• Title/Summary/Keyword: inducers

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Histone Deacetylase Inhibitor Stimulate CYP3A4 Proximal Promoter Activity in HepG2 Cells

  • Kim Ja Young;Ahn Mee Ryung;Kim Dae-Kee;Sheen Yhun Yhong
    • Archives of Pharmacal Research
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    • v.27 no.4
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    • pp.407-414
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    • 2004
  • The expression of CYP3A4 gene is induced by a variety of structurally unrelated xenobiotics including the antibiotic rifampicin, pregnenolone 16-carbonitrile (PCN), and endogenous hormones, that might mediate through steroid and xenobiotic receptor (SXR) system. The molecular mechanisms underlying regulation of CYP3A4 gene expression have not been understood. In order to gain the insight of the molecular mechanism of CYP3A4 gene expression, study has been undertaken to investigate if the histone deacetylation is involved in the regulation of CYP3A4 gene expression by proximal promoter in human hepatoma HepG2 cells. Also we have investigated to see if SXR is involved in the regulation of CYP3A4 proximal promoter activity in human hepatoma HepG2 cells. HepG2 cells were transfected with a plasmid PCYP3A4-Luc containing ${\~}1kb$ of the CYP3A4 proximal promoter region (-863 to +64 bp) in front of a reporter gene, luciferase, in the presence or absence of pSAP-SXR. In HepG2 cells, CYP3A4 inducers, such as rifampicin, PCN and RU486 showed minimal stimulation of CYP3A4 proximal promoter activity in the absence of SXR and histone deacetylase (HDAC) inhibitors. 4-Dimethylamino-H-[4-(2-hydroxycarbamoylvinyl)benzyl]benzamide (IN2001), a new class HDAC inhibitor significantly increased CYP3A4 proximal promoter activity over untreated control cells and rifampicin concomitant treatment with IN2001 increased further CYP3A4 proximal promoter activity that was stimulated by IN2001 The results of this study demon-strated that both HDAC inhibitors and SXR are essential to increase of CYP3A4 proximal promoter activity by CYP3A4 inducers such as PCN, rifampicin, and RU486. Especially SXR seems to be important for the dose dependent response of CYP3A4 inducing chemicals to stimulate CYP3A4 proximal promoter activity. Also this data suggested that HDAC inhibitors seemed to facilitate the CYP3A4 proximal promoter to be activated by chemicals.

The Effect of Neuroactive Compounds on Settlement of Pacific Oyster, Crassostrea gigas Pediveliger Larvae (굴, Crassostrea gigas 부착기 유생의 부착에 미치는 신경전달물질종의 영향)

  • Hur, Youngbaek;Jo, Qtae;Byun, Soongyu;Mun, Tesek
    • The Korean Journal of Malacology
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    • v.30 no.4
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    • pp.343-351
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    • 2014
  • We determined the effects of neuroactive compounds known as synthetic larval settlement inducers on the settlement of the Pacific oyster C. gigas pediveliger on the larval collector. Six types of the inducers, serotonin (5-HT), ${\gamma}$-amino butyric acid (GABA), L-3,4-dihydroxyphenylalanine (L-DOPA), norepinephrine, epinephrine and methyl bromide (MB) were tested. All the chemicals induced larval settlement, MB being the most effective with settlement rate of $42.7{\pm}2.7%$, followed by GABA ($35.4{\pm}2.0%$), 5-HT ($29.1{\pm}2.2%$), L-DOPA ($19.2{\pm}2.1%$), epinephrine ($15.2{\pm}0.9%$), and norepinephrine ($11.0{\pm}1.2%$). The chemicals ${\gamma}$-amino butyric acid and methyl bromide were also better in terms of settled density on the collector with their respective density of $1.97{\pm}1.42$ and $2.37{\pm}1.86ind/cm^2$, reminiscent of being most effective candidates for a larval settlement inducer in the oyster hatchery.

