• Title/Summary/Keyword: F9 cells

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Anti-cancer effects of kelp extract in mouse melanoma B16-F0 cell line through apoptosis (마우스 흑색종 세포주 B16-F0에서 다시마 추출물의 세포사멸을 통한 항암 효과)

  • Lee, Seong-Uk;Kim, Yoon Hee
    • Korean Journal of Food Science and Technology
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    • v.54 no.2
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    • pp.134-140
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    • 2022
  • Kelp belongs to the brown algae family and has been reported to exert anti-cancer effects on some cancer types, however studies have not been reported on the anti-cancer effects of kelp extracts on melanoma. In this study, the anti-cancer effects of kelp extract in B16-F0 cells were investigated, and the underlying molecular mechanisms were assessed. Kelp extract was found to inhibit the proliferation of B16-F0 cells, induce cytotoxicity, inhibit cell colony formation, and induce DNA fragmentation and apoptosis. The molecular mechanism was found to involve kelp extract increasing the expression of cytochrome-c and activated caspase-9 in the intrinsic apoptotic pathway. In addition, kelp extract upregulated the expression of Fas-associated protein with death domain and activated caspase-8 in the extrinsic apoptosis pathway. Activation of caspase-9 and caspase-8 by kelp extract induced activation of caspase-3 and cleaved poly adenosine diphosphate-ribose polymerase, consequently inducing apoptosis. These data suggest that kelp extract represents a potential therapeutic agent for melanoma.

Production of Extracellular Polysaccharide by Perfusion Culture of Angelica gigas Nakai Suspension Cells (배지교환식 고농도 배양에 의한 참당귀 현탁세포 유래 ECP 생산)

  • Kim, Young-Hwa;Kim, Ik-Hwan;Kim, Dong-Il
    • KSBB Journal
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    • v.21 no.5
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    • pp.336-340
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    • 2006
  • High-density perfusion cultivation was performed to produce extracellular polysaccharide(ECP) as immunostimulating agents in suspension cell cultures of Angelica gigas Nakai. In batch culture, the maximum cell density was 16.8 gDCW/L at day 6 and 0.9 g/L of ECP was obtained at day 8. When the medium exchange was started at the fifth day after inoculation for the perfusion culture, high concentration of the cells at 23.8 gDCW/L could be achieved with continuous production of ECP. Treatments of ultrasound and Pluronic F-68 were found to be helpful for the secretion of intracellular ECP into the culture medium.

Comparison of Characteristics and Performance of Membrane in Proton Exchange Membrane Fuel Cells (고분자전해질 연료전지 고분자막의 특성 및 성능 비교)

  • Lee, Daewoong;Lim, Daehyun;Oh, Sohyeong;Chung, Hoi-Bum;Yoo, Seung-Eul;Ku, Young-Mo;Park, Kwonpil
    • Korean Chemical Engineering Research
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    • v.58 no.2
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    • pp.171-175
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    • 2020
  • In the proton exchange membrane fuel cells (PEMFC), the development of a reinforced membrane with improved durability by a support is actively in progress in Korea. In this study, the initial performance and characteristics of four types of reinforced membranes were compared. Reinforced membranes with higher amounts of C-F chains in the polymer membrane showed lower water diffusion coefficients due to the hydrophobicity of the C-F chains. The thicker the polymer membrane, the more the hydrogen permeability decreased and the higher the OCV. Membrane with short resistance below 1.5 Ωcm2 showed OCV below 0.9 V and the lowest performance, so short resistance should be above 3.0 Ωcm2. Compared with the current standard membrane, there was a similar domestic membrane, which could confirm the possibility of localization of PEMFC polymer membrane.

Anti-oxidant, anti-inflammatory and Whitening effect of Benincasa hispida seed extract (동과자 추출물의 항산화, 항염 및 미백 효능)

  • Park, Gyu-Ri;Lee, Ji-An
    • Journal of Convergence for Information Technology
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    • v.10 no.7
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    • pp.249-256
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    • 2020
  • To investigate the natural cosmetic ingredients of Benincasa hispida seed extract on skin care, we measured anti-oxidant and anti-inflammatory, and whitening effect. DPPH free radical scavenging activity was increased in a dose-dependent manner. The total phenolic content was higher in methanol extract (22.42±0.002 mgGAE/g) than water extract (9.77±0.002 mgGAE/g). MTT assay was demonstrated that the seed extract did not have a cytotoxic effect in RAW264.7 and B16F10 cell lines. We also examined to find out the inhibitory activity on NO production and secretion of TNF-α cytokine in LPS-induced RAW264.7 cells. In B16F10 melanoma cells, the seed extract significantly suppressed α-MSH induced melanin synthesis. Furthermore, westernblot analysis revealed that methanol extract dramatically downregulated the expression level of MITF, TRP-1 and TRP-2. Taken together, the B. hispida seed extract posses anti-oxidant, anti-inflammatory and skin whitening activities, which might provided its functional efficacy in cosmetic materials.

