• Title/Summary/Keyword: cell concentration

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A New Method for Determination of Enzyme Reaction and Activity of Lysozyme with UV-Spectrophotometer (UV-분광광도계를 이용한 새로운 Lysozyme의 효소반응 및 활성측정법 연구)

  • Kim, Woon-Soo;Kim, Yong-Wook;Kim, Woo-Sik
    • Applied Chemistry for Engineering
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    • v.9 no.6
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    • pp.857-863
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    • 1998
  • A simple and new experimental method for determination of lysozyme-M. lysodeikticus cell lysis reaction and lysozyme activity was suggested using Beer's law. The UV transmittance of the solution changed with the concentration of M. lysodeikticus and the relationship between the UV transmittance and M. lysodeikticus cell concentration followed Beer's Law. In addition, it was experimentally proven that the UV transmittance of the solution was not influenced by the lysozyme concentration and product of the lysis reaction. During the lysozyme-M. lysodeikticus cell lysis reaction, thus, M. lysodeikticus cell concentration in the solution could be measured in-situ by UV-spectrophotometer. By using these experimental data, kinetic Parameters of the Michaelis-Menten equation for the lysozyme-M. lysodeikticus cell 1ysis reaction was simply determined The maximum reaction rate constant ($k_3$) and Michaelis-Menten constants were $0.1734sec^{-1}$ and $9.83{\times}10^{-6}M$ respectively. The activity of the lysozyme could also be obtained with this experiment because the lysis reaction rate of the 1ysozyme depended on its activity.

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Bioremediation of Diesel-Contaminated Soil by Bacterial Cells Transported by Electrokinetics

  • LEE, HYO-SANG;KISAY LEE
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1038-1045
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    • 2001
  • The electrokinetic technology was applied in bioremediation for the purpose of supplying a Pseudomonas strain capable of degrading diesel to contaminated soil bed, and their biodegradation of diesel was carried out after a desired cell distribution was obtained. Electrokinetic injection of the strain was made possible because the cells acted as negatively charged particles at neutral pH, and thus the cells were transported with a precise directionality through the soil mostly by the mechanism of electrophoresis and in part by electroosmosis. A severe pH change in the soil bed was formed due to the penetration of electrolysis products, which was harmful to the cell viability and cell transport. To achieve a desirable cell transport and distribution, the control of pH in soil bed by a recirculating buffer solution in electrode chambers was essential during the appliation of an electric field. The judicious selections of electrolyte concentration and conductivity were also important for achieving an efficient electrokinetic cell transport since a higher electrolyte concentration favored the maintenance of pH stability in soil bed, but lowered electrophoretic mobility on the other hand. With electrolyte solution of pH 7 phosphate buffer, a 0.05 M concentration showed a better cell transport buffer, a 0.05 M concentration showed a better cell transport than 0.02 M and 0.08 M. The cell under pH 8 were obtained, compared to the cells under pH 7 or pH 9 in a given time period Up to $60\%$ of diesel was degraded in 8 days by the Pseudomonas cell, which were distributed electrokinetically under the conditions of pH 8 ($1,800{\mu}S/cm$, a mixture of phosphate and ammonia buffers) and 40 mA in a soil bed of 15 cm length.

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Ethanol Elicits Inhibitory Effect on the Growth and Proliferation of Tongue Carcinoma Cells by Inducing Cell Cycle Arrest

  • Le, Thanh-Do;Do, Thi Anh Thu;Yu, Ri-Na;Yoo, Hoon
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.3
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    • pp.153-158
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    • 2012
  • Cellular effects of ethanol in YD-15 tongue carcinoma cells were assessed by MTT assay, caspase activity assay, Western blotting and flow cytometry. Ethanol inhibited the growth and proliferation of YD-15 cells in a dose- and time-dependent manner in an MTT assay. The effects of ethanol on cell cycle control at low percent range of ethanol concentration (0 to 1.5%), the condition not inducing YD-15 cell death, was investigated after exposing cells to alcohol for a certain period of time. Western blotting on the expression of cell cycle inhibitors showed that p21 and p27 was up-regulated as ethanol concentration increases from 0 to 1.5% whilst the cell cycle regulators, cdk1, cdk2, and cdk4 as well as Cyclin A, Cyclin B1 and Cyclin E1, were gradually down-regulated. Flow cytometric analysis of cell cycle distribution revealed that YD-15 cells exposed to 1.5% ethanol for 24 h was mainly arrested at G2/M phase. However, ethanol induced apoptosis in YD-15 cells exposed to 2.5% or higher percent of ethanol. The cleaved PARP, a marker of caspase-3 mediated apoptosis, and the activation of caspase-3 and -7 were detected by caspase activity assay or Western blotting. Our results suggest that ethanol elicits inhibitory effect on the growth and proliferation of YD-15 tongue carcinoma cells by mediating cell cycle arrest at G2/M at low concentration range and ultimately induces apoptosis under the condition of high concentration.

