Daily requirement for and splanchnic uptake of leucine in healthy adult Indians.

BACKGROUND: The 1985 FAO/WHO/UNU requirement for leucine is too low according to tracer-derived estimates of leucine oxidation and balance in adults from developed regions. OBJECTIVE: The leucine requirement in populations in developing countries was assessed with use of the 24-h tracer balance method and on the basis of nitrogen balances. DESIGN: Twenty healthy Indian men were studied during their consumption for 6 d of 2 L-amino acid diets that supplied either 14 and 30 (n = 10) or 22 and 40 (n = 10) mg leucine x kg(-1) x d(-1) in random order. At 1800 on day 7, a 24-h constant intravenous [13C]leucine tracer-infusion protocol was conducted to determine leucine oxidation and daily leucine balance. During the intake of 40 mg leucine/d, [2H3]leucine was given orally to assess the splanchnic uptake of leucine. RESULTS: Mean 24-h leucine oxidation rates were 29.8, 30.6, 33.6, and 39.3 mg x kg(-1) x d(-1) at leucine intakes of 14, 22, 30, and 40 mg x kg(-1) x d(-1), respectively; daily leucine balances were -16.5, -9.0, -3.3, and 0.5 mg x kg(-1) x d(-1), respectively. Mixed-models linear regression of balance against leucine intake resulted in a zero balance at a leucine intake of 37.3 mg x kg(-1) x d(-1). Nitrogen balances were -12.7, -17.9, -3.9, and 1.0 mg x kg(-1) x d(-1) at leucine intakes of 14, 22, 30, and 40 mg x kg(-1) x d(-1). Regression of nitrogen balance against intake resulted in a zero balance at a leucine intake of 37.6 mg x kg(-1) x d(-1). The first-pass splanchnic uptake of leucine was 45.7% and 33.9% in the fasted and fed periods, respectively. CONCLUSION: A tentative mean leucine requirement of 40 mg x kg(-1) x d(-1) is proposed for healthy Indian adults, as it is for Western subjects.

Lysine requirements of healthy adult Indian subjects, measured by an indicator amino acid balance technique.

BACKGROUND: In an earlier study, using a modification of the indicator amino acid oxidation approach, we concluded that the 1985 FAO/WHO/UNU-proposed lysine requirement of 12 mg x kg(-1) x d(-1) is likely inadequate to maintain body amino acid homeostasis in apparently healthy south Asian subjects and that our proposed requirement of 30 mg x kg(-1) x d(-1) is more appropriate. OBJECTIVE: We assessed the lysine requirement in a similar population by using 4 test lysine intakes (12, 20, 28, and 36 mg x kg(-1) x d(-1)) with an indicator amino acid balance approach. DESIGN: Sixteen healthy male Indians were studied during each of 2 randomly assigned 8-d L-amino acid diets that supplied either 12 and 28 or 20 and 36 mg lysine. At 1800 on day 8, a 24-h intravenous [(13)C]leucine tracer-infusion protocol was conducted to assess leucine oxidation and daily leucine balance at each lysine intake. RESULTS: Mean 24-h leucine oxidation rates decreased significantly (P = 0.005) across different lysine intakes and were 104.1, 97.8, 87.3, and 87.3 mg x kg(-1) x d(-1) at intakes of 12, 20, 28, and 36 mg x kg(-1) x d(-1), respectively; mean 24-h leucine balances were 3.3, 9.1, 19.7, and 20.7 mg x kg(-1) x d(-1), respectively (P = 0.015, mixed-model analysis of variance). Oxidation and balances differed significantly between the lower and higher lysine intakes but were not significantly different between the 12- and 20-mg and 28- and 36-mg test intakes. Two-phase regression analysis indicated a mean breakpoint at 29 mg lysine x kg(-1) x d(-1) in the relation between lysine intake and leucine oxidation or balance. CONCLUSION: We propose a mean lysine requirement of 30 mg x kg(-1) x d(-1) for healthy Indian adults, which is the same amount we proposed previously for Western populations.

Cysteine metabolism and whole blood glutathione synthesis in septic pediatric patients.