Effect of Ulmus macrocapa Ethanolic Extracts on Anti-oxidant Activity and Melanin Synthesis in B16F1 Cells (B16F1세포에서 항산화 활성 및 멜라닌 합성에 대한 유백피 에탄올 추출물의 효능)

  • Kwon, Eun-Jeong;Park, Hye-Jung;Kim, Moon-Moo;Lee, Kyeong Rok;Hong, Il;Lee, Do Gyeong;Oh, Yunghee
    • Journal of Life Science
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    • v.24 no.9
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    • pp.946-951
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    • 2014
  • Melanin plays a key role in the protection of skin from ultraviolet light that generates reactive oxygen species (ROS), such as superoxide, hydroxyl radical, singlet oxygen and hydrogen peroxide. However, the ROS leading to the oxidation of lipids, proteins and DNA are involved in the overproduction of melanin that is known to cause melasma, age spots and freckles. Among the herb medicines, Ulmus macrocarpa used in this study was reported to contain flavonoids as a main component. The aim of this study is to investigate the whitening and anti-oxidant effects of Ulmus macrocarpa ethanolic extracts (UMEE) in B16F1 cells. UMEE below $3.12{\mu}g/ml$ did not show cytotoxicity. In an anti-oxidant experiment, UMEE showed not only high reducing power and scavenging activity on DPPH, but it was also observed that UMEE exhibit an inhibitory effect on lipid peroxidation. UMEE did not display an inhibitory effect on tyrosinase activity in vitro. However, UMEE inhibited melanin synthesis in B16F1 cells. In addition, UMEE reduced the expression levels of tyrosinase and tyrosinase-related protein-2 (TRP-2), which are key enzymes in melanogenesis. These results indicate that UMEE exert a whitening effect through the inhibition of both tyrosinase and TRP-2 expressions as well as anti-oxidant activity, suggesting that UMEE could have the functional potential for a whitening effect on the skin.

Carotenoids Components of Tunicata, Shellfishes and Its Inhibitory Effects on Mutagenicity and Growth of Tumor Cell (미색동물 및 패류의 Carotenoids 색소성분과 돌연변이 및 종양세포 증식의 억제효과)

