Kinetics of Antiamebic Compound Activity

To determine the kinetics of our compounds activity against E. histolytica, we adapted the viability assay we developed in our previous studies [6 (link)] and performed a time course to measure changes in the number of live trophozoites over 48 hours. E. histolytica trophozoites (10,000 parasites in 150μl media) were seeded into 96-well plates and allowed to grow overnight in an anaerobic chamber. The next day, a single plate was removed for analysis, and drug was added to remaining plates at 2x the previously calculated EC₅₀ concentration. Parasite viability was assayed at 10, 24 and 48h using the live cell marker fluorescein diacetate (Sigma) as previously described [6 (link)]. Three independent biological replicates were performed.