• Title/Summary/Keyword: 3-D Mechanism

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Cellular activities and docking studies of eckol isolated from Ecklonia cava (Laminariales, Phaeophyceae) as potential tyrosinase inhibitor

  • Lee, Seung-Hong;Kang, Sung-Myung;Sok, Chang Hyun;Hong, Jin Tae;Oh, Jae-Young;Jeon, You-Jin
    • ALGAE
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    • v.30 no.2
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    • pp.163-170
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    • 2015
  • Tyrosinase inhibitors are an important component of cosmetic products. Our previous studies have proposed that eckol isolated from the brown alga Ecklonia cava, can be explored as a tyrosinase inhibitor. However, cellular activities and mechanism of action of eckol remain unknown. Therefore, the current study analyzed the eckol binding modes using the crystal structure of Bacillus megaterium tyrosinase. The effects of eckol on melanin synthesis induced by ${\alpha}$-melanocyte stimulating hormone in B16F10 melanoma cells were also investigated. We predicted the 3D structure of tyrosinase and used a docking algorithm to simulate binding between tyrosinase and eckol. These molecular modeling studies were successful (calculated binding energy value, $-115.84kcal\;mol^{-1}$) and indicated that eckol interacts with Asn205, His208, and Arg209. Furthermore, eckol markedly inhibited tyrosinase activity and melanin synthesis in B16F10 melanoma cells. We also found that eckol decreased the expression of tyrosinase, tyrosinase-related protein (TRP) 1, and TRP2. These results indicate that eckol is a potent inhibitor of melanogenesis, and this finding may be useful for the development of novel pharmaceutical and cosmetic agents.

A Study about the Mechanism of $Ca^{2+}$ and Pi Homeostasis by Estradiol 17$\beta$ in Proximal Tubule Cells in the Osteoporosis

  • Han Ho-jae;Park Soo-Hyun
    • Biomedical Science Letters
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    • v.10 no.4
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    • pp.375-383
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    • 2004
  • It has been reported that osteoporosis induced by the deficiency of estrogens in menopause is associated with the unbalance of Ca/sup 2+/ and Pi levels. Proximal tubule is very important organ to regualte Ca/sup 2+/ and Pi level in the body. However, the effect of estrogens on Ca/sup 2+/ and Pi regulation was not elucidated. Thus, we examined the effect of 17-β estradiol (E₂) on Ca/sup 2+/ and Pi uptake in the primary cultured rabbit renal proxiaml tubule cells. In the present study, E₂(> 10/sup -9/M) decreases Ca/sup 2+/uptake and stimulates Pi uptake over 3 days. E₂-induced decrease of Ca/sup 2+/ uptake and stimulation of Pi uptake were blocked by actinomycin D (a gene transcription inhibitor), cycloheximide (a protein synthesis inhibitor). tamoxifen, and progesterone (estrogen receptor antagonists). E₂-induced decrease of Ca/sup 2+/ uptake and stimulation of Pi uptake were blocked by SQ22536 (an adenylate cyclase inhibitor), Rp-cAMP (a cAMP antagonist), and PKI (a protein kinase A inhibitor). Indeed, E₂ increased cAMP formation. In addition, E₂-induced decrease of Ca/sup 2+/ uptake and stimulation of Pi uptake were blocked by staurosporine, H-7, and bisindolylmaleimide I (protein kinase C inhibitors) and E₂ translocated PKC from cytoslic fraction to membrane fraction. In conclusion, E₂ decreased Ca/sup 2+/ uptake and stimulated Pi uptake via cAMP and PKC pathway in the PTCs.

