• Title/Summary/Keyword: A431 cells

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Production of Red Pigment by Serratia sp. KH-95 and its Cultural Properties (Serratia sp. KH-95에 의한 적색 색소 생산 및 배양학적 특성)

  • 김창호;김승욱;홍석인
    • KSBB Journal
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    • v.13 no.4
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    • pp.431-437
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    • 1998
  • Optimal media and cultural conditions for the production of prodigiosin-like pigment were established using Serratia sp. KH-95. Glucose and phosphate(K2PO4) stimulated the cell growth, but inhibited the production of pigment at concentration levels of above 10 g/L and 2.0 g/L, respectively. Addition of soy been oil or rice oil to the production medium accelerated cell growth up to more than 2-3 times, but the production of prodigiosin increased about 15-20% in spite of the good cell growth. The effect of pH on the production of pigment was investigated in a 5 liter-bioreactor. When the pH of culture broth was maintained below 8.0, most of pigment was attached to the surface of cells. When the pH of culture broth was above 8.5, however, about 70% of total pigment was suspended in the supernatant of the broth. The cell growth and production of pigment were inhibited at dissolved oxygen concentration of below 10% of air-saturation.

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CRISPR/Cas9-mediated knockout of the Vanin-1 gene in the Leghorn Male Hepatoma cell line and its effects on lipid metabolism

  • Lu Xu;Zhongliang Wang;Shihao Liu;Zhiheng Wei;Jianfeng Yu;Jun Li;Jie Li;Wen Yao;Zhiliang Gu
    • Animal Bioscience
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    • v.37 no.3
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    • pp.437-450
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    • 2024
  • Objective: Vanin-1 (VNN1) is a pantetheinase that catalyses the hydrolysis of pantetheine to produce pantothenic acid and cysteamine. Our previous studies have shown that the VNN1 is specifically expressed in chicken liver which negatively regulated by microRNA-122. However, the functions of the VNN1 in lipid metabolism in chicken liver haven't been elucidated. Methods: First, we detected the VNN1 mRNA expression in 4-week chickens which were fasted 24 hours. Next, knocked out VNN1 via CRISPR/Cas9 system in the chicken Leghorn Male Hepatoma cell line. Detected the lipid deposition via oil red staining and analysis the content of triglycerides (TG), low-density lipoprotein-C (LDL-C), and high-density lipoprotein-C (HDL-C) after VNN1 knockout in Leghorn Male Hepatoma cell line. Then we captured various differentially expressed genes (DEGs) between VNN1-modified LMH cells and original LMH cells by RNA-seq. Results: Firstly, fasting-induced expression of VNN1. Meanwhile, we successfully used the CRISPR/Cas9 system to achieve targeted mutations of the VNN1 in the chicken LMH cell line. Moreover, the expression level of VNN1 mRNA in LMH-KO-VNN1 cells decreased compared with that in the wild-type LMH cells (p<0.0001). Compared with control, lipid deposition was decreased after knockout VNN1 via oil red staining, meanwhile, the contents of TG and LDL-C were significantly reduced, and the content of HDL-C was increased in LMH-KO-VNN1 cells. Transcriptome sequencing showed that there were 1,335 DEGs between LMH-KO-VNN1 cells and original LMH cells. Of these DEGs, 431 were upregulated, and 904 were downregulated. Gene ontology analyses of all DEGs showed that the lipid metabolism-related pathways, such as fatty acid biosynthesis and long-chain fatty acid biosynthesis, were enriched. KEGG pathway analyses showed that "lipid metabolism pathway", "energy metabolism", and "carbohydrate metabolism" were enriched. A total of 76 DEGs were involved in these pathways, of which 29 genes were upregulated (such as cytochrome P450 family 7 subfamily A member 1, ELOVL fatty acid elongase 2, and apolipoprotein A4) and 47 genes were downregulated (such as phosphoenolpyruvate carboxykinase 1) by VNN1 knockout in the LMH cells. Conclusion: These results suggest that VNN1 plays an important role in coordinating lipid metabolism in the chicken liver.