Effect of CLA (Conjugated Linoleic Acid) on the Anti-Atherosclerotic actors in Human Hepatoma HepG2 Cells (간암세포 (HepG2 Cell)에서의 식이성 CLA(Conjugated Linoleic Acid)가 항동맥경화성 인자에 미치는 영향)

  • 오현희;문희정;이명숙
    • Journal of Nutrition and Health
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    • v.37 no.3
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    • pp.182-192
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    • 2004
  • Conjugated linoleic acid (CLA) is the mixture of positional and geometric isomers of linoleic acid (LA), which is found abundantly in dairy products and meats. This study was performed to investigate the anticarcinogenic effect of CLA in HepG2 hepatoma cells. HepG2 cell were treated with LA and CLA at the various concentrations of 10, 20, 40, 80 uM each at different incubation times. After each incubation times, cell proliferation, fatty acids incorporation into cell, peroxidation and postaglandin E$_2$ (PGE$_2$) and thromboxane $A_2$ (TXA$_2$) for the eicosanoid metabolism were measured. LA treated HepG2 cells were increased cell growth 6 - 70% of control whereas CLA increased cell death the half of those in LA group (p 〈 0.001). LA and CLA were incorporated very well into the cellular membranes four times higher than in control according to concentration and longer incubation times. Moreover, LA synthesized significantly arachidonic acids corresponding with LA concentration compared to CLA supplementation. The supplementation with LA increased intracellular lipid peroxides concentration corresponding with LA concentration and five times higher than those in CLA significantly at any incubation times (p 〈 0.001). PGE$_2$ and TXA$_2$ levels were three to twenty times lower in condition of CLA treatments than LA, respectively. Overall, the dietary CLA might change the HepG2 cell growth by the changes of cell composition, production of lipid peroxide. Since CLA have not changed the levels of arachidonic acid of cell membrane, which was sources of eicosanoids, eicosanoid synthesis was not increased in CLA compared to LA. Our results was suggest CLA has a possibility to protect the progress of atherosclerosis because CLA does not produce lipid production and endothelial contraction factors in liver.

The Effect of High Concentration of Ascorbic Acid on the Growth of Primary Cultured Cells of Chondrocytes (고농도 비타민 C첨가가 연골 초대배양세포의 증식에 미치는 영향)

  • 김미향
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.5
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    • pp.797-802
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    • 2004
  • L-Ascorbic acid (AsA), commonly known as vitamin C, which is one of the antioxidant vitamins, plays a role in cellular oxidant quenching. Some of the biochemical reactions in which it takes part have been traced through organ culture technique. But in cell cultured system, views on stimulatory and inhibitory action of AsA on cell growth are conflicting. Therefore, this study aimed to clarify the inhibitory action of high concentration AsA on the cell growth in Primary chondrocyte isolated from rat ribs. Cells were exposed to ascorbate at various concentrations. Supplement of AsA induced stimulation of cell growth in primary cultured cells of chondrocytes. Most remarkable stimulation of cell growth by AsA was found in primary cultured chondrocytes. However, it showed that they were dead in the medium which contained AsA at the concentration higher than 1.0 mM. This lethal effect of AsA causing the cell death was inhibited by the addition of catalase in the medium. This supposed that hydroxyl radical (ㆍOH) induced from $H_2O$$_2$ was actively cytotoxic agent. Based on the results, when AsA was added in medium at normal concentrations, the cell growth was stimulated by inducing the formation of extracellular matrix. On the contrary, if added in medium at excess concentrations, the cell growth was inhibited because $H_2O$$_2$ were generated from AsA in medium. Therefore, addition of AsA at the normal concentrations stimulates cell growth, but excess concentrations of AsA induces cell death.

Effects of 2,4-D, BA, and Sucrose on Growth, Production of Anthocyanin, pH, and Sugar Content in 'Sheridan' Grape Cell Suspension Cultures

  • Kim, Seung-Heui;Kim, Seon-Kyu
    • Journal of Plant Biotechnology
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    • v.4 no.2
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    • pp.77-82
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    • 2002
  • To elucidate the effect of sucrose on cell growth and anthocyanin production, 1, 3, 5, and 7% sucrose were applied to liquid MS basal medium supplemented with 0.5 mg/L BA + 0.1 and 1 mg/L 2,4-D. Higher sucrose concentration decreased the cell growth regardless of the hormonal composition. Cain in fresh weight was gradual, showing the peak at day 12 in culture, and then decreased. Anthocyanin content increased with sucrose concentration in the medium, and practically there was no difference in anthocyanin content between the two media differing in 2,4-D content. Sucrose concentration for appropriate anthocyanin production was 7%, while 5% was more suitable for increase in total anthocyanin content. At higher sucrose levels, anthocyanin content was high due to the cessation of the cell growth. Medium pH decreased at the early stage and gradually increased thereafter.

