• Title/Summary/Keyword: B16

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Cloning and Expression of A Bacillus licheniformis Cellulase Gene (Bacillus licheniformis WL-12의 cellulase 유전자 클로닝과 발현)

  • Yoon, Ki-Hong
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.313-318
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    • 2006
  • A thermophilic bacterium producing the extracellular cellulase was isolated from soybean paste, and the isolate WL-12 has been identified as Bacillus licheniformis on the basis on its 16S rRNA sequence, morphology and biochemical properties. A gene encoding the cellulase of B. licheniformis WL-12 was cloned and its nucleotide sequence was determined. This cellulase gene, designated celA, consisted of 1,551 nucleotides, encoding a polypeptide of 517 amino acid residues. The gene product contained catalytic domain and cellulose binding domain. The deduced amino acid sequence was highly homologous to those of cellulases of B. licheniformis, B. subtilis and B. amytoliquefaciens belonging to the glycosyl hydrolase family 5. When the celA gene was highly expressed using a strong B. subtilis promoter, the extracellular cellulase was produced up to 7.0 units/ml in B. subtilis WB700.

Characteristics of Magnetic Tunnel Junctions Comprising Ferromagnetic Amorphous NiFeSiB Layers (강자성 비정질 NiFeSiB 자유층을 갖는 자기터널접합의 스위칭 특성)

  • Hwang, J.Y.;Rhee, S.R.
    • Journal of the Korean Magnetics Society
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    • v.16 no.6
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    • pp.279-282
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    • 2006
  • Magnetic tunnel junctions (MTJs), which consisted of amorphous ferromagnetic NiFeSiB free layers, were investigated. The NiFeSiB layers were used to substitute for the traditionally used CoFe and/or NiFe layers with the emphasis being given to obtaining an understanding of the effect of the amorphous free layer on the switching characteristics of the MTJs. $Ni_{16}Fe_{62}Si_{8}B_{14}$ has a lower saturation magnetization ($M_{s}:\;800\;emu/cm^{3}$) than $Co_{90}Fe_{10}$ and a higher anisotropy constant ($K_{u}:\;2700\;erg/cm^{3}$) than $Ni_{80}Fe_{20}$. The $Si/SiO_{2}/Ta$ 45/Ru 9.5/IrMn 10/CoFe $7/AlO_{x}/CoFeSiB\;(t)/Ru\;60\;(in\;nanometers)$structure was found to be beneficial for the switching characteristics of the MTJ, leading to a reduction in the coercivity ($H_{c}$) and an increase in the sensitivity resulted from its lower saturation magnetization and higher uniaxial anisotropy. Furthermore, by inserting a very thin CoFe layer at the tunnel barrier/NiFeSiB interface, the TMR ratio and switching squareness were improved more with the increase of NiFeSiB layer thickness up to 11 nm.

MDP-Lys (L18), a Synthetic Muramyl Dipeptide Derivative, Enhances Antitumor Activity of an Inactivated Tumor Vaccine

  • Yoo, Yung-Choon;Park, Seung-Yong;Lee, Kyung-Bok;Azuma, Ichiro
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.399-404
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    • 2000
  • The adjuvant effect of a muramyl dipeptide (MDP) derivative, MDP-Lys(L18), on enhancing of antitumor immunity induced by X-irradiated tumor cells against highly metastatic B16-BL6 melanoma cells was examined in mice. Mice immunized intradermally (i.d.) with a mixture of X-irradiated B16-BL6 cells and MDP-Lys (L18) [Vac+MDP-Lys (L18)] followed by an intravenous (i.v.)inoculation of $10^4$ viable tumor cells 7 days after immunization, showed a significant inhibition of experimental lung metastasis of B16-BL6 melanoma cells. The most effective immunization for the prophylactic inhibition of tumor metastasis was obtained from the mixture of $100{\;}\mu\textrm{g}$ of MDP-Lys (L18) and $10^4$ X-irradiatied tumor vaccine. Furthermore, immunization of mice with Vac+MDP-Lys(L18), 3 days after tumor challenge, resulted in a significant inhibition of lung metastasis of B16-BL6 melanoma cells in an experimental lung metastasis model. Similarly, the administration of Vac+MDP-Lys(L18), 1 or 7 days after tumor removal, markedly inhibited tumor metastasis of B16-BL6 in a spontaneous lung metastasis model. When Vac+MDP-Lys (L18) was i.d. administered 3 days after subcutaneous (s.c.) inoculation of tumor cells ($5{\times}10^5/site$) on the back, mice treated with Vac+MDP-Lys(L18) showed inhibition of significantly tumor growth on day 20. These results suggest that MDP-Lys (L18) is able to enhance antitumor activity induced by X-irradiated tumor vaccine to reduce lung metastasis of tumor cells, and is a potent immunomodulating agent which may be applied prophylactically as well as therapeutically to treatment of cancer metastasis.

