Ebselen Case Study

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1. Chemical reagents Ebselen was purchased from Sigma-Aldrich (USA). Stock solution of 100 mM in dimethyl sulfoxide (DMSO) was prepared and deposited at −30 °C until use. 2. In vitro cultivation of Babesia parasites Ebselen was valued for its chemotherapeutic effect against B. bovis (Texas strain) (Hines et al., 1995), B. bigemina (Argentina strain (Bork et al., 2004b), B. caballi (Bork et al., 2004b), and T. equi (U.S. Department of Agriculture) (AbouLaila et al., 2010b). Parasites were cultured in bovine or equine red blood cells by means of a continuous micro-aerophilous stationary phase culture system (AbouLaila et al., 2010b). The culture medium, M199, applied to B. bovis, B.bigemina, and T. equi (acquired from Sigma-Aldrich, Tokyo, Japan), was supplemented with 40 % bovine or equine serum and 60 U/ml of penicillin G, 60 μg/ml of streptomycin, and 0.15 μg/ml of amphotericin B (Sigma-Aldrich). TEShemisodium salt (229 mg/ml) N-tris- (hydroxymethyl)-methyl-2-aminoethansulfonic acid, 2-[(2-hydroxy-1,1-bis-…show more content…
B. bigemina, B. caballi, B. bovis, and T. equi were gained from cultures with parasitemia of 5%, which was diluted with suitable uninfected red blood cells to a starting parasitemia of 1% for the assays. The growth inhibition assay was done in 96-well plates containing 20 μl of packed red blood cell inoculum and 200 μL of an appropriate culture medium containing 5, 10, 25, 50, and 100 μM of ebselen. The concentrations used were based on a initial study. Positive control cultures received 5, 10, 50, 100, 1000 or 2000 nM of diminazene aceturate (Aboulaila et al., 2010a). For negative experimental control, cultures without the drug and cultures containing DMSO 0.01 % for Ebselen and DDW 0.02 % for diminazene aceturate were arranged. The experiments were carried out in triplicate wells per drug concentration for each parasite species and in three isolated trials. Cultures were incubated at 37 °C in an atmosphere of 5 %
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