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DURING recent years two groups of workers have reported a notable increase in urinary cholesterol excretion in cancer. Bloch and Sobotka (1938) worked with pooled urine specimens from cancer wards and compared them with pooled specimens from cardiac and tuberculosis wards, and found approximately ten times as much cholesterol in the cancer urine as compared with the cardiac and tuberculosis urine. In this study the cholesterol was isolated in relatively large quantities (0.35 g. per 100 litres of urine), and its identity was proved by determination of melting-point, optical rotation and preparation of the acetate.This work was later extended by Sobotka, Bloch and Rosenbloom (1940) to individual cases of cancer using a less specific colorimetric method, depending on the Liebermann-Burchard reaction. They reported an increase of urinary cholesterol in 19 out of 92 cancer patients. Later, Bruger and Ehrlich (1943), using a similar method of estimation, found a higher incidence of hypercholesteroluria, 11 of their 32 cases giving results above normal limits. However, the actual concentration of cholesterol reported by Bruger and Ehrlich in normal urine was approximately twice as great as that found by Sobotka, Bloch and Rosenbloom. It appeared, therefore, that the methods used in these two studies must have been significantly different, and we felt that further work on the technique of estimation was needed. While admitting the possible non-specificity of the Liebermann-Burchard reaction, we felt that this type of method was the only one likely to find immediate clinical application, and it does appear to have given results consistent with the original work of Bloch and Sobotka. The present work was undertaken with the main object of finding out whether refinements of this colorimetric method of estimation would be of any value in the diagnosis of cancer.EXPERIMENTAL.Extraction method.The chloroform extraction method of Neustadt, Howard and Myerson (1946) was first tried, but the amount of colour obtained at the final stage from the normal urines was too small for accurate measurement. Adaptation of this method, using the whole of the extract from 200 ml. of urine (in place of 50 ml.), gave a brown rather than a green colour with the Liebermann-Burchard reagent, as shown in Fig. 1. An attempt was made to correct for this error by the introduction of a blank reading, which was obtained by adding to half the urinary extract in 5 ml. of chloroform 0-9 ml. of ethanol and 0-1 ml. of concentrated H2S04 in place of the Liebermann-Burchard reagent. This sometimes gave
THiE application of the methods of urinary cholesterol estimation given by Burchell and Maclagan (1949) to cancer and control cases will now be described. BMATERIALUrine specimens were obtained from 13 normal subjects and 116 patients admitted to Westminster Hospital or to Westminster Hospital (.All Saints') Urological Centre. Twenty-five of these patients were suffering from diseases thought to be unlikely to affect cholesterol excretion as shown in the tables, and 73 were suffering from various forms of cancer. The remaining 18 were patients suffering from haematuria or pyuri3 of non-malignant origin. The diagnosis in the cancer group was established in most caes by laparotomy (3), biopsy (38) or autopsy (12), but in 20 cases the diagnosis was based on unequivocal X-ray or other clinical data.Collection and treatment of specimens.Preliminary results having shown that a considereble amount of cholesterol might be associated with the urinary sediment in certain cases, a routine procedure was adopted to standardize the treatment of the sediment.Twelve-hour specimens of urine were collected into clean winchesters, the time of the collection being usually between the hours of 6 p.m. and 6 a.m.The volume of specimen was measured, the urine adjusted to pHE 5-0 with acetic acid and, if urates were present, warmed to 37°C. Cell counts on the urinary depo/it (UD) were carried out by the method of Addis (1925) on a 10 ml. portion of the well-mixed specimen. For cholesterol estimation on the deposit a 500 ml. sample of the mixed specimen, or a smaller aliquot in the case of a concentrated urine, was centrifuged, the deposit washed once with normal saline and extracted directly with three portions of boiling acetone. The acetone extract was evaporated to dryness, and the residue extracted with petroleum ether for the determination of cholesterol as above.The cholesterol content of the supernatant urine (SU) was determined on 200 ml. portions by the aluminium tungstate method described by Burchell and Maclagan (1949 Similarly, any patient with haematuria or pyuria may be expected to show an increase in the UD cholesterol as shown in Table I. In this group of cases an attempt was made to correlate the cholesterol content of the UD) with quantitative cell counts, and although no exact correlation could be established, it will be noted that the three highest cholesterol values were associated with the highest cell counts. Pyuria was the commonest accompaniment of raised UD cholesterol in this series, but increase in red cells and epithelial cells appear to be significant in cases No. 137 and 152. It is evident from these results that a careful examination of the urinary deposit must accompany any cholesterol estimation if the result is to be Fig. 4, and full details of these patients are given in Table II The UD cholesterol results in 4 cases of cancer with haematuria or pyuria (leucocyte counts above 100 millions per 12 hours) were shown in Table I, but it will be seen that the high values here were equally distribut...
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