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Commelina communis Ledeb Inhibits Melanin Synthesis in Alpha-MSH-stimulated B16F10 Cells (압척초추출물의 Alpha-MSH 유도성 멜라닌합성 억제 효과)

  • Kang, Moon Kyung;Lee, Young Eun;Woo, Won Hong;Mun, Yeun Ja
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.5
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    • pp.506-511
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    • 2014
  • Commelina communis Ledeb is a widely used medication for the treatment of antidiabetic, antioxidant and hypoglycemic agent in Korea. Alpha-melanocyte stimulating hormone (${\alpha}$-MSH) is a major factor to stimulate melanin synthesis in the skin. The purposes of this study was to investigate the inhibitory effects of extract from Commelina communis Ledeb (ECC) on ${\alpha}$-MSH-stimulated melanogenesis in B16F10 cells. ECC suppressed melanin synthesis and intracellular tyrosinase activity in B16F10 cells or ${\alpha}$-MSH-induced B16F10 cells in a dose dependent manner. In study on the melanogenic protein expressions, it had especially influence on expressions of tyrosinase and tyrosinase-related protein (TRP-1). Tyrosinase and TRP-1 expressions were gradually decreased in a dose-dependent. Additionally, the extract also decreased the ${\alpha}$-MSH-induced over-expression of tyrosinase and TRP-1. This results show that the anti-melanogenic activity of ECC is correlated with the suppression of tyrosinase and TRP-1 protein expressions in B16F10 cells.

Effect of Nigella sativa Oil on Melanogenesis (니겔라 사티바 오일의 미백 효능에 관한 연구)

  • Lee, Su-Yeon;Lee, Sae-Mi;Heo, Woo-Beom;Kim, Jin-Guk;Kim, Young-Heui
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.37 no.4
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    • pp.319-326
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    • 2011
  • In order to investigate the potential of Nigella sativa (N. sativa) oil as an active ingredient for whitening cosmetics, we prepared N. sativa oil. We measured its inhibitory effects on mushroom tyrosinase activity, cellular tyrosinase activity, and melanin synthesis inhibitory activity in B16 melanoma cells. N. sativa oil and its components showed inhibitory activity against mushroom tyrosinase and melanin synthesis. In a melanin synthesis inhibition assay using mouse B16-F10 melanoma cell, it reduced melanin production up to 86 % at a concentration of 10 mg/mL without cytotoxicity. In the study on the melanogenic protein expressions by using RT-PCR and Western blot, N. sativa oil and its components inhibited expression of tyrosinase protein, which is a well-known key protein on melanogenesis, and tyrosinase expression was gradually decreased in a dose-dependent manner. Therefore, this result suggests that N. sativa oil could be used as an active ingredient for whitening cosmetics.

