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      Ileal digestibility of intrinsically labeled hen's egg and meat protein determined with the dual stable isotope tracer method in Indian adults

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          ABSTRACT

          Background

          Protein quality assessment through the Digestible Indispensable Amino Acid Score requires accurate measurements of true ileal protein and amino acid digestibility, for which a dual isotope technique was recently developed. However, the ileal digestibility of indispensable amino acids (IAA) in humans from high-quality proteins is not well known.

          Objective

          The aim of this study was to intrinsically label hen's egg and meat protein by the use of uniformly 2H-labeled amino acids, and to measure their true ileal indispensable amino acid (IAA) digestibility via the dual isotope method in humans.

          Design

          2H-labeled lyophilized boiled egg white protein, whole boiled egg, and cooked meat were obtained from layer hens (BV-300) administered a uniformly 2H-labeled amino acid mix orally for 35 d with their daily feed. The ileal IAA digestibility of these proteins was determined with reference to digestibility of previously characterized [U- 13C]spirulina in a dual tracer method in healthy Indian subjects whose intestinal health was measured by the plasma kynurenine-to-tryptophan (KT) ratio.

          Results

          All subjects had normal KT ratios. The mean ± SD true ileal IAA digestibility of 2H-labeled egg white protein, whole boiled egg, and cooked meat was 86.3% ± 4.6%, 89.4% ± 4.5%, and 92.0% ± 2.8%, respectively. Leucine digestibility correlated with the KT ratio ( r = −0.772; P = 0.009).

          Conclusions

          Uniformly 2H-labeled hen's egg and meat protein can be used to measure ileal IAA digestibility by the dual isotope tracer approach in humans. The mean IAA digestibility values for these high-quality proteins in the healthy Indians studied were similar to values obtained in earlier human and animal experiments. Leucine digestibility in these meal matrices correlated with the KT ratio, but this aspect needs further evaluation. This trial was registered at the Clinical Trials Registry of India ( http://ctri.nic.in) as CTRI/2018/03/012265.

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          Most cited references33

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          IDO expression by dendritic cells: tolerance and tryptophan catabolism.

          Indoleamine 2,3-dioxygenase (IDO) is an enzyme that degrades the essential amino acid tryptophan. The concept that cells expressing IDO can suppress T-cell responses and promote tolerance is a relatively new paradigm in immunology. Considerable evidence now supports this hypothesis, including studies of mammalian pregnancy, tumour resistance, chronic infections and autoimmune diseases. In this review, we summarize key recent developments and propose a unifying model for the role of IDO in tolerance induction.
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            Plasma Tryptophan and the Kynurenine–Tryptophan Ratio Are Associated with the Acquisition of Statural Growth Deficits and Oral Vaccine Underperformance in Populations with Environmental Enteropathy

            Early childhood enteric infections have adverse impacts on child growth and can inhibit normal mucosal responses to oral vaccines, two critical components of environmental enteropathy. To evaluate the role of indoleamine 2,3-dioxygenase 1 (IDO1) activity and its relationship with these outcomes, we measured tryptophan and the kynurenine–tryptophan ratio (KTR) in two longitudinal birth cohorts with a high prevalence of stunting. Children in rural Peru and Tanzania (N = 494) contributed 1,251 plasma samples at 3, 7, 15, and 24 months of age and monthly anthropometrics from 0 to 36 months of age. Tryptophan concentrations were directly associated with linear growth from 1 to 8 months after biomarker assessment. A 1-SD increase in tryptophan concentration was associated with a gain in length-for-age Z-score (LAZ) of 0.17 over the next 6 months in Peru (95% confidence interval [CI] = 0.11–0.23, P < 0.001) and a gain in LAZ of 0.13 Z-scores in Tanzania (95% CI = 0.03–0.22, P = 0.009). Vaccine responsiveness data were available for Peru only. An increase in kynurenine by 1 μM was associated with a 1.63 (95% CI = 1.13–2.34) increase in the odds of failure to poliovirus type 1, but there was no association with tetanus vaccine response. A KTR of 52 was 76% sensitive and 50% specific in predicting failure of response to serotype 1 of the oral polio vaccine. KTR was associated with systemic markers of inflammation, but also interleukin-10, supporting the association between IDO1 activity and immunotolerance. These results strongly suggest that the activity of IDO1 is implicated in the pathophysiology of environmental enteropathy, and demonstrates the utility of tryptophan and kynurenine as biomarkers for this syndrome, particularly in identifying those at risk for hyporesponsivity to oral vaccines.
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              Digestibility of cooked and raw egg protein in humans as assessed by stable isotope techniques.

