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Manish Kumar Aneja,Johannes Geiger,Rabea Imker,Senta Üzgün,Michael Kormann,Guenther Hasenpusch,Christof Maucksch,Carsten Rudolph 생화학분자생물학회 2009 Experimental and molecular medicine Vol.41 No.12
φC31 integrase has emerged as a potent tool for achieving long-term gene expression in different tissues. The present study aimed at optimizing elements of φC31 integrase system for alveolar type II cells. Luciferase and β-galactosidase activities were measured at different time points post transfection. 5-Aza-2’deoxycytidine (AZA) and trichostatin A (TSA) were used to inhibit DNA methyltransferase and histone deacetylase complex (HDAC) respectively. In A549 cells, expression of the integrase using a CMV promoter resulted in highest integrase activity, whereas in MLE12 cells, both CAG and CMV promoter were equally effective. Effect of polyA site was observed only in A549 cells, where replacement of SV40 polyA by bovine growth hormone (BGH) polyA site resulted in an enhancement of integrase activity. Addition of a C-terminal SV40 nuclear localization signal (NLS) did not result in any significant increase in integrase activity. Long-term expression studies with AZA and TSA, provided evidence for post-integrative gene silencing. In MLE12 cells, both DNA methylases and HDACs played a significant role in silencing, whereas in A549 cells, it could be attributed majorly to HDAC activity. Donor plasmids comprising cellular promoters ubiquitin B (UBB), ubiquitin C (UCC) and elongation factor 1α (EF1α) in an improved backbone prevented post-integrative gene silencing. In contrast to A549 and MLE12 cells, no silencing could be observed in human bronchial epithelial cells, BEAS-2B. Donor plasmid coding for murine erythropoietin under the EF1α promoter when combined with φC31 integrase resulted in higher long-term erythropoietin expression and subsequently higher hematocrit levels in mice after intravenous delivery to the lungs. These results provide evidence for cell specific post integrative gene silencing with φC31 integrase and demonstrate the pivotal role of donor plasmid in long-term expression attained with this system. φC31 integrase has emerged as a potent tool for achieving long-term gene expression in different tissues. The present study aimed at optimizing elements of φC31 integrase system for alveolar type II cells. Luciferase and β-galactosidase activities were measured at different time points post transfection. 5-Aza-2’deoxycytidine (AZA) and trichostatin A (TSA) were used to inhibit DNA methyltransferase and histone deacetylase complex (HDAC) respectively. In A549 cells, expression of the integrase using a CMV promoter resulted in highest integrase activity, whereas in MLE12 cells, both CAG and CMV promoter were equally effective. Effect of polyA site was observed only in A549 cells, where replacement of SV40 polyA by bovine growth hormone (BGH) polyA site resulted in an enhancement of integrase activity. Addition of a C-terminal SV40 nuclear localization signal (NLS) did not result in any significant increase in integrase activity. Long-term expression studies with AZA and TSA, provided evidence for post-integrative gene silencing. In MLE12 cells, both DNA methylases and HDACs played a significant role in silencing, whereas in A549 cells, it could be attributed majorly to HDAC activity. Donor plasmids comprising cellular promoters ubiquitin B (UBB), ubiquitin C (UCC) and elongation factor 1α (EF1α) in an improved backbone prevented post-integrative gene silencing. In contrast to A549 and MLE12 cells, no silencing could be observed in human bronchial epithelial cells, BEAS-2B. Donor plasmid coding for murine erythropoietin under the EF1α promoter when combined with φC31 integrase resulted in higher long-term erythropoietin expression and subsequently higher hematocrit levels in mice after intravenous delivery to the lungs. These results provide evidence for cell specific post integrative gene silencing with φC31 integrase and demonstrate the pivotal role of donor plasmid in long-term expression attained with this system.
Elizabeth Minton,Stephanie Geiger-Oneto 글로벌지식마케팅경영학회 2018 Global Marketing Conference Vol.2018 No.07
While extant research examines the consumption of luxury products, the disposal behaviors of such products and business’ means for promoting this behavior through social media has yet to be examined. This research builds on belief congruence theory and the anticonsumption literature to understand how religiosity (with prescriptions against material possessions and performing actions just for show) influences disposal method of luxury goods and disposal behavior on social media. Specifically, findings show that extrinsically (intrinsically) religious consumers are more likely to throw away (donate) luxury products after use. The moderating influence of emotions is also explored to show that intrinsically (extrinsically) religious consumers are more (less) likely to use sustainable methods of product disposal for luxury and non-luxury products alike after being primed to feel shame/guilt in comparison to a control condition. A separate study manipulates product type (luxury vs. non-luxury) and product state (used vs. new), showing that extrinsically religious consumers are most (least) likely to use sustainable disposal methods when a product is used (new) and non-luxury (luxury). Additionally, findings show that identity mediates this relationship and has clear outcomes on social media behavior regarding product disposal and end-consumption behavior with luxury products. Implications for belief congruence theory and advertising practitioners are provided (with a specific emphasis on advertisers of luxury products using social media).
