• Title/Summary/Keyword: Over-expression

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Measurement of Human Cytochrome P450 Enzyme Induction Based on Mesalazine and Mosapride Citrate Treatments Using a Luminescent Assay

  • Kim, Young-Hoon;Bae, Young-Ji;Kim, Hyung Soo;Cha, Hey-Jin;Yun, Jae-Suk;Shin, Ji-Soon;Seong, Won-Keun;Lee, Yong-Moon;Han, Kyoung-Moon
    • Biomolecules & Therapeutics
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    • v.23 no.5
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    • pp.486-492
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    • 2015
  • Drug metabolism mostly occurs in the liver. Cytochrome P450 (CYP) is a drug-metabolizing enzyme that is responsible for many important drug metabolism reactions. Recently, the US FDA and EU EMA have suggested that CYP enzyme induction can be measured by both enzymatic activity and mRNA expression. However, these experiments are time-consuming and their interassay variability can lead to misinterpretations of the results. To resolve these problems and establish a more powerful method to measure CYP induction, we determined CYP induction by using luminescent assay. Luminescent CYP assays link CYP enzyme activity to firefly luciferase luminescence technology. In this study, we measured the induction of CYP isozymes (1A2, 2B6, 2C9, and 3A4) in cryopreserved human hepatocytes (HMC424, 478, and 493) using a luminometer. We then examined the potential induction abilities (unknown so far) of mesalazine, a drug for colitis, and mosapride citrate, which is used as an antispasmodic drug. The results showed that mesalazine promotes CYP2B6 and 3A4 activities, while mosapride citrate promotes CYP1A2, 2B6, and 3A4 activities. Luminescent CYP assays offer rapid and safe advantages over LC-MS/MS and qRT-PCR methods. Furthermore, luminescent CYP assays decrease the interference between the optical properties of the test compound and the CYP substrates. Therefore, luminescent CYP assays are less labor intensive, rapid, and can be used as robust tools for high-throughput CYP screening during early drug discovery.

Flower bud development and the influence of sex expression of summer pruning in Schizandra chinensis Baillon (오미자의 화아(花芽) 발달과 성 발현에 미치는 하계전정의 영향)

  • Kim, Sun;Kim, Tai-Su;Park, Moon-Soo;Park, Ho-Ki
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.2
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    • pp.97-101
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    • 2003
  • The purpose of this experiment is to improve the ratio of female flower in Schizandra chinensis Baillon. The experiment was carried out in two purposes. One was to understand the development stages on bud formation and determining sex after bud formation by stage. The other was to investigate characteristics of flowers forming on spring when summer pruning was taken place in different cutting part and timing in S. chinensis Baillon. The outcome was that on 7th of June the flower bud was found next to leaf primordium in branch of this year and no changes were found in shape for two weeks by 26th July. On 26th of June, an ovule that forms organ was observed and then, an ovule was found after sex was determined on 6th of July. Second experiment was carried out to find out when the sex was determined by adapting different pruning methods; cutting the branch at 6th node on June 1, June 15, and July 1; cutting the branch at 10th node on June 15, and July 1. The result was that the re-growth ratio on the branch 6th node was cut on June 1 and June 15. July 1 was 34-56% and, however, the number of setting flowers and mail flowers were fewer than the normal branches. More over, even though the re-growth ratio of branches were 34-25% when the cutting was taken place at 10th node on June 15 and July 1, the number of setting flowers and mail flowers were the same as the non treated branch. As a result, it is assumed that cutting controlling in this plant to increase the number of female flowers has no effect.

Characterization of Transgenic Tall Fescue Plants Expressing Two Antioxidant Genes in Response to Environmental Stresses (두 가지 항산화유전자를 동시에 발현시킨 형질전환 톨 페스큐 식물체의 환경스트레스에 대한 내성 특성 해명)

  • Lee, Sang-Hoon;Lee, Ki-Won;Kim, Ki-Yong;Choi, Gi-Jun;Seo, Sung;Kwak, Sang-Soo;Kwon, Suk-Yoon;Yun, Dae-Jin;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.27 no.2
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    • pp.109-116
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    • 2007
  • Environmental stress is the major limiting factor in plant productivity. As an effort to solve the global food and environmental problems using the plant biotechnology, we have developed transgenic tall fescue (Festuca arundinacea Schreb.) plants via Agrobacterium-mediated gene transfer method. To develop transgenic tall fescue plants with enhanced tolerance to the environmental stresses, both CuZn superoxide dismutase (CuZnSOD) and ascorbate peroxidase (APX) genes were incorporated in a pIG121 binary vector and the both of the genes were controlled separately by an oxidative stress-inducible sweet potato peroxidase 2 (SWPA2) premoter expressed in chloroplasts. Leaf discs of transgenic plants showed 10-30% less damage compared to the wild-type when they exposed to a wide range of environmental stresses including methyl viologen (MV), $H_2O_2$ and heavy metals. In addition, when $200{\mu}M$ MV was sprayed onto the whole plants, transgenic plants showed a significant reduction of visible damage compared to wild-type plants that were almost damaged. These results suggest that over expression of CuZnSOD and APX genes in transgenic plants might be a useful strategy to protect the crops against a wide range of environmental stresses.

