• Title/Summary/Keyword: Cell growth and division

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Effect of Nitrogen on Cell Dynamics at Leaf Growth Zone in Two Rice Varieties

  • Sung, Jwa-Kyung;Lee, Chul-Won;Kim, Tae-Wan;Hwang, Seon-Woong;Song, Beom-Heon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.2
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    • pp.121-125
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    • 2004
  • In plants, nitrogen is the major component for growth and development. Leaf growth is based on the division, elongation and maturation of cells, which are used for making of epidermis, mesophyll, bundle sheath, xylem, phloem and so on. Dynamics of these tissues with respect to nitrogen are required for better understanding. This experiment was conducted to evaluate effect of nitrogen on the elongation of epidermal and guard cell of two rice (Oryza sativa L.) varieties, Seoanbyeo and Dasanbyeo on May 2000 at Chungbuk national university in Cheongju. After transplaning the 20-day-old seedlings into a/5000 pots, the main characteristics related with cell elongation were investigated and evaluated. A maximum. leaf length reached at 7 or 8 days after emerging from the collar, and also the leaf elongation rates were greatly affected by the increase of N application rate. The initial and final cell length were about $17\mu\textrm{m}$ and $130\mu\textrm{m}$, respectively. Cell divisions occurred within 1.0mm from leaf base. With die higher nitrogen application rate of 22 kg-N $10\textrm{a}^{-1}$, cell division per hour was greater 1.5 to 1.9 and 1.2 to 1.3 fold as compared to the N application rate of 0 and 11 kg-N $10\textrm{a}^{-1}$, respectively. Cell enlargement of epidermal and guard cell under higher N application rate (22kg-N $10\textrm{a}^{-1}$) was finished within about 20 (Seoanbyeo) and 15 hours (Dasanbyeo), while it took much time, about 30 hours.

Optimization of Betacyanin Production by Red Beet (Beta vulgaris L.) Hairy Root Cultures. (Red Beet의 모상근 배양을 이용한 천연색소인 Betacyanin 생산의 최적화)

  • Kim, Sun-Hee;Kim, Sung-Hoon;Lee, Jo-No;An, Sang-Wook;Kim, Kwang-Soo;Hwnag, Baik;Lee, Hyeong-Yong
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.435-441
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    • 1998
  • Optimal conditions for the production of natural color, betacyanin were investigated by varying light intensity, C/N ratio, concentrations of phosphate and kinds of elicitors. Batch cultivation was employed to characterize cell growth and betacyanin production of 32 days. The maximum specific growth rate, ${\mu}$$\sub$max/, was 0.3 (1/day) for batch cultivation. The maximum specific production rate, q$\^$max/$\sub$p/, was enhanced 0.11 (mg/g-cell/day) at 3 klux. A light intensity of 3 klux was shown to the best for both cell growth and betacyanin production. The maximum specific production rate was 0.125 (mg/g-cell/day) at 0.242 (1/day), the maximum specific growth rate. The dependence of specific growth rate on the light lintensity is fit to the photoinhibition model. The correlation between ${\mu}$ and q$\sub$p/ showed that the product formation parameters, ${\alpha}$ and ${\beta}$$\sub$p/ were 0.3756 (mg/cell) and 0.001 (mg/g-cell/day), respectively. The betacyanin production was partially cell growth related process, which is different from the production of a typical product in plant cell cultures. In C/N ratio experiment, high carbon concentration, 42.1 (w/w) improved cell growth rate while lower concentration, 31.6 (w/w) increased the betacyanin production rate. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.26 (1/day) and 0.075 (mg/g-cell/day), respectively. Beta vulgaris L. cells under 1.25 mM phosphate concentration produced 10.15 mg/L betacyanin with 13.46 (g-dry wt./L) of maximum cell density. The production of betacyanin was elongated by adding 0.1 ${\mu}$M of kinetin. This also increased the cell growth. Optimum culture conditions of light intensity, C/N, phosphate concentration were obtained as 5.5 klux, 27 (w/w), 1.25 mM, respectively by the response surface methodology. The maximum cell density, X$\sub$max/, and maximum production, P$\sub$max/, in optimized conditions were 16 (g-dry wt./L), 12.5 (mg/L) which were higher than 8 (g-dry wt./L), 4.48 (mg/L) in normal conditions. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.376 (1/day) and 0.134 (mg/g-cell/day) at the optimal condition. The overall results may be useful in scaling up hairy root cell culture system for commercial production of betacyanin.

