Total Sulfur Amino Acid Requirements Are Not Altered in Children with Chronic Renal Insufficiency, but Minimum Methionine Needs Are Increased.

Background: The total sulfur amino acid (TSAA) and minimum Met requirements have been previously determined in healthy children. TSAA metabolism is altered in kidney disease. Whether TSAA requirements are altered in children with chronic renal insufficiency (CRI) is unknown.Objective: We sought to determine the TSAA (Met in the absence of Cys) requirements and minimum Met (in the presence of excess Cys) requirements in children with CRI.Methods: Five children (4 boys, 1 girl) aged 10 ± 2.6 y with CRI were randomly assigned to receive graded intakes of Met (0, 5, 10, 15, 25, and 35 mg · kg-1 · d-1) with no Cys in the diet. Four of the children (3 boys, 1 girl) were then randomly assigned to receive graded dietary intakes of Met (0, 2.5, 5, 7.5, 10, and 15 mg · kg-1 · d-1) with 21 mg · kg-1 · d-1 Cys. The mean TSAA and minimum Met requirements were determined by measuring the oxidation of l-[1-13C]Phe to 13CO2 (F13CO2). A 2-phase linear-regression crossover analysis of the F13CO2 data identified a breakpoint at minimal F13CO2 Urine samples collected from all study days and from previous studies of healthy children were measured for sulfur metabolites.Results: The mean and population-safe (upper 95% CI) intakes of TSAA and minimum Met in children with CRI were determined to be 12.6 and 15.9 mg · kg-1 · d-1 and 7.3 and 10.9 mg · kg-1 · d-1, respectively. In healthy school-aged children the mean and upper 95% CI intakes of TSAA and minimum Met were determined to be 12.9 and 17.2 mg · kg-1 · d-1 and 5.8 and 7.3 mg · kg-1 · d-1, respectively. A comparison of the minimum Met requirements between healthy children and children with CRI indicated significant (P < 0.05) differences.Conclusion: These results suggest that children with CRI have a similar mean and population-safe TSAA to that of healthy children, suggesting adequate Cys synthesis via transsulfuration, but higher minimum Met requirement, suggesting reduced remethylation rates.

Protein requirement of healthy school-age children determined by the indicator amino acid oxidation method.

BACKGROUND: The current Dietary Reference Intake (DRI) recommendations for protein requirements in children are based on a factorial estimate and have not been directly determined. OBJECTIVE: The objective of the current study was to determine the protein requirement in healthy, school-age children by measuring the oxidation of L-[1-(13)C]-phenylalanine to (13)CO(2) [label tracer oxidation (F(13)CO(2))] in response to graded intakes of protein. DESIGN: Seven healthy children (6-11 y old) each randomly received a minimum of 7 protein intakes (range: 0.1-2.56 g · kg(-1) · d(-1)) for a total of 56 studies. The diets provided energy at 1.7 times the resting energy expenditure and were made isocaloric by using carbohydrate. Protein was given as an amino acid mixture on the basis of the egg-protein pattern, except for phenylalanine and tyrosine intakes, which were maintained constant across intakes. The mean protein requirement was determined by applying a 2-phase linear regression crossover analysis on F(13)CO(2) data, which identified a breakpoint (requirement) at minimal F(13)CO(2) in response to graded amounts of protein intake. RESULTS: Mean and population-safe (upper 95% CI) protein requirements were determined to be 1.3 and 1.55 g · kg(-1) · d(-1), respectively. These results are significantly higher than the mean and population-safe protein requirements currently recommended by the DRI 2005 for macronutrients (0.76 and 0.95 g · kg(-1) · d(-1), respectively). CONCLUSION: To our knowledge, this study was the first to directly estimate protein requirements in children by using stable isotopes and indicated that current recommendations are severely underestimated.

Evidence that protein requirements have been significantly underestimated.

PURPOSE OF REVIEW: This review discusses recent evidence that suggests a significant underestimation of protein requirements in adult humans. RECENT FINDINGS: Traditionally, total protein requirements for humans have been determined using nitrogen balance. The recent Dietary Reference Intake recommendations for mean and population-safe intakes of 0.66 and 0.8 g/kg/day, respectively, of high-quality protein in adult humans are based on a meta-analysis of nitrogen balance studies using single linear regression analysis. We reanalyzed existing nitrogen balance studies using two-phase linear regression analysis and obtained mean and safe protein requirements of 0.91 and 0.99 g/kg/day, respectively. The two-phase linear regression analysis is considered more appropriate for biological analysis of dose-response curves. Considering the inherent problems associated with the nitrogen balance method, we developed an alternative method, the indicator amino acid oxidation technique, to determine protein requirements The mean and population-safe requirements in adult men were determined to be 0.93 and 1.2 g/kg/day and are 41 and 50%, respectively, higher than the current Dietary Reference Intakes recommendations. SUMMARY: The indicator amino acid oxidation-based requirement values of 0.93 and 1.2 g protein/kg/day and the reanalysis of existing nitrogen balance studies are significantly higher than current recommendations. Therefore, there is an urgent need to reassess recommendations for protein intake in adult humans.

