YDC (yttria doped ceria : Ce0.8Y0.2O1.9) with mixed conductivity have been investigated in order to develop the low-temperature solid oxide fuel cell. The thickness (t) of spin-coated YSZ thin films after the heat-treatment at 600$^{\circ}C$ was increased proportionally to the sol con-centrations (C) while the decrease in its thickness with the spin rate ($\omega$) could be expressed in the e-quation of ln t=9.49-0.53 ln $\omega$(0.99mol//s sol conc.) When the sol concentration and the spin rate being less than 0.99 mol/l and higher than 1000 rpm respectively reliable YSZ/YDC composite electrolytes could be obtained by multi-coating although several micro-cracks were observed in singly coated YSZ film surfaces. The dense YSZ film with a 1$\mu\textrm{m}$ thickness was prepared by coating of 0.99 mol/l YSZ sol five-times at 2000 rpm followed by heat-treatment at 1400$^{\circ}C$ for 2h, The adhesion between YSZ film and YDC substrate was found to be very good. The open circuit voltages of H2/O2 single cell with YSZ/YDC composite electrolytes were 0.79∼0.82 V at 800$^{\circ}C$ and 0.75∼0.77V at 900$^{\circ}C$ The open circuit voltage was inversely proportioned to the thickness ratio of YSZ thin film (1$\mu\textrm{m}$) to YDC substrate(0.28-2.22 mm)

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Developmental Competence of Porcine NT Embryos Constructed by Microinjection of Fibroblast Cells into Vitrified Porcine Oocytes

  • Kim, Y.H.;Seok, H.B.;Kim, S.K.
    • Journal of Embryo Transfer
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    • v.22 no.4
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    • pp.265-269
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    • 2007
  • This study was conducted to investigate the efficacy of vitrification procedure for the cryopreservation of porcine oocytes and the utilization of vitrified oocytes as recipient cytoplasts for somatic cell nuclear transfer (NT), and observed that porcine oocytes are evaluated by pronuclear formation, and parthenogenetic development. Single fetal donor cells were deposited into the perivitelline space of vitrified enucleation oocytes, followed by electrical fusion and activation. NT embryos were cultured in NCSU-23 medium supplemented with 5% FBS, at $38.5^{\circ}C$ in 5% $CO_2$ and air. 1. When the developmental rates of the oocytes after being culture for $0{\sim}10$ hours vitrified with EDS and ETS were 42.0%, 38.0%, respectively. This results were lower than the control group(62.2%). 2. When the developmental rates of the oocytes after being culture for $0{\sim}10$ hours vitrified-thawed with sucrose and glucose, 5% PVP, NCSU-23 supplemented with 10% FBS were 33.3%, 25.9%, respectively. This results were lower than the control group(55.6%). 3. The fusion and development to the blastocyst stage between the NT embryos constructed with the vitrified and non-vitrified oocytes were significant differences. Developmental rate of oocytes and NT embryos constructed with the vitrified or non-vitrified oocytes were $13.0{\pm}2.4%\;and\;23.2{\pm}2.4%$, respectively.

Identification of 5-Hydroxy-3,6,7,8,3',4'-Hexamethoxyflavone from Hizikia fusiforme Involved in the Induction of the Apoptosis Mediators in Human AGS Carcinoma Cells

  • Kim, Min Jeong;Lee, Hye Hyeon;Seo, Min Jeong;Kang, Byoung Won;Park, Jeong Uck;Kim, Kyoung-Sook;Kim, Gi-Young;Joo, Woo Hong;Choi, Yung Hyun;Cho, Young-Su;Jeong, Yong Kee
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1665-1672
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    • 2012
  • An 80% ethanol extract of Hizikia fusiforme was obtained and followed by successive fractionation using the organic solvents n-hexane, ethyl acetate, and n-butanol to identify the antioxidative substance. The aqueous part of the nbutanol fractionation step, showing high antioxidative activity, was subjected to reverse-phase liquid chromatography. As a result, a substance purified from a BB-2 fraction showed high antioxidative activity. The m/z 419 [M+H] molecular ion peak in the fraction was observed by the analysis of the ESI-LC/MS spectrum. By the analysis of 1H NMR (500 MHz, DMSO-$d_6$) and $^{13}C$ NMR (125 MHz, DMSO-$d_6$) spectra, a unique compound of the fraction was biochemically identified as a 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5HHMF). We also investigated the effect of 5HHMF on human gastric AGS carcinoma cells. Western blot analysis suggested that the flavone substantially increased the levels of the death receptor-associated apoptosis mediators Fas, Fas L, FADD, TRADD, and DR4 in a concentration-dependent manner. The levels of Fas, Fas L, TRADD, and DR4 in the cells treated with 5HHMF ($5{\mu}g/ml$) were approximately 26.4-, 12.8-, 6.7-, and 9.8-times higher than those of non-treated cells, respectively. Of note, the level of FADD protein in the cells exposed to 5HHMF ($1{\mu}g/ml$) increased approximately 9.6-times. In addition, the cleavage of caspase-3, -8, and -9 in cultured AGS cells treated with 5HHMF was significantly confirmed. Therefore, our results suggest that 5HHMF from H. fusiforme is involved in the induction of death receptor-associated apoptosis mediators in human gastric AGS carcinoma cells.

