Tissue segments were homogenized with T-PER (ThermoFisher Scientific, Massachusetts, USA) and centrifuged at 10,000 rpm at 4°C. Then the homogenates were incubated in wells of a 96-well plate at 37°C with 4.2 mM tetramethylbenzidine (Serva, Heidelberg, Germany), 2.5 mM citrate, 5 mM NaH2PO4 and 1.18 mM H2O2 pH 5.0 at a final volume of 150 μl. After 5 minutes the reaction was terminated by adding 50 ml 0.18M H2SO4. Absorbance was read at 450 nm against blank wells. Results were adjusted for tissue sample protein content on Bradford assay (Sigma-Aldrich) and they were expressed as MPO units/mg protein/g.

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