Anti-proliferation Effect of Coscinoderma sp. Extract on Human Colon Cancer Cells (Coscinoderma sp.의 대장암세포 증식 억제 효과)

  • Choi, Ki Heon;Jung, Joohee
    • Journal of Food Hygiene and Safety
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    • v.31 no.4
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    • pp.294-298
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    • 2016
  • Natural products are attractive as the source of new drug development. Especially, numerous unknown marine bioresources are an object of attention because the ocean occupies three fourth of the earth. Survival of marine bioresources in extreme environment may induce the production of biological active compounds. As previous study, we examined over 40 specimens of marine sponges collected from Micronesia and screened their anti-proliferative activities in various cancer cell lines. Among them, we investigated Coscinoderma sp.'s activity and mechanism in human colon carcinoma HCT116 and RKO cells. Furthermore, we also used the p53-knockout of HCT116 cells and the p53 loss of RKO cells for elucidating the role of p53. Coscinoderma sp. inhibited cellular viability independently of the p53 status. Therefore, we compared the expression level of cell death-related proteins by Coscinoderma sp. in HCT16 and in HCT116 p53KO cells. Coscinoderma sp. increased p53 level and NOXA levels and induced apoptosis under the condition of p53 existence. On the other hand, Coscinoderma sp. increased p21 and mTOR levels in HCT116 p53KO cells. These results suggest that Coscinoderma sp. induced anti-proliferation effect through different pathway depending on p53 status.

Study on Expression and Characterization of HRD3 Gene Related DNA Repair from Eukaryotic Cells (진핵세포에서 DNA 회복에 관련된 HRD3 유전자의 분리, 발현 및 특성 연구)

  • Shin, Su-Hwa;Park, In-Soon
    • Journal of Life Science
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    • v.14 no.2
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    • pp.325-330
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    • 2004
  • The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA and DNA-RNA helicase activities. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, the RAD3 homolog gene was isolated by screening of genomic DNA library. The isolated gene was designated as HRD3 (Homologue of RAD3 gene). The over-expressed HRD3 protein was estimated to be a 75 kDa in size which is in good agreement with the estimated by the nucleotide sequence of the cloned gene. Two-dimensional gel electrophoresis showed that a number of other protein spots dramatically disappeared when the HRD3 protein was overexpressed. The overexpressed RAD3 protein showed a toxicity in E. coli host, suggesting that this protein may be involved in the inhibition of protein synthesis and/or degradation of host protein. To determine which part of HRD3 gene contributes to the toxicity in E. coli, various fusion plasmids containing a partial sequence of HRD3 and lac'Z gene were constructed. These results suggest that the C-terminal domain of HRD3 protein may be important for both toxic effect in E. coli and for its role in DNA repair in S. pombe.

Studies of Inhibitory Mechanism on Melanogenesis by Partially Purified Asiasari radix in α-MSH Stimulated B16F10 Melanoma Cells (세신추출물이 α-MSH 자극에 의한 B16F10 세포의 멜라닌생성에 미치는 영향)

  • Jang, Ji-Yeon;Kim, Ha-Neui;Kim, Yu-Ri;Kim, Byung-Woo;Choi, Yung-Hyun;Choi, Byung-Tae
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1617-1624
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    • 2010
  • Recently, it has been found that Asiasari radix showed a hypopigmenting effect on melanogenesis through activation of mitogen-activated protein kinase (MEK)/extracellular signal-activated kinase (ERK) in B16F10 melanoma cells. However, the hypopigmenting effect of A. radix on the $\alpha$-melanocyte stimulating hormone ($\alpha$-MSH)-stimulated melanogenesis has remained unknown. The purpose of this study was to investigate the inhibitory mechanism of the partially purified A. radix (PPAR)-induced hypopigmentating effects on $\alpha$-MSH-stimulated melanogenesis in B16F10 mouse melanoma cells. PPAR strongly inhibited tyrosinase activity and leads to decreased melanin synthesis in $\alpha$-MSH-stimulated B16F10 melanoma cells. PPAR also decreased the $\alpha$-MSH-induced over-expression of the melanogenic enzymes, tyrosinase, tyrosinase-related protein (TRP)-1, dopachrome tautomerase (Dct) and microphthalmia-associated transcription factor (MITF). We further showed that PPAR inhibits $\alpha$-MSH-induced melanogenesis via phosphorylation of MEK/ERK and PI3K/Akt, and that their activation was blocked by MEK inhibitors, PD98059 and PI3K inhibitors, LY294002 in $\alpha$-MSH-stimulated B16F10 melanoma cells. These results suggest that PPAR inhibits $\alpha$-MSH-induced melanogenesis by activation of MEK/ERK and PI3K/Akt through MITF degradation, which may lead to down-regulation of tyrosinase.

