• Title/Summary/Keyword: biological cells

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Rhythmic Gene Expression in Somite Formation and Neural Development

  • Kageyama, Ryoichiro;Niwa, Yasutaka;Shimojo, Hiromi
    • Molecules and Cells
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    • v.27 no.5
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    • pp.497-502
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    • 2009
  • In mouse embryos, somite formation occurs every two hours, and this periodic event is regulated by a biological clock called the segmentation clock, which involves cyclic expression of the basic helix-loop-helix gene Hes7. Hes7 expression oscillates by negative feedback and is cooperatively regulated by Fgf and Notch signaling. Both loss of expression and sustained expression of Hes7 result in severe somite fusion, suggesting that Hes7 oscillation is required for proper somite segmentation. Expression of a related gene, Hes1, also oscillates by negative feedback with a period of about two hours in many cell types such as neural progenitor cells. Hes1 is required for maintenance of neural progenitor cells, but persistent Hes1 expression inhibits proliferation and differentiation of these cells, suggesting that Hes1 oscillation is required for their proper activities. Hes1 oscillation regulates cyclic expression of the proneural gene Neurogenin2 (Ngn2) and the Notch ligand Delta1, which in turn lead to maintenance of neural progenitor cells by mutual activation of Notch signaling. Taken together, these results suggest that oscillatory expression with short periods (ultradian oscillation) plays an important role in many biological events.

A Sphingosine Kinase-1 Inhibitor, SKI-II, Induces Growth Inhibition and Apoptosis in Human Gastric Cancer Cells

  • Li, Pei-Hua;Wu, Jin-Xia;Zheng, Jun-Nian;Pei, Dong-Sheng
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10381-10385
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    • 2015
  • SKI-II has been reported as an inhibitor of sphingosine kinase 1 and has been extensively used to prove the involvement of sphingosine kinase and sphingosine-1-phosphate (Sphk1) in cellular processes. In the current study, we investigated the effects of SKI-II and its potential mechanisms in human gastric cancer SGC7901 cells. After treatment with SKI-II, cell growth, cell cycle distribution, apoptosis, expression of Sphk1, NF-${\kappa}B$, Bcl-2, Bax and p27 were assessed by MTT assay, flow cytometry, electron microscopy, immunocytochemistry and Western-blot assay, respectively. Our results showed that SKI-II markedly inhibited SGC7901 cell survival in a dose-dependent manner, reduced cell proliferation with accumulation of cells in the G0/G1 phase and induced apoptosis in the tumor cells. Furthermore, Western blotting and immunocytochemistry showed that the expression of p27 and Bax was increased significantly, but the expression of NF-${\kappa}B$, Bcl-2 and Sphk1 decreased by different degrees. These results indicate that SKI-II induced cell growth arrest and apoptosis. The increased apoptotic sensitivity of SGC7901 was correlated with NF-${\kappa}B$ or Bcl-2/Bax activation.

Overexpression and Purification of PreS Region of Hepatitis B Virus Antigenic Surface Protein adr Subtype in Escherichia coli

  • Abbas, Naaz;Ahmad, Aftab;Shakoori, Abdul Rauf
    • BMB Reports
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    • v.40 no.6
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    • pp.1002-1008
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    • 2007
  • PreS domain of Hepatitis B virus (HBV) surface antigen is a good candidate for an effective vaccine as it activates both B and T cells besides binding to hepatocytes. This report deals with overexpression and purification of adr subtype of surface antigen that is more prevalent in Pakistan. PreS region, comprising 119 aa preS1 region plus a 55 aa preS2 region plus 11 aa from the N-terminal S region, was inserted in pET21a+ vector, cloned in E. coli $DH5\alpha$ cells and expressed in E. coli BL21 codon+ cells. The conditions for over expression were optimized using different concentrations of IPTG (0.01-5 mM), and incubating the cells at different temperatures (23-$41^{\circ}C$) for different durations (0-6 h). The cells were grown under the given optimized conditions (0.5 mM IPTG concentration at $37^{\circ}C$ for 4 h), lysed by sonication and the protein was purified by ion exchange chromatography. On the average, 24.5 mg of recombinant protein was purified per liter of culture. The purified protein was later lyophilized and stored at $-80^{\circ}C$.

