• Title/Summary/Keyword: Colony formation

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Combination of Colony Formation and Congo Red Reaction for Detecting Intra- and Extra-Cellular Cellulolytic Activities (세포내외 섬유소 분해능 검출을 위한 Colony 형성과 Congo Red반응의 병용)

  • Lee, D.S.;Kim, H.K.;Pack, M.Y.
    • Microbiology and Biotechnology Letters
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    • v.12 no.4
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    • pp.305-309
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    • 1984
  • A selective medium which allows growth of only cellulolytic bacteria was developed. The medium composed of 0.5% carboxymethylcellulose (CMC), 0.005% yeast extract, minerals and agar. Colony formation on this medium indicates overall activities of cellulose utilization. A subsequent test with Congo Red dye could distinguish extracellular cellulolysis from intracellular type.

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The Radioprotective Effects of Bu-Zhong-Yi-Qi-Tang as a Prescriptions of Traditional Chinese Medicine in Irradiated Mice

  • Kim, Sung-Ho;Kim, Se-Ra;Heon Oh;Yang, Jung-Ah;Jo, Sung-Kee;Byun, Myung-Woo;Yee, Sung-Tae
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2000.09a
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    • pp.21-21
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    • 2000
  • We performed this study to determine the effect of Bu-Zhong-Yi-Qi-Tang, as a prescription of traditional Oriental medicine, and its major ingredients on jejunal crypt survival, endogenous spleen colony formation, and apoptosis in jejunal crypt cells of mice irradiated with high and low dose of r-radiation. Bu-Zhong-Yi-Qi-Tang administration before irradiation protected the jejunal crypts (p<0.0001), increased the formation of endogenous spleen colony (p<0.05) and reduced the frequency of radiation-induced apoptosis (p<0.05). (omitted)

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Modification of Radiation Response in Mice by Panax ginseng and Diethyldithiocarbamate

  • Kim, Sung-Ho;Son, Chang-Ho;Nah, Seung-Yeol;Jo, Sung-Kee;Byun, Myung-Woo;Shin, Dong-Ho
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2001.09a
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    • pp.22-22
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    • 2001
  • We performed this study to determine the effect of Panax ginseng and its fractions on jejunal crypt survival, endogenous spleen colony formation, and apoptosis in jejunal crypt cells of mice irradiated with high and low dose of $\gamma$-irradiation. The radioprotective effect of ginseng was compared with the effect of diethyldithiocarbamate (DDC). Ginseng administration before irradiation protected the jejunal crypts (p<0.005), increased the formation of endogenous spleen colony (p<0.005) and reduced the frequency of radiation-induced apoptosis (p<0.005). (omitted)

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Induction of Apoptosis by IGFBP3 Overexpression in Hepatocellular Carcinoma Cells

  • Han, Jian-Jun;Xue, De-Wen;Han, Qiu-Rong;Liang, Xiao-Hong;Xie, Li;Li, Sheng;Wu, Hui-Yong;Song, Bao
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10085-10089
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    • 2015
  • Background: The insulin-like growth factor (IGF) system comprises a group of proteins that play key roles in regulating cell growth, differentiation, and apoptosis in a variety of cellular systems. The aim of this study was to investigate the role of insulin-like growth factor binding protein 3 (IGFBP3) in hepatocellular carcinoma. Materials and Methods: Expression of IGF2, IGFBP3, and PTEN was analyzed by qRT-PCR. Lentivirus vectors were used to overexpress IGFBP3 in hepatocellular carcinoma cell (HCC) lines. The effect of IGFBP3 on proliferation was investigated by MTT and colony formation assays. Results: Expression of IGF2, IGFBP3, and PTEN in several HCC cell lines was lower than in normal cell lines. After 5-aza-2'-deoxycytidine/trichostatin A treatment, significant demethylation of the promoter region of IGFBP3 was observed in HCC cells. Overexpression of IGFBP3 induced apoptosis and reduced colony formation in HUH7 cells. Conclusions: Expression of IGF2, IGFBP3, and PTEN in several HCC cell lines was lower than in normal cell lines. After 5-aza-2'-deoxycytidine/trichostatin A treatment, significant demethylation of the promoter region of IGFBP3 was observed in HCC cells. Overexpression of IGFBP3 induced apoptosis and reduced colony formation in HUH7 cells.

