During the first 12 h period, the animals displayed blood in the abdominal cavity, signs of lung hemorrhage (hemorrhagic spots), spleen and kidney enlargement and congestion (Fig. 1). The kidneys also seemed to have darkened slightly and had black spots on their surface (Fig. 1). The bladder was often edematous and enlarged. The brain and gastrointestinal system appeared to be macroscopically normal (not shown). To evaluate the acute systemic physiopathological effects of the venom, several biochemical and hematological markers of tissue BGB324 in vitro lesions were measured (Table 1). Subcutaneous injection of L. obliqua venom caused a marked increase in serum AST, peaking between 12 and

48 h. Although less markedly than AST, serum ALT also increased rapidly after the first 2 h, reaching a maximum at 12 h. Serum levels of γ-GT increased over the first 6 h and remained elevated until 48 h. In comparison to the controls, high levels of plasma free hemoglobin, LDH and bilirubin were detected at 6 and 12 h, indicating that intravascular hemolysis had occurred. Markers of renal damage, such as creatinine, BUN and uric acid, also displayed important

alterations. Serum creatinine increased mainly between 6 and 96 h, reaching maximal values at 48 h, whereas BUN increased 12, 24 and 48 h after venom injection, returning to normal levels thereafter. The animals had hyperuricemia throughout the time of envenomation, with the levels of uric acid reaching 8 times the control values (p < 0.001) ( Table 1). Hematological parameters were evaluated at 6, 12 and 48 h post-envenomation. HIF inhibitor The obtained results are summarized in Table 2. LOBE injection caused a statistically significant O-methylated flavonoid decrease in red blood cell count and hemoglobin at 12 and 48 h, whereas the platelet count decreased slightly at 12 h and returned to normal after 48 h. Hematocrit values were lower when compared

to the controls at all of the time points evaluated. The reticulocyte number (immature red cells) increased in the blood stream as a result of hemolysis and anemia. The hematimetric indices, MCV and MCH, also increased at 48 h, whereas MCHC and total protein remained unchanged. Envenomed rats displayed leukocytosis between 6 and 12 h, mainly due to high neutrophil (6–48 h) and lymphocyte (6–12 h) counts. Compared to control values, a 15-fold increase was observed only in neutrophil numbers at 6 h. A less expressive increase in monocytes and eosinophil counts was also observed at the same time. Under light microscopy, the blood smears revealed fragmented erythrocytes, spherocytes and significant anisocytosis. The leukocytes appeared to have normal morphology (data not shown). Evidence of tissue damage was observed mainly between 6 and 48 h of envenomation. Skin microscopy, at the site of LOBE injection, showed hemorrhagic lesions, muscle necrosis and focal inflammatory infiltration that was associated with edema of varying intensities (Fig. 2A and B).