The Effect of Neuroactive Compounds on Settlement of Pacific Oyster, Crassostrea gigas Pediveliger Larvae (굴 Crassostrea gigas 부착기 유생의 부착에 미치는 신경전달물질종의 영향)

  • Hur, Young Baek;Cho, Kyu Tae;Byun, Soon Gyu;Jeon, Chang Young;Cho, Kee Chae
    • The Korean Journal of Malacology
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    • v.29 no.2
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    • pp.139-146
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    • 2013
  • We determined the effects of neuroactive compounds known as synthetic larval settlement inducers on the settlement of the Pacific oyster C. gigas pediveliger on the larval collector. Six types of the inducers, serotonin (5-HT), ${\gamma}$-amino butyric acid (GABA), L-3,4-dihydroxyphenylalanine (L-DOPA), norepinephrine, epinephrine and methyl bromide (MB) were tested. All the chemicals induced larval settlement, MB being the most effective with settlement rate of $42.7{\pm}2.7%$, followed by GABA ($35.4{\pm}2.0%$), 5-HT ($29.1{\pm}2.2%$), L-DOPA ($19.2{\pm}2.1%$), epinephrine ($15.2{\pm}0.9%$), and norepinephrine ($11.0{\pm}1.2%$). The chemicals ${\gamma}$-amino butyric acid and methyl bromide were also better in terms of settled density on the collector with their respective density of $1.97{\pm}1.42$ and $2.37{\pm}1.86\;ind/cm^2$, reminiscent of being most effective candidates for a larval settlement inducer in the oyster hatchery.

PROBLEMS IN OSTEOGENIC DIFFERENTIATION OF RAT BONE MARROW STROMAL CELLS (쥐의 골수로부터 추출한 줄기세포를 이용한 조골세포로의 분화 유도과정에서 나타난 문제점에 관한 분석 연구)

  • Kim, In-Sook;Cho, Tae-Hyung;Zhang, Yu-Lian;Lee, Kyu-Back;Park, Yong-Doo;Rho, In-Sub;Weber, F.;Lee, Jong-Ho;Kim, Myung-Jin;Hwang, Soon-Jung
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.1
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    • pp.1-8
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    • 2005
  • This study was aimed to characterize osteogenic potential of rat bone marrow stromal cells (BMSC) isolated with standard flushing method and investigate the plasticity of transdifferentiation between osteoblastic and adipocytic lineage of cultured BMSC. Unlike aspiration method in human, rat bone marrow was extracted by means of irrigation with culture media that elevates the possibility of co-extraction of committed osteoprogenitor, or preosteoblast or other progenitor cells of several types present inside bone marrow. The cultured stromal cells showed high ALP activity which is representative marker of osteoblast without any treatment. Osteogenic inducers such as Dex and BMP-2 were examined for the evaluation of their effect on osteogenic and adipocytic differentiation of stromal cells, because they function as osteoinductive agent in stromal cells, but simultaneously induce adipogenic differentiation. Osteogenic differentiation was evaluated by measuring alkaline phosphatase activity or mRNA expression of osteoblast markers such as osteopontin, bone sialoprotein, collagen type I and CbfaI, and in vitro matrix mineralization by von Kossa staining. Oil red staining method was used to detect adipocyte and adipocytic marker, aP2 and $PPAR{\gamma}2$ expression was examined using RT-PCR. It can be supposed that irrigation procedure resulted in high portion of already differentiation-committed osteoprogenitor cell showing elevated ALP activity and strong mineralization only under the supplement of $100{\mu}M$ ascorbic 2-phosphate and 10mM ${\beta}$-glycerophosphate without any treatment of osteogenic inducers such as Dex and BMP-2. Dex and BMP-2 seemed to transdifferentiate osteoprogenitor cells having high ALP activity into adipocytes temporarily, but continuous treatment redifferentiated into osteoblast and developed in vitro matrix mineralization. This property must be considered either in tissue engineering for bone regeneration, or in research of characterization of osteogenic differentiation, with rat BMSC isolated by the standard irrigation method.

