A method for determination of carbon dioxide specific radioactivity and concentration in blood, rumen fluid, and expired air.
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A method for measuring the unsaturated iron-binding capacity of serum using radioactive iron with magnesium carbonate as the adsorbing substance is described. The radioactive iron-magnesium carbonate method, compared with the methods of Caraway (1963), Herbert, Gottlieb, Lau, Fisher, Grevirtz, and Wasserman (1966), and Bothwell, Jacobs, and Kamener (1959), was shorter and simpler and equally reproducible.
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Radioactively labeled carbon monoxide (CO) dehydrogenase has been obtained in good yield and purity from Pseudomonas carboxydoflava grown in the presence of [32P]phosphate. One enzyme molecule contained an average of 8.32 molecules of phosphate. The entire phosphate content was confined to 2 molecules of FAD and 2 molecules of a pterin. These were noncovalently bound. Molybdoenzyme cofactors could be extracted into N-methyl formamide; pterins were isolated by thin-layer chromatography. CO dehydrogenase contained a novel pterin, different from molybdopterin, which was also resolved in other bacterial molybdoenzymes. Therefore, it was tentatively named bactopterin. The characteristic features of bactopterin were as follows. A relative molecular mass, Mr, of 730 which was much greater than that of molybdopterin (330) (Mr values refer to molybdenum-free forms of the cofactors; presumably, the latter were also devoid of the sulfhydryl groups contained in the native compounds). A content of 2 molecules of phosphate/molecule compared to only 1 phosphate in molybdopterin. Bactopterin was three times less susceptible to air oxidation than molybdopterin. Native bactopterin was cleaved by perchloric acid into two phosphorous-containing fragments with Mr of 330 and 420. The smaller one is believed to be very similar to molybdopterin, the larger one was not a pterin but probably contained an aromatic structure.
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The information content of citrate carbon specific radioactivity ratios in steady-state 14C isotopic studies has been analyzed. Sixteen steady-state equations resulted containing five unknowns, 14 equations in terms of citrate carbon specific radioactivity ratios, and two modified forms of the "CO2 ratio" equation. Although each of the 16 equations is not independent, there is more than one independent equation for each variable. These additional equations may be used to test the assumptions on which the model is based. Each of the five unknowns is defined as the probability of flux around a complete cycle, either the tricarboxylic acid (TCA) cycle or a futile cycle such as pyruvate--oxaloacetate--pyruvate. To solve these equations for the five unknowns, an investigator need only measure the specific radioactivity of various citrate carbons and the 14CO2 production rate. The study did not yield a direct expression for net flux between pyruvate and 4-carbon TCA cycle intermediates. However, these equations do place certain constraints on the net flux through this important pathway.
Tests based on the analysis of the gaseous components of expired air have been developed to study intestinal absorption and intermediary metabolism of various nutrients. This paper reviews the breath-analysis tests based on the measurement of CH4, H2, and isotopically-labelled CO2 for studying the intestinal absorption of carbohydrates, fats, and bile salts, and intrahepatic metabolism. New technology employing mass spectrometry allows the use of the stable isotope, carbon-13, instead of the radioactive isotope, carbon-14, for CO2 breath tests. The nutritional application of the breath-analysis tests is discussed, and the advantages of the non-radioactive, non-invasive procedures, especially for use in children and pregnant women in whom standard investigational methods represent a discomfort or a radioactive hazard, are emphasized.