Chemical Imaging Analysis of the Micropatterns of Proteins and Cells Using Cluster Ion Beam-based Time-of-Flight Secondary Ion Mass Spectrometry and Principal Component Analysis

  • Shon, Hyun Kyong;Son, Jin Gyeong;Lee, Kyung-Bok;Kim, Jinmo;Kim, Myung Soo;Choi, Insung S.;Lee, Tae Geol
    • Bulletin of the Korean Chemical Society
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    • v.34 no.3
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    • pp.815-819
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    • 2013
  • Micropatterns of streptavidin and human epidermal carcinoma A431 cells were successfully imaged, as received and without any labeling, using cluster $Au_3{^+}$ ion beam-based time-of-flight secondary ion mass spectrometry (TOF-SIMS) together with a principal component analysis (PCA). Three different analysis ion beams ($Ga^+$, $Au^+$ and $Au_3{^+}$) were compared to obtain label-free TOF-SIMS chemical images of micropatterns of streptavidin, which were subsequently used for generating cell patterns. The image of the total positive ions obtained by the $Au_3{^+}$ primary ion beam corresponded to the actual image of micropatterns of streptavidin, whereas the total positive-ion images by $Ga^+$ or $Au^+$ primary ion beams did not. A PCA of the TOF-SIMS spectra was initially performed to identify characteristic secondary ions of streptavidin. Chemical images of each characteristic ion were reconstructed from the raw data and used in the second PCA run, which resulted in a contrasted - and corrected - image of the micropatterns of streptavidin by the $Ga^+$ and $Au^+$ ion beams. The findings herein suggest that using cluster-ion analysis beams and multivariate data analysis for TOF-SIMS chemical imaging would be an effectual method for producing label-free chemical images of micropatterns of biomolecules, including proteins and cells.

Licochalcone H Induces Cell Cycle Arrest and Apoptosis in Human Skin Cancer Cells by Modulating JAK2/STAT3 Signaling

  • Park, Kyung-Ho;Joo, Sang Hoon;Seo, Ji-Hye;Kim, Jumi;Yoon, Goo;Jeon, Young-Joo;Lee, Mee-Hyun;Chae, Jung-Il;Kim, Woo-Keun;Shim, Jung-Hyun
    • Biomolecules & Therapeutics
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    • v.30 no.1
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    • pp.72-79
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    • 2022
  • Licochalcone H (LCH) is a phenolic compound synthetically derived from licochalcone C (LCC) that exerts anticancer activity. In this study, we investigated the anticancer activity of LCH in human skin cancer A375 and A431 cells. The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) cell viability assay was used to evaluate the antiproliferative activity of LCH. Cell cycle distribution and the induction of apoptosis were analyzed by flow cytometry. Western blotting assays were performed to detect the levels of proteins involved in cell cycle progression, apoptosis, and the JAK2/STAT3 signaling pathway. LCH inhibited the growth of cells in dose- and time-dependent manners. The annexin V/propidium iodide double staining assay revealed that LCH induced apoptosis, and the LCH-induced apoptosis was accompanied by cell cycle arrest in the G1 phase. Western blot analysis showed that the phosphorylation of JAK2 and STAT3 was decreased by treatment with LCH. The inhibition of the JAK2/STAT3 signaling pathway by pharmacological inhibitors against JAK2/STAT3 (cryptotanshinone (CTS) and S3I-201) simulated the antiproliferative effect of LCH suggesting that LCH induced apoptosis by modulating JAK2/STAT3 signaling.

Effects of reversible metastable defect induced by illumination on Cu(In,Ga)Se2 solar cell with CBD-ZnS buffer layer

  • Lee, Woo-Jung;Yu, Hye-Jung;Cho, Dae-Hyung;Wi, Jae-Hyung;Han, Won-Seok;Yoo, Jisu;Yi, Yeonjin;Song, Jung-Hoon;Chung, Yong-Duck
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.431-431
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    • 2016
  • Typical Cu(In,Ga)Se2 (CIGS)-based solar cells have a buffer layer between CIGS absorber layer and transparent ZnO front electrode, which plays an important role in improving the cell performance. Among various buffer materials, chemical bath deposition (CBD)-ZnS is being steadily studied to alternative to conventional CdS and the efficiency of CBD-ZnS/CIGS solar cell shows the comparable values with that of CdS/CIGS solar cell. The intriguing thing is that reversible changes occur after exposure to illumination due to the metastable defect states in completed ZnS/CIGS solar cell, which induces an improvement of solar cell performance. Thus, it implies that the understanding of metastable defects in CBD-ZnS/CIGS solar cell is important issue. In this study, we fabricate the ITO/i-ZnO/CBD-ZnS/CIGS/Mo/SLG solar cells by controlling the NH4OH mole concentration (from 2 M to 3.5 M) of CBD-ZnS buffer layer and observe their conversion efficiency with and without light soaking for 1 hr. From the results, NH4OH mole concentration and light exposure can significantly affect the CBD-ZnS/CIGS solar cell performance. In order to investigate that which layer can contain metastable defect states to influence on solar cell performance, impedance spectroscopy and capacitance profiling technique with exposure to illumination have been applied to CBD-ZnS/CIGS solar cell. These techniques give a very useful information on the density of states within the bandgap of CIGS, free carriers density, and light-induced metastable effects. Here, we present the rearranged charge distribution after exposure to illumination and suggest the origin of the metastable defect states in CBD-ZnS/CIGS solar cell.

