To assay possible hemag glutination by EF itself, 50 ?l of EF in PBS at the indicated concentrations were incubated with 50 ?l CES for 60 min or 4 hours at 4 C. All assays were performed in quadrupli cate. Virus Resistance Assay MDCK cells grown over night at 37 C and 5% CO2 were pre incubated with 2 ml complete medium with or without EF, at 37 C and U0126 price 5% CO2 for 60 min. In parallel virus in PBSBA was incubated with EF or left untreated for 60 Inhibitors,Modulators,Libraries min. After the pre incubation Inhibitors,Modulators,Libraries period the cells were washed and infected with 500 ?l virus suspension Echinaforce. Cells were then incubated for 60 min in the dark at room temperature after which the inoculum was removed. Cells were further incubated in 2 ml medium, Tamiflu or without test sub stances at 37 C, 5% CO2 for 24 hours.

Samples of the supernatants were collected, which were then assayed by focus forming assay for further determination of infec tious virus. Inhibitors,Modulators,Libraries Inhibitors,Modulators,Libraries Following the assays, these supernatants Inhibitors,Modulators,Libraries were used to infect another set of http://www.selleckchem.com/products/Roscovitine.html cultures under the same con ditions as described above. This process of sequential infection with supernatants was repeated once more to yield in total three rounds of infection and replication. Experiments done in duplicates were stopped when the Tamiflu sample reached titers of the untreated control. Biosafety All experiments with infectious virus were performed according to German and Canadian regulations for the propagation of influenza A viruses. All experiments involving highly pathogenic influenza A viruses and the pandemic S OIV were performed in a biosafety level 3 containment laboratory approved for such use by the local authorities.

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