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Relationship between Differential Expression of Estrogen Receptor and Follicle Stimulating Hormone Receptor Genes in Ovary and Heterosis of Egg Number Traits in Chickens

  • Wang, Hui;Sun, Dongxiao;Yu, Ying;Wang, Dong;Zhang, Yi;Zhang, Yuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.3
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    • pp.325-330
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    • 2008
  • In order to understand the molecular mechanism of heterosis of reproduction traits in chickens, we used the quantitative real-time reverse transcriptional polymerase chain reaction (Quantitative real-time RT-PCR) technique to investigate the differential expression of estrogen receptor (ESR) and follicle stimulating hormone receptor (FSHR) genes in 32-week-old ovaries of inbred chickens and their hybrid offspring in $4{\times}4$ diallel crosses, which involved White Plymouth Rock (E), CAU Brown (D), Silkies (C) and White Leghorn (A). We found that there were significant differences in mRNA expression of ESR and FSHR genes not only between hybrids and their parental lines (p<0.01), but also among different crosses (p<0.01). Furthermore, positive correlations between differential expression of both ESR and FHSR in hybrids and heterosis percentages of 32-week-old and 42-week-old egg number traits were significant at p<0.05. Our results suggested that differential expression of ESR and FSHR genes in the ovaries of inbred chickens and their hybrids could play roles in the formation of heterosis of egg number traits to some extent.

Mechanism of Action of Nigella sativa on Human Colon Cancer Cells: the Suppression of AP-1 and NF-κB Transcription Factors and the Induction of Cytoprotective Genes

  • Elkady, Ayman I;Hussein, Rania A;El-Assouli, Sufian M
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7943-7957
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    • 2015
  • Background and Aims: Colorectal cancer is one of the leading causes of death in the world. The aim of this study was to investigate the growth-suppression potentiality of a crude saponin extract (CSENS) prepared from medicinal herb, Nigella sativa, on human colon cancer cells, HCT116. Materials and Methods: HCT116 cells were subjected to increasing doses of CSENS for 24, 48 and 72 h, and then harvested and assayed for cell viability by WST-1. Flow cytometry analyses, cell death detection ELISA, fluorescent stains (Hoechst 33342 and acridine orange/ethidium bromide), DNA laddering and comet assays were carried out to confirm the apoptogenic effects of CSENS. Luciferase reporter gene assays, quantitative reverse transcription-polymerase chain reaction and Western blot analyses were performed to assess the impact of CAERS and CFEZO on the expression levels of key regulatory proteins in HCT116 cells. Results: The results demonstrated that CSENS inhibited proliferation and induced apoptosis. Apoptosis was confirmed by flow cytometry analyses, while CSENS-treated cells exhibited morphological hallmarks of apoptosis including cell shrinkage, irregularity in cellular shape, cellular detachment and chromatin condensation. Biochemical signs of apoptosis, such as DNA degradation, were observed by comet assay and gel electrophoresis. The pro-apoptotic effect of CSENS was caspase-3-independent and associated with increase of the Bax/Bcl-2 ratio. CSENS treatment down-regulated transcriptional and DNA-binding activities of NF-${\kappa}B$ and AP-1 proteins, associated with down-regulation of their target oncogenes, c-Myc, cyclin D1 and survivin. On the other hand, CSENS up-regulated transcriptional and DNA-binding activities of Nrf2 and expression of cytoprotective genes. In addition, CSENS modulated the expression levels of ERK1/2 MAPK, p53 and p21. Conclusions: These findings suggest that CSENS may be a valuable agent for treatment of colon cancer.

Molecular mechanism underlying Arabidopsis root architecture changes in response to phosphate starvation

  • Chun, Hyun Jin;Lee, Su Hyeon;Kim, Min Chul
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.174-174
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    • 2017
  • To cope with phosphate (Pi) deficient stress, plants modulate various physiological and developmental processes, such as gene expression, Pi uptake and translocation, and root architecture changes. Here, we report the identification and characterization of novel activation-tagged mutant involved in Pi starvation signaling in Arabidopsis. The hpd (${\underline{h}ypersensitive}$ to ${\underline{P}i}$ $ {\underline{d}eficiency}$) mutant exhibits enhanced phosphate uptake and altered root architectural change under Pi starvation compared to wild type. Expression analysis of auxin-responsive DR5::GUS reporter gene in hpd mutant indicated that auxin translocation in roots under Pi starvation are suppressed in hpd mutant plants. Impaired auxin translocation in roots of hpd mutant was attributable to abnormal root architecture changes in Pi starvation conditions. Our results indicated that abnormal auxin translocation in hpd mutant might be due to mis-regulation of auxin efflux carrier proteins, PIN-FORMED (PIN) 1, and 2 under Pi starvation conditions. Not only expression levels but also expression domains of PIN proteins were altered in hpd mutant in response to Pi starvation. Molecular genetic analysis of hpd mutant revealed that the mutant phenotype is caused by the lesion in ENHANCED SILENCING PHENOTYPE4 (ESP4) gene whose function is proposed in mRNA 3'-end processing. The results suggest that mRNA processing plays crucial roles in Pi homeostasis as well as developmental reprograming in response to Pi deprivation in Arabidopsis.