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Inhibitory Effect of the Methanolic Extract of Symphyocladia latiuscula on the Growth of HT-29 Human Colon Cancer Cells (보라우무 메탄올추출물의 HT-29 대장암세포 증식 억제 효과)

  • Kim, Eun-Ji;Park, So-Young;Hong, Ji-Eun;Shin, Min-Jeong;Lim, Soon-Sung;Shin, Hyun-Kyung;YoonPark, Jung-Han
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.4
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    • pp.431-438
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    • 2007
  • In the present study, twenty eight marine algae species were evaluated for their antiproliferative effect on HT-29 human colon cancer cells. Among these, the methanolic extract of Symphyocladia latiuscula (SL Ex) showed the highest inhibitory activity on HT-29 cell growth. In this study, we examined the mechanism by which SL Ex inhibited the HT-29 cell growth. Cells were cultured with various concentrations of $(0{\sim}20{\mu}g/mL)$ SL Ex. The SL Ex substantially decreased the viable cell numbers and induced apoptosis of HT-29 cells in a dose-dependent manner Western blot analyses of total cell lysates revealed that SL Ex increased the levels of cleaved caspase-8, -9, -7, and -3, and poly (ADP-ribose) polymerase in HT-29 cells. In addition, SL Ex increased truncated Bid levels but moderately decreased Bax levels at only $20{\mu}g/mL$. Furthermore, SL Ex did not affect Bcl-2 protein levels but increased the levels of Fas in HT-29 cells. The present results indicate that SL Ex inhibits cell growth via inducing apoptosis in human colon cancer cells. The mechanism of apoptosis induction by SL Ex involves caspase-8 activation leading to changes in mitochondrial events and subsequent activation of the caspase-7/caspase-3 cascade. Our finding may lead to the development of new therapeutic strategies for the treatment of colon cancer.

The role of calpain in skeletal muscle

  • Pandurangan, Muthuraman;Hwang, Inho
    • Animal cells and systems
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    • v.16 no.6
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    • pp.431-437
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    • 2012
  • Calpains are a class of proteins that belong to the calcium-dependent, non-lysosomal cysteine proteases. There are three major types of calpains expressed in the skeletal muscle, namely, ${\mu}$-calpain, m-calpain, and calpain 3, which show proteolytic activities. Skeletal muscle fibers possess all three calpains, and they are $Ca^{2+}$-dependent proteases. The functional role of calpains was found to be associated with apoptosis and myogenesis. However, calpain 3 is likely to be involved in sarcomeric remodeling. A defect in the expression of calpain 3 leads to limb-girdle muscular dystrophy type 2A. Calpain 3 is found in skeletal muscle fibers at the N2A line of the large elastic protein, titin. A substantial proportion of calpain 3 is activated 24 h following a single bout of eccentric exercise. In vitro studies indicated that calpain 3 can be activated 2-4 fold higher than normal resting cytoplasmic [$Ca^{2+}$]. Characterization of the calpain system in the developing muscle is essential to explain which calpain isoforms are present and whether both ${\mu}$-calpain and m-calpain exist in differentiating myoblasts. Information from such studies is needed to clarify the role of the calpain system in skeletal muscle growth. It has been demonstrated that the activation of ubiquitous calpains and calpain 3 in skeletal muscle is very well regulated in the presence of huge and rapid changes in intracellular [$Ca^{2+}$].

