• Title/Summary/Keyword: SK2

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Inhibitory Effect of Artemisiae Annuae Herba Extracts on Melanin Synthesis, Tyrosinase Activities and Production Levels of Tyrosinase, MMP-1 and MMP-9 in SK-MEL-2 Cells (SK-MEL-2 세포에서 청호(靑蒿) 추출물의 Melanin 생성, Tyrosinase 활성과 생성, MMP-1 및 MMP-9 생성 억제 효과)

  • Lee, Eun-Kyu;Kim, Soo-Hyeon;Park, Kyung-Mi;Yang, Seung-Jeong;Cho, Seong-Hee
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.2
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    • pp.37-48
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    • 2017
  • Objectives: Artemisiae Annuae Herba is the dried aerial part of Artemisia annua L. (AAL). In Oriental medicine, Artemisiae Annuae Herba (AAH) is traditionally used to treat fever. AAH clears summerheat or damp-Heat, clears deficiency fevers, cools the blood and stops bleeding, stops malarial disorders and relieves heat, clears liver heat and brightens the eyes. Recently, there were many studies about effects of AAH on anti-oxidative, anti-inflammatory, anti-cancer, hair growth and plasma lipid composition. So, we expected AAH has an availability that can effect on skin whitening and elasticity. Methods: The present study was designed to investigate the effects of AAH on skin whitening and elasticity in SK-MEL-2 cells. In this experiment, the effects of AAH on proliferation rates, melanin synthesis, tyrosinase activities and production levels of tyrosinase, MMP-1 and MMP-9 in vitro were examined. Results: AAH did not affect viability of SK-MEL-2 cells and inhibited melanin synthesis induced by ${\alpha}$-Melanocyte-stimulating hormone (${\alpha}$-MSH) significantly. In addition, AAH also inhibited tyrosinase activity and lowered tyrosinase level in SK-MEL-2 cells. Finally, AAH inhibited productions of Matrix metalloproteinase-1 (MMP-1) and Matrix metalloproteinase-9 (MMP-9). Conclusions: These data suggest that AAH can be used to treat patients with skin diseases such as freckled face and also used as skin whitening agent.

Cytotoxic Activity of Leguminous Seed Extracts against Human Tumor Cell Lines

  • Lee, Hoi-Seon;Lee, Jeong-Ock;Lee, Hee-Kwon;Oh, Jong-Hwan;Ahn, Young-Joon
    • Applied Biological Chemistry
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    • v.41 no.4
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    • pp.246-250
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    • 1998
  • The cytotoxic activity of methanol extracts of 25 leguminous seeds in vitro was evaluated by sulforhodamine B assay, using the five human solid A549 lung, SK-OV-2 ovarian, SK-MEL-2 melanoma, XF-498 CNS and HCT-15 colon tumor cell lines. The responses varied with both cell line arid leguminous seed used. Extracts of Canavalia lineata and Glycine soja revealed potent cytotoxic activity against A549 arid SK-MEL-2 cell lines. Moderate activity was observed in the extracts of Cassia obtusifolia and Glyeine max var. chungtae, and C. lineata and Vigna angulasis against SK-MEL-2 and HCT-15 cell lines, respectively. The other seed extracts were ineffective against model tumor cell lines. Because of their potent cytotoxic activities, the activity of each solvent fraction from C. lineata and G. soja was determined and the potent activity was produced from their chloroform fractions. As a naturally occurring therapeutic agent, leguminous seeds described could be useful for developing new types of anti-tumor agents.