Anti-diabetic mechanism of melania snail (Semisulcospira libertina) protamex hydrolysates (다슬기 protamex 가수분해물(MPH)의 항당뇨 기작 연구)

  • Pyo, Sang-Eun;Choi, Jae-Suk;Kim, Mi-Ryung
    • Food Science and Preservation
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    • v.24 no.7
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    • pp.1007-1016
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    • 2017
  • Melania snail (Semisulcospira libertina) was traditionally used as the healthy food in Korea. It was generally known to improve liver function and heal a diabetes. The aim of this study was to elucidate the anti-diabetic mechanism of melanian snail hydrolysates treated with protamex (MPH) by investigating the inhibitory action on protein tyrosine phosphatase 1B (PTP1B), the improving effect on the insulin resistance in C2C12 myoblast and the protective effect for pancreatic beta-cell (INS-1) under the glucose toxicity. The melania snail hydrolysates treated with protamex (MPH), which showed the highest degree of hydrolysis (43%), and inhibited effectively PTP1B activity ($IC_{50}=15.42{\pm}1.1{\mu}g/mL$), of which inhibitory effect was higher than usolic acid, positive control ($IC_{50}=16.65{\mu}g/mL$). MPH increased the glucose uptake in C2C12 myoblast treated with palmitic acid. In addition, MPH increased insulin mRNA expression level by over 160% with enhanced cell viability in INS-1 cell under the high glucose concentration (30 mM). These results suggest that MHP may improve the diabetic symptom by the inhibiting the PTP1B activity, increasing the glucose uptake in muscle cell and protecting the pancreatic beta-cell from glucose toxicity.

Role of Iridin Isolated from Iris koreana Nakai on Doxorubicin-induced Necrosis in HK-2 Cells, and Effect on Cancer Cells (노랑붓꽃에서 분리된 Iridin의 독소루비신 유도 HK-2 세포 괴사에 대한 역할 및 암세포에 대한 작용)

  • Nho, Jong Hyun;Lee, Ki Ho;Jung, Ho Kyung;Lee, Mu Jin;Jang, Ji Hun;Sim, Mi Ok;Jung, Ja Kyun;Jung, Da Eun;Cho, Hyun Woo
    • Korean Journal of Plant Resources
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    • v.31 no.2
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    • pp.95-101
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    • 2018
  • Doxorubicin is a anti-cancer drugs that interferes with the growth and spread of cancer cells in human body. Doxorubicin is used to treat different types of cancers that affect the ovary, thyoid and lungs, but induced side effect such as nephrotoxicity and cardiotoxicity. Thus, we investigated that the effect of iridin on doxorubicin-induced necrosis in HK-2 cells, a human proximal tubule cell. To confirm effect of iridin on doxorubicin-induced necrosis, HK-2 cells are treated with $10{\mu}M$ doxorubicin and $80{\mu}M$ iridin. $80{\mu}M$ iridin reduced $10{\mu}M$ doxorubicin-induced necrosis, the mitochondrial over activation and caspase-3 activation. However, iridin reduces anti-cancer effect of doxorubicin such as PARP1 and caspase-3 activation, checkpoint proteins (CDK4 and CDK6) in NCI-H1129 cells (Human non-small cell lung cancer cell). In HCT-116 cells (Human colorectan cancer cell), iridin do not increased protein expression of CDK4 and CDK6 decreased by doxorubicin. Results indicate that treatment of iridin was diminished doxorubicin-induced necrosis in HK-2 cells. However, iridin was decreased anti-cancer effect of doxorubicin on NCI-H1229, but not HCT-116. Thus, further experiment are required to iridin treatment on various cancer cells and animal models because effect of iridin different cell type.