A PROMISING NEW ANTI-WRINKLE INGREDIENT: Pericarpium castaneae extracts

  • Kim, Beom-Jun;Jo, Byoung-Kee;Kim, Jeong-Ha
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.25 no.4 s.34
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    • pp.57-63
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    • 1999
  • Pericarpium castaneae extracts have variously potent activities, such as anti-oxidative activity and free radical scavenging activity. in vivo and in vitro studies both indicate that pericarpium castaneae extracts acts as a free radical scavenger($IC_{50}:7.6{\mu}g/ml$) stronger than gallic acid($IC_{50}:12.5{\mu}g/ml$) and ellagic acid($IC_{50}:15{\mu}g/ml$) which could prevent cutaneous UV damages and skin aging. The extracts showed a good effect as a anti-oxidant($IC_{50}:50{\mu}g/ml$). It was shown that the appearance of wrinkle in human skin was reduced by topical application of pericarpium castaneae extracts. And the treatment of human skin with the extracts increased the elasticity and moisture of the skin. We investigated the effect of the pericarpium castaneae extracts on production of extracellular matrix using cultured A431 fibroblast cells. The results indicated that the extracts had no detectable effect on collagen synthesis. But synthesis of cell adhesion protein was increased by the extracts. The results suggest that increase of cell adhesion protein synthesis by pericarpium castaneae extracts has closely related to reduction of wrinkle in skin.

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A PROMISING NEW ANTI-WRINKLE INGREDIENT : Pericarpium castaneae extracts

  • Kim, Beom-Jun;Jo, Byoung-Kee;Kim, Jeong-Ha
    • Proceedings of the SCSK Conference
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    • 1999.10a
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    • pp.57-64
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    • 1999
  • Pericarpium castaneae extracts have variously potent activities, such as anti-oxidative activity and free radical scavenging activity. in vivo and in vivo studies both indicate that pericarpium castaneae extracts acts as a flee radical scavenger ($IC_{50}$/: 7.6$\mu\textrm{g}$/ml) stronger than gallic acid($IC_{50}$/: 12.5$\mu\textrm{g}$/ml) and ellagic acid($IC_{50}$/: 15$\mu\textrm{g}$/ml) which could prevent cutaneous UV damages and skin aging. The extracts showed a good effect as a anti-oxidant ($IC_{50}$/: 50$\mu\textrm{g}$/ml). It was shown that the appearance of wrinkle in human skin was reduced by topical application of pericarpium castaneae extracts. And the treatment of human skin with the extracts increased the elasticity and moisture of the skin. We investigated the effect of tile pericarpium castaneae extracts on production of extracellular matrix using cultured A431 fibroblast cells. The results indicated that the extracts had no detectable effect on collagen synthesis, But synthesis of cell adhesion protein was increased by the extracts. The results suggest that increase of cell adhesion protein synthesis by pericarpium castaneae extracts has closely related to reduction of wrinkle in skin.

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Intracellular Signaling Pathways for Type II IgE Receptor (CD23) Induction by Interleukin - 4 and Anti - CD40 Antibody

  • Kim, Hyun-Il;Park, Hee-Jeoung;Lee, Choong-Eun
    • BMB Reports
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    • v.30 no.6
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    • pp.431-437
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    • 1997
  • Since the role of CD40 on the interleukin-4(IL-4) -induced B cell activation has been strongly implicated in the agumentation of IgE production and response, we have investigated the intracelluar signaling pathways utilized by IL-4 and CD40 for type II IgE receptor (CD23) expression. IL-4 and anti-CD40 antibody treatment of human B cells, independently caused a rapid induction of CD23 gene activation within 2 h. There was a noticeable synergism between the action of the two agents inducing CD23 expression: the addition of anti-CD40 to the IL-4-treated culture significantly agumented the IL-4-induced CD23 on both mRNA and surface protein levels, and the inclusion of IL-4 in the anti-CD40-treated cells caused a further increase of CD23 expression far above the maximal level induced by anti-CD40. Protein tyrosine kinase (PTK) inhibitors effectively suppressed the both IL-4- and anti -CD40-induced CD23 expression. whereas protein kinase C (PKC) inhibitors had no effects. Electrophoretic mobility shift assays (EMSA) have shown that IL-4 and anti-CD40 induce the activation of NF-IL-4 and $NF-_{K}B$, respectively, binding to the CD23 promoter, both in a PKC-independent and PTK-dependent manner. These data suggest that the synergistic activation of CD23 gene expression by IL-4 and anti-CD40 is mediated by co-operative action of distinct nuclear factors. each of which is rapidly activated via PKC-independent and PTK-dependent process.

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