By Gregory Pincus
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Additional resources for Hormones and Atherosclerosis. Proceedings of the Conference Held in Brighton, Utah, March 11–14, 1958
I think your data on the half-life corroborates our work on bile fistula rats. If you give a dose of tracer cholesterol to normal rats, it is excreted with a half-life of maybe 25 days. In the fistula rat, we find a half-life of 6 days. That checks very nicely with yours. Don't you think your data might just as well be explained by some of the newly formed cholesterol being directly oxidized to bile acid? It is all microsomal reactions. KENDALL: Wouldn't you expect the cholesterol being excreted by the liver to be diluted with the freshly synthesized cholesterol as much as the bile acids being diluted by the oxidation of the freshly synthesized cholesterol?
Relative rates of hepatic cholesterol synthesis were obtained by incubating 1 g. liver* slices in 10 ml. Krebs-Henseleit-Ringer containing 1 4 14 C H 3 C 0 0 ~ N a + (acetate-C ). The incubations were performed under 100% oxygen for 3 hours at 37 ° C , shaking at 110 oscillations per minute. After incubation, the tissues were saponified by alcoholic potassium hydroxide, and the sterol was then extracted with petroleum ether. This extract was washed with water, 1% sodium acetate solution, water washed again and dried over anhydrous sodium sulfate, and taken to dryness.
As far as isoprene is concerned, we have in fact prepared labeled isoprene and found it to be totally inactive as a precursor. We do not think that this result is necessarily in conflict with our postulated scheme, provided we do not insist on free isoprene as an intermediate. What I have presented is— or so it appears to us—the simplest mechanism to fit the isotopic data. For example, the D 2 0 data force us to conclude that one of the two phosphates is split off simultaneously with the decarboxylation and before condensation occurs.