ABSTRACT
BACKGROUND: A stable isotope tracer method to quantify the synthesis of proteins of hepatic origin in response to feeding is described. The response of albumin synthesis on one mixed meal in a piglet model was investigated and the intragastric and intravenous administration modes of 13C-valine were compared. METHODS: The fasting and postprandial fractional synthesis rates (FSRs) of albumin in 15 piglets were measured while infusion rates of 13C-valine were changed in anticipation of the increased appearance of the tracee after a single liquid food bolus (30 mL/kg infant formula). 13C-valine enrichments in albumin hydrolysates at regular time intervals were determined with gas chromatography-combustion isotope ratio mass spectrometry. RESULTS: The intravenous mode (n = 8) showed constant plasma alpha-ketoisovalerate tracer-to-tracee ratios (coefficient of variation range: 1-8%), and a 27% increase in albumin FSR after the food bolus (mean FSR +/- standard error [SE]: fasting 14.4% +/- 1.6% vs. postprandial 18.3% +/- 2.2% per day; P < 0.005). In the intragastric mode (n = 7), albumin FSR calculated from the mean precursor values increased 32% after feeding (fasting 14.6% +/- 1.5% vs. postprandial 19.3% +/- 1.6% per day; P = 0.005), despite absence of constant alpha-ketoisovalerate enrichment (coefficient of variation range: 15-31%). The FSRs were not significantly different between both infusion modes. CONCLUSIONS: A mixed food bolus increases albumin FSR in growing piglets by approximately 30%, irrespective of the tracer administration route. The concept of anticipated precursor steady state is applicable to study changes of hepatic protein synthesis after a single meal. The intragastric mode of tracer administration can be applied as a less invasive method to measure tissue specific protein synthesis in children.
Subject(s)
Eating/physiology , Liver/metabolism , Protein Precursors/metabolism , Serum Albumin/biosynthesis , Animals , Carbon Isotopes , Dietary Proteins/pharmacology , Fasting/physiology , Female , Infusions, Intravenous , Intubation, Gastrointestinal , Isotope Labeling/methods , Models, Animal , Postprandial Period/physiology , Random Allocation , Serum Albumin/metabolism , Swine , Time Factors , Valine/administration & dosage , Valine/pharmacokineticsABSTRACT
A gas chromatographic-combustion isotope ratio mass spectrometric (GC-C-IRMS) method for the determination of [1-13C]valine enrichments in protein hydrolysates is described. Using a quick derivatization method, delta13C values of the N-methoxycarbonyl methyl ester of valine can be determined from baseline separated GC peaks. Evaluation studies with respect to precision, accuracy, linearity, reduction capacity of the CuO combustion furnace and isotope dilution as a result of derivatization, showed that our GC-C-IRMS system allows robust measurement of enrichments of [1-13C]valine in the range 0 to 1.5 MPE (S.D.+/-0.01 MPE, n=3). Therefore this method is suited to determine fractional synthetic rates (FSRs) of proteins as low as one-tenth of the FSR of human albumin, in studies using a primed, continuous (6 h) infusion with [1-13C]valine plasma enrichments of approximately 15 MPE and an hourly sampling schedule.
Subject(s)
Chromatography, Gas/methods , Mass Spectrometry/methods , Serum Albumin/chemistry , Valine/analysis , Carbon Isotopes , Evaluation Studies as Topic , Gases , Humans , Hydrolysis , Nitrogen Oxides/chemistry , Reproducibility of ResultsABSTRACT
Variation in crassulacean acid metabolism (CAM) was investigated in thirtyAeonium species under controlled, well-watered conditions. Carbon isotope ratios and the specific activities of the CAM enzymes phosphoenolpyruvate carboxylase (PEPC) and malic enzyme ME_ are highly correlated and indicate a large variation in CAM. The δ13C values range from -13.6 to -27.1; PEPC and ME activities vary seven- and five-fold, respectively. Since these observations were made under controlled conditions, the differences are genetically determined and represent adaptations to different habitats. The CAM activity of the species is correlated with the characteristics of their natural habitats. CAM activity is also highly correlated with the growth- and life-forms of the species and accordingly agrees well with the infrageneric classifications which are partly based on these characters. Our results support the hypothesis that the Sempervivoideae have evolved from an EurasiaticSedum, which used C3 photosynthesis or weak CAM, and that evolution withinAeonium was from undifferentiatedSedum-like dwarf shrubs with C3 photosynthesis to woody candelabrum-shaped shrubs exhibiting strong CAM. The monocarpic, rosulate species with intermediate CAM may have evolved via an independent secondary route.