In Vivo Expression of the PTB-deleted Odin Mutant Results in Hydrocephalus

  • Park, Sunjung;Lee, Haeryung;Park, Soochul
    • Molecules and Cells
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    • v.38 no.5
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    • pp.426-431
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    • 2015
  • Odin has been implicated in the downstream signaling pathway of receptor tyrosine kinases, such as the epidermal growth factor and Eph receptors. However, the physiologically relevant function of Odin needs to be further determined. In this study, we used Odin heterozygous mice to analyze the Odin expression pattern; the targeted allele contained a ${\beta}$-geo gene trap vector inserted into the 14t intron of the Odin gene. Interestingly, we found that Odin was exclusively expressed in ependymal cells along the brain ventricles. In particular, Odin was highly expressed in the subcommissural organ, a small ependymal glandular tissue. However, we did not observe any morphological abnormalities in the brain ventricles or ependymal cells of Odin null-mutant mice. We also generated BAC transgenic mice that expressed the PTB-deleted Odin (dPTB) after a floxed GFP-STOP cassette was excised by tissue-specific Cre expression. Strikingly, Odin-dPTB expression played a causative role in the development of the hydrocephalic phenotype, primarily in the midbrain. In addition, Odin-dPTB expression disrupted proper development of the subcommissural organ and interfered with ependymal cell maturation in the cerebral aqueduct. Taken together, our findings strongly suggest that Odin plays a role in the differentiation of ependymal cells during early postnatal brain development.

In vitro Growth Inhibition and Apoptotic Effects of Hang-baek-Tang on HL-60 Cells

  • Park Jun-Ho;Ju Sung-Min;Kim Kun-Jung;Jeon Byung-Hoon;Oh Jung-Mi;Lee Chae-Ho;Han Dong-Min;Kim Won-Sin
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.6
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    • pp.1636-1639
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    • 2005
  • To develop novel anti-leukemic medicine, we have prepared a Korean traditional medicine, named Hang-baek-Tang, which is composed of 8 kinds of anti-leukemic medicinal plants. The water extracts was examined anti-leukemic activity using the human leukemia cell line, HL-60 cells. HL-60 cells showed the growth inhibition and several apoptotic features, including DNA ladders, morphological changes, by treatment of the cells with Hang-Daek-Tang. We have observed that Hang-baek-Tang induced the activation of caspase-3, caspase-8 and caspase-9. Further molecular analysis demonstrated that Hang-baek-Tang induced cleavage of PARP and increase of hypodiploid (Sub-G1) population in flow cytometric analysis. These results indicate that Hang-baek-Tang has been considered to exert anti-leukemic activity through the procaspase-3 activation pathway.

Development of Biological Cell Manipulation System using Visual Tracking Method

  • Lee, Geunho;Kang, Hyun-Jae;Kwon, Sang-Joo;Park, Gwi-Tae;Kim, Byungkyu
    • Proceedings of the IEEK Conference
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    • 2003.07c
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    • pp.2911-2914
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    • 2003
  • Conventionally, biological manipulations have been performed manually with long training and pretty low success rates. To overcome this problem, a novel biological manipulation system has been developed to manipulate biological cells without any interference of a human operator, In this paper, we demonstrate a development of tole-autonomous Cell Manipulation System (CMS) using an image processing at a remote site. The CMS consists of two manipulators, a plane stage, and an optical microscope. We developed deformable template-model-matching algorithm for micro objects and pattern matching algorithm of end effect for these manipulators in order to control manipulators and the stage. Through manipulation of biological cells using these algorithms, the performance of the CMS is verified experimentally.

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Chemical Genomics and Medicinal Systems Biology: Chemical Control of Genomic Networks in Human Systems Biology for Innovative Medicine

  • Kim, Tae-Kook
    • BMB Reports
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    • v.37 no.1
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    • pp.53-58
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    • 2004
  • With advances in determining the entire DNA sequence of the human genome, it is now critical to systematically identify the function of a number of genes in the human genome. These biological challenges, especially those in human diseases, should be addressed in human cells in which conventional (e.g. genetic) approaches have been extremely difficult to implement. To overcome this, several approaches have been initiated. This review will focus on the development of a novel 'chemical genetic/genomic approach' that uses small molecules to 'probe and identify' the function of genes in specific biological processes or pathways in human cells. Due to the close relationship of small molecules with drugs, these systematic and integrative studies will lead to the 'medicinal systems biology approach' which is critical to 'formulate and modulate' complex biological (disease) networks by small molecules (drugs) in human bio-systems.

Identification of to Hexapeptides that Render C2 Myoblasts the Resistant Menadione-induced Cell Death

  • Hwang, Sung-Ho;Kim, Min-Jeong;Lim, Jeong-A;Woo, Joo-Hong;Kim, Hye-Sun
    • Animal cells and systems
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    • v.12 no.1
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    • pp.35-39
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    • 2008
  • Menadione induced cell death in cultured C2 myoblasts. By screening synthetic peptide libraries composed of random sequence of hexapeptides, we identified the hexa-peptides pool of(Ala/Ile)-(Ile/Met)-Val-Ile-Asp-(Met/Ser)-$NH_2$ that protected the myoblasts against menadioneinduced cell death. Pre-incubation with the hexapeptide pool reduced the number of cells detached from culture dish substrate and increased the ratio of relative viability against menadione. In addition, the peptides strongly increased the expression of Bcl-2, an anti-apoptotic protein. These results suggest that the hexapeptides might enhance the resistance to cell death against menadione by increasing the expression of Bcl-2.