OBJECTIVE: To investigate whole body in vivo cysteine kinetics and its relationship to whole blood glutathione (GSH) synthesis rates in septic, critically ill pediatric patients and controls. DESIGN: Prospective cohort study. SETTING: Multidisciplinary intensive care unit and pediatric inpatient units at a children's hospital. PATIENTS: Ten septic pediatric patients and ten controls (children admitted to the hospital for elective surgery). INTERVENTIONS: Septic patients (age, 31 months to 17 yrs) and controls (age, 24 months to 21 yrs) received a 6-hr primed, constant, intravenous tracer infusion of l-[1-13C]cysteine. Blood samples were obtained to determine isotopic enrichment of plasma cysteine and whole blood [1-13C]cysteinyl-glutathione by gas-chromatography mass spectrometric techniques. The plasma flux and oxidation rate of cysteine and the fractional and absolute synthesis rates of GSH were determined. Septic patients received variable protein and energy intake, as per routine clinical management, and controls were studied in the early postabsorptive state. MEASUREMENTS AND MAIN RESULTS: Plasma cysteine fluxes were increased in the septic patients when compared with the controls (68.2 +/- 17.5 [sd] vs. 48.7 +/- 8.8 micromol x kg(-1) x hr(-1); p <.01), and the fraction of plasma cysteine flux associated with oxidative disposal was similar among the groups. The absolute rates of GSH synthesis in whole blood were decreased (p <.01) in the septic patients (368 +/- 156 vs. 909 +/- 272 micromol x L(-1) x day(-1)). The concentration of whole blood GSH also was decreased in the septic group (665.4 +/- 194 vs. 1059 +/- 334 microM; p <.01) CONCLUSIONS: Whole blood glutathione synthesis rates are decreased, by about 60%, in critically ill septic children receiving limited nutritional support. Plasma cysteine fluxes and concentration of cysteine were increased in the septic patients, suggesting a hypermetabolic state with increased protein breakdown. The mechanisms whereby GSH synthesis rates are decreased in these patients are probably multifactorial, presumably involving an inflammatory response in the presence of limited nutritional support. The role of nutritional modulation and the use of cysteine prodrugs in maintaining GSH concentration and synthesis remain to be established.

Total energy expenditure & physical activity level in chronically energy deficient Indian males measured by the doubly labelled water technique.

Total energy expenditure (TEE) and basal metabolic rate (BMR) in the free-living state were measured in healthy south Indians, by the doubly labelled water (DLW) method. From these measurements, estimates of physical activity level (PAL) could be calculated. A total of 18 subjects were studied in 3 groups of 6 subjects each: Group I (urban controls), Group II (urban slum, chronically undernourished) and Group III (rural). The urban slum recruits were chronically energy deficient (CED), and had a low BMI (average 17.0 kg/m2), while the rural subjects had an average BMI of 18.1 kg/m2. The TEE measured by the doubly labelled water technique was 11.2, 7.1 and 12.2 MJ/day for Groups I, II and III respectively. The TEE was significantly lower in the urban CED group as obtained by the one way ANOVA. The estimates of TEE by the DLW technique were compared with estimates made by whole body calorimetry, in the urban groups of subjects. The TEE obtained by calorimetry was 10.3 +/- 1.6 and 7.3 +/- 0.2 MJ/day in Groups I and II respectively. There were no significant differences between the TEE measured by the two methods, in both the groups. The two methods also correlated well, and the mean difference between the methods, in both groups was -0.5 +/- 1.1 MJ/day, which was about 6 per cent of the value of TEE measured by the DLW method. The calculated PAL (by using TEE measured by DLW/BMR) was 1.79, 1.54 and 1.90 for Groups I, II and III respectively. The results showed that the activity of chronically energy deficient subjects, in urban slums, was reduced, and that this may be their method of conserving energy to maintain a stable body weight. Rural males had a high activity level, even though they had a low BMI.

Nitrogen and amino acid requirements: : the Massachusetts Institute of Technology amino acid requirement pattern.

