Journal Articles

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    The kinetics of amino acid disappearance in the small intestine is related to the extent of amino acids absorbed in growing pigs
    (Cambridge University Press on behalf of The Nutrition Society, 2024-03-14) Montoya CA; van Bemmel M; Kreutz K; Hodgkinson SM; Stroebinger N; Moughan PJ
    This study evaluated the importance of a correction for amino acids (AA) released into the hindgut on a measure of AA absorption kinetics and tested whether AA absorption kinetics are related to the extent of AA absorption using the growing pig as a model for humans. Thirty-six nine-week-old pigs (22·3 kg) received a diet containing whey protein as the sole protein source for 8 d. Pigs received their last meal containing the indigestible marker titanium dioxide before being euthanised at 1, 2, 3, 4, 6 and 12 h post-feeding. The entire content of each gastrointestinal tract (GIT) region was collected to determine AA released into the hindgut, and the kinetics and extent of AA absorption (uncorrected and corrected for AA entering the hindgut). Amounts of AA released into the hindgut increased over time (e.g. 33 and 180 mg of Glu for 4 and 6 h post-feeding). The corrected apparent amount of each AA absorbed from the GIT lumen after 4 h post-feeding was generally lower (P ≤ 0·05) than the uncorrected counterpart. Differences in both the kinetics and extent of AA absorption were observed across AA. For example, the time to reach half of the apparent AA absorption (T50) was 1·5 and 3·4 h for Met and Arg, respectively, whereas their extent of apparent absorption was 93 and 73 %. Negative correlations between parameters related to kinetics and the extent of apparent absorption were observed (e.g. for T50 r = -0·81; P < 0·001). The kinetics of AA absorption is related to the extent of AA absorption.
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    Ileal Digestibility of Nitrogen and Amino Acids in Human Milk and an Infant Formula as Determined in Neonatal Minipiglets
    (Elsevier Inc. on behalf of American Society for Nutrition, 2023-04) Charton E; Henry G; Cahu A; Le Gouar Y; Dahirel P; Moughan PJ; Montoya CA; Bellanger A; Dupont D; Le Huërou-Luron I; Deglaire A
    BACKGROUND: Infant formula (IF) has to provide at least the same amount of amino acids (AAs) as human milk (HM). AA digestibility in HM and IF was not studied extensively, with no data available for tryptophan digestibility. OBJECTIVES: The present study aimed to measure the true ileal digestibility (TID) of total nitrogen and AAs in HM and IF to estimate AA bioavailability using Yucatan mini-piglets as an infant model. METHODS: Twenty-four 19-day-old piglets (males and females) received either HM or IF for 6 days or a protein-free diet for 3 days, with cobalt-EDTA as an indigestible marker. Diets were fed hourly over 6 h before euthanasia and digesta collection. Total N, AA, and marker contents in diets and digesta were measured to determine the TID. Unidimensional statistical analyses were conducted. RESULTS: Dietary N content was not different between HM and IF, while true protein was lower in HM (-4 g/L) due to a 7-fold higher non-protein N content in HM. The TID of total N was lower (P < 0.001) for HM (91.3 ± 1.24%) than for IF (98.0 ± 0.810%), while the TID of amino acid nitrogen (AAN) was not different (average of 97.4 ± 0.655%, P = 0.272). HM and IF had similar (P > 0.05) TID for most of the AAs including tryptophan (96.7 ± 0.950%, P = 0.079), except for some AAs (lysine, phenylalanine, threonine, valine, alanine, proline, and serine), with small significant difference (P < 0.05). The first limiting AA was the aromatic AAs, and the digestible indispensable AA score (DIAAS) was higher for HM (DIAASHM = 101) than for IF (DIAASIF = 83). CONCLUSION: HM, compared to IF, had a lower TID for total N only, whereas the TID of AAN and most AAs, including Trp, was high and similar. A larger proportion of non-protein N is transferred to the microbiota with HM, which is of physiological relevance, although this fraction is poorly considered for IF manufacturing.