  • 하봉석;백승한;김수영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.5
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    • pp.922-934
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    • 2000
  • To investigate the composition of carotenoids present in marine organisms and the biological activity of the carotenoids, carotenoids of the muscles and tunic of tunicates and shellfishes were isolated and identified. Anitmutagenic activities of the carotenoids for S. typhimurium TA 98 and cytotoxic activity for cancer cell lines were determined. Total carotenoid contents in the muscle of tunicata ranged from 18.65 mg% to 2.39 mg%. The highest amount of the total carotenoid was found in the muscle of Halocynthia aurantium, followed by Styela clava (HERDMAN), H. roretzi, H. hilgendorfi f. igaboya, H. hilgendorfi f. retteri, S. plicata (LESUEUR) in order. Interestingly, total carotenoid content in the muscle of S. clava (HERDAMAN) was higher than that of H. roretzi. Total carotenoid content of all tunicata, other than H. aurantium and H. roretzi, were higher in muscle than tunic. The major carotenoids in H. roretzi, H. aurantium, S. plicata (LESUEUR), and S. clava (HERDAMAN) were cynthiaxanthin (25.1∼42.2%), halocynthiaxanthin (9.7∼26.3%), diatoxanthin (8.0∼18.7%) and β-carotene (7.7%∼21.7%). Similarly, cantaxanthin (19.6%), cynthiaxanthin (15.4%), halocynthiaxanthin (14.8%), and (3R, 3'R), (3S, 3'S)-astaxanthin (22.6%) in H. hilgendorfi f. retteri and fucoxanthin (26.6%), cynthiaxanthin (21.8%), halocynthiaxanthin (15.2%), and β-carotene (9.3%) in H. hilgendorfi f. igaboya were major carotenoids in both tunicate. However, the composition of carotenoids in muscle and tunic of tunicata was similar each other. Among the shellfishes examined, total carotenoid content of the muscle of Peronidia venulosa (Schrenck) and Corbicula fluminea, and of the gonad of Atrina pinnata and Chlamys farreri, was ranged from 2.51 to 6.83 mg% which were relatively higher than that of other shellfishes. The composition of the carotenoids of shellfishes, which might depend upon their living environments, was varied. But cynthiaxanthin (15.9∼39.0%) and zeaxanthin (9.6∼21.9%) in gonad of C. farreri, and muscles of Buccinum Volutharpa perryi (JAY) and Crassostrea gigas, cynthiaxanthin (21.5∼48.6%) and mytiloxanthin (14.6%) in muscle of C.fluminea and gonad of A. pinnata, and canthaxanthin (60.6%) and isozeaxanthin (20.5%) in muscles of P. venulosa (Schrenck), and β-carotene (23.7%∼37.8%) and zeaxanthin (18.2∼20.4) in muscles of Semisulcospira libertina and Meretrix lusoria were major carotenoids. Interestingly, diester type-carotenoids were present along with free type-carotenoids in muscles of C. gigas. antimutagenic effect of the carotenoids isolated from tunicata and shellfishes against 2-amino-3-methylimidazol [4,5-f]quinoline (IQ) for S. typhimurium TA 98 was proportional to the amount (20, 50 and 100㎍/plate) treated. Mutagenicity of IQ was significantly reduced by astaxanthin, isozeaxanthin, mytiloxanthin and halocynthiaxanthin, whereas the mutagenicity of aflatoxin B₁(AFB₁) was significantly reduced by β-carotene, isozeaxanthin, and mytiloxnthin. Growth inhibition effect of carotenoids isolated from tunicata and shellfishes for cancer cell was proportional to the amount (5, 10, and 20㎍/plate) treated. The growth of HeLa cell by β-carotene, cynthiaxanthin, astaxanthin and halocynthiaxanthin, NCI-H87 cell by β-carotene, astaxanthin, cynthiaxanthin, and halocynthiaxanthin, HT-29 cell by β-carotene, cynthiaxanthin, mytiloxanthin and halocynthiaxanthin, and MG-63 cells by β-carotene, cynthiaxanthin, astaxanthin, canthaxanthin and halocynthiaxanthin were statistically reduced.

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Effects of Synthetic Pseudoceramides on Sphingosine Kinase Activity in F9-12 Cells

  • Jin, You-Xun;Shin, Kyong-Oh;Park, Myung-Yong;Lee, Shin-Hee;Park, Byeong-Deog;Oh, Sei-Kwan;Yoo, Hwan-Soo;Lee, Yong-Moon
    • Biomolecules & Therapeutics
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    • v.19 no.1
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    • pp.134-139
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    • 2011
  • Sphingosine kinase (SPHK) has a central role to control cell death and cell proliferation, which is suggested as a sphingolipid rheostat by regulating the levels between ceramide and sphingosine 1-phosphate (S1P). Therefore, physiological regulators of SPHK will be a good candidate to develop a new targeted drug. For this purpose, a series of synthetic pseudoceramides were tested by SPHK assay either cell-based or cell-free system. K10PC-5 strongly inhibited SPHK, while K6PC-5 activated SPHK in cell-free system. Specifically, K6PC-5 activated SPHK under the co-treatment with $50\;{\mu}M$ dimethylsphingosine (DMS), a SPHK inhibitor. Collectively, we developed a simple SPHK assay system to find SPHK regulatory pseudoceramide compounds, K10PC-5 and K6PC-5 which may be useful to cancer treatment or immune regulation like FTY720, a synthetic sphingolipid mimetic compound.

Anti-Melanogenic Effect of Dendropanax Morbiferus and Its Active Components via Protein Kinas e A/Cyclic Adenos ine Monophosphate-Responsive Binding Protein-and p38 Mitogen-Activated Protein Kinase-Mediated Microphthalmia-Associated Transcription Factor Downregulation

