| Popis: |
With a direct fluorescence assay, the levels of mixed-function oxidase activity were determined in mitogen-activated human lymphocytes. The O-deethylation of ethoxyresorufin to resorufin was used to quantitate mixed-function oxidase activity. Ethoxyresorufin O-deethylase activity was low to nondetectable in noninduced, mitogen-activated cells, but it was readily detected in 3-methylcholanthrene, mitogen-activated lymphocytes. The activity was: (a) dependent on assay time and number of lymphocytes; (b) dependent on the presence of reduced nicotinamide adenine dinucleotide phosphate; (c) stable to freezing at -80 degrees for at least 2 weeks; (d) reproducibly detected in duplicate samples of blood from one individual when cultured and assayed at the same time; but (e) quite variable in samples of blood from one individual at different times. Since in hepatic and pulmonary tissue of model animal systems ethoxyresorufin is a specific substrate for cytochrome P-448-associated monooxygenases, the use of this chemical could proffer an assay that specifically measures human cytochrome P-448-associated activity. |
