TO DEVELOP TEST METHODS WHICH REPLACE OR REDUCE THE USE OF EXPERIMENTAL VERTEBRATE ANIMALS.
The kinetic characteristics of aniline hydroxylase and N-methylaniline demethylase were determined in different tissue preparations after optimising experimental conditions. While liver S9 and red blood cell supernatants were characterised by comparing enzymic efficacy, the use of whole red blood cells was more effective for N-methylaniline. This suggested that membrane or cellular integrity was important. The roles of reduced nicotamide adenine dinuceotide (NADH) and reduced nicotamide adenine dinuceotide phosphate (NADPH) were investigated and both cofactors were found to be essential for maximum reactivity.
TWO ALTERNATIVE SYSTEMS ARE UTILIZED, WHICH DO NOT INVOLVE THE USE OF LABORATORY ANIMALS. THEY ARE HUMAN RED CELLS (ERYTHROCYTES) AND EXTRACTS FROM PLANTS.
A NUMBER OF END POINTS ARE STUDIED, IN PARTICULAR IN BACTERIA (FORWARD MUTATIONS, RESERVE MUTATIONS, SOS FUNCTIONS) AND IN MAMMALIAN CELLS CULTURE ( CHROMOSOMAL ABERRATIONS, SISTER CHROMATID EXCHANGE). VARIOUS CLASSES OF CHEMICALS, REQUIRING DIFFERENT MODES OF ACTIVATION ARE SCREENED.
THE RESULTS OBTAINED ARE COMPARED WITH THOSE PROVIDED BY THE SAME TESTS USING THE CLASSICAL S9 ACTIVATION SYSTEM.
Funding SchemeCSC - Cost-sharing contracts