Detection of Chlorotoluene and Nitrotoluene Compounds by Recombinant Microbial Biosensors (재조합 미생물 바이오센서를 이용한 chlorotoluene과 nitrotoluene 화합물의 검출)

  • Lee, Da Young;Cho, Jae Ho;Lim, Woon Ki;Shin, Hae Ja
    • Journal of Life Science
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    • v.24 no.1
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    • pp.54-60
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    • 2014
  • Aromatic hydrocarbons are toxic environmental pollutants that are detrimental to the ecosystem and human health. Among them, chlorotoluene and nitrotoluene are toxic to hydrobios and irritate the skin, eyes, and respiratory organs of humans. We herein report the development of recombinant microbial biosensors for cheap and rapid monitoring of chlorotoluene and nitrotoluene compounds. Plasmids were constructed by inserting the xylR regulatory gene for BTEX (benzene, toluene, ethylbenzene, and xylene) degradation into upstream of Po' (the DmpR activator promoter Po with the deletion of its own upstream activating sequences) or Pu (the cognate promoter of XylR)::lacZ (the ${\beta}$-galactosidase gene) and transformed into Escherichia coli $DH5{\alpha}$. In the presence of inducers, the biosensor cells immobilized in agarose developed a red color in 1-2 h due to the hydrolysis of chlorophenol red ${\beta}$-D-galactopyranoside (CPRG), a substrate of ${\beta}$-galactosidase that was expressed by the inducers. Among BTEX, high responses were specifically observed with o-, m-, p-chlorotoluene ($0.1{\mu}M-100 mM$) and o-, m-, p-nitrotoluene (0.1 mM-100 mM). Po' demonstrated higher responses than those with Pu. The biosensors immobilized in agarose showed good stability after 21 days' storage at $4^{\circ}C$, and responses in untreated wastewater spiked with chlorotoluene and nitrotoluene, suggesting they can be used to detect compounds in wastewater.

Preliminary Studies on Production of Porcine Leucocyte Interferon (돼지 Leucocyte Interferon 생산(生産)에 대한 기초연구(基礎硏究))

  • Jun, Moo Hyung;Chung, Un Ik;Park, Bong Kyun;Kim, Kyo Joon;Lee, Hun Jun
    • Korean Journal of Agricultural Science
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    • v.12 no.1
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    • pp.153-161
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    • 1985
  • As a preliminary step for production of large quantities of porcine leucocyte interferon(Por IFN-${\alha}$), various factors such as the sources of leucocytes, inducers and culture conditions were investigated. In addition, an assay system for the potency of porcine interferon was developed in a micro-system by using porcine kidney cell line, PK-15, and vesicular stomatitis virus. By the experiments, it was found that the porcine leucocyte interferon was synthesized more effectively in the leucocyte suspension with the higher viability and the lesser erythrocyte contamination. Regarding cell sources, the peripheral leucocytes produced a consistently higher unit of interferon than the spleen cells. When the efficiency of inducers was compared, the Sendai virus at the concentration of 125 HA unit/ml was found to be more effective for porcine leucocyte interferon production than the Newcastle disease virus. Average potency of 9 batches of the crude interferon measured by the established assay system was 1,200 unit/ml.

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Detection of m-toluate in Soils using Bioluminescence Producing Recombinant Bacteria (유전자 재조합 발광균주를 이용한 토양 오염원 m-toluate 탐지)

  • Kong, In-Chul;Kim, Myung-Hee;Jung, Yun-Ho;Ko, Kyung-Seok;Kim, Jae-Gon;Shin, Sung-Chun
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.5
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    • pp.507-512
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    • 2005
  • This research focuses on the development and application of a method for the detection of m-toluate in soils using a genetically engineered bioluminescent bacteria, Pseudomonas putida mt-2 KG1206. KG1206 produces light by direct (m-toluate and benzoate) and indirect (toluene analogs) inducers. For detection of m-toluate in soil system, 9.9 mL strain was amended with 0.1 mL soil ethanol extractant. A high correlation ($r^2>0.97$) was observed between bioluminescence and m-toluate concentration. The unknown concentrations of m-toluate in soil samples were pre-determined using a method developed based on bioluminescence activity of strain with extracted inducers. Values between by LC analysis and bioluminescence activity show moderate statistical results. These results demonstrate the feasibility of recombinant bioluminescent microorganism, engineered to generate a quantifiable bioluminescence signal in response to specific pollutants, may serve as combined sensing and reporting tools in environmental monitoring.

