Simplified turbidimetric assay for lipase activity.
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Biomedical subjects
Publications and source records attributed to C Bishop.
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Free or combined (3)H-labeled fatty acids are converted to their methyl-(14)C esters or, if labeled with (14)C, to their methyl-(3)H esters. For a given specific radioactivity of the methyl group, the nuclide ratio in the esters separated by GLC is a direct measure of the specific radioactivity of the fatty acids, and quantitative collection is unnecessary. Methods of methylation with minimum quantities of labeled methanol, and of deriving nuclide ratios from channel ratios in a scintillation spectrometer, are given.
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1. Tritium-labelled palmitic acid combined in olive-oil triglycerides was introduced into the rumen of a lactating cow and the specific radioactivity of the lipids of milk and of the lipoproteins of both jugular and mammary venous serum was measured. 2. As previously found in a similar experiment with [(3)H]stearic acid, the specific radioactivity of the triglyceride fraction of the dextran sulphate-precipitable lipoproteins reached a maximum earlier and greater than that of the milk fat. 3. This fraction was the only lipid separated that had a significant arteriovenous difference in concentration, and is therefore identified as the main circulating lipid precursor of milk fat. 4. Although the non-esterified fatty acids showed no arteriovenous difference in concentration, they showed a negative difference in specific radioactivity that could have occurred only at the expense of the triglycerides of the precipitable lipoproteins. 5. The mean specific radioactivity of the triglycerides immediately after removal from the blood is calculated and shown to be very close in value to that of the corresponding fraction in mammary venous serum. 6. By comparison of the mean specific radioactivities of milk fat and of this precursor, its contribution is calculated as 36%. 7. This value is discussed with reference to the concentration of C(16) and C(18) fatty acids in milk fat and it is concluded that substantial amounts of these acids must have been derived from a source other than preformed circulating lipids.
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1. Tritium-labelled olive-oil triglycerides were introduced into the rumens of lactating cows and the specific activities of the lipids of milk and plasma and of serum lipoproteins were measured. 2. On treatment of serum with dextran sulphate it was found that the lipid of the precipitated beta-lipoproteins consistently had a specific activity-time curve with a maximum comparable in value with, and occurring earlier than, that of the milk fat. 3. On fractionation of the lipids of these lipoproteins it was found that only the triglycerides and diglycerides had specific activity-time curves with maxima greater than that of milk fat, and on radioactivity data alone they are the only blood constituents studied that meet the requirements for being the precursor of milk fat. 4. From a consideration of abundances and the mean specific activities over the period of the experiment it is shown that the contribution of the diglycerides to the radioactivity in the milk fat must have been negligible and that only the triglycerides could have been responsible for all the radioactivity found in it. 5. Although no other fraction could alone have been responsible for all the radioactivity in the milk fat, at least one, the phospholipids, could have made some contribution. 6. It is calculated that 35-48% by weight of milk fat was derived from the beta-lipoprotein triglycerides, according to whether the phospholipids made an improbably large contribution or none at all.
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Subjective estimates of sleep onset among patients with a variety of sleep disorders have been shown to be inaccurate. This inability in perceiving sleep onset is potentially dangerous for this population, in particular, for individuals who are required to drive long distances or operate heavy machinery as part of their daily activities. This study evaluated the perception of sleep among 237 consecutive patients diagnosed with obstructive sleep apnea. Patients completed an overnight sleep-laboratory evaluation followed by an objective evaluation of sleep propensity. The latter was done using the Multiple Sleep Latency Test (MSLT). Patients with an accurate perception of having slept on the MSLT were found to have shorter sleep latencies when compared with those with an inaccurate perception of their sleep. The results of this study suggest that the rapidity with which sleep is manifested is an important contributor to the accurate perception of sleep.
In humans, XX maleness is the best known example of a sex reversal syndrome occurring with an incidence of one XX male among approximately 20,000 to 30,000 newborn boys. The karyotypes of the majority of these individuals are apparently normal, with respect to the numbers and structure of the chromosomes, but is in contradiction with the phenotypic sex which they display. XX maleness may be either a non Y-related mechanism triggered by a mutation on another chromosome or could be the result of the expression of some cytogenetically undetectable Y chromosome material present in the genome of such individuals. Recently, a number of human Y-specific single copy probes have been isolated. In this study, using several of these Y-specific probes we definitively demonstrate the presence of Y-chromosomal material in the genome of some 46,XX human males. These XX males carry only a fraction of the human Y chromosome. In the three positive cases reported here, presence of inclusive overlapping chromosomal fragments has been detected, implying a genetic heterogeneity of these patients.