Inhibitory Effects of Saposhnikoviae Radix Extracts on the Melanin Production and Elastase Activity in B16F10 cells (흑색종 세포주에서 멜라닌 생성과 엘라스타제 활성 억제에 미치는 방풍의 효과)

  • Choi, Chan Hun;Wang, Kung The;Cho, Hye Rin;Jeong, Jong Gil;Jeong, Hyun Woo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.3
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    • pp.296-302
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    • 2014
  • Saposhmikoviae Radix can treat various skin disease by anti-pruitus and anti-inflammatory effects. This study was designed to investigate effects of Saposhmikoviae Radix Extracts(SRE) on skin elasticity and whitening using B16F10 cell lines. In this experiment, We observed effect of SRE on cell viability, inhibition of melanin synthesis and inhibitory effect on tyrosinase and elastase. In results, SRE treated group showed lowered proliferation rates significantly compared to non-treated group. More than SRE $125{\mu}g/m{\ell}$ of treated groups were lower levels of melanin synthesis respectively. SRE did not show inhibitory effect on tyrosinase activities in vitro and in B16F10 cells. Finally, SRE suppressed elastse type I and IV activities in dose-dependent manner in vitro. And SRE also slightly suppressed elastase activities in B16F10 cells. In conclusion, these results suggest that SRE can inhibit melanin synthesis and inhibt elastase activity. So, We suggest that SRE can be maintained skin elasticity or whitening.

Barbigerone Inhibits Tumor Angiogenesis, Growth and Metastasis in Melanoma

  • Yang, Jian-Hong;Hu, Jia;Wan, Li;Chen, Li-Juan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.167-174
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    • 2014
  • Tumor angiogenesis, growth and metastasis are three closely related processes. We therefore investigated the effects of barbigerone on all three in the B16F10 tumor model established in both zebrafish and mouse models, and explored underlying molecular mechanisms. In vitro, barbigerone inhibited B16F10 cell proliferation, survival, migration and invasion and suppressed human umbilical vascular endothelial cell migration, invasion and tube formation in concentration-dependent manners. In the transgenic zebrafish model, treatment with $10{\mu}M$ barbigerone remarkably inhibited angiogenesis and tumor-associated angiogenesis by reducing blood vessel development more than 90%. In vivo, barbigerone significantly suppressed angiogenesis as measured by H and E staining of matrigel plugs and CD31 staining of B16F10 melanoma tumors in C57BL/6 mice. Furthermore, it exhibited highly potent activity at inhibiting tumor growth and metastasis to the lung of B16F10 melanoma cells injected into C57BL/6 mice. Western blotting revealed that barbigerone inhibited phosphorylation of AKT, FAK and MAPK family members, including ERK, JNK, and p38 MAPKs, in B16F10 cells mainly through the MEK3/6/p38 MAPK signaling pathway. These findings suggested for the first time that barbigerone could inhibit tumor-angiogenesis, tumor growth and lung metastasis via downregulation of the MEK3/6/p38 MAPK signaling pathway. The findings support further investigation of barbigerone as a potential anti-cancer drug.

Inhibition of Tumor Invasion and Metastasis by Calcium Spirulan(Ca-SP), a Novel Sulfated Polysaccharide Derived from a Blue-Green Alga Spirulina Platensis

  • Saiki, Ikuo;Murata, Jun;Fujii, Hideki;Kato, Toshimitsu
    • Nutritional Sciences
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    • v.7 no.3
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    • pp.144-150
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    • 2004
  • We have investigated the effect of calcium spirulan(Ca-SP) isolated from a blue-green alga Spirulina platensis, which is a sulfated polysaccharide chelating calcium and mainly composed of rhamnose and fructose, on invasion of both B16- BL6 melanoma cells, Colon 26 carcinoma and HT-1080 fibrosarcoma cells through reconstituted basement membrane (Matrigel). Ca-SP significantly inhibited the invasion of these tumor cells through Matrigel/fibronectin-coated filters in a concentration-dependent manner. Ca-SP also inhibited the haptotactic migration of tumor cells to laminin, but it had no inhibitory effect on tumor cell migration to fibronectin-coated filters. Ca-SP prevented the adhesion of B16-BL6 cells to Matrigel- and laminin-substrates but did not affect the adhesion to fibronectin. The pretreatment of tumor cells with Ca-SP inhibited the adhesion to laminin in a concentration-dependent fashion, while the pretreatment of laminin-substrates did not. Ca-SP had no effect on the production and activation of type IV collagenase in gelatin zymography. In contraset, Ca-SP significantly inhibited degradation of heparan sulfate by purified heparanase. The experimental lung metastasis was significantly reduced by co-injection of B16-BL6 cells with Ca-SP in a dose-dependent manner. Seven intermittent ⅰ.ⅴ. injection of 100$\mu\textrm{g}$ of Ca-SP caused a marked decrease of lung tumor colonization of B16-BL6 cells in a spontaneous lung metastasis model. These results suggest that Ca-SP, a novel sulfated polysaccharide, could reduce the lung colonization of B16-BL6 melanoma cells in experimental metastasis model, by inhibiting the tumor invasion of basement membrane Matrigel, probably through the prevention of the adhesion and migration of tumor cells to laminin-substrate and of the heparanase activity.