Indicator amino acid oxidation is not affected by period of adaptation to a wide range of lysine intake in healthy young men.

The number of days of adaptation to a specific amino acid intake required prior to the determination of amino acid requirements using the indicator amino acid oxidation method (IAAO) is still in debate. In this study, our objective was to determine whether adaptation for 8 h, 3 d, and 7 d to a wide range of lysine intakes had any effect on the oxidation of the indicator amino acid, l-[1-(13)C]phenylalanine, to (13)CO(2) (F(13)CO(2)). Five healthy young men randomly received each of 4 levels of lysine (5, 20, 35, and 70 mg x kg(-1) x d(-1)) along with an amino acid mixture to achieve a protein intake of 1.0 g x kg(-1) x d(-1) and energy intake of 1.5x resting energy expenditure during 4 separate 7-d study periods. IAAO studies were conducted on d 1, 3, and 7. During each study day, oral consumption of l-[1-(13)C]phenylalanine was followed by collection of breath for F(13)CO(2) and plasma for measurement of phenylalanine enrichment. F(13)CO(2) was affected by lysine intake but did not differ among adaptation periods of 8 h, 3 d, or 7 d. Phenylalanine flux was not significantly affected by period of adaptation. These results suggest that the minimally invasive IAAO model, where participants are adapted prior to protein intake for 2 d followed by study day adaptation to the test amino acid intake for 8 h, may be sufficient to estimate individual amino acid requirements in healthy young men.

Reevaluation of the protein requirement in young men with the indicator amino acid oxidation technique.

BACKGROUND: The current estimated protein requirements are based on the nitrogen balance method, which has many limitations. An alternate approach is needed to permit a reevaluation of protein requirements. OBJECTIVE: The objective was to determine protein requirements in men by using the indicator amino acid oxidation technique. DESIGN: Eight healthy men randomly received graded protein intakes (0.10, 0.30, 0.60, 0.90, 1.2, 1.5 and 1.8 g kg(-1) d(-1)) as a crystalline amino acid mixture along with L-[1-(13)C]phenylalanine. The mean protein requirement was determined by applying a biphase linear regression crossover analysis on F(13)CO(2) data, which identified a breakpoint at the minimal rate of appearance of (13)CO(2) to graded protein intakes. RESULTS: The mean and population-safe (recommended dietary allowance; RDA) protein requirements were found to be 0.93 and 1.2 g kg(-1) d(-1), respectively. These requirements are comparable with those estimated by the application of a biphase linear regression model to the data from nitrogen balance studies (0.91 and 1.0 g kg(-1) d(-1), respectively). These requirements are 41% and 50% higher than the current recommendations for the estimated average requirement (EAR) of 0.66 g kg(-1) d(-1) and the RDA of 0.80 g kg(-1) d(-1), as determined by applying a linear regression model where it intersects the zero balance line. CONCLUSION: The indicator amino acid oxidation technique defined a protein requirement that is comparable with that estimated by the application of a biphase linear regression model to nitrogen balance data in the literature. Our data and the reanalysis of the preexisting nitrogen balance data suggest that the current recommended protein requirements are too low and require reassessment.

Lysine requirement of healthy school-age children determined by the indicator amino acid oxidation method.

BACKGROUND: The current Dietary Reference Intake (DRI) recommendations for lysine requirements in children are based on a factorial estimate. OBJECTIVE: The objective of the current study was to determine the lysine requirement in healthy school-age children by measuring the oxidation of l-[1-(13)C]phenylalanine to (13)CO(2) (F(13)CO(2)) in response to graded intakes of lysine. DESIGN: Five healthy school-age children randomly received each of 7 lysine intakes (5, 15, 25, 35 50, 65, and 80 mg x kg(-1) x d(-1)) along with an amino acid mixture to give a final calculated protein intake of 1.5 g x kg(-1) x d(-1) and an energy intake of 1.7 x resting energy expenditure (REE). The mean lysine requirement was determined by applying 2-phase linear regression crossover analysis on F(13)CO(2) data, which identified a breakpoint (requirement) at minimal F(13)CO(2) in response to graded lysine intakes. RESULTS: The mean and population-safe (upper 95% CI) lysine requirements were determined to be 35 and 58 mg x kg(-1) x d(-1), respectively. CONCLUSIONS: The mean and population-safe lysine requirements for children are similar to those for adults (36 and 52 mg x kg(-1) x d(-1), respectively), which suggests that the findings from the current study reflect predominantly the maintenance lysine requirements in children and not all requirements for growth. Therefore, to ensure age-appropriate growth in school-age children, we propose the addition of the requirement of lysine for growth (approximately 6 mg x kg(-1) x d(-1)) to the mean estimate. The new mean and population-safe lysine requirements are 41 and 58 mg x kg(-1) x d(-1), respectively; these values are significantly higher than the current DRIs of 37 and 46 mg x kg(-1) x d(-1), respectively.

