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Inflammation may play a role in the progression of venous disease and contribute to complications such as blood clots and leg ulcers

Measure of competitive binding titers by ELISA Coating, blocking, and gH/gL immobilization steps were performed as described under Measurement of serum antibody endpoint binding titers by anti-His capture ELISA. Following the capture of monomeric gH/gL, equal amounts of sera from each mouse in a group were pooled and diluted in blocking buffer and 2-fold serial dilutions were performed, followed by a 1 h incubation at 37 C

The next day, cells were diluted in two parallel cultures (control and iron) to a final OD 600nm of 0.05 and then grown at 37 C and 160 rpm

Finally, minimum inhibitory concentrations (MIC) against silver acetate were determined using standard Clinical and Laboratory Institute (CLSI) broth-microdilution method