Fluids produced by the uterus of pregnant sheep (OUF-ovine uterine fluids) were assayed for mitogenic activity in a thymidine incorporation assay. A dose-dependent mitogenic activity was observed in OUF which exceeded that of adult ovine plasma or fetal bovine serum. Uterine fluids were capable of stimulating thymidine incorporation in mouse 3T3 fibroblasts, rat L6 myoblasts, ovine trophoblast-derived cells, HeLa S3 cells, and bovine aortic endothelial cells. The greatest stimulation was observed in L6 myoblasts. The name ovine uterine-derived growth factor (ovine UDGF) has been suggested for this activity.
The kinetics of the reaction between the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and methylated urates was studied. Urates that had methyl groups on the 1,3,9, or on the 1 and 3 or 1 and 9 nitrogens reacted with DPPH 15 to 77% faster than uric acid. Urates substituted with methyl groups on the 7 nitrogen or on both the 3 and 9 nitrogens reacted with DPPH at rates that were less than 0.1 that of uric acid. 3,7,9-Trimethyluric acid and 1,3,7,9-tetramethyluric acid reacted with DPPH at barely detectable rates. DPPH reacted with uric acid, the monomethylated urates, and some of the dimethylated urates in a ratio of 2:1. DPPH reacted with other dimethylated and trimethylated urates in a ratio of 1:1. Semiempirical MNDO calculations indicate that the most stable radical of uric acid is formed by hydrogen abstraction from the 3, 7 or 9 position. The most stable species resulting from loss of a second hydrogen lack hydrogens at the 3 and 7 positions or the 7 and 9 positions. For maximum reactivity with DPPH, methylated uric acid derivatives must have a hydrogen at nitrogen 7 and one of the hydrogens at either the 3 or 9 position.