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A Permeation Behavior for the Pervaporation of Aqueous Ethanol Solution (에탄올 수용액의 Pervaporation에서의 투과거동)

  • Bae, Seong-Youl;Lee, Han-Sun;Hwang, Seong-Min;Kim, Hee-Taik;Kumazawa, Hidehiro
    • Applied Chemistry for Engineering
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    • v.5 no.1
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    • pp.127-138
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    • 1994
  • In the process of pervaporation separation for aqueous ethanol solution through cellulose tai-acetate(CTA) membrane, the modelling on the solution-diffusion permeation mechanism was built up on the basis of sorption and permeation experimental results. Also its function type and parameter were examined. The composition of sorption equilibrium in three component system(Ethanol/Water/CTA) were compared with the calculated value by Flory-Huggins' equation using the pure component sorption data. In order to apply the thermodynamic equilibrium relationship between the membrane free composition in the membrane and the equilibrium composition in the liquid phase, the apparent activity this system, however, the results were not satisfied. Diffusion equations were expressed with the concentration gradient considering permeate alone, and a concentration-dependent diffusion coefficient which includes a parameter was used. And this model was fitted with the measured permeation rates. If the permeation rate and the amount of sorption of one component were much larger than those of the other, the bulk flow term could not be negligible. The flux and selectivity were increased with increasing temperature, and with decreasing downstream pressure.

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Methanol Extracts of Codium fragile Induces Apoptosis through G1/S Cell Cycle Arrest in FaDu Human Hypopharynx Squamous Carcinoma Cells

  • Lee, Seul Ah;Park, Bo-Ram;Moon, Sung Min;Kim, Do Kyung;Kim, Chun Sung
    • International Journal of Oral Biology
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    • v.43 no.2
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    • pp.61-68
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    • 2018
  • Codium fragile (Suringar) Hariot is an edible green seaweed that belong to the Codiaceae family and has been used in Oriental medicine for the treatment of enterobiasis, dropsy, and dysuria. Methanol extract of codium fragile has anti-oxidant, anti-inflammatory and anti-cancer properties, although the anti-cancer effect on oral cancer has not yet been reported. In this study, we investigated the anti-cancer activity and the mechanism of cell death by methanol extracts of Codium fragile (MeCF) on human FaDu hypopharyngeal squamous carcinoma cells. Our data showed that MeCF inhibits cell viability in a dose-dependent manner, and markedly induced apoptosis, as determined by the MTT assay, Live/Dead assay, and DAPI stain. In addition, MeCF induced the proteolytic cleavage of procaspase -3, -7, -9 and poly(ADP-ribose) polymerase(PARP), and upregulated or downregulated the expression of mitochondrial-apoptosis factor, Bax(pro-apoptotic factor), and Bcl-2(anti-apoptotic factor). Futhermore, MeCF induced a cell cycle arrest at the G1/S phase through suppressing the expression of the cell cycle cascade proteins, p21, CDK4, CyclinD1, and phospho-Rb. Taken together, these results indicated that MeCF inhibits cell growth, and this inhibition is mediated by caspase- and mitochondrial-dependent apoptotic pathways through cell cycle arrest at the G1/S phase in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, methanol extracts of Codium fragile can be provided as a novel chemotherapeutic drug due to its growth inhibition effects and induction of apoptosis in human oral cancer cells.