We review the current international recommendations concerning the protein (nitrogen) and amino acid requirements of healthy individuals, from infancy to the later years of adult life and describe the changes in the recommendations for protein that have been made, since those issued in 1985 by Food and Agriculture Organization/World Health Organization/United Nations University (FAO/WHO/UNU), by the International Dietary Energy Consultative Group. The current international requirements for the specific indispensable amino acids are critiqued briefly, and the rationale and basis for the proposed Massachusetts Institute of Technology (MIT) amino acid requirement pattern are presented. The evidence is then summarized that supports its use in practical considerations of protein nutrition. It is suggested that this MIT amino acid requirement pattern provides the best current estimates of the minimum physiological requirements for the indispensable amino acids in children and adults. It is further concluded that it would be difficult to argue for the continued use of the amino acid requirement values proposed by FAO/WHO/UNU in 1985 in the planning and assessment of dietary protein intakes for population groups worldwide. The MIT amino acid requirement pattern supports and strengthens the relevance of dietary protein quality as an important factor in human protein and amino acid nutrition.

Twenty-four-hour oral tracer studies with L-[1-13C]lysine at a low (15 mg.kg (-1).d (-1) and intermediate (29 mg.kg (-1).d(-1)) lysine intake in healthy adults.

BACKGROUND: We proposed previously that the mean lysine requirement value is approximately 30 mg * kg(-)(1) * d(-)(1) rather than the proposed 1985 FAO/WHO/UNU estimate of the upper range of the requirement, which is 12 mg * kg(-)(1) * d(-)(1). OBJECTIVE: Our objective was to explore the 24-h pattern and rate of whole-body lysine [l-(13)C]oxidation and status of whole-body lysine balance in healthy, young adults given an L-amino acid diet supplying either a low lysine intake (14-15 mg * kg(-)(1) * d(-)(1)) or an intermediate lysine intake (29 mg * kg(-)(1) * d(-)(1)) for 6 d before a continuous tracer study with L-[1-(13)C]lysine. DESIGN: Five subjects received the low lysine intake, 6 subjects received the intermediate intake, and all were studied by using a standard 24-h oral tracer protocol that was described earlier for studies at a generous lysine intake. RESULTS: The rate of lysine oxidation was not significantly different between the 12-h fasted and 12-h fed states. The daily oxidation rate (f1.gif" BORDER="0"> +/- SD) was 27. 9 +/- 8.8 and 27.3 +/- 17.6 mg lysine * kg(-)(1) * d(-)(1) for the low- and intermediate-intake groups, respectively (NS). Daily lysine balance was -12.4 +/- 92 and 1.8 +/- 17.7 mg * kg(-)(1) * d(-)(1), respectively (P < 0.025), for the low and intermediate intakes. The balance was significantly less than zero (P < 0.001) for the low intake. CONCLUSION: The FAO/WHO/UNU lysine requirement value is not sufficient to maintain lysine homeostasis in healthy adults. From the results of this and tracer studies done by others, the mean lysine requirement of healthy adults was determined to be 30 mg * kg(-)(1) * d(-)(1).

Proteins, peptides and amino acids in enteral nutrition: overview and some research challenges.

We have selectively reviewed various aspects of protein and amino acid metabolism and nutrition which we feel are relevant and important in any comprehensive consideration of proteins, peptides, and amino acids in enteral nutrition. These, together with various clinical aspects of the problem that we have not addressed, are covered in more specific detail elsewhere in this volume. Our hope is that by the time this workshop is at an end a reasonable picture of the state of the art will emerge in terms of an understanding of the physiology and biochemistry of protein, peptide, and amino acids in enteral nutrition. Simultaneously, we will need to know, through careful clinical investigation, how this knowledge can best be exploited for optimizing the design of the protein/peptide/amino acid component of enteral nutrition formulations. In doing so it should be possible to achieve a more effective enteral use of proteins, peptides, and amino acids in the comprehensive clinical management of individuals under various pathophysiological states.

Adult human amino acid requirements.

This brief review is concerned with the status of our current understanding of the quantitative needs for dietary amino acids in healthy adults. The nutritional significance of the conditionally indispensable amino acids is assessed and the requirements for the indispensable amino acids are discussed. Recent research involving the use of the tracer balance approach is reviewed, and the importance of resolving the current uncertainties regarding adult human amino acid needs is highlighted.

A minimally invasive tracer protocol is effective for assessing the response of leucine kinetics and oxidation to vaccination in chronically energy-deficient adult males and children.

