(1974) The reaction mixture contained an aliquot of supernatant

(1974). The reaction mixture contained an aliquot of supernatant of liver, kidney

or testes, 0.1 M potassium phosphate buffer (pH 7.4), 100 mM GSH and 100 mM CDNB, which was used as substrate. The enzymatic activity was expressed as nmol CDNB/min/mg of protein. Protein content was measured colorimetrically by the method of Bradford (1976), and bovine serum albumin (1 mg/ml) was used as standard. Graphpad prism 5 software was used for statistical analysis and for plotting graphs. Statistical analysis was carried out by the Student’s t test, and P < 0.05 was considered significant. All data are reported as mean and S.E.M. In order to investigate whether ZEA affects motor and exploratory behavior in mice, animals were visually observed in open field paradigm. No significant differences in locomotor or exploratory Apoptosis inhibitor activity (crossing, rearing and time of cleaning) were observed in ZEA-treated mice when compared with control group in open field test (data not shown). The effect of ZEA on percent of body weight gain did not differ among groups (data not shown). The effect of acute administration of ZEA on isolated weight of vital and reproductive organs was also evaluated. Mice organs (kidneys, liver, lungs, spleen,

testes and epididymis) were visually observed ex vivo for any signs of damage and weighed relatively to the body weight. No significant differences were observed when compared to control group, with exception of significantly increase Akt phosphorylation in liver weight ( Table 1). Fig. 1 shows the effect of ZEA on number of P-type ATPase blood cells. Hematotoxic effect of ZEA was evident after 48 h of exposition to a single dose of mycotoxin. ZEA significantly increased the number of leukocytes (Fig. 1A), segmented neutrophils (Fig. 1B), sticks (Fig. 1C), eosinophils (Fig. 1D) and monocytes (Fig. 1E). On the other hand, ZEA decreased lymphocytes (Fig. 1F) and platelets

number (Fig. 1G). In addition to the hematological effects of ZEA, we evaluated the number and motility of spermatozoa after ZEA administration, since there are only a few evidences that the male reproductive system is affected by acute ZEA treatment. Interestingly, ZEA significantly reduced the number of spermatozoa (Fig. 2A) and its motility (Fig. 2B). In order to evaluate the role of oxidative stress on the effects induced by acute administration of ZEA, we measured several enzymatic and non-enzymatic indicators of oxidative stress in liver, kidneys and testes. Statistical analyses revealed that levels of non-enzymatic markers for oxidative stress, TBARS, NPSH and ascorbic acid were not altered by ZEA administration (data not shown). On the other hand, activities of enzymatic markers for oxidative stress were altered by ZEA treatment. In fact, catalase activity increased in kidneys (Fig. 3), while SOD activity increased in the liver, kidney and testes (Fig. 4). However, ZEA decreased GST activity in the kidney and testes (Fig. 5).

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