Injections were performed daily for 8 days using a 1-ml syringe and a polyethylene catheter (PE-50). With the mouse immobilized, the catheter was inserted 3 to 4 cm intrarectally, and 200 μl of either vehicle (50% dimethyl sulfoxide), GA (17.5 mM), or MPA (1.75 or 17.5 mM) was slowly injected by hand. After removal of the catheter, the rectum was held closed for a minimum of 2 min. Body weight of each animal was recorded daily. After 8 days, animals were sacrificed. Colonic tissue was homogenized in hexadecyltrimethylammonium buffer using a Polytron homogenizer (Kinematica). O-dianisidine (Sigma-Aldrich) was added to the cell lysate, and its oxidation was measured by absorbance at 450 nm using a SpectraMax Plus 384 microplate reader. MPO activity (U/mg tissue) was calculated from absorbance based on a standard curve.

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