In disadvantaged populations, recurrent infections lead to a loss of body nitrogen and worsen nutritional status. The resulting malnutrition, in its turn, produces a greater susceptibility to infection. This study aimed to examine the ability of a new minimally invasive tracer protocol to measure leucine oxidation, and then to use it to quantify the effect of vaccination on leucine kinetics and oxidation. Undernourished men (n = 5; body mass index 16.3 +/- 0.9 kg/m(2)) and children (n = 9; age 4.1 +/- 0.6 y; weight-for-age Z-score -2.3 +/- 0.7) underwent metabolic studies 6 d before and 1 d after vaccination with diphtheria, pertussis and tetanus (DPT). The tracer protocol was performed in the fed state and involved two 3-h sequential periods of frequent (20 min) oral doses of NaH(13)CO(3) or [1-(13)C] leucine. Frequent breath samples and urine collections were made. Blood samples were obtained from the men and used for the determination of the isotopic enrichment of alpha-ketoisocaproic acid. The prevaccination oxidation of leucine (percentage of dose +/- SD) was 18.1 +/- 2.3 (men) and 16.7 +/- 3.8 (children). One day after vaccination, these values had risen to 19. 9 +/- 1.9 (P < 0.05) in the men and to 19.5 +/- 4.6 (P < 0.01) in the children. In the adults, vaccination was associated with a rise in whole-body protein breakdown [mg protein/(kg.h)] from 200 +/- 40 to 240 +/- 10 (P < 0.05). A minor simulated infection increases leucine catabolism in undernourished humans and this new, minimally invasive protocol is sufficiently sensitive to measure these changes.

An initial assessment, using 24-h [13C]leucine kinetics, of the lysine requirement of healthy adult Indian subjects.

The international 1985 FAO/WHO/UNU upper dietary requirement for lysine of 12 mg.kg-1.d-1 may be inadequate for healthy Indian adults. To test this, we used a modified indicator amino acid oxidation technique to assess the adequacy of lysine intake of 12 and 28 mg.kg-1.d-1. Seven healthy, male, Indian subjects were studied during each of two randomly assigned 6-d periods while receiving an otherwise adequate diet based on an L-amino acid mixture. Beginning at 1800 on day 6 of the diet, a 24-h infusion protocol in which a [13C]leucine tracer was administered intravenously was used to assess leucine oxidation and daily leucine balance at each test lysine intake. Mean 24-h leucine oxidation was 54.7 compared with 46.9 mg.kg-1.d-1 (P < 0.05) and mean 24-h leucine balances were -4.1 and 3.5 mg.kg-1.d-1 (P < 0.05) for lysine intakes of 12 and 28 mg, respectively. Leucine balances were significantly negative (0.025 < P < 0.05) with the 12-mg lysine intake and not significantly different (P > 0.10) from zero or equilibrium with the 28-mg intake. These findings indicate that the international requirement for lysine appears to be inadequate to maintain body amino acid homeostasis and function in apparently healthy subjects characteristic of the south Asia region. They further indicate that our previously proposed, tentative lysine requirement of 30 mg.kg-1.d-1 is probably adequate for this population.

Prediction of total body water in Indian men from anthropometry and bioelectrical impedance using deuterium dilution as reference.

Total body water (TBW) obtained by deuterium dilution (TBWD) in 45 adult Indian males of low body mass index was compared with values obtained from bioelectrical impedance (TBWImp) and two anthropometry-based prediction equations. Bioelectrical impedance significantly underestimated TBW when compared with deuterium dilution, by 2.75 SD 2.86 kg. Comparisons of the anthropometry-based prediction equations demonstrated that TBW was overestimated by 0.87 SD 2.49 kg, and 2.47 SD 2.57 kg, respectively. Since all parameters in the bioelectrical impedance method were standardized, the difference in the values could have been because of the equation used. Therefore, a fresh equation for the measurement of TBW by the bioelectrical impedance method was derived using the variables of height2/impedance and weight. New prediction equations for TBW were also derived based on anthropometric variables of weight and height. Body weight has the greatest influence in the prediction equations for TBW, and the equation derived was TBW (kg) = 0.533 x body weight (kg) + 3.77, SEE = 2.4 kg. Using height2/impedance gave the relationship: TBW (kg) = 0.568 body weight-0.04 height2/impedance + 4.35, SEE = 1.9 kg.

