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The homogenate was centrifuged at 12,000 rpm for 15 min at 4C and the supernatant was collected for assay MPO activity as determined by measuring the H 2 O 2 -dependent oxidation of o -dianisidine solution (3,3-dimethoxybenzidine dihydrochloride in potassium phosphate buffer, pH 6.0) at 450 nm using an Epoch microplate spectrophotometer (BioTek, VT, USA)

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The expedition led by Portol, which included Father Junpero Serra, the President of the Missions, along with a combination of missionaries, settlers, and leather-jacket (leather jackets made of several layers of leather could stop most Indian arrows) soldiers, including Jos Raimundo Carrillo, left Velicata on 15 May 1769, accompanied by about 46 mules, 200 cows and 140 horsesall that could be spared by the poor Baja Missions

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