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Functional significance of rSK2 N-terminal region revealed by electrophysiology and Preliminary Structural Studies

  • Narae Shin;Kang, Gil-boo;Eom, Soo-Hyun;Park, Chul-Seung
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.41-41
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    • 2003
  • Small conductance calcium-activated potassium channels (or SKCa channels) are potassium selective, voltage-independent, and activated by intracellular calcium concentration. These channels play important roles in excitable cells such as neuron in the central nervous system (Vergara et al., 1998). The activity of SKCa channels underlies the slow afterhyperpolarization that inhibits neuronal cell firing (Hille, 1991; Vergara et al.,1998). Until now, N-terminal region of rSK2 isn't characterized. To study the role of N-terminus, we constructed the N-terminal deletion mutant and characterized by electrophysiological means. Interestingly, N-terminal deletion mutant be trafficked to membrane couldn't evoke any ionic currents. Thus, N-terminal region has a role in functional rSK2 channel formation. To elucidate the function of N-terminal region, (His)6-conjugated protein was purified and filtrated by affinity column chromatography. Surprisingly, N-terminal region was shown in tetramer size that was supported by cross-linking result. Thus, we predicted that N-terminal region might be involved in the tetramerization of rSK2.

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Apoptosis of Human Hepatocarcinoma (HepG2) and Neuroblastoma (SK-N-SH) Cells Induced by Polysaccharides-Peptide Complexes Produced by Submerged Mycelial Culture of an Entomopathogenic Fungus Cordyceps sphecocephala

  • Oh, Jung-Young;Baek, Yu-Mi;Kim, Sang-Woo;Hwang, Hye-Jin;Hwang, Hee-Sun;Lee, Sung-Hak;Yun, Jong-Won
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.512-519
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    • 2008
  • Three different polysaccharide-peptide complexes (PPC, named as Fr-I, Fr-II, and Fr-III) were produced by submerged mycelial culture of an entomopathogenic fungus Cordyceps sphecocephala, and their anticancer activities were investigated in human hepatocarcinoma (HepG2) and neuroblastoma (SK-N-SH) cells. The highest inhibitory effects of PPC on both HepG2 and SK-N-SH cells were achieved with Fr-I, whereas Fr-III with low molecular mass showed lower inhibition effects. Interestingly, the inhibitory effects of the three fractions were increased after protease digestion, suggesting that the inhibitory effects resulted mainly from the carbohydrate moiety, at least in the case of Fr-II and Fr-III, of PPC. The results of DNA fragmentation in PPC-induced apoptotic cells were confirmed by both DNA ladder assay and comet assay. Our investigation also showed that PPC-induced apoptosis of both cancer cells was associated with intracellular events including DNA fragmentation, activation of caspase-3, and modulation of Bcl-2 and Bax. We conclude that PPC has potential as a novel therapeutic agent for the treatment of both HepG2 and SK-N-SH cancer cells without any cytotoxicity against normal cells.

Anti-apoptotic Effects of Red Ginseng on Oxidative Stress Induced by Hydrogen Peroxide in SK-N-SH Cells

  • Kim, Eun-Hye;Lee, Mi-Jeong;Kim, In-Hye;Pyo, Suhk-Neung;Choi, Kwang-Tae;Rhee, Dong-Kwon
    • Journal of Ginseng Research
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    • v.34 no.2
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    • pp.138-144
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    • 2010
  • Ginseng (Panax ginseng C.A. Meyer) has been shown to have anti-stress effects in animal studies. However, most studies have only managed to detect altered levels of biomarkers or enzymes in blood or tissue, and the actual molecular mechanisms by which ginseng exerts these effects remain unknown. In this study, the anti-oxidative effect of Korean red ginseng (KRG) was examined in human SK-N-SH neuroblastoma cells. Incubation of SK-N-SH cells with the oxidative stressor hydrogen peroxide resulted in significant induction of cell death. In contrast, pre-treatment of cells with KRG decreased cell death significantly. To elucidate underlying mechanisms by which KRG inhibited cell death, the expression of apoptosis-related proteins was examined by Western blot analysis. KRG pre-treatment decreased the expression of the pro-apoptotic gene caspase-3, whereas it increased expression of the anti-apoptotic gene Bcl-2. Consistent with this, immunoblot analysis showed that pre-treatment of the SK-N-SH cells with KRG inhibited expression of the pro-inflammatory gene cyclooxygenase 2 (COX-2). RT-PCR analysis revealed that the repression of COX-2 expression by KRG pre-treatment occurred at the mRNA level. Taken together, our data indicate that KRG can protect against oxidative stress-induced neuronal cell death by repressing genes that mediate apoptosis and inflammation.

