23 research outputs found
Last exit for peace: the struggle for a Palestinian State has entered a new stage
Eberhard Rhei
Euro-Mediterranean Co-operation: Enlarging and Widening the Perspective. ZEI Discussion Paper: 2004, C 131
[Table of Contents]. Introduction, by Andreas Jacobs; The Future of Euro-Mediterranean Relations: the Vision of Malta, by H.E. President Guido de Marco; The Euro-Med Partnership Needs a Strong Push, by Stephen Calleya/Eberhard Rhein; European Enlargement and its Impact on the Barcelona Process, by Bechir Chourou; European Enlargement and the Barcelona Process, by Krzysztof Bobinski; Mapping the Impact of Enlargement on the Euro-Mediterranean Partnership, by Zlatko Ć abiĂš/Sabina KajnĂš; Under Full Sail in a Millennium of Migration? Enlargement in the East and âPush and Pull Factorsâ in the South, by Babak Khalatbari/Marc Lauterfeld; Impact of the Iraq War on Mediterranean Security and Dialogue, by Ian O. Lesser; Lâimpact de la guerre contre lâIrak sur le processus MĂ©diterranĂ©en, by Abdallah Saaf; From the Mediterranean to a Greater Middle East: Challenges for European Policy Formulation, by Jean-François Daguzan; System-Opening and Cooperative Transformation of the Greater Middle East: Elements of a New Common Transatlantic Project, by Ludger KĂŒhnhardt
Strategisches Krisenmanagement, Ăkonomische Krisenresistenz durch risikobasiertes Kapitalkostenmanagement (Teil B)
<p><b>Binding of recombinant CHIKV E2 protein domains A, B, and C, and E2ex to 293T, CHO-K1, and pgsA-745 cells in the presence of soluble glycosaminoglycans (GAGs).</b> 10 Όg of the indicated recombinant proteins were incubated with the indicated soluble GAGs (500 Όg/ml) for 30 minutes at 4°C. 293T (top), CHO-K1 (middle), and pgsA-745 (bottom) cells were then incubated with this mixture. Binding was measured by flow cytometry using an anti-His-tag and an anti-mouse FITC conjugated antibody. The results are shown as relative values to the control (incubation of cells with the respective recombinant protein alone). A value of one represents the mean FITC value of the control and is labeled by the dashed line. Values above this line indicate inhibition of binding. Data represent the average of three independent experiments. * and ** indicate significant differences to the GAG-free controls. n.s. means not significant. * (P †0.05), ** (P †0.01), *** (P †0.001) and **** (P †0.0001).</p
Binding of Fc-fusion proteins containing variants of the CHIKV E2 domain A to CHO-K1 and pgsA-745 cells.
<p>A: Fc-E2 domain A-fusion proteins and Fc protein were expressed from HEK293T cells, affinity purified by protein-A chromatography and separated by SDS-PAGE. The Western blot was detected with an HRP-labeled anti-human IgG antibody. The calculated molecular weights are: Fc-CHIKV-E2-A, E79K and E166K 53 kDa; Fc-CHIKV-E2-A-Ă 43,4 kDa. B: Separation of the Fc-fusion proteins under native conditions. The Western blot was detected with an HRP-labeled anti-human IgG antibody. C: CHO-K1 (black) and pgsA-745 (grey) cells were incubated with the indicated recombinant Fc-fusion proteins. Binding was measured by flow cytometry using an anti-human IgG FITC conjugated antibody. The results are shown as fold induction compared to Fc binding. A value of one represents the mean FITC value of the control and is labeled by the dashed line. Values above this line indicate binding. Data represent an average experiment of three independent experiments performed.</p
Infection and cell entry by CHIKV-mCherry-490 in the presence of soluble GAGs.
<p>A: 293T cells were seeded in 24-well plates and 500 Όg/ml of the indicated GAG was added. Afterwards, cells were infected with CHIKV-mCherry-490 using an MOI of 1. The viral replication was determined 6 hrs post-infection by flow cytometry detecting mCherry. B: 500 Όg/ml of the indicated GAG and CHIKV-mCherry-490 (MOI 1) were incubated together at 4°C for 30 minutes. After addition to 293T cells, another incubation of 30 minutes at 4°C followed. Then, the unbound virus together with the respective GAG were washed away and fresh medium was added to the cells. The viral replication was determined 6 hrs post-infection by flow cytometry detecting mCherry. Data represent the average of three independent experiments. *** and **** indicate significant differences in infection rates to untreated control cells. ** (P †0.01) and *** (P †0.001).</p
Binding of Fc-fusion proteins containing variants of the CHIKV E2 domain A to CHO-K1 and pgsA-745 cells.
<p>A: Fc-E2 domain A-fusion proteins and Fc protein were expressed from HEK293T cells, affinity purified by protein-A chromatography and separated by SDS-PAGE. The Western blot was detected with an HRP-labeled anti-human IgG antibody. The calculated molecular weights are: Fc-CHIKV-E2-A, E79K and E166K 53 kDa; Fc-CHIKV-E2-A-Ă 43,4 kDa. B: Separation of the Fc-fusion proteins under native conditions. The Western blot was detected with an HRP-labeled anti-human IgG antibody. C: CHO-K1 (black) and pgsA-745 (grey) cells were incubated with the indicated recombinant Fc-fusion proteins. Binding was measured by flow cytometry using an anti-human IgG FITC conjugated antibody. The results are shown as fold induction compared to Fc binding. A value of one represents the mean FITC value of the control and is labeled by the dashed line. Values above this line indicate binding. Data represent an average experiment of three independent experiments performed.</p
Transduction of cells with CHIKV Env pseudotyped vectors in the presence of soluble GAGs.
<p>293T (left), CHO-K1 (right), and pgsA-745 (bottom) cells were seeded in 384-well plates and transduced with CHIKV Env-pseudotyped vectors transferring a <i>luciferase</i> gene. Before addition to the cells, the vector particles were incubated with DX or one of the indicated GAGs for 30 minutes at 4°C (DX and GAGs in five 3-fold dilutions, ranging from 500.0 to 6.2 Όg/Όl). One day after transduction, the luciferase expression of the cells was detected by a luminometer [<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0005318#pntd.0005318.ref020" target="_blank">20</a>]. The results are given as percentages of the untreated control values. The experiment was carried out twice in triplicate, and one representative triplicate result is shown.</p