Deng, Pan-Yue,Xiao, Zhaoyang,Jha, Archana,Ramonet, David,Matsui, Toshimitsu,Leitges, Michael,Shin, Hee-Sup,Porter, James E,Geiger, Jonathan D,Lei, Saobo The Society 2010 The Journal of neuroscience Vol.30 No.15
<P>Cholecystokinin (CCK), a neuropeptide originally discovered in the gastrointestinal tract, is abundantly distributed in the mammalian brains including the hippocampus. Whereas CCK has been shown to increase glutamate concentration in the perfusate of hippocampal slices and in purified rat hippocampal synaptosomes, the cellular and molecular mechanisms whereby CCK modulates glutamatergic function remain unexplored. Here, we examined the effects of CCK on glutamatergic transmission in the hippocampus using whole-cell recordings from hippocampal slices. Application of CCK increased AMPA receptor-mediated EPSCs at perforant path-dentate gyrus granule cell, CA3-CA3 and Schaffer collateral-CA1 synapses without effects at mossy fiber-CA3 synapses. CCK-induced increases in AMPA EPSCs were mediated by CCK-2 receptors and were not modulated developmentally and transcriptionally. CCK reduced the coefficient of variation and paired-pulse ratio of AMPA EPSCs suggesting that CCK facilitates presynaptic glutamate release. CCK increased the release probability and the number of readily releasable vesicles with no effects on the rate of recovery from vesicle depletion. CCK-mediated increases in glutamate release required the functions of phospholipase C, intracellular Ca(2+) release and protein kinase Cgamma. CCK released endogenously from hippocampal interneurons facilitated glutamatergic transmission. Our results provide a cellular and molecular mechanism to explain the roles of CCK in the brain.</P>
Wu, Kan,Park, Ji Young,Al-Saadon, Rachael,Nam, Hyerim,Lee, Yujin,Top, Siden,Jaouen, Gé,rard,Baik, Mu-Hyun,Geiger, William E. American Chemical Society 2018 Organometallics Vol.37 No.12
<P>The oxidative electrochemical behavior of 1,1′-diphenyl-2-cymantrenylbutene (<B>1</B>), a cymantrene analogue of the breast cancer drug ferrocifen, was shown to involve the sequential electron-transfer series <B>1</B>/<B>1</B><SUP>+</SUP>/<B>1</B><SUP>2+</SUP> in dichloromethane/0.05 M [NBu<SUB>4</SUB>][B(C<SUB>6</SUB>F<SUB>5</SUB>)<SUB>4</SUB>] (<I>E</I><SUB>1/2</SUB> values 0.78 and 1.18 V vs ferrocene). By a combination of spectroscopic and computational techniques, it was shown that the cymantrene functionality plays an important role in dissipating the positive charges in the oxidized compounds and is therefore an active participant in the redox events. The redox-active orbital goes from roughly equal degrees of organometallic and π-organic (diphenylolefin) makeup in <B>1</B> to increasingly organic based fractions in <B>1</B><SUP>+</SUP> and <B>1</B><SUP>2+</SUP>. Structural changes mimicking those of oxidized tetrakis(aryl)ethylenes accompany the one-electron oxidations. There is sufficient unpaired electron density on the manganese center in <B>1</B><SUP>+</SUP> to allow for oxidatively induced ligand exchange of one or more of the carbonyl ligands with donor ligands, including phosphites and pyridine. The complex Mn(CO)<SUB>2</SUB>P(OPh)<SUB>3</SUB>(η<SUP>5</SUP>-C<SUB>5</SUB>H<SUB>4</SUB>(Et)C═C(C<SUB>6</SUB>H<SUB>5</SUB>)<SUB>2</SUB>) was prepared by the “electrochemical switch” method, wherein [Mn(CO)<SUB>2</SUB>P(OPh)<SUB>3</SUB>(η<SUP>5</SUP>-C<SUB>5</SUB>H<SUB>4</SUB>(Et)C═C(C<SUB>6</SUB>H<SUB>5</SUB>)<SUB>2</SUB>)]<SUP>+</SUP>, produced by the oxidation of <B>1</B> in the presence of P(OPh)<SUB>3</SUB>, was reduced back to the neutral CO-substituted complex.</P> [FIG OMISSION]</BR>
Classen, I G J,Lauber, Ph,Curran, D,Boom, J E,Tobias, B J,Domier, C W,Luhmann Jr, N C,Park, H K,Garcia Munoz, M,Geiger, B,Maraschek, M,Van Zeeland, M A,da Graç,a, S Published jointly by The Institute of Physics and 2011 Plasma physics and controlled fusion Vol.53 No.12
<P>Detailed measurements of the 2D mode structure of Alfvén instabilities in the current ramp-up phase of neutral beam heated discharges were performed on ASDEX Upgrade, using the electron cyclotron emission imaging (ECEI) diagnostic. This paper focuses on the observation of reversed shear Alfvén eigenmodes (RSAEs) and bursting modes that, with the use of the information from ECEI, have been identified as beta-induced Alfvén eigenmodes (BAEs). Both RSAEs with first and second radial harmonic mode structures were observed. Calculations with the linear gyro-kinetic code LIGKA revealed that the ratio of the damping rates and the frequency difference between the first and second harmonic modes strongly depended on the shape of the <I>q</I>-profile. The bursting character of the BAE type modes, which were radially localized to rational <I>q</I> surfaces, was observed to sensitively depend on the plasma parameters, ranging from strongly bursting to almost steady state.</P>