Phase Diagrams and Stable Structures of Stranski-Krastanov Structure Mode for III-V Ternary Quantum Dots

  • Nakaima, Kazuno;Ujihara, Toru;Miyashita, Satoru;Sazaki, Gen
    • Proceedings of the Korea Association of Crystal Growth Conference
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    • 1999.06a
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    • pp.81-114
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    • 1999
  • The strain, surface and inerfacial energies of III-V ternary systems were calculated for three kinds of structure modes: the Frank-van der Merwe (FM) mode, the Stranski-Krastanov (SK) mode an the Volmer-Wever (VW) mode. The free energy for each mode was estimated as functions of the thickness and composition or lattice misfit. Through comparison of the free energy of each mode, it was found that the thickness-composition phase diagrams of III-V ternary systems can be determined only by considering the balance of the free energy and three kinds of structure modes appear in the phase diagrams. The SK mode appears only when the lattice misfit is large and/or the lattice layer is thick. The most stable structure of the SK mode is a cluster with four lattice layers or minimum thickness on a wetting layer of increasing lattice layers. The VW mode appears when the lattice misfit is large and the lattice layer is thin and only in the InPSb/InP and GaPSb/GaP systems which have the largest lattice misfit of III-V ternary systems. The stable region of the SK mode in the GaPSb/GaP and InPSb/InP phase diagrams is largest of all because the composition dependence of the strain energy of these systems is stronger than that of the other systems. The critical number of lattice layers below which tow-dimensional (2D) layers precede the three-dimensional (3D) nucleation in the SK mode at x=1.0 depnds on the lattice misfit.

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Effect on Alzheimer's Disease by Sesim-tang in CT105-overexpressed SK-N-SH Cell Lines (CT105로 유도된 신경모세포종 세포주에서 세심탕의 항치매 효과)

  • 권형수;박치상;박창국
    • The Journal of Korean Medicine
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    • v.25 no.2
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    • pp.138-150
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    • 2004
  • Objectives : Alzheimer's disease (AD) is a geriatric dementia that is widespread in old age. In the near future AD will be the biggest problem in public health service. Although a variety of oriental prescriptions, including Sesim-tang, have been traditionally utilized for the treatment of AD, their pharmacological effects and action mechanisms have not yet been fully elucidated. The present study investigated the effects of Sesim-tang on apoptotic cell death induced by CT105 (carboxy terminal 105 amino acid peptide fragment of APP) overexpression in SK-N-SH neuroblastoma cell lines. Methods: We studied the regenerative and inhibitory effects on Alzheimer's disease in CT105-induced SK-N-SH cell lines by Sesim-tang water extract. We examined for cell morphological pattern, DNA fragmentation, LDH activity assay, zymography assay, and immunohistochemistric analysis. Additionally, we investigated the association between the CT105 and neurite degeneration caused by CT105-induced apoptotic response in neurone cells. Results: Findings from our experiments have shown that Sesim-tang inhibits the synthesis or activities of CT105, which has neurotoxicities and apoptotic activities in the cell line. In addition, pretreatment with Sesim-tang ($>50\mu\textrm{g}/ml$ for 12 hours) partially prevented CT105-induced cytotoxicity in SK-N-SH cell lines. SK-N-SH cell lines overexpressed with CT105 exhibited remarkable apoptotic cell damage. Based on morphological observations by phase-contrast microscope and LDH activity measurements in the culture media, the CT105-induced cell death was significantly inhibited by Sesim-tang water extract. Sesim-tang was found to reduce the expression of APP and caspase-3 induced by CT105 in SK-N-SH cell lines and in rat hippocampus. Conclusions: As the result of this study, in the Sesim-tang group, apoptosis in the nervous system is inhibited, the repair against the degeneration of SK-N-SH cell lines by CT105 expression is promoted. Hence, Sesim-tang may be beneficial for the treatment of AD.