Synergistic effect of ionizing radiation and $\beta$-lapachone against tumor in vitro and in vivo

  • Park, Eun-Kyung;Kim, Young-Seok;Lee, Sang-wook;Ahn, Seung-Do;Shin, Seong-Soo;Park, Heon-Joo;Song, Chang-Won
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.80-80
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    • 2003
  • ${\beta}$-lapachone(${\beta}$-Lap), a natural o-naphthoquinone, presents in the bark of the Lapacho tree. ${\beta}$-Lap is cytotoxic against a variety of human cancer cells and it potentiates the anti-tumor effect of Taxol. In addition, ${\beta}$-Lap has been reported to radiosensitize cancer cells by inhibiting the repair of radiation-induced DNA damage.In the present study, we investigated the cytotoxicity of ${\beta}$-Lap against RKO human colorectal cancer cells as well as the combined effect of ${\beta}$-LaP and ionizing radiation. An incubation of RKO cells with 5 ${\mu}$M of ${\beta}$-Lap for 4 h killed almost 90% of the clonogenic cells. An incubation of RKO cells with 5 ${\mu}$M of ${\beta}$-Lap for 4 h or longer also caused massive apoptosis. Unlike other cytotoxic agents, ${\beta}$-Lap did not increase the expression of p53 and p21 and it suppressed the NFkB expression. The expression of Caspase 9 and 3 was minimally altered by ${\beta}$-Lap. Radiation and ${\beta}$-Lap acted synergistically in inducing clonogenic cell death and apoptosis in RKO cells when ${\beta}$-Lap treatment was applied after but not before the radiation exposure of the cells. Interestingly, a 4 h treatment with 5 ${\mu}$M of ${\beta}$-Lap starting 5 h after irradiation was as effective as that starting immediately after irradiation. The mechanisms of ${\beta}$-Lap-induced cell killing is controversial but a recent hypothesis is that ${\beta}$-Lap is activated by NAD(P)H: quinone-onidoreductase (NQO1) in the cells followed by an elevation of cytosolic Ca$\^$2+/ level and activation of proteases leading to apoptosis. It has been reported that NQO1 level in cells is markedly up-regulated for longer than 10 h after irradiation. Indeed, using immunological staining of NQO1, we observed a significant elevation of NQO1 expression in RKO cells 5h after 2-4 Gy irradiation. Such a prolonged elevation of NQO1 level after irradiation may be the reasons why the ${\beta}$-Lap treatment applied S h after irradiation was as effective as that applied immediately after irradiation in killing the cells. In view of the fact that the repair of radiation-induced damage is usually completed within 1-2 h after irradiation, it is highly likely that the ${\beta}$-Lap treahment applied 5 h after irradiation could not inhibit the repair of radiation-induced damage. For in vivo study, RKO cells were injected S.C. into the hind-leg of Nu/Nu mice, and allowed to grow to 130 mm3 tumor. The mice were i.p. injected with ${\beta}$-lapachone or saline 2 h after irradiation of tumors with 10 Gy of X-rays. The radiation induced growth delay was increased by 2.4 $\mu\textrm{g}$/g of ${\beta}$-lapachone. Taken together, we may conclude that the synergistic interaction of radiation and ${\beta}$-Lap in killing cancer cells is not due to radiosensitization by ${\beta}$-Lap but to an enhancement of ${\beta}$-Lap cytotoxicity by radiation through an upregulation of NQO1. The fact that NQO1 is elevated in tumors and that radiation causes prolonged increase of the NQO1 expression may be exploited to preferentially kill tumor cells using ${\beta}$-Lap in combination with radiotherapy.

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RNA Production from the Hydrolyzate of Sliced and Dried Sweet Potatoes. (절간고구마를 이용한 RNA 생산에 관한 연구)

  • 이종임;송재철;조원대;양한철
    • Microbiology and Biotechnology Letters
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    • v.8 no.1
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    • pp.9-18
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    • 1980
  • During an extensive screening tests of yeasts for their RNA formation, it was found that Cryptococcus laurentii had especially high RNA content and high dry cell weight, when hydrolyzate of sliced and dried sweet potatoes was used as a carbon source. Growth conditions of this strain were examined, and the most desirable results were obtained at 48 hours of cultivation on a reciprocal shaker at 3$0^{\circ}C$ with initial pH 6.0. Under the above conditions, the RNA content and yield of dry cells were investigated using various media compositions. Ammonium sulfate 0.40%, peptone 0.6 %, and yeast extract 0.4% were appeared to be favorable as a nitrogen sources. The optimum concentrations of K $H_2$P $O_4$, M $n^{++}$, C $O^{++}$ were 0.05 %, 0.1 %, and 0.001 %, respectively. Ca-pantothenate, 400$\mu\textrm{g}$/$m\ell$, showed relatively favorable effects as a growth factor. The maximum RNA content obtained in this study was 16.8 % of the total dry cell weight.t.t.