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Autophagy Inhibition with Monensin Enhances Cell Cycle Arrest and Apoptosis Induced by mTOR or Epidermal Growth Factor Receptor Inhibitors in Lung Cancer Cells

  • Choi, Hyeong Sim;Jeong, Eun-Hui;Lee, Tae-Gul;Kim, Seo Yun;Kim, Hye-Ryoun;Kim, Cheol Hyeon
    • Tuberculosis and Respiratory Diseases
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    • v.75 no.1
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    • pp.9-17
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    • 2013
  • Background: In cancer cells, autophagy is generally induced as a pro-survival mechanism in response to treatment-associated genotoxic and metabolic stress. Thus, concurrent autophagy inhibition can be expected to have a synergistic effect with chemotherapy on cancer cell death. Monensin, a polyether antibiotic, is known as an autophagy inhibitor, which interferes with the fusion of autophagosome and lysosome. There have been a few reports of its effect in combination with anticancer drugs. We performed this study to investigate whether erlotinib, an epidermal growth factor receptor inhibitor, or rapamycin, an mammalian target of rapamycin (mTOR) inhibitor, is effective in combination therapy with monensin in non-small cell lung cancer cells. Methods: NCI-H1299 cells were treated with rapamycin or erlotinib, with or without monensin pretreatment, and then subjected to growth inhibition assay, apoptosis analysis by flow cytometry, and cell cycle analysis on the basis of the DNA contents histogram. Finally, a Western blot analysis was done to examine the changes of proteins related to apoptosis and cell cycle control. Results: Monensin synergistically increases growth inhibition and apoptosis induced by rapamycin or erlotinib. The number of cells in the sub-$G_1$ phase increases noticeably after the combination treatment. Increase of proapoptotic proteins, including bax, cleaved caspase 3, and cleaved poly(ADP-ribose) polymerase, and decrease of anti-apoptotic proteins, bcl-2 and bcl-xL, are augmented by the combination treatment with monensin. The promoters of cell cycle progression, notch3 and skp2, decrease and p21, a cyclin-dependent kinase inhibitor, accumulates within the cell during this process. Conclusion: Our findings suggest that concurrent autophagy inhibition could have a role in lung cancer treatment.

Myogenic Differentiation of p53- and Rb-deficient Immortalized and Transformed Bovine Fibroblasts in Response to MyoD

  • Jin, Xun;Lee, Joong-Seub;Kwak, Sungwook;Jung, Ji-Eun;Kim, Tae-Kyung;Xuo, Chenxiong;Hong, Zhongshan;Li, Zhehu;Kim, Sun-Myoung;Whang, Kwang Youn;Hong, Ki-Chang;You, Seungkwon;Choi, Yun-Jaie;Kim, Hyunggee
    • Molecules and Cells
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    • v.21 no.2
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    • pp.206-212
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    • 2006
  • We have established in culture a spontaneously immortalized bovine embryonic fibroblast (BEF) cell line that has lost p53 and $p16^{INK4a}$ functions. MyoD is a muscle-specific regulator capable of inducing myogenesis in a number of cell types. When the BEF cells were transduced with MyoD they differentiated efficiently to desmin-positive myofibers in the presence of 2% horse serum and 1.7 nM insulin. The myogenic differentiation of this cell line was more rapid and obvious than that of C2C12 cells, as judged by morphological changes and expression of various muscle regulatory factors. To confirm that lack of the p53 and $p16^{INK4a}$ pathway does not prevent MyoD-mediated myogenesis, we established a cell line transformed with SV40LT (BEFV) and introduced MyoD into it. In the presence of 2% horse serum and 1.7 nM insulin, the MyoD-transduced BEFV cells differentiated like the MyoD-transduced BEFS cells, and displayed a similar pattern of expression of muscle regulatory proteins. Taken together, our results indicate that MyoD overexpression overcomes the defect in muscle differentiation associated with immortalization and cell transformation caused by the loss of p53 and Rb functions.