Effects of axial distance between inducer and impeller on the performance of the turbopump (인듀서와 임펠러 축방향 간극이 터보펌프 성능에 미치는 영향)

  • Choi, Chang-Ho;Kim, Dae-Jin;Hong, Soon-Sam;Kim, Jin-Han
    • The KSFM Journal of Fluid Machinery
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    • v.11 no.5
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    • pp.37-43
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    • 2008
  • An inducer is employed in a modern rocket feed system because it allows a turbopump system to operate at a high speed with low inlet pressures so as to minimize the weight and the size of the system. Cavitation performance can be improved by installing an inducer to the pump, enabling to increase the operational speed of the pump. The main purpose of an inducer is to increase the static pressure prior to an impeller to enable the impeller to operate satisfactorily under cavitation environments. In the present study the effects of axial distance between the inducer and the impeller on the performance of the pump were studied using both experimental and computational methods. Two inducers with different axial length were used for the experiments and the pump performances were measured. The experimental results show that the suction performance decreases as the axial gap between the inducer and impeller is increased.

Numerical Simulation of Unsteady Cavitating Flow Around 2D Hydrofoil (수중익 주위의 2차원 비정상 공동 현상 해석)

  • Lee, Se-Young;Park, Soo-Hyung;Lee, Chang-Jin
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.31 no.8
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    • pp.653-662
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    • 2007
  • Due to the difficulty raised from the coupling of cavitation modeling with turbulent flow, numerical simulation for two phase flow remains one of the challenging issues in the society. This research focuses on the development of numerical code to deal with incompressible two phase flow around 2D hydrofoil by combing the cavitation model suggested by Kunz et al. with $k-{\varepsilon}$ turbulent model. The simulation results are compared to experimental data to verify the validity of the developed code. Also, the comparison of the calculation results is made with LES results to evaluate the capability of $k-{\varepsilon}$ turbulence model. The calculation results show very good agreement with experimental observations even though this code can not grasp the small scaled bubbles in the calculation wheres LES can hold the real physics. This code will be extended to 3D compressible two phase flow for the study on the fluid dynamics in the inducers and impellers.

Effect of Platycodin D on Airway MUC5AC Mucin Production and Gene Expression Induced by Growth Factor and Proinflammatory Factor

  • Lee, Hyun-Jae;Lee, Su-Yel;Jeon, Byeong-Kyou;Lee, Jae-Woo;Kim, Young-Sik;Lee, Mi-Nam;Lee, Choong-Jae
    • Biomolecules & Therapeutics
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    • v.18 no.3
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    • pp.294-299
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    • 2010
  • In this study, we tried to investigate whether platycodin D significantly affects MUC5AC mucin production and gene expression induced by epidermal growth factor (EGF), phorbol ester (PMA) and tumor necrosis factor-$\alpha$ (TNF-$\alpha$) from human airway epithelial cells. Confluent NCI-H292 cells were pretreated with varying concentrations of platycodin D for 30 min and then stimulated with EGF, PMA and TNF-$\alpha$ for 24h, respectively. MUC5AC mucin gene expression and mucin protein production were measured by RT-PCR and ELISA. The results were as follows: (1) Platycodin D was found to inhibit the production of MUC5AC mucin protein induced by EGF, PMA, and TNF-$\alpha$, respectively. (2) It also inhibited the expression of MUC5AC mucin gene induced by the same inducers. These results suggest that platycodin D can regulate mucin gene expression and production of mucin protein, by directly acting on human airway epithelial cells.