Aryl Phosphate Derivatives of Bromo-Methoxy-Azidothymidine: MATERIALS AND METHODS(4)
To test the effect of duration of incubation on SIA in the presence of WHI compounds (01, 03, 05, and 07), a motile fraction of sperm (107/ml) was incubated at 37°C in 0.5 ml of BWW-0.3% BSA in the presence of 200 |xM WHI-01, or 100 |xM each of WHI-03, WHI-05, WHI-07, and N-9 or 1% DMSO alone. At timed intervals, duplicate aliquots (4-^l) were transferred to two 20-^m Microcell (Conception Technologies, San Diego, CA) chambers, and sperm motility was assessed by CASA. For WHI-01, CASA was performed every 20 min for 150 min; for N-9, every 15 min for 120 min; for WHI-05, every 10 min for 70 min; and for WHI-03 and WHI-07, every 5 min for 40 min.
The possible effects of seminal plasma on SIA of WHI compounds were studied either in the presence of 10% cell-free seminal plasma in the assay medium or by direct addition (200 |xM) of WHI-03, WHI-05, and WHI-07 to dilutions (1:2-1:6) of liquefied donor semen (n = 3) in PBS and incubation at 37°C. After 3 h of incubation, duplicate aliquots (4-^l) were used for CASA. ventolin inhalers
To assess the persistence of SIA after removal of WHI compounds, pooled motile sperm (5 X 106) were added to 0.5 ml of assay medium in the presence (100 ^M) or absence of WHI-01, WHI-05, and WHI-07 in 1% DMSO.