Induction of apoptosis by methanol extracts of Ficus carica L. in FaDu human hypopharynx squamous carcinoma cells

  • Lee, Seul Ah;Park, Bo-Ram;Kim, Chun Sung
    • International Journal of Oral Biology
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    • v.45 no.3
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    • pp.99-106
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    • 2020
  • Ficus carica L. (fig) is one of the first cultivated crops and is as old as humans. This plant has been extensively used as a traditional medicine for treating diseases, such as cough, indigestion, nutritional anemia, and tuberculosis. However, the physiological activity of fig leaves on oral cancer is as yet unknown. In this study, we investigated the anticancer effect of methanol extracts of Ficus carica (MeFC) and the mechanism of cell death in human FaDu hypopharyngeal squamous carcinoma cells. MeFC decreased the viability of oral cancer (FaDu) cells but did not affect the viability of normal (L929) cells, as determined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay and Live and Dead assay. In addition, MeFC induced apoptosis through the proteolytic cleavage of procaspase-3, -9, poly (ADP-ribose) polymerase (PARP), downregulation of Bcl-2, and upregulation of Bax, as determined by 4′,6-diamidino-2-phenylindole dihydrochloride staining and western blot analysis. Moreover, a concentration of MeFC without cytotoxicity (0.25 mg/mL) significantly suppressed colony formation, a hallmark of cancer development, and completely inhibited the colony formation at 1 mg/mL. Collectively, these results suggest that MeFC exhibits a potent anticancer effect by suppressing the growth of oral cancer cells and colony formation via caspase- and mitochondrial-dependent apoptotic pathways in FaDu human hypopharyngeal squamous carcinoma cells. Therefore, the methanol extract of Ficus carcica leaves provide a natural chemotherapeutic drug for human oral cancer.

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.

The effects of cytokinin and plating density on protoplast culture of sunflower

  • Chitpan Kativat;Witsarut Chueakhunthod;Piyada Alisha Tantasawat
    • Journal of Plant Biotechnology
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    • v.49 no.4
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    • pp.331-338
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    • 2022
  • Sunflower (Helianthus annuus L.) protoplasts were isolated from seven-day-old etiolated hypocotyls of 10 A line and four-week-old fully expanded young leaves of PI 441983 line in vitro seedlings using an enzymatic method. Purified protoplasts were collected by filtration and floatation in sucrose solution. Semi-solid protoplast culture was performed using the L4 regeneration protocol with various culture media and plating densities to achieve the highest efficiencies for protoplast culture of hypocotyl and mesophyll protoplasts of 10 A and PI 441983 lines, respectively. The concentrations in liquid L'4M medium and different plating densities were evaluated in two types of cytokinins, the adenine-type 6-benzyladenine (BA) and the phenylurea-type thidiazuron (TDZ). The highest colony formation was achieved in both sunflower lines when 0.5 mgL-1 BA and 0.5 mgL-1 TDZ were applied with a high plating density (3 × 105 protoplasts mL-1). These conditions led to 38.45% and 39.40% colony formation for hypocotyl protoplasts of the 10 A line and mesophyll protoplasts of the PI 441983 line, respectively. Moreover, many hypocotyl protoplast-derived colonies developed into micro-calli. In addition, superior development of both sunflower protoplasts was observed with all plating densities when BA was used in combination with TDZ. This finding will be applicable to future sunflower hybrid production via somatic hybridization.

