• Title/Summary/Keyword: F9 EC cells

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Histone Methylation Regulates Retinoic Acid-induced Hoxc Gene Expression in F9 EC Cells (F9 EC 세포에서 레티노산에 의해 유도되는 Hoxc 유전자의 발현에 히스톤 메틸화가 미치는 영향)

  • Min, Hyehyun;Kim, Myoung Hee
    • Journal of Life Science
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    • v.25 no.6
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    • pp.703-708
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    • 2015
  • Hox genes encode a highly conserved family of homeodomain-containing transcription factors controlling vertebrate pattern formation along the anteroposterior body axis during embryogenesis. Retinoic acid (RA) is a key morphogen in embryogenesis and a critical regulator of both adult and embryonic cellular activity. Specifically, RA regulates Hox gene expression in mouse- or human-derived embryonic carcinoma (EC) cells. Histone modification has been reported to play a pivotal role in the process of RA-induced gene expression and cell differentiation. As histone modification is thought to play an essential role in RA-induced Hox gene expression, we examined RA-induced initiation of collinear expression of Hox genes and the corresponding histone modifications in F9 murine embryonic teratocarcinoma (EC) cells. Hox expression patterns and histone modifications were analyzed by semiquantitative RT-PCR, RNA-sequencing, and chromatin immuno-precipitation (ChIP)-PCR analyses. The Hoxc4 gene (D0) was initiated earlier than the Hoxc5 to –c10 genes (D3) upon RA treatment (day 0 [D0], day 1 [D1], and day 3 [D3]). The Hox nonexpressing D0 sample had a strong repressive marker, H3K27me3, than the D1 and D3 samples. In the D1 and D3 samples, reduced enrichment of the H3K27me3 marker was observed in the whole cluster. The active H3K4me3 marker was closely associated with the collinear expression of Hoxc genes. Thus, the Hoxc4 gene (D1) and all Hoxc genes (D3) expressed H3K4me3 upon transcription activation. In conclusion, these data indicated that removing H3K27me3 and acquiring H3K4me3 regulated RA-induced Hoxc gene collinearity in F9 cells.

In Vitro Antioxidant and Antiproliferative Activities of Novel Orange Peel Extract and It's Fractions on Leukemia HL-60 Cells

  • Diab, Kawthar AE;Shafik, Reham Ezzat;Yasuda, Shin
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7053-7060
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    • 2015
  • In the present work, novel orange peel was extracted with 100%EtOH (ethanol) and fractionated into four fractions namely F1, F2, F3, F4 which were eluted from paper chromatographs using 100%EtOH, 80%EtOH, 50%EtOH and pure water respectively. The crude extract and its four fractions were evaluated for their total polyphenol content (TPC), total flavonoid content (TFC) and radical scavenging activity using DPPH (1,1-diphenyl-2-picrylhydrazyl) assay. Their cytotoxic activity using WST assay and DNA damage by agarose gel electrophoresis were also evaluated in a human leukemia HL-60 cell line. The findings revealed that F4 had the highest TPC followed by crude extract, F2, F3 and F1. However, the crude extract had the highest TFC followed by F4, F3, F2, and F1. Depending on the values of $EC_{50}$ and trolox equivalent antioxidant capacity, F4 possessed the strongest antioxidant activity while F1 and F2 displayed weak antioxidant activity. Further, incubation HL-60 cells with extract/fractions for 24h caused an inhibition of cell viability in a concentration-dependent manner. F3 and F4 exhibited a high antiproliferative activity with a narrow range of $IC_{50}$ values ($45.9-48.9{\mu}g/ml$). Crude extract exhibited the weakest antiproliferative activity with an $IC_{50}$ value of $314.89{\mu}g/ml$. Analysis of DNA fragmentation displayed DNA degradation in the form of a smear-type pattern upon agarose gel after incubation of HL-60 cells with F3 and F4 for 6 h. Overall, F3 and F4 appear to be good sources of phytochemicals with antioxidant and potential anticancer activities.

Effect of Substrate Size on Activities of Thiocarbamides with the Human Flavin-containing Monooxygenase 3 (사람 Flavin-containing Monooxygenase 3의 Thiocarbamide 화합물의 기질 크기에 따른 효소활성에 관한 연구)

  • 김영미
    • Environmental Analysis Health and Toxicology
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    • v.16 no.2
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    • pp.97-102
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    • 2001
  • The flavin-containing monooxygenases(FMOs) (EC1.14.13.8) are NADPH0dependent flavoenzymes that catalyze oxidation of soft nucleophilic heteroatom centers in a range of structurally diverse compounds, including foods, drugs, pesticides, and other xenobiotics. In humans, FMO3 is quantitatively a major human liver monooxygenase. In the present study, the baculovirus expression vector system was used to overexpress human FMO3 in sect cells for stalytic studies. Microsomes isolated from Spodoptera frugiperda(Sf)9 cells infected with human FMO3 recombinant baculovirus catalyzed the NADPH-and O$_2$-dependent oxidation of methimazole, thiourea, and phenylthiourea. However there was no detectable activity with 1, 3-diphenylthiourea or larger thiocarbamides. Microsomes from control Sf9 cells were devoid of methimazole or thiourea S-oxygenase activity. 1, 3-diphenylthiourea is apparently completely excluded from the catalytic site, these amines drugs are probably approaching the upper size limits of xenobiotics accepted by human FMO3. The substrate specificity of this iosform in humans appears considerably more restriceted than that of pig, guinea pig, rat or rabbit FMO3.