Improvement of Polishing Characteristics Using with and without Oxidant ($H_2O_2$) of Ti/FiN Layers (산화제($H_2O_2$)의 첨가 유무에 따른 Ti/TiN막의 CMP 연마 특성)

  • Lee, Kyoung-Jin;Seo, Yong-Jin;Park, Chang-Jun;Kim, Gi-Uk;Park, Sung-Woo;Kim, Sang-Yong;Lee, Woo-Sun
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2003.07a
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    • pp.88-91
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    • 2003
  • Tungsten is widely used as a plug for the multi-level interconnection structures. However, due to the poor adhesive properties of tungsten (W) on $SiO_2$ layer, the Ti/TiN barrier layer is usually deposited onto $SiO_2$ for increasing adhesion ability with W film. Generally, for the W-CMP (chemical mechanical polishing) process, the passivation layer on the tungsten surface during CMP plays an important role. In this paper, the effect of oxidants controlling the polishing selectivity of W/Ti/TiN layer were investigated. The alumina ($Al_2O_3$) abrasive containing slurry with $H_2O_2$ as the oxidizer, was studied. As our preliminary experimental results, very low removal rates were observed for the case of no-oxidant slurry. This low removal rate is only due to the mechanical abrasive force. However, for Ti and TiN with $H_2O_2$ oxidizer, different removal rate was observed. The removal mechanism of Ti during CMP is mainly due to mechanical abrasive, whereas for TiN, it is due to the formation of metastable soluble peroxide complex.

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Evaluation of Fatigue Behavior for Laser Welded High Strength Steel Sheets (SPFC590) (고장력 강판(SPFC590)의 레이저 용접부 피로거동 평가)

  • Heo, Cheol;Kwon, Jong-Wan;Cho, Hyun-Deog;Choi, Sung-Jong;Chung, Woo-Young
    • Transactions of the Korean Society of Automotive Engineers
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    • v.20 no.5
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    • pp.53-64
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    • 2012
  • Deep and narrow welds can be produced by laser welding at high welding speeds with a narrow heat-affected zone (HAZ) and little distortion of the workpiece. This study aims to evaluate the usefulness of laser welding at automobile component manufacture. Microstructure observation, hardness test, tensile test and fatigue life test are performed by using the fiber laser welded SPFC590 steel sheets which is used widely in the manufacture of automotive seat frame. Three kinds of specimens are only a SPFC590 steel plate, quasi-butt joint plate and lap joint plate by laser welding. The following results that will be helpful to understand the static strength, fatigue crack initiation and growth mechanism were obtained. (1) The tensile strength of quasi butt joint specimens nearly equal to base metal specimens, but lap joint specimens fractured in shear area of weld metal. (2) The fatigue strength of quasi-butt joint specimen was approximately 8 percent lower than that of the base metal specimens. Furthermore, the lap joint specimens were less than 86 percent of the base metal specimens. (3) The lap joint fatigue specimens fractured at shear area in high level stress amplitude, while fractured at normal area in low level stress amplitude. From these results, the applicability of the laser welding to the automobile component is discussed.

Anti-allergic Activity of Dojuk-San Ethanol Extract (도적산(導赤散) 에탄올 추출물의 알레르기 억제효과)

  • Sung, Sun-Heui;Lee, Su-Kyoung;Kang, Ok-Hwa;Kwon, Dong-Yeul;Chong, Myong-Soo;Lee, Ki-Nam
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.3
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    • pp.438-444
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    • 2011
  • Dojuk-San is known to be effective for treating a urinary diseases and stomatitis. However, its effects on the bone marrow-derived mast cell(BMMC) mediated allergy and inflammation mechanism remain unknown. In this study, the biological effects of Dojuk-San ethanol extract(DJS) were evaluated while focusing on its effects on the allergic mediator in phorbol 12-myristate 13-acetate(PMA) plus calcium ionophore A23187(A23187)-stimulated BMMCs. An investigation was also conducted to determine its effects on the production of several allergic mediators including interleukin-6(IL-6), prostaglandin D2($PGD_2$), leukotrieneC4(LTC4) and ${\beta}$-Hexosaminidase(${\beta}$-Hex). The results revealed that DJS inhibited the PMA plus A23187 induced production of IL-6, PGD2, LTC4 and ${\beta}$-Hex. Taken together, these findings indicate that DJS has the potential using in the treatment of allergy.