An Optimization of Flavonoid Production from the Suspension Culture of Scutellaria baicalensis Georgi Cells

  • SEO, WEON-TAEK;YOUNG-HOON PARK;TAE-BOO CHOE
    • Journal of Microbiology and Biotechnology
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    • v.6 no.5
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    • pp.347-351
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    • 1996
  • Flavonoid production by suspended cells of Scutellaria baicalensis Georgi was studied and the medium was optimized for cell growth and baicalin production. In SH medium the flavonoid production was not closely associated with the cell growth. A modified SH medium, FPM, was therefore designed for enhanced baicalin production. In FPM, both cell growth and baicalin production were increased by 1.5 times and 1.67 times than in the original SH medium, respectively. The increases could be attributed to the increased metabolic activities involved in the flavonoid biosynthesis as represented by enhanced activities of phenylalanine ammonia-lyase.

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Purification and Characterization of CDMHK, a Growth Inhibitory Molecule Against Cancer Cell Lines, from Myxobacterium sp. HK1 Isolated from Korean Soil

  • LEE HAN-KI;LEE IN-HYE;YIM JEE-SUN;KIM YONG-HO;LEE SANG-HEE;LEE KISAY;KOO YOON-MO;KIM SANG-JIN;JEONG BYEONG-CHUL
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.734-739
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    • 2005
  • Myxobacterium sp. HK1, isolated from Korean soil, degrades cellulose, differentiates to fruiting body, and its 16s rDNA has $95\%$ similarity to Polyangium sp. An anticancer molecule, CDMHK, was identified from culture broth of Myxobacterium sp. HK1, and purified by Diaion HP20, Silica gel, Sephadex LH-20 chromatography, and preparative HPLC using an YMC OSD-A C18 column. The molecular structure and formula were determined to be $C_{l2}H_{l9}N_3O_2$ (M.W 237) by MS spectrometry, 300 MHz $^{1}H\;and\;^{13}C$ NMR. The CDMHK was not active against Escherichia coli, Staphylococcus aureus, and Candida albicans. However, this molecule inhibited the growth of various cancer cell lines. The $ED_{50}$ values of CDMHK were determined to be 0.147, 0.086, 0.18, 0.166, and 0.142 $\mu$g/ml against A549, SK-OV-3, SK-MEL-2, VF498, and HCTl5 cancer cell lines, respectively. In addition, the CDMHK was able to induce apoptosis of the CCRF-CEM cancer cell line, evidenced by DNA fragmentation assay and DAPI staining.

Ceramide-Mediated Cell Death Was Accompanied with Changes of c-Myc and Rb Protein

  • Moon, Soon-Ok;Lee, Jin-Woo
    • BMB Reports
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    • v.31 no.4
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    • pp.333-338
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    • 1998
  • The sphingomyelin cycle and ceramide generation have been recognized as potential growth suppression signals in mammalian cells. Ceramide has been shown to induce differentiation, cell growth arrest, senescence, and apoptosis. Although the intracelluar target for the action of ceramide remains unknown, recent studies have demonstrated the role of cytosolic ceramideactivated protein phosphatase(CAPP). In this study, the cytotoxic effect of C2-ceramide, a synthetic cellpermeable ceramide analog, on HEp-2 cells and the mechanism by which ceramide induces cell death were investigated. The addition of exogenous C2-ceramide resulted in a concentration dependent cell death. Okadaic acid, a potent inhibitor of CAPP, enhanced ceramide-mediated cell death, which suggests that CAPP is not involved in this process. To understand the mechanism of action of ceramide, we studied the relationship between ceramide and c-Myc and pRb which are defined components of cell growth regulation. Western blot analyses revealed that C2-ceramide (10${\mu}M$) induced c-Myc down-regulation, but there were no significant changes in pRb. However, treatment of okadaic acid (10 nM) enhanced c-Myc and pRb down-regulation. Reduction of the amount of c-Myc and pRb occurred during HEp-2 cell death. These results suggest that the cytotoxic effect of ceramide in HEp-2 cells may not be mediated through the action of CAPP and that the downstream target for ceramide is c-Myc and pRb.

