5% reduction, respectively, Fig. 1B). When mouse survival was evaluated, it was noted that the first animal died on the 9th day in EAT-inoculated group of mice. At the 10th day a 20% death toll was noted, and only 50% of the animals were alive on the 15th day. In the R-954-treated group of mice, only one animal died during the experimental period and this death occurred only after 14 days of consecutive treatment (Fig. 1C). On the 10th day after EAT cell inoculation into mice, a 12.6-fold increase of total blood cell count was observed (0.5 ± 0.2 × 107

cells in control group vs. 6.3 ± 2.1 × 107 cells in EAT-inoculated mice). This effect was accompanied by a proportional increase in total bone marrow cell count (0.12 ± 0.02 × 107 cells in selleck chemicals llc control

group vs. 1.3 ± 0.02 × 107 cells in EAT-inoculated mice) and on cells from ascitic lavage (0.8 ± 0.3 × 107 cells in control group vs. 11.7 ± 1.1 × 107 cells in EAT-inoculated mice). SCH772984 mw Vincristine (0.5 mg/kg, i.p.) a well known carcinostatic agent used for comparison purpose reduced total blood cell counts by 55.5% (6.3 ± 2.1 × 107 cells in EAT-inoculated mice vs. 2.8 ± 0.8 × 107 cells in vincristine-treated EAT inoculated mice), total bone marrow cell count by 76.9% (1.3 ± 0.02 × 107 cells in EAT-inoculated mice vs. 0.3 ± 0.2 × 107 cells in vincristine-treated EAT inoculated mice), and total cells in ascitic fluid by 71.8% (11.7 ± 1.1 × 107 cells in EAT-inoculated mice vs. 3.3 ± 1 × 107 cells in vincristine-treated EAT inoculated mice). Treatment of animals previously inoculated with EAT cells with B1 antagonist R-954 reduced total blood, bone marrow, and ascitic cell counts to values similar to vincristine-treated mice (3.2 ± 0.9; 0.5 ± 0.1; 4.8 ± 1.1 × 107 cells, respectively) (Fig. 2). In order to evaluate inflammatory mediator release after EAT inoculation, mice were sacrificed on the 10th day after tumor cell injection. Peritoneal ascitic fluid was collected and total protein, nitric oxide, PGE2, and TNFα were measured. In mice inoculated with EAT cells, marked increases

of the total proteins (from 13.8 ± 3.1 to 493.5 ± 33.8 mg/ml), O-methylated flavonoid nitric oxide (from 2.8 ± 3.3 to 76.9 ± 12.7 μM), PGE2 (from 28.4 ± 5.9 to 344.9 ± 45.8 pg/ml), and TNFα (from 31.7 ± 9.9 to 792.3 ± 113.4 U/ml) were noted when compared to the levels in the fluids of non-inoculated animals. Treatment of mice with R-954 reduced significantly the total protein extravasation (57.3%) as well as the production of nitric oxide (56%), PGE2 (82%) and TNFα (85.7%). The antitumoral drug vincristine also significantly reduced by 92% the protein extravasation, by 84.5% nitric oxide, by 94.7% PGE2, and by 92.2% TNFα levels (Table 1). When Ehrlich cells are inoculated intraperitoneally, the tumor process develops in an ascitic form but when the cells are inoculated subcutaneously, the tumor develops in a solid form.