The Cytotoxic Effect of Oral Wet Wipes on Gingival Cells (시판 중인 구강청결티슈의 세포 독성 관찰)

  • Jung, Im-hee;Park, Ji Hyeon;Lee, Min Kyeng;Hwang, Young Sun
    • Journal of dental hygiene science
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    • v.18 no.2
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    • pp.76-84
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    • 2018
  • Wet wipes are being increasingly used because of their convenience. Particularly, oral wet wipes are useful for regular cleaning of a baby's mouth after birth. Therefore, the consumption of oral wet wipes has increased over the past few years and a variety of products are commercially available. However, product information on safety is not sufficiently provided and still raises doubts regarding adverse effects. To confirm the safety of wet wipes as an oral hygiene item and provide information for their use, we investigated the cytotoxicity of oral wet wipes and verified the underlying mechanism. The anti-bacterial effect of oral wet wipes was analyzed using the disk diffusion method. The cytotoxic effects of oral wet wipes were observed based on morphological changes using microscopy and determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in gingival epithelial cells and gingival fibroblasts. Evaluation of apoptosis by oral wet wipes was explored using propidium iodide flow cytometric analysis and a terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL) assay. Apoptosis-related molecules were also analyzed using western blotting. Five types of oral wet wipes were tested, and two products from Fisher-Price and Dr. Kennedy revealed strong cytotoxic effects on gingiva epithelial cells and gingiva fibroblasts, although they also showed intense anti-bacterial effects on oral bacteria. Cell cycle arrest in the G2/M phase and apoptosis were observed based on treatment of extracts from Fisher-Price and Dr. KENNEDY. Relatively high TUNEL levels, reduction of proliferating cell nuclear antigen and cyclin-dependent kinase 4 expression, and fragmentation of poly (ADP-ribose) polymerase were also elucidated. These results suggest that commercial oral wet wipes could exert cytotoxic influences on oral tissue, although there are anti-bacterial effects, and careful attention is required, especially for infants and toddlers.

The Symbols of the Body Image Expressed in Modern Fashion Design (현대 패션디자인에 표현된 신체이미지의 상징성)

  • 권기영;조필교
    • The Research Journal of the Costume Culture
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    • v.8 no.5
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    • pp.681-706
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    • 2000
  • This study is intended to understand an implication system and significance in the current community which a body image symbolizes by correlating it with fashion that is on the rise as a principal issue in various fields. For this study, the symbolism of the body image was contemplated in terms of philosophy and art, and then on the basis of it, the symbolism of the body image shown in modern fashion design was analyzed through fashion works. The research results are as follows, 1. The manifestation of sex can be taken as the symbolism of the body image which is expressed in modern fashion design. Recently sexual chaos and vagueness such as homosexuality and bisexuality are expressed through a dress and its ornaments. Though displaying sexual characteristics of male and female as they are exposing a sign or a diagram, decorating a part of body or representing sex in garments, uncertain sex identity in modern society is manifested in dress and its ornaments. It is to deny absoluteness regarding sex and emphasize diversity indwelt in human beings, and after all it shows to pursue the human essence. 2. Another symbolism of the body image is body expression as the human race and an ethnic group. The discriminating situations and the restoration of their status appeared in modern fashion too. Moreover, their cultures and issues came to alter the aesthetic standard of body made from a view of the Western white supremacist. Hereupon, fashion trends like ethnic fashion, Orientalism and African look etc. appear according to this tendency, which represents race and national identity and in addition, which signifies to present transcendental human conception embracing alienated human conception. 3. The symbolism of the body image expressed in a body, and a dress and its ornaments as nature can be considered in terms of the concern on environmental contamination and the respect of echo system. Getting away from reigning over, developing and stamping down nature at their will, the human beings pursue unity with nature, which is described in fashion. They are stressing that natural materials and objects such as animal, plant and soil etc. should activily be introduced into fashion and humans are a communal fate group and should reframe their status in nature at last. 4. The body image shown in a body, a dress and its ornaments as technology is transformed and recreated by modern scientific techniques and medical science to show post human conception namely, forthcoming future human conception as a cyborg which loses individual identity. This presents a perfect future human conception with high level of preternatural power but after all, leaves us a task to seek the meaning of human existence in alienation caused by the loss of human identity and existence. In this manner, the moderns crave for perceiving the identity of a natural human being in the current thoughts tendency of the modern times such as postmodernism, post structuralism, deconstructionism, feminism and so on, which build discussions affecting the art and fashion worlds. The categories, like sexual characteristics indwelt in a human body, racial classifications, the natural environment surrounding human beings and development of science, bring out the importance of the internal and external meaning in today's fashion which a human body contains, and present sew human conception in the coming future society.

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