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Cyclin-Dependent Kinase Inhibitor 2A is a Key Regulator of Cell Cycle Arrest and Senescence in Endothelial Colony-Forming Cells in Moyamoya Disease

  • Seung Ah Choi;Youn Joo Moon;Eun Jung Koh;Ji Hoon Phi;Ji Yeoun Lee;Kyung Hyun Kim;Seung-Ki Kim
    • Journal of Korean Neurosurgical Society
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    • v.66 no.6
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    • pp.642-651
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    • 2023
  • Objective : Endothelial colony-forming cells (ECFCs) have been reported to play an important role in the pathogenesis of moyamoya disease (MMD). We have previously observed stagnant growth in MMD ECFCs with functional impairment of tubule formation. We aimed to verify the key regulators and related signaling pathways involved in the functional defects of MMD ECFCs. Methods : ECFCs were cultured from peripheral blood mononuclear cells of healthy volunteers (normal) and MMD patients. Low-density lipoproteins uptake, flow cytometry, high content screening, senescence-associated β-galactosidase, immunofluorescence, cell cycle, tubule formation, microarray, real-time quantitative polymerase chain reaction, small interfering RNA transfection, and western blot analyses were performed. Results : The acquisition of cells that can be cultured for a long time with the characteristics of late ECFCs was significantly lower in the MMD patients than the normal. Importantly, the MMD ECFCs showed decreased cellular proliferation with G1 cell cycle arrest and cellular senescence compared to the normal ECFCs. A pathway enrichment analysis demonstrated that the cell cycle pathway was the major enriched pathway, which is consistent with the results of the functional analysis of ECFCs. Among the genes associated with the cell cycle, cyclin-dependent kinase inhibitor 2A (CDKN2A) showed the highest expression in MMD ECFCs. Knockdown of CDKN2A in MMD ECFCs enhanced proliferation by reducing G1 cell cycle arrest and inhibiting senescence through the regulation of CDK4 and phospho retinoblastoma protein. Conclusion : Our study suggests that CDKN2A plays an important role in the growth retardation of MMD ECFCs by inducing cell cycle arrest and senescence.

Effect of Dietary Supplementation with Yeast Cell Suspension (Saccharomyces cerevisiae) on Nutrient Utilisation and Growth Response in Crossbred Calves

  • Singh, Rameshwar;Chaudhary, L.C.;Kamra, D.N.;Pathak, N.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.3
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    • pp.268-271
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    • 1998
  • Twenty crossbred calves of $88{\pm}5.5kg$ initial live weight and 3-4 month of age were divided into two groups and fed wheat straw and concentrate to support a 500 g daily gain in body weight. Calves in the experimental group (YC) were given a daily dose of 10 ml yeast cell suspension (YC) containing live cells $(5{\times}10^9 cells/ml)$ of Saccharomyces cerevisiae ITCCF 2094. After a growth study of 122 days metabolism trials were conducted. The calves in the YC group recorded a daily weigt gain of $492{\pm}27.8g$ as compared to $476{\pm}20.1g$ in control group. There were no significant differences in feed intake, nutrient digestibility, feed/gain ratio and nitrogen retention between the YC supplemented and control groups.

Effects of Ulmi Pumilae Cortex on AGS Gastric Cancer Cells

  • Lim, Bora;Lee, Hee Jung;Kim, Min Chul;Kim, Byung Joo
    • Journal of Pharmacopuncture
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    • v.16 no.2
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    • pp.55-61
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    • 2013
  • Objective: Ulmi Pumilae Cortex (UPC) is a deciduous tree with uneven pinnate leaves and is classified as a subfamily of Ulmuceae and contains many pharmacologically active constituents. The aim of this study was to investigate the effects of UPC on the growth and survival of AGS cells, the most common human gastric adenocarcinoma cell lines. Methods: The AGS cells were treated with varying concentrations of UPC. Analyses of the sub G1, caspase-3 activity, and mitochondrial depolarization were conducted to determine whether AGS cell death occured by apoptosis. Furthermore, to identify the role of the transient receptor potential melastatin (TRPM) 7 channels in AGS cell growth and survival, we used human embryonic kidney (HEK) 293 cells overexpressed with TRPM7 channels. Results: The addition of UPC to a culture medium inhibited AGS cell growth and survival. Experimental results showed that the sub G1, caspase-3 activity, and mitochondrial depolarization were increased. Furthermore, TRPM7 channel overexpression in HEK 293 cells exacerbated UPC-induced cell death. Conclusion: These findings indicate that UPC inhibits the growth and survival of gastric cancer cells due to a blockade of the TRPM7 channel activity. Therefore, UPC is a potential drug for treatment of gastric cancer, and TRPM7 channels may play an important role in survival in cases of gastric cancer.