Application of the indicator amino acid oxidation technique for the determination of metabolic availability of sulfur amino acids from casein versus soy protein isolate in adult men.

Our objective was to determine the metabolic availability (MA) of sulfur amino acids in dietary proteins using the indicator amino acid oxidation (IAAO) technique. Five to seven men received graded levels (20, 40, 60, and 70%) of the mean total sulfur amino acid (TSAA) requirement of 13 mg x kg(-1) x d(-1) as a crystalline AA mixture, casein, and soy protein isolate (SPI) (40, 50, 60, and 70%), respectively. Five of these subjects received 40% of TSAA requirement from SPI supplemented with methionine to the level of 40% of requirement. These 5 subjects also repeated the level of 60% TSAA requirements from both casein and SPI to assess repeatability. The mean MA of TSAA from SPI (71.8 +/- 3.6%) was lower than from casein (87.4 +/- 3.8%, P < 0.05). Supplementation of SPI with methionine decreased the IAAO (11.5 +/- 0.3% administered dose) compared with unsupplemented SPI (12.8 +/- 0.5% administered dose, P < 0.05). IAAO was similar for repeated measurements of casein and SPI, respectively, at the 60% TSAA intake level (10.8 +/- 1.0 vs. 10.7 +/- 1.2% for casein; 12.7 +/- 1.3 vs. 12.9 +/- 2.6% for SPI). In conclusion, the IAAO technique can be used to determine the MA of AA for protein synthesis in test proteins for humans.

Minimum methionine requirement and cysteine sparing of methionine in healthy school-age children.

BACKGROUND: Cysteine can provide a portion of the sulfur amino acid requirement in adults. Whether this is true in children-and, if so, to what extent-is not known. OBJECTIVES: The objectives were to determine minimum methionine requirements in healthy, school-age children when excess cysteine is provided and to subsequently determine the cysteine-sparing effect by comparing these methionine requirements with those determined previously in the same children when no cysteine was provided. DESIGN: Six healthy, school-age children randomly received graded intakes of methionine (0, 2.5, 5, 7.5, 10, and 15 mg . kg(-1) . d(-1)) along with 21 mg cysteine . kg(-1) . d(-1) in the diet. The mean methionine requirement was determined by using a biphasic linear regression crossover analysis of measurements of the rate of appearance of (13)CO(2) in the breath (F(13)CO(2)), which identified a breakpoint at the minimal F(13)CO(2) in response to graded levels of methionine intake. RESULTS: The mean and population-safe minimum methionine requirements, in the presence of excess dietary cysteine, were found to be 5.8 and 7.3 mg . kg(-1) . d(-1), respectively. The mean and population-safe (upper 95% CI) methionine requirements, in the absence of dietary cysteine, were previously determined to be 12.9 and 17.2 mg . kg(-1) . d(-1), respectively. These values represent a cysteine-sparing effect of 55% and 58% in comparison with mean and population-safe methionine requirements, respectively. CONCLUSION: Excess intake of dietary cysteine results in the reduction in the requirements for methionine to a minimum obligatory requirement level.

Total sulfur amino acid requirement of healthy school-age children as determined by indicator amino acid oxidation technique.

BACKGROUND: Current total sulfur amino acid (TSAA) requirements of children are based on a factorial estimate that involves several assumptions. OBJECTIVE: The objective was to determine the TSAA requirement (methionine alone) of healthy school-age children by measuring the appearance of 13CO2 (F13CO2) in breath after the oxidation of l-[1-13C]phenylalanine in response to graded methionine intakes. DESIGN: Six healthy school-age children randomly received each of 6 methionine intakes (0, 5, 10, 15, 25, and 35 mg.kg(-1).d(-1)) along with an amino acid mixture to give a final protein intake of 1.5 g.kg(-1).d(-1) and an energy intake of 1.7 x resting energy expenditure. The diet was devoid of cysteine. The mean TSAA requirement was determined by applying a biphase linear regression crossover analysis on F13CO2 data, which identified a breakpoint at minimal F13CO2 in response to graded methionine intakes. RESULTS: The mean and population-safe (upper 95% CI) intakes of TSAA (as methionine) were determined to be 12.9 and 17.2 mg.kg(-1).d(-1), respectively. CONCLUSIONS: The current study suggests that children of this age group have a mean TSAA requirement similar to that of adults (12.6 mg.kg(-1).d(-1)). Therefore, it is valid to use a factorial approach, which assumes that maintenance requirements in childhood are similar to adult requirements, to estimate TSAA requirements in school-age children.