  • Bohyun Yun;Ji Soo Kim;Jung Up Park
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.104-104
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    • 2022
  • Dendropanax morbiferus H. Lev has been reported to have some pharmacologic activities and also interested in functional cosmetics. We found that the water extract of D. morbiferus leaves significantly inhibited tyrosinase activity and melanin formation in α-melanocyte stimulating hormone (MSH)-induced B16-F10 cells. D. morbiferus reduced melanogenesis-related protein levels, such as microphthalmia? associated transcription factor (MITF), TRP-1, and TRP-2, without any cytotoxicity. Two active ingredients of D. morbiferus, (10E)-9,16-dihydroxyoctadeca-10,17-dien-12,14-diynoate (DMW-1) and (10E)-(?)-10,17-octadecadiene-12,14-diyne-1,9,16-triol (DMW-2) were identified by testing the anti-melanogenic effects and then by liquid chromatography-tandem mass spectrometry (LC/MS/MS) analysis. DMW-1 and DMW-2 significantly inhibited melanogenesis by the suppression of protein kinase A (PKA)/cyclic AMP (cAMP)-responsive binding protein (CREB) and p38 MAPK phosphorylation. DMW-1 showed a better inhibitory effect than DMW-2 in α-MSH-induced B16-F10 cells. D. morbiferus and its active component DMW-1 inhibited melanogenesis through the downregulation of cAMP, p-PKA/CREB, p-p38, MITF, TRP-1, TRP-2, and tyrosinase. These results indicate that D. morbiferus and DMW-1 may be useful ingredients for cosmetics and therapeutic agents for skin hyperpigmentation disorders.

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NICKEL INCORPORATION INTO Klebsiella aerogenes UREASE (Klebsiella aerogenes Urease로의 닉켈의 도입)

  • Lee, Mann-Hyung-
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.11a
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    • pp.69-80
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    • 1994
  • Although ureases play important roles in microbial nitrogen metabolism and in the pathogenesis of several human diseases, little is known of the mechanism of metallocenter biosynthesis in this Ni-Containing enzyme. Klebsiella aerogenes urease apo-protein was purified from cells grown in the absence of Ni. The purified apo-enzyme showed the same native molecular weight, charge, and subunit stoichiometry as the holo-enzyme. Chemical modification studies were consistent with histidinyl ligation of Ni. Apo-enzyme could not be activated by simple addition of Ni ions suggesting a requirement for a cellular factor. Deletion analysis showed that four accessory genes (ureD, ureE, ureF, and ureG) are necessary for the functional incorporation of the urease metallocenter. Whereas the $\Delta$ureD, $\Delta$ureF, and $\Delta$ureG mutants are inactive and their ureases lack Ni, the $\Delta$ureE mutants retain partial activity and their ureases possess corresponding lower levels of Ni. UreE and UreG peptides were identified by SDS-polyacrylamide gel comparisons of mutant and wild type cells and by N-terminal sequencing. UreD and UreF peptides, which are synthesized at ve교 low levels, were identified by using in vitro transcription/translation methods. Cotransformation of E. coli cells with the complementing plasmids confirmed that ureD and ureF gene products act in trans. UreE was purified and characterized. immunogold electron microscopic studies were used to localize UreE to the cytoplasm. Equilibrium dialysis studies of purified UreE with $^{63}$ NiC1$_2$ showed that it binds ~6 Ni in a specific manner with a $K_{d}$ of 9.6 $\pm$1.3 $\mu$M. Results from spectroscopic studies demonstrated that Ni ions are ligated by 5 histidinyl residues and a sixth N or O atom, consistent with participation of the polyhistidine tail at the carboxyl termini of the dimeric UreE in Ni binding. With these results and other known features of the urease-related gene products, a model for urease metallocenter biosynthesis is proposed in which UreE binds Ni and acts as a Ni donor to the urease apo-protein while UreG binds ATP and couples its Hydrolysis to the Ni incorporation process.ouples its Hydrolysis to the Ni incorporation process.s.

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Transmission electron microscopic ultrastructures of the male germinal cells of Fibricola seoulensis (Fibricola seoulensis 웅성생식세포의 투과전자현미경적 미세구조)

  • 손원목;이순형
    • Parasites, Hosts and Diseases
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    • v.31 no.3
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    • pp.183-192
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    • 1993
  • A transmission electron microscopic study was performed to observe the ultrastructures of the male germinal cells and spermatozoa of Fibricola seoulensis. Spermatogonia were found in the periphery of the testis and characterized by large nuclei and comparatively little cytoplasms. Spermatocytes contained an oval to spherical nucleus. Their nuclear volume was little larger in comparative to that of cytoplasm, and the chromatin was comparatively little. The early spermatids were characterized by a great amount of cytoplasm, and numerous mitochondria encircled the nucleus. In a more advanced spermatids the electron-dense strands of chromatin appeared in the nucleus, and a pair of rootlet of the axoneme and a microtubule-organizing center (MTOC) were observed near the nucleus. The sectioned spermatozoa were found in the testis and the seminal vesicle. Their cross sectional views were divided into 6 types when they were distinguished on the basis of the morphology and components. The spermatozoa of F. seoulensis showed two flagella of 9+1 type axoneme.

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