Effects of Nutrients and Cell Aggregate Size on the Biosynthesis of Carotenoid in Daucus carota Suspension Culture (Daucus carota 현탁 배양에서 배지 조성 및 세포 응집 크기가 Carotenoid 생합성에 미치는 영향)

  • 윤정원;김지현유영제변상요
    • KSBB Journal
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    • v.5 no.4
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    • pp.347-353
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    • 1990
  • The effects of nutrients and cell aggregate size on the cell growth of D. carota and the biosynthesi of carotenoid were investigated. Highest carotenoid content was obtained with sucrose as a carbon source and the equal ratio of ammonium to nitrate. High phosphate concentration stimulated the carotenoid biosynthesis in D. corota. 2,4-D inhibited the cell growth but stimulated the specific carotenoid content at high concentration. By modifying the medium composition based on these findings, three times higher specific carotenoid content and 2.5 times higher total carotenoid content were obtained as compared with the results obtained with basic MS media. Biosynthesis of carotenoid was found to be affected by cell aggregate size; high carotenoid production was obtained from the large aggregated cells resulted from high sucrose concentration.

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Concentration of cell-free fermentation broth containing poly (${\gamma}-glutamic acid$) by ultrafiltration

  • Do, Jin-Hwan;Jang, Ho-Nam;Lee, Sang-Yeop
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.45-46
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    • 2000
  • After cell removal from fermentation broth the supernatant containing poly (${\gamma}-glutamic acid$) (${\gamma}-PGA$) was concentrated by ultrafiltration in order to reduce the amount of organic solvents such as methanol, ethanol, propanol required for the recover of ${\gamma}-PGA$ with precipitation. The concentration and volume reduction of cell-free solution by ultrafiltration could reduce 3 times the amount of ethanol for the recover of ${\gamma}-PGA$ from cell-free fermentation broth.

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Selective in vitro cytotoxicity of Hypericum hookerianum towards cancer cell lines

  • P., Vijayan;S., Vinod Kumar;Badami, Shrishailappa;PK, Mukherjee;SA, Dhanaraj;B., Suresh
    • Advances in Traditional Medicine
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    • v.3 no.3
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    • pp.141-146
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    • 2003
  • The methanol extracts of the aerial parts, leaves and stem of Hypericum hookerianum were tested for in vitro cytotoxicity on selected normal and cancer cell lines and anti tumor activity using DLA cells. Cell viability and morphological changes were assessed. Among the three extracts tested, the stem extract of Hypericum hookerianum showed potent cytotoxicity against HEp-2 and RD cell lines. The $CTC_{50}$(concentration required to reduce viability by 50%) of this extract was found to be $2.02\;{\mu}g/ml$ for RD cell line, $10.25\;{\mu}g/ml$ for HEp-2 cell line and $100.06\;{\mu}g/ml$ for Vero cell line. In the clonogenic assay, no colony formation was observed up to a concentration of $100\;{\mu}g/ml$. In the short term cytotoxicity studies using DLA cells, 50% viability was observed in the concentration range of $50-100\;{\mu}g/ml$ for aerial parts, $100-200\;{\mu}g/ml$ for stem and more than $200\;{\mu}g/ml$ for leaf extracts of Hypericum hookerianum. In the long-term activity using HEp-2 cell line, no colony formation was observed over a concentration of 200 mg/ml for the stem extract. Hypericum hookerianum stem extract was fractionated into petroleum ether, chloroform, ethyl acetate and methanol soluble fractions. The petroleum ether and chloroform soluble fractions showed higher cytotoxic activity against HEp-2 cell line when compared to the other two fractions. The methanol stem extract of Hypericum hookerianum has the potential for further investigation in animal models to determine its anti-tumor activity and to identify its active principles.

Preventive Effect of Lysis in the Cadmium-Tolerant Hansenula anomala B-7 Cells by Metal Ions (금속 이온에 의한 카드늄 내성균주 Hansenula anomala B-7 세포의 용균 방지 효과)

  • Song, Hyung-Ik;Yu, Tae-Shick
    • The Korean Journal of Mycology
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    • v.19 no.4
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    • pp.282-284
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    • 1991
  • The yeast, Hansenula anomala B-7, isolated from the $Cd^{2+}$ rich soils and determined to be tolerant in the high concentration of $Cd^{2+}$ were employed in this work. Its intact cells grown in high concentration of $Cd^{2+}$ were observed to be Iysed at the early stage when transferred to a cadmium deficient broth. Its intact cells found to be not Iysed and grow well under the high concentration of $Cd^{2+}$. The Iysis of the intact cells grown at the high concentration of $Cd^{2+}$ ion was not found when the metal ions were replaced with $Cd^{2+}$ ion in the same concentration. This result indicated that Iysis of yeast cells, at least in this isolate, would be related to cell osmosis with the mineral ions added.

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