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Anti-inflammatory and PPAR Subtypes Transactivational Activities of Phenolics and Lignans from the Stem Bark of Kalopanax pictus

  • Quang, Tran Hong;Ngan, Nguyen Thi Thanh;Minh, Chau Van;Kiem, Phan Van;Nhiem, Nguyen Xuan;Tai, Bui Huu;Thao, Nguyen Phuong;Luyen, Bui Thi Thuy;Song, Seok-Bean;Kim, Young-Ho
    • Bulletin of the Korean Chemical Society
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    • v.32 no.11
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    • pp.4049-4054
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    • 2011
  • A new compound, kalopanaxin F (3), and 11 known compounds (1, 2, 4-12), were isolated from the stem bark of Kalopanax pictus. Their structures were elucidated on the basis of chemical and spectroscopic methods. Five of the compounds (2, 3, 5, 6, and 12) significantly inhibited $TNF{\alpha}$-induced NF-${\kappa}B$ transcriptional activity in HepG2 cells in a dose-dependent manner, with $IC_{50}$ values ranging from 6.2 to 9.1 ${\mu}M$. Furthermore, the transcriptional inhibitory function of these compounds was confirmed based on decreases in COX-2 and iNOS gene expression in HepG2 cells. Compounds 3-7, 9, and 12 significantly activated the transcriptional activity of PPARs dose-dependently, with $EC_{50}$ values ranging from 4.1-$12.7{\mu}M$. Compounds 4 and 5 exhibited $PPAR{\alpha}$, $PPAR{\gamma}$, and $PPAR{\beta}({\delta})$ transactivational activities in a dose-dependent manner, with $EC_{50}$ values of 16.0 and 17.0, 8.7 and 16.5, 26.2 and 26.3 ${\mu}M$, respectively.

Effect of Cigarette Filter on Cytotoxicity Potential of Mainstream Smoke (담배주류연의 세포독성에 대한 담배필터의 영향)

  • Shin Han-Jae;Sohn Hyung-Ok;Han Jung-Ho;Park Chul-Hoon;Hur Jae-Yeon;Lee Dong-Wook;Hwang Keon-Joong;Hyun Hak-Chul
    • Journal of the Korean Society of Tobacco Science
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    • v.27 no.1 s.53
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    • pp.51-58
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    • 2005
  • The objective of this study was to evaluate the effect of cigarette filter on in vitro cytotoxicity of cigarette mainstream smoke from the cigarette. In this work, we used 3 types of cigarettes included non-filtered 2R4F cigarette, cellulose acetate-filtered 2R4F cigarette, and carbon dual-filtered 2R4F cigarette which was made from original 2R4F by replacing with an acetate filter containing carbon. The cytotoxicity of both the cigarette smoke condensate (CSC), which was collected in Cambridge filter pad, and the gas/vapor phase (GVP), which was bubbled through in phosphate-buffered saline in a gas-washing bottle, was determined using a neutral red uptake assay with CHO-K1 cells. With regard to cytotoxicity when calculated on an equal puff basis, the cytotoxicity of CSC from the filtered cigarettes was lower than that of the non filtered cigarette. Also, $EC_{50}$ vlaue of GVP from carbon filter cigarette was 40.9 puff/L, indicating the cytotoxicity to be $20\%$ lower than that of the CA filter cigarette. The cytotoxicity of the GVP was correlated to the several vapor phase components (formaldehyde, acetaldehyde, acetone, acrolein, crotonaldehyde and MEK). In conclusion, carbon filter, which significantly reduced the amount of carbonyl compounds in mainstream cigarette smoke, results in significant reductions in the cytotoxicity potential of the smoke.