Effect of gacS and gacA Mutations on Colony Architecture, Surface Motility, Biofilm Formation and Chemical Toxicity in Pseudomonas sp. KL28

  • Choi, Kyung-Soon;Veeraragouda, Yaligara;Cho, Kyoung-Mi;Lee, Soo-O;Jo, Geuk-Rae;Cho, Kyung-Yun;Lee, Kyoung
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.492-498
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    • 2007
  • GacS and GacA proteins form a two component signal transduction system in bacteria. Here, Tn5 transposon gacS and gacA (Gac) mutants of Pseudomonas sp. KL28, an alkylphenol degrader, were isolated by selecting for smooth colonies of strain KL28. The mutants exhibited reduced ability to migrate on a solid surface. This surface motility does not require the action of flagella unlike the well-studied swarming motility of other Pseudomonas sp. The Gac mutants also showed reduced levels of biofilm and pellicle formation in liquid culture. In addition, compared to the wild type KL28 strain, these mutants were more resistant to high concentrations of m-cresol but were more sensitive to $H_2O_2$, which are characteristics that they share with an rpoS mutant. These results indicate that the Gac regulatory cascade in strain KL28 positively controls wrinkling morphology, biofilm formation, surface translocation and $H_2O_2$ resistance, which are important traits for its capacity to survive in particular niches.

Growth and Differentiation of Mammary Epithelial Cells in Extracellular Maxtrix Culture (세포외 기질 세포 배양법에 의한 유선상피세포의 성장 및 분화 유도)

  • Paik, Kee-Joo;Yoon, Jeong-Hyun;Kim, Dong-Yeom;Jeon, Seong-Shil;Yang, Han-Suk;Kim, Nam Deuk
    • Journal of Life Science
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    • v.5 no.1
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    • pp.1-7
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    • 1995
  • Mammary orgamoids(ductal and endbud fragments) were cultured in a complete hormone medium(CHM) with 10%FBS, estradiol, progesterone, hydrocortisone, insulin, and prolactin, Several types of colonies were observed: stellate(14$$\pm$5.5%), duct(41$\pm$5.6%), web(35$\pm$3.6%), squamous(6$\pm$2.1%), and lobuloduct(4$\pm$1.2%), Squamous colony was typical squamous metaplasia(SM) with several layers of squamous epithlia and keratin pearls. At the immunocytochemical study, casein proteins were predominantly localized near the apical surfaces of the cells or in the lumina of ductal or lobuloductal colonies. To inhibit the formation of SM, we treated organoids with all-trans retinoic acid(RA) from 10$^{-6}$ to 10$^{-17}$ M in CHM. Formation of SN was completely inhibited at 10$^{-9}$M RA in CHM. The frequency of lobuloductal colony formation was increased with the augmentation of RA concentration.

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Effects of Tobacco-Specific Carcinogen on Protein Kinase C Isoforms (흡연특이성 발암물질이 특정 Protein Kinase C Isoform에 미치는 영향)

  • Kang, Hyung-Seok;Ko, Moo-Sung;Park, Ki-Sung;Lee, Sub;Jheon, Sang-Hoon;Kwon, Oh-Choon
    • Journal of Chest Surgery
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    • v.36 no.9
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    • pp.666-673
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    • 2003
  • Cigarette smoking is the leading cause of the lung cancer. However, mechanism of action underlying the carcinogenesis in the lung still remains to be elucidated. The present study attempted to look into the carcinogenic potential of tobacco-specific nitrosamine, NNK (4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone) and the effects of protein kinase C (PKC) isoforms in an immortalized human epithelial cell model. Material and Method: Immortalized human epithelial cells were exposed with NNK and examined for its carcinogenic potential as measured by saturation density, soft-agar colony formation, and cell aggregation assay. The specific isoform of PKCs involved in the cellular transformation was analysed through western blot with monoclonal antibody and measured separately in cytosolic fraction and membrane fraction. Result: Human epithelial cells exposed with NNK showed prominent carcinogenic potential in saturation density, soft agar colony formation, and cell aggregation assay. PKC isoform analysis results are as follows: PKC- $\alpha$ showed significant translocation of protein levels from cytosolic fraction to membrane fraction, as analyzed by immunoblot. PKC- $\varepsilon$ showed a dose-dependent increase of translocation. PKC- λ was not affected by NNK treatment. Conclusion: The study demonstrated that there was a certain specificity in the patterns of isoform induction following chemical carcinogen exposure. Thus, it is suggested that identification of specific isoform be a clue to find target molecules in the carcinogenesis.