A Method for the Histochemical Differentiation of Cholesterol and Its Esters
40
Citation
0
Reference
10
Related Paper
Citation Trend
Abstract:
Free cholesterol is demonstrated in formalin-fixed frozen sections when treated successively by digitonin, alcohol-ether, and the Schultz technique, in which circumstances cholesterol esters are not visualized. Cholesterol esters and free cholesterol are both demonstrated in comparable sections treated by the Schultz method alone, so that the difference between such sections indicates the sites at which cholesterol esters may be considered present.Keywords:
Digitonin
Digitonin
Hexokinase
Cite
Citations (7)
Abstract In the classic Schoenheimer—Sperry method for estimating cholesterol [J. Biol. Chem. 106, 745 (1934)] it is selectively precipitated with digitonin, and interference by other substances is minimized. The standards used in the color development are pure cholesterol. We suggest that the standard cholesterol should also be converted to cholesterol digitonide and treated identically to the test samples. When this is done, the values for cholesterol in test samples are significantly higher, both for free and total cholesterol in blood serum and for total cholesterol in tissues. Moreover, recovery experiments showed that error is diminished. Values for serum total cholesterol obtained when cholesterol digitonide standards were used were closer to those obtained by the method of Abell et al. than when pure cholesterol standards were used.
Digitonin
Blood cholesterol
Cite
Citations (7)
Primary alcohol
Cite
Citations (92)
From the study of the inhibitory action of the sera of rabbits fed on diets containing varying amounts of cholesterol on the hæmolysis of blood by saponin we were led to the conclusion, in an earlier paper of this series, that when cholesterol, free or in the form of esters, is given with the food of rabbits some is absorbed and finds its way into the blood stream as free cholesterol; and also that when phytosterol is used instead of cholesterol it behaves similarly, and some appears in the blood either itself or in the form of cholesterol. The method adopted in these experiments only gave information as to the amount of free cholesterol under various conditions; but not as to us to the amount of cholesterol esters that might be present. It seemed to us desirable to confirm these results by a chemical method, and also, if possible, to estimate both the free cholesterol and the esters present at the same time in the blood under various conditions. The usual methods for the chemical estimation of cholesterol are not sufficiently accurate to give reliable information in the case of a fluid such as blood, which contains but a small percentage of cholesterol, and which is only available in relatively small quantities. The discovery of Windaus in 1909 that cholesterol, but not cholesterol esters, readily combines quantitatively with digitonin to form a highly insoluble compound, digitonin cholesteride, according to the following equation- C 55 H 94 O 28 +C 27 H 46 O = C 82 H 140 O 29 , afforded a means of solving the problem.
Digitonin
Blood cholesterol
Phytosterol
Cite
Citations (13)
The colorimetric method for the determination of total cholesterol in dog bile is consistently accurate as checked by the oxidative digitonin method. This method has the further advantages of being simple, rapid and economical. A method for the determination of esterified cholesterol in bile is described and it is shown that there are no esters of cholesterol in normal dog bile.
Digitonin
Cite
Citations (8)
Digitonin
Serum cholesterol
Cite
Citations (101)
Digitonin
Cite
Citations (30)
Digitonin
Cite
Citations (23)
Free cholesterol is demonstrated in formalin-fixed frozen sections when treated successively by digitonin, alcohol-ether, and the Schultz technique, in which circumstances cholesterol esters are not visualized. Cholesterol esters and free cholesterol are both demonstrated in comparable sections treated by the Schultz method alone, so that the difference between such sections indicates the sites at which cholesterol esters may be considered present.
Digitonin
Cite
Citations (40)
As is well known, digitonin exerts a hemolytic action on red blood corpuscles, and cholesterol has the power to inhibit this activity. In view of the fact that our experiments have shown that the chemical properties of cholesterol are altered in various ways as the result of irradiation with ultra-violet light, it seemed worth investigating whether its property of inhibiting the action of digitonin remained unchanged after irradiation. A comparison was made of the effect, in relation to hemolysis, of ordinary cholesterol with that of cholesterol which had been irradiated for periods of 1/2, 2 and 10 hours with the radiation of a mercury vapor lamp, set at a distance of 1 foot. For this purpose a suspension of 1 per cent ethereal solution of cholesterol and 0.1 per cent digitonin was tested on the red cells of the dog and of the sheep. Without going into detail at this time, it may be stated that it was found that the rate of speed with which the cholesterol bound the digitonin had been altered as the result of irradiation—that the irradiated cholesterol bound digitonin more readily than ordinary cholesterol, and brought about a comparative delay in its hemolytic action. For example, whereas under controlled and constant conditions ordinary cholesterol allowed complete hemolysis to take place immediately, when cholesterol was used which had been irradiated for 2 hours, complete hemolysis did not occur until after an interval of 1 1/2 to 3 minutes. The sterol-digitonin was incubated for 3 hours before it was added to the red cells. When, however, the cholesterol was irradiated for a prolonged period, for 10 hours, this delay did not take place and complete heniolysis occurred immediately.
Digitonin
Cite
Citations (7)