Synthesis of Polyurethane Foam/Organonanoclay/Phosphates Composites and its Characterization (폴리우레탄폼/유기나노점토/포스페이트 복합체의 합성과 그 특성)

  • Park, Kyeong-Kyu;Lee, Sang-Ho
    • Elastomers and Composites
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    • v.46 no.4
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    • pp.343-351
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    • 2011
  • We prepared polyurethane foam/cloisite30B/phosphates composites and characterized their rise time, density, cell morphology, and thermal properties. The composites were synthesized with polyadipatediol-cloisite30B composite (f=2.0), polyether-polyol (f=4.6), polymeric 4,4-diphenyl methane diisocyanate (f=2.5), and D-580 (phenyl polyoxyalkenyl phosphate). As a blowing agent, cyclopentane and distilled water were used at various concentrations of D-580 from 0 to 2.81 wt%. The rise times of PUF/Closite30B/Phosphate composites blown with distilled water were faster than those blown with cyclopentane by 30%. The composites blown with cyclopentane had spherical-shape cells and the cell diameter was decreased with increasing D-580 wt%. While $T_g$ of the composites blown with cyclopentane linearly decreased with increasing the D-580 content, the $T_g$ of the composites blown with distilled water increased with the D-580 content. All PUF/Closite30B/Phosphate composites began to decompose from $250^{\circ}C$. The composites blown with cyclopentane showed the second thermal decomposition at temperatures higher than $500^{\circ}C$. The thermal stability of all composites increased with the D-580 content. The effect of D-580 on the thermal stability of the composites was measured higher at the composites blown with distilled water.

Effects of Environmental Factors on the Bacterial Community in Eutrophic Masan Reservoir (이화학적 수질인자가 부영양화된 마산저수지의 세균분포에 미치는 영향)

  • 남귀숙;손형식;차미선;조순자;이광식;이상준
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.95-101
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    • 2003
  • The total bacterial numbers, Eubacterial community structures and environmental factors which affect bacterial community were estimated monthly using DAPI and fluorescent in situ hybridization monthly, from June to November 2000 to evaluate the correlation between the bacterial community and environmental factors in eutrophic agricultural Masan reservoir in Asan. Average water temperatures varied from 12.3 to $27.5^{\circ}C$, pH 7.5 to 9.0, DO 7. I~12.8 mg/L, COD 6.4~13.0 mg/L, chlorophyll a 30.5~99.0 mg/㎥, SS 7.S~25.7 mg/L, TN 1.748~3.543 mg/L., and TP 0.104~0.581 mg/L, respectively. Total bacterial numbers showed high ranges from 0.4 to 9.6$\times$ $10^{6}$ cells/ml, and these indicated the mesotrophic or eutrophic state. The ratio of Eubacteria to total bacteria was 67.6-88.0%, which was higher than that in other reservoir. The relationships of total bacteria and Eubacteria community were more significant with organic nitrogen (Org-N), and organic phosphorus (Org-P) than with water temperature. Proteobacteria groups showed strongly significant relationships with Org-P and Org-N and significant relationships with water temperature, conductivity, COD, and inorganic nitrogen. C-F group was the most significant with Org-N, and HGC group with water temperature. However, relationships of Chl-a, pH, DO and SS showed no significance with any bacterial community. These results were different from other studies, because of the specific characteristics of Masan reservoir such as old, shallow and eutrophic states. The seasonal variation of bacterial community in Masan reservoir does not seem to depend on phytoplankton dynamics but on storm event and organic materials from watershed and the sediment of reservoir.

[ $SiO_2$ ] Effect on the Electrochemical Properties of Polymeric Gel Electrolytes Reinforced with Glass Fiber Cloth ($SiO_2$가 유리섬유로 보강된 고분자 겔 전해질의 전기 화학적 특성에 미치는 영향)

  • Park Ho Cheol;Kim Sang Heon;Chun Jong Han;Kim Dong Won;Ko Jang Myoun
    • Journal of the Korean Electrochemical Society
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    • v.4 no.1
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    • pp.6-9
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    • 2001
  • [ $SiO_2$ ] effect on the electrochemical properties of polymeric gel electrolytes(PGEs) reinforced with glass fiber cloth(GFC) was investigated . PGEs were composed of polyacrylronitrile(PAN), poly(vinylidenefluoride-co-hexafluoropropylene) (P(VdF-co-HFP)), $LiClO_4$ and three kind of plasticizer(ethylene carbonate, dietyl carbonate, propylene carbonate). $SiO_2$ was added to PGEs in the weight fraction of 10, 20, $30\%$ respectively. PGEs containing $SiO_2$ showed conductivity of over $10^{-3}S/cm\;at\;23^{\circ}C$ and electrochemical stability window to 4.8V. In the impedance spectra of the cells, which were constructed by lithium metals as electrodes, interfacial resistance increased due to growth of passivation layer during storage time and remarkable difference was not observed with content of $SiO_2$. In the impedance spectra of the lithium ion polymer batteries consisted of $LiClO_2$ and mesophase pitch-based carbon fiber(MCF), ohmic cell resistance of $SiO_2-free$ PGE was changed continuously with number of cycle, but those of $SiO_2-dispersed$ PGEs were not. Discharge capacity of the PGE containing $20wt\%\;SiO_2$ showed 132 mAh/g at 0.2C rate and $85\%$ of discharge capacity was retained at 2C rate.