The Antimicrobial Effects Of Compound 48/80 On Tetrahymena Pyriformis In Vivo

Lau, Clifford
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University of Hawaii at Manoa
Compound 48/80, a well-known and potent mast cell degranulator, has recently been found to possess antimicrobial activity. This thesis describes studies on the mechanism of the antimicrobial action of compound 48/80 on the protozoan Tetrahymena pyriformis GL. T. pyriformis cell division was synchronized using the heat treatment method developed by Scherbaum and Zeuthen. When compound 48/80 was added at different times following the end of heat treatment (EHT), normally effective concentrations had no significant effect on cell division if added 60 minutes after EHT. A lapse of 60 minutes after EHT correlates to the G2 period in the cell cycle, indicating that compound 48/80 acts somewhere within the G2 phase. Synchronized cells treated with this compound at 25 ug/ml showed a 10 minute lag for the first division and a 30 minute lag for the second division when compared to cells without the drug. Incorporation studies of amino acids (14C), thymidine (methyl-14c), and uridine (2-14c) into proteins, DNA, and RNA showed that compound 48/80 affected the incorporation of all three precursors at concentrations that inhibited cell division. At 80 minutes after EHT, 25 ug/ml of compound 48/80 inhibited incorporation of precursors into proteins by 38%, DNA by 47%, and RNA by 32%. The surface-active property of compound 48/80 was compared to the surfactant Triton X-100, (Octyl Phenoxy Polyethoxyethanol) and distilled water using a volumetric method. When compared to the volume of distilled water, Triton X-100 (0.01%) reduced drop volume by 55% whereas compound 48/80 (0.01% and 0.05%) gave volumes of 99.5% and 99% respectively. These results indicate that compound 48/80's mode of action is not as a surfactant such as Triton X-100.
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