In all animals exposed to alumina dust the presence of alumina crystals in the lung (alveolar spaces and airways) was qualitatively evaluated under polarized light (Axioplan, Zeiss, Oberkochen, Germany) at

1000× magnification. The right lungs were homogenized in 1 mL of PBS with protease inhibitors (1 μg/mL leupeptin and 1 μg/mL pepstatin). Homogenates were centrifuged (Centrifuge 5415R, Hamburg, Germany, 4 °C, 6700 × g, 15 min) and then, the supernatant was collected for transforming growth factor beta (TGF-β) and interleukin-1beta (IL-1β) assays by ELISA (R&D Systems Inc., Minneapolis, MN, USA), according to the manufacturer’s protocol. Total protein concentration in lung homogenates was determined by Bradford’s method ( Bradford, 1976). Concentration of cytokines in lung homogenates Ivacaftor in vitro was further normalized to protein concentration in the samples and expressed Baf-A1 chemical structure as picograms per milligram of protein. Optical density was measured at 450 nm by a microplate reader (SpectraMax 190, Molecular Devices, Sunnyvale, CA, USA). The normality of the data and the homogeneity of variances were tested by Kolmogorov–Smirnov test with Lilliefors’ correction and Levene median test, respectively. In all instances both conditions were satisfied and parametric

tests were run. One-way ANOVA was used to compare the values of body weight measured every 7 days, throughout 4 weeks in each group. Weight differences between control and exercise groups at every 7 days were

evaluated by Student’s t-test. Two-way ANOVA was applied to the remaining parameters (factors: exercise and alumina). For all ANOVAs, the Student–Newman–Keuls learn more was used as a post hoc test. The morphometric data, originally expressed as percent, underwent an arcsine transformation, in order to generate a normal distribution. The statistical analyses were carried out by the SigmaStat 9.0 software (SYSTAT, Point Richmond, CA, USA). In all instances p < 0.05 was considered a statistically significant difference. Metal composition of alumina dust is presented in Table 1. A high concentration of the element Al, followed by Fe and Hg was found. Scanning electron micrographs of particles are shown in Fig. 1, demonstrating the frequency distribution of diameters of particle sample. 90% of particles diameter are under 150 μm, being 50% below 100 μm and 10% smaller than 57 μm. A progressive increase in body weight was observed along time in animals not submitted to physical exercise. In exercising group, a decrease in body weight occurred during the first week of aquatic training, but thereafter the values did not differ from those in control mice (Fig. 2). All mechanical parameters (ΔP2, ΔE and Est) but ΔP1 were higher after alumina dust exposure in animals not submitted to physical exercise. Additionally, exercise training before particle exposure caused no changes in resistive and viscoelastic components, but Est increased in this group ( Fig. 3).