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Heterologous Production of Streptokinase in Secretory Form in Streptomyces lividans and in Nonsecretory Form in Escherichia coli

  • Kim,, Mi-Ran;Choeng, Yong-Hoon;Chi, Won-Jae;Kang, Dae-Kyung;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.132-137
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    • 2010
  • The skc gene encoding streptokinase (SK) with a molecular mass of approximately 47.4 kDa was cloned from Streptococcus equisimilis ATCC 9542 and heterologously overexpressed in Streptomyces lividans TK24 and E. coli using various strong promoters. When the promoter for sprT [Streptomyces griseus trypsin (SGT)] was used in the host S. lividans TK24, a 47.4-kDa protein was detected along with a smaller hydrolyzed protein (44 kDa), suggesting that posttranslational hydrolysis had occurred as has been reported in other expression systems. The casein/plasminogen plate assay revealed that the plasmid construct containing the SGT signal peptide was superior to that containing the SK signal peptide in terms of SK production. Maximal production of SK was calculated to be about 0.25 unit/ml of culture broth, a value that was five times higher than that obtained with other expression systems using ermE and tipA promoters in the same host. When the skc gene was expressed in E. coli BL21(${\Delta}DE3$)pLys under the control of the T7 promoter, a relatively large amount of SK was expressed in soluble form without hydrolysis. SK activity in E. coli/pET28a-$T7_pSK_m$ was more than 2 units/ml of culture broth, even though about half of the expressed protein formed an inactive inclusion body.

New Cytotoxic Sulfated Saponins from the Starfish Certonardoa semiregularis

  • Wang Wei Hong;Jang Hyo Jin;Hong Jong Ki;Lee Chong Ok;Bae Song Ja;Shin Sook;Jung Jee H.
    • Archives of Pharmacal Research
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    • v.28 no.3
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    • pp.285-289
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    • 2005
  • Two new sulfated saponins designated as certonardosides P$_{2}$ and I$_{3}$ (1 and 2) were isolated from the brine shrimp active fraction of the MeOH extract of the starfish Certonardoa semiregularis. The structures were determined on the basis of spectral analysis. Compounds 1 and 2 were tested for cytotoxicity against five human tumor cell lines (A549, SK-OV-3, SK-MEL-2, XF498, and HCT15), and compound 1 displayed significant cytotoxicity against the SK-MEL-2 skin cancer cell.

Purification, Characterization, and Cloning of Trimethylamine Dehydrogenase from Methylophaga sp. Strain SK1

  • Kim, Hee-Gon;Kim, Yan;Lim, Heon-Man;Shin, Hyun-Jae;Kim, Si-Wouk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.337-343
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    • 2006
  • Trimethylamine dehydrogenase (TMADH, EC 1.5.99.7), an iron-sulfur flavoprotein that catalyzes the oxidative demethylation of trimethylamine to form dimethylamine and formaldehyde, was purified from Methylophaga sp. strain SK1. The active TMADH was purified 12.3-fold through three purification steps. The optimal pH and temperature for enzyme activity was determined to be 8.5 and $55^{\circ}C$, respectively. The $V_{max}\;and\;K_m$ values were 7.9 nmol/min/mg protein and 1.5 mM. A genomic DNA of 2,983 bp from Methylophaga sp. strain SK1 was cloned, and DNA sequencing revealed the open reading frame (ORF) of the gene coding for TMADH. The ORF contained 728 amino acids with extensive identity (82%) to that of Methylophilus methylotrophus $W_3A_1$.