Inhibitory Effect of Belamcandae Rhizoma on the Melanogenesis in MSH-stimulated B16F10 cells (MSH에 의해 자극된 B16F10세포에서 사간(射干)의 멜라닌 합성 억제 효과)

  • Kim, Dae-Sung;Sung, Byung-Gon;Lee, Jang-Cheon;Lee, Boo-Kyun;Woo, Won-Hong;Lim, Kyu-Sang
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.24 no.1
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    • pp.25-35
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    • 2011
  • Objective : The present study was designed to assess the potential inhibitory activity of an ethanol extract of Belamcandae Rhizoma (EBR) on the alpha-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis signal pathway in B16F10 melanoma cells. Methods : Several experiments were performed in B16F10 melanoma cells. We studied tyrosinase activity, melanin content, cell-free tyrosinase activity and DOPA stain, and performed Western blots and RT-PCR for proteins and mRNA involved in melanogenesis. Results : ${\alpha}$-MSH-induced tyrosinase activity and melanin content were inhibited significantly by EBR. EBR markedly suppressed the protein expression level of tyrosinase in B16F10 melanoma cells. On the other hand, the expression of tyrosinase-related protein-1 (TRP-1) and -2 (TRP-2; DCT) were not affected by EBR. To elucidate the mechanism of the depigmenting property of EBR, we examined the involvement EBR in cAMP response element binding (CREB) protein phosphorylation and microphthalmia-associated transcription factor (MITF) signalling induced by ${\alpha}$-MSH. EBR did not regulate CREB phosphorylation and MITF expression by ${\alpha}$-MSH. Nevertheless, the mRNA expression of tyrosinase was significantly attenuated by EBR treatment without changes in the expression of TRP-1 and -2 mRNA. Conclusion : Our study suggested that EBR inhibits ${\alpha}$-MSH-induced melanogenesis by suppressing tyrosinase mRNA.

The effects of green tea (Camellia sinensis) flower extract on melanin synthesis in B16-F10 melanoma cells

  • Dissanayake, Chanuri-Yashara;Moon, Hae-Hee;Yang, Kyeong-Mi;Lee, Younjae;Han, Chang-Hoon
    • Korean Journal of Veterinary Research
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    • v.58 no.2
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    • pp.65-72
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    • 2018
  • The present study observed the effects of a green tea (Camellia sinensis) flower extract (GTFE) on melanin synthesis in B16-F10 melanoma cells. GTFE exhibited antioxidant activity on 2,2-diphenyl-1-picrylhydrazyl and inhibited mushroom tyrosinase activity in a dose-dependent manner. Furthermore, GTFE significantly diminished ${\alpha}-melanocyte$ stimulating hormone (${\alpha}-MSH$) stimulated cellular melanin content and tyrosinase activity throughout the concentration range evaluated. Based on RNA sequencing analysis, differential gene expression patterns observed in ${\alpha}-MSH$ stimulated B16-F10 melanoma cells were normalized by the addition of GTFE. In particular, the expression levels of melanoregulin and tyrosinase genes which are key regulating genes in melanin synthesis were up-regulated by 3.5 and 3 fold respectively by ${\alpha}-MSH$, and were normalized to control levels by the addition of GTFE. The results suggest that GTFE inhibits melanin synthesis in ${\alpha}-MSH$ stimulated B16-F10 melanoma cells by normalizing expression of genes that are essential for melanin synthesis. Overall, the results suggest that GTFE could be applied in the development of a whitening agent for the treatment of dermal hyperpigmentation.