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    Presence of Unabsorbed Free Amino Acids at the End of the Small Intestine Indicates the Potential for an Increase in Amino Acid Uptake in Humans and Pigs
    (Elsevier Inc on behalf of American Society for Nutrition, 2023-03) van der Wielen N; de Vries S; Gerrits WJ; Lammers-Jannink K; Moughan PJ; Mensink M; Hendriks W
    BACKGROUND: Unabsorbed free amino acids (AAs) at the end of the small intestine result in a potential preventable nutritional loss. OBJECTIVES: This study aimed to quantify free AAs in terminal ileal digesta of both humans and pigs to investigate its relevance for the nutritional value of food proteins. METHODS: Two studies with three diets were performed: a human study-ileal digesta from eight adult ileostomates were collected over 9 h after ingestion of a single meal unsupplemented or supplemented with 30 g zein or whey; pig study-12 cannulated pigs were fed for 7 d with a diet containing whey or zein or no-protein diet, and ileal digesta were collected on the last 2 d. Digesta were analyzed for total and 13 free AAs. True ileal digestibility (TID) of AAs was compared with and without free AAs. RESULTS: All terminal ileal digesta samples contained free AAs. The TID of AAs in whey was 97% ± 2.4% (mean ± SD) in human ileostomates and 97% ± 1.9% in growing pigs. If the analyzed free AAs would have been absorbed, TID of whey would increase by 0.4%-units in humans and 0.1%-units in pigs. The TID of AAs in zein was 70% ± 16.4% in humans and 77% ± 20.6% in pigs and would increase by 2.3%-units and 3.5%-units, respectively, if the analyzed free AAs would have been fully absorbed. The largest difference was observed for threonine from zein: if free threonine was absorbed, the TID would increase by 6.6%-units in both species (P < 0.05). CONCLUSIONS: Free AAs are present at the end of the small intestine and can potentially have a nutritionally relevant effect for poorly digestible protein sources, whereas the effect is negligible for highly digestible protein sources. This result provides insight into the room for improvement of a protein's nutritional value if all free AAs are to be absorbed. J Nutr 2023;xx:xx-xx. This trial was registered at clinicaltrials.gov as NCT04207372.
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    In vitro digestibility of dietary proteins and in vitro DIAAS analytical workflow based on the INFOGEST static protocol and its validation with in vivo data
    (Elsevier Ltd, 2023-03-15) Sousa R; Recio I; Heimo D; Dubois S; Moughan PJ; Hodgkinson SM; Portmann R; Egger L
    The FAO recommends the digestible indispensable amino acid score (DIAAS) to determine protein quality in foods, preferably tested in vivo. Here, the INFOGESTin vitrodigestion protocol was applied and supplemented with an analytical workflow allowing the assessment of protein digestibility and DIAAS calculation. The protocol was applied to selected samples WPI, zein, collagen, black beans, pigeon peas, All-Bran®, and peanuts. The total protein digestibility, digestibility of individual amino acids (AA), and DIAAS values were established and compared with in vivo data for the same substrates. Total protein digestibility (total Nitrogen, r = 0.7, P < 0.05; primary amines (OPA), r = 0.6, P < 0.02; total AA, r = 0.6, P < 0.02) and digestibility of individual AA (r = 0.6, P < 0.0001) were in good agreement, between in vitro and in vivo, with a mean difference of 1.2 %. In vitro DIAAS was highly correlated with DIAAS obtained from in vivo true ileal digestibility values (r = 0.96, R2 = 0.89, P < 0.0001) with a mean difference of 0.1 %.
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    Comparison of True Ileal Amino Acid Digestibility between Adult Humans and Growing Pigs
    (Oxford University Press on behalf of the American Society for Nutrition, 2022-07) Hodgkinson SM; Stroebinger N; van der Wielen N; Mensink M; Montoya C; Hendriks WH; de Vries S; Stein HH; Moughan PJ
    BACKGROUND: It is not feasible to determine the true ileal amino acid (AA) digestibility of protein sources in humans on a routine basis, and the growing pig has been recommended as an animal model for this purpose but requires further validation. OBJECTIVES: To determine and compare true ileal AA digestibility between adult human ileostomates and growing cannulated pigs for a range of food proteins. METHODS: Seven protein sources (black beans, bread, collagen, pigeon peas, wheat bran, whey protein isolate, and zein) that spanned the range of digestibilities typically seen in foods were evaluated. Six female growing pigs received each of the protein sources, as well as a protein-free diet, and digesta were collected via ileal T-cannula. Adult human ileostomates consumed the same protein sources (5-8 ileostomates, depending on the protein source), as well as a protein-free diet, and digesta were collected. Titanium dioxide and celite were included in the diets as indigestible markers. True ileal AA digestibility coefficients were determined. RESULTS: There was a significant effect of protein source (P ≤ 0.001) for all AAs. The effect of species was not significant (P > 0.05) except for total lysine (but not for available lysine). When analyzed within diets, the statistically significant species effect for true lysine digestibility was found for black beans only. Pig and human digestibility values were generally highly and significantly (P ≤ 0.05) correlated. A linear regression equation derived for true ileal AA digestibility (given as coefficients) determined in the human and pig for the overall mean of all AAs was (y = human, x = pig) y = 1.00x - 0.010, with the slope not statistically significant (P > 0.05) from unity and the intercept not different (P > 0.05) from zero. CONCLUSIONS: True ileal AA digestibility values determined in the growing pig can be directly used for predicting digestibility in adult humans.
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    Gut luminal endogenous protein: Implications for the determination of ileal amino acid digestibility in humans
    (Cambridge University Press, 2012) Moughan PJ; Rutherfurd S
    The true ileal digestibility assay provides the most informative measure of digestibility to assess bioavailability of amino acids in foods for humans. To determine ‘true’ estimates of ileal amino acid digestibility, requires that endogenous amino acids present in digesta at the terminal ileum be quