Tracer methods underestimate short-term variations in urea production in humans

Urea production rate (Ra) is commonly measured using a primed continuous tracer urea infusion, but the accuracy of this method has not been clearly established in humans. We used intravenous infusions of unlabeled urea to assess the accuracy of this technique in normal, postabsorptive men under the following four different conditions: 1) tracer [13C]urea was infused under basal conditions for 12 h (control); 2) tracer [13C]urea was infused for 12 h, and unlabeled urea was infused from hours 4 to 12 at a rate twofold greater than the endogenous Ra (“step” infusion); 3) tracer [13C]urea was infused for 12 h, and unlabeled urea was infused from hours 4 to 8 (“pulse” infusion); and 4) tracer [13C]urea was infused for 9 h, and unlabeled alanine was infused at a rate of 120 mg ⋅ kg−1 ⋅ h−1(1.35 mmol ⋅ kg−1 ⋅ h−1) from hours 4 to 9. Urea Ra was calculated using the isotopic steady-state equation (tracer infusion rate/tracer-to-tracee ratio), Steele’s non-steady-state equation, and urinary urea excretion corrected for changes in total body urea. For each subject, endogenous urea Ra was measured at hour 4 of the basal condition, and the sum of this rate plus exogenous urea input was considered as “true urea input.” Under control conditions, urea Ra at hour 4 was similar to that measured at hour 12. After 8-h step and 4-h pulse unlabeled urea infusions, Steele’s non-steady-state equation underestimated true urea input by 22% (step) and 33% (pulse), whereas the nonisotopic method underestimated true urea input by 28% (step) and 10% (pulse). Similar conclusions were derived from the alanine infusion. These results indicate that, although Steele’s non-steady-state equation and the nontracer method more accurately predict total urea Ra than the steady-state equation, they nevertheless seriously underestimate total urea Ra for as long as 8 h after a change in true urea Ra.

Composition of liquid-and particle-associated bacteria and their contribution to the rumen outflow

A method is described to estimate the composition and rumen outflow of microbes associated with liquid (LAB) and solid (SAB) digesta. Four rumen-cannulated Rasa Aragonesa ewes were given, in random order, the following 4 diets: (1) NaOH-treated barley straw, as a sole diet (700 g/day, TS); (2) NaOH-treated barley straw mixed (50 : 50) with 400 g/day of rolled barley grain (BS); (3) Diet 1 with addition of 8 g/day of urea; and (4) Diet 2 with addition of 16 g/day of urea. Co-EDTA was used as a marker for the liquid phase to estimate rumen outflow of liquid-associated purine bases (PB), and urinary purine derivatives were used as an indirect marker of total duodenal flow of PB. Solid-associated PB were calculated by the difference between both estimates. Urea infusion increased ammonia-N concentration in the rumen fluid from 4·8 to 15·9 mg/100 mL (P < 0· 05) and enhanced dry matter intake of TS diets (from 343±63· 5 to 556±41·2 g/day, P < 0·001). Significant differences were observed in the PB/N ratio of bacteria harvested from the liquid phase compared with that isolated from the solid phase (1·89±0·25 v. 1·66±0·32 mol/mg in LAB and SAB, respectively). Because of the differences observed between the liquid- and solid-associated bacteria, estimated values of bacterial N supply varied depending on which bacterial extract was used as reference. The fractional contribution of LAB and SAB to the postruminal bacteria was significantly influenced by the experimental diets, mainly through variations in the amount of LAB flowing out of the rumen.

NaCl and/or urea infusion fails to increase renal inner medullary myo-inositol in protein-deprived rats

As we recently demonstrated that in potassium depletion a decrease in inner medullary organic osmolytes might precede and cause a renal concentrating defect, we hypothesized that in the protein deprivation the same mechanism may occur. To clarify the relationship between renal medullary organic osmolytes and urine concentration defects during protein deprivation, we examined the effect of protein malnutrition on organic osmolyte content after water deprivation or sodium and/or urea infusion. Water deprivation did not restore urine urea and osmolality or tissue sodium and urea in protein-deprived rats to control levels. NaCl infusion only increased urinary and medullary Na. Urea infusion increased medullary urea but not urine urea. NaCl plus urea infusion increased only urinary sodium and urea. Regardless of the protocols of hyperosmolality used, protein deprivation significantly decreased the medullary contents of myo-inositol and taurine and the level of sodium-dependent myo-inositol transporter mRNA. We conclude that factors other than NaCl and urea but associated with protein feeding are responsible for the decreased accumulation of organic osmolytes.

Effects of intraruminal infusion of propionate on the concentrations of ammonia and insulin in peripheral blood of cows receiving an intraruminal infusion of urea

SUMMARYTo test the hypothesis that propionate can reduce hepatic capacity to detoxify ammonia, effects of the inclusion of propionate in intraruminal infusions of urea on the concentrations of ammonia, other metabolites and insulin in peripheral blood were investigated in two experiments with non-lactating dairy cows. Both experiments were of a 4 × 4 Latin square design with four animals, four treatments and four experimental periods of 7 d; feed was given in two equal meals each day, all intraruminal infusions were given for 1 h at the time of the morning feed, and propionic acid was partly neutralized with NaOH. In Expt 1, the treatments were a basal diet of pelleted lucerne and chopped hay alone or with the following infusions (g/d): urea 80, propionic acid 350, urea 80 plus propionic acid 350. The inclusion of propionate in the urea infusion markedly increased (P < 0·001) the concentration of ammonia in plasma compared with infusion of urea alone. Moreover, the inclusion of urea with the propionate infusion abolished (P < 0·01) the increase in blood insulin level seen with the infusion of propionate alone. In Expt 2, less severe treatments were imposed, the aim being to reproduce metabolic loads of propionate and ammonia that might be expected from a diet of high-protein grass silage rich in lactic acid. The treatments were a basal diet of grass silage alone or with the following infusions (g/d): NaCl 145, NaCl 145 plus urea 50, propionic acid 200, urea 50 plus propionic acid 200. Effects were less pronounced than in Expt 1 but, in the period immediately after infusion, similar effects were seen. It is concluded that propionate–ammonia interactions may have potentially important effects on milk production especially for diets with high proportions of grass silage containing high levels of protein and lactic acid.