              Egg proteins contribute substantially to the daily nitrogen allowances in Western countries and are generally considered to be highly digestible. However, information is lacking on the true ileal digestibility of either raw or cooked egg protein. The recent availability of stable isotope-labeled egg protein allowed determination of the true ileal digestibility of egg protein by means of noninvasive tracer techniques. Five ileostomy patients were studied, once after ingestion of a test meal consisting of 25 g of cooked 13C- and 15N-labeled egg protein, and once after ingestion of the same test meal in raw form. Ileal effluents and breath samples were collected at regular intervals after consumption of the test meal and analyzed for 15N- and 13C-content, respectively. The true ileal digestibility of cooked and raw egg protein amounted to 90.9 +/- 0.8 and 51.3 +/- 9.8%, respectively. A significant negative correlation (r = -0.92, P < 0.001) was found between the 13C-recovery in breath and the recovery of exogenous N in the ileal effluents. In summary, using the 15N-dilution technique we demonstrated that the assimilation of cooked egg protein is efficient, albeit incomplete, and that the true ileal digestibility of egg protein is significantly enhanced by heat-pretreatment. A simple 13C-breath test technique furthermore proved to be a suitable alternative for the evaluation of the true ileal digestibility of egg protein.
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                Author and article information

                Journal
                Am J Clin Nutr
                Am. J. Clin. Nutr
                ajcn
                The American Journal of Clinical Nutrition
                Oxford University Press
                0002-9165
                1938-3207
                November 2018
                01 October 2018
                01 October 2018
                : 108
                : 5
                : 980-987
                Affiliations
                [1 ]Division of Nutrition, St. John's Medical College, St. John's National Academy of Health Sciences, Bangalore, India
                [2 ]Departments of Physiology, St. John's Medical College, St. John's National Academy of Health Sciences, Bangalore, India
                [3 ]Departments of Biostatistics, St. John's Medical College, St. John's National Academy of Health Sciences, Bangalore, India
                [4 ]Indian Council of Agricultural Research-National Institute of Animal Nutrition and Physiology, Adugodi, Bangalore, India
                [5 ]Scottish Universities Environmental Research Centre, East Kilbride, Scotland, United Kingdom
                Author notes
                Address correspondence to AVK (e-mail: a.kurpad@ 123456sjri.res.in ).
                Author information
                http://orcid.org/0000-0001-7940-5454
                http://orcid.org/0000-0002-1786-6076
                http://orcid.org/0000-0002-9900-5682
                http://orcid.org/0000-0002-8344-6897
                http://orcid.org/0000-0001-7998-2438
                Article
                nqy178
                10.1093/ajcn/nqy178
                6250983
                30272112
                9f66bd03-2089-4ea3-bd67-4eab7887a8c9
                © 2018 American Society for Nutrition.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 04 April 2018
                : 04 July 2018
                Page count
                Pages: 8
                Funding
                Funded by: Baylor College of Medicine 10.13039/100007915
                Categories
                Original Research Communications

                Nutrition & Dietetics
                intrinsic labeling,egg protein,meat protein,dual stable isotope tracer method,kynurenine-tryptophan ratio,environmental enteric dysfunction,digestible indispensable amino acid score (diaas)

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