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Effect of Puromycin Aminonucleoside on Podocyte P-Cadherin (Puromycin aminonucleoside의 사구체 족세포 P-cadherin에 대한 영향)

  • Ha, Tae-Sun
    • Childhood Kidney Diseases
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    • v.17 no.2
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    • pp.79-85
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    • 2013
  • Purpose: To test whether the expression of P-cadherin, a component of slit diaphragms between podocyte foot processes, would be altered by puromycin aminonucleoside (PAN) in a cultured podocyte in vitro. Methods: Rat glomerular epithelial cells (GEpC) were cultured with various concentrations of PAN. The distribution of P-cadherin was examined with a confocal microscope. Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to measure the change in P-cadherin expression. Results: This study found that P-cadherin was concentrated in the inner and peripheral cytoplasm with high concentrations of PAN under immunofluorescence views. Western blotting of GEpC revealed that PAN induced a decrease of P-cadherin in dose- and time-dependent manners. A high dose ($50{\mu}g/mL$) of PAN decreased P-cadherin expression by 21.9% at 24 h (P <0.05) and 31.9% at 48 h (P <0.01) compared to those without PAN. In RT-PCR, high concentrations ($50{\mu}g/mL$) of PAN also decreased P-cadherin mRNA expression, similar to protein suppression, by 23.5% at 48 h (P <0.05). Conclusion: Podocytes exposed to PAN in vitro concentrated P-cadherin internally, and reduced P-cadherin mRNA and protein expression. This could explain the development of proteinuria in experimental PAN-induced nephropathy.

Composting of Livestock Waste and Development of Operating Parameters I. Development of Optimum Process Parameters in Cow Manure Composting (축산 폐기물의 퇴비화 및 운용지표 개발 I. 우분의 퇴비화에 있어서 최적 공정운용지표의 개발)

  • Chung, Jae-Chun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.1 no.1
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    • pp.69-84
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    • 1993
  • In order to determine the optimum operational paramsters in cow manure composting, 4 laboratory scale composters were established. The cow manure was mixed with certain amount of saw dust to adjust the initial C/N ratio to 24, initial pH to 6.9 and composting was performed with varying operational conditions. It was found that the optimum aeration rate was 1000 ml/min kg. VS, the optimum moisture content 50% and no significant difference was found with different initial pH condition. Microorganisms were counted under the optimum conditions determined in this study. At the end of the experimental period, the number of bacteria, actinomycetes and fungi was $1.5{\times}10^9$ cells, $1.1{\times}10^8$ cells and $3.0{\times}10^8$ cells/g dry compost, respectively. At day 0, the number of coliforms, fecal coliforms and fecal streptococci was $3.1{\times}10^3$ cells, $7.5{\times}10^2$ cells and $5.6{\times}103$ cells/g dry composting material, respectively. Their population was decreased with time lapse, However, their survival time was longer than those reported by other researchers. Microorganisms were identified at the end of the experiment. Genus Bacillus was the most dominant comprising 89.3% of the total population. Among the Genus Bacillus, B. circulans compoex was the most abundant, followed by B. Stearothermophilus, B. Sphericus, B. licheniformis and B, brevis.

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Effect of green tea on the survival of Escherichia coli O157:H7 and Salmonella typhimurium in mayonnaise (마요네즈에 첨가한 녹차가 Escherichia coli O157:H7과 Salmonella typhimurium의 생존에 미치는 영향)

  • 박찬성;박금순
    • Korean journal of food and cookery science
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    • v.18 no.1
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    • pp.57-62
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    • 2002
  • The purpose of this study was to investigate the antibacterial activity of green tea in mayonnaise against pathogenic bacteria (Escherichia coli O157:H7 and Salmonella typhimurium). Mayonnaise was prepared with salad oil, egg yolk, sugar, salt and vinegar, and green tea powder was added to the mayonnaise at 0.1, 0.3 and 0.5% for the experiment. Then, the mayonnaise samples with various levels of green tea were inoculated with about 10$\^$6/ cells/g of E. coli and S. typhimurium per 1 gram of mayonnaise and stored at 5$^{\circ}C$, 15$^{\circ}C$ and 25$^{\circ}C$ for 3∼9 days. Antibacterial activity of green tea was tested by the colony counting method for E. coli on violet red bile agar(VRBA) and S. typhimurium on xylose lysine desoxycholate agar(XLDA). The D-values of E. coli controls were 2.89, 2.73 and 2.13 days while those of S. typhimurium controls were 0.58, 0.53 and 0.52 days at 5$^{\circ}C$, 15$^{\circ}C$ and 25$^{\circ}C$, respectively Inhibitory effect of green tea in mayonnaise on the survival of E. coli and S. typhimurium was increased with increasing concentration of green tea and/or increasing storage temperature. The most effective antibacterial activity of green tea was shown against E. coli in mayonnaise during storage at 25$^{\circ}C$.