Body composition by a three compartment model in adult Indian male and female subjects.

The body composition of 10 adult Indian male and female subjects was investigated by a three compartment model, using measurements of Total Body Water (TBW) by deuterium dilution, and of body density by hydrodensitometry. The three compartment model yielded significantly different (P < 0.005) estimates of percent body fat of 15.9+/-3.8 and 19.7+/-4.2% and of the Fat Free Mass (FFM) of 41+/-3.3 kg and 33.9+/-4.1 kg in the male and female subjects respectively. The hydration of the FFM was 0.704+/-0.032 in the males and 0.719+/-0.024 in the females; this difference was not signifcant between groups. The density of the FFM, measured from estimates of percent body fat by the 3 compartment approach and of body density by hydrodensitometry, was 1.107+/-0.014 in the males and 1.101+/-0.001 in the females with no significant differences between the groups. This study demonstrates differences in body composition between BMI matched healthy adult male and female subjects. Although there are significant differences for % Fat and FFM between the sexes, there are no significant differences in the hydration fraction and the density of the FFM.

Measurement of total energy expenditure by the doubly labelled water technique in free living Indians in Bangalore city.

Doubly labelled (2H2(18)O) water was used to determine the daily total energy expenditure (TEE) in the free living state of 6 adult, healthy, weight stable, male volunteers over a period of 21 days. The body weights of the subjects ranged from 42.3-70.4 kg. Isotope pool sizes and elimination rates were calculated from 18O and 2H enrichments in basal and daily (21 days) post dose urine samples using the multipoint slope intercept method after corrections for isotope fractionation. The physical activity level (PAL) of the subjects was also measured during the experiment as the ratio of measured TEE to measured basal metabolic rate (BMR). Simultaneous prediction of the total energy expenditure was also carried out by combining the measurements of BMR by indirect calorimetry, and daily physical activity level by 7 day recall. TEE calculated by the isotopic technique was 9.35 +/- 2.00 MJ/day, with an inter individual variation of 21.4 per cent. The measured BMRs in the subjects along with PALs obtained by recall, gave a total daily energy expenditure of 8.66 +/- 2.20 MJ/day with an inter individual variation of 25.4 per cent. The average BMR was 5.59 +/- 0.99 MJ/day and the average PAL (by recall) was 1.54 +/- 0.12. The inter individual variation of the BMR was 17.7 per cent and that of the recalled PAL was 7.9 per cent; the latter increased to 12.2 per cent when the PAL was calculated from the ratio of the measured TEE to the BMR. There was no significant differences between the methods (isotopic and predicted by BMR), although, the TEE obtained by the isotopic method was higher, by about 0.7 MJ/day, or 7.9 per cent, than the TEE predicted by BMR.

The 13C urea breath test to assess Helicobacter pylori infection in school children.

BACKGROUND: The 13C urea breath test was used in this study to establish it as a diagnostic tool as well as to assess the prevalence of Helicobactor pylori in a group of school children. METHODS AND RESULTS: In a group of 50 children studied, 82% were found to be positive for H. pylori by this test. The influence of diet in modifying the results of the test was also assessed. Relatively small errors were seen if adequate precautions were taken. CONCLUSION: Epidemiological studies are required to further quantify the magnitude of the prevalence of H. pylori in the Indian setting.

Total body water measurements by deuterium dilution in adult Indian males and females.

Total Body Water (TBW) was measured in a group of 20 healthy adult Indian men and 10 women by the deuterated water dilution technique and their body composition was determined by applying a hydration factor of 0.7194 for fat free mass (FFM). The TBW in the male subjects whose mean body weight was 49.8 +/- 6.7 kg, was 60.6 +/- 3.2% of body weight (range 55.8%-65.4%), from which a FFM of 41.9 +/- 6.1 kg (range 31.8 kg-51.3 kg) was obtained. Total body water in the group of 10 female subjects whose mean body weight was 42.7 +/- 4.9 kg, was 57.0 +/- 4.5% of body weight (range 52.5%-64.2%) from which a FFM of 34.0 +/- 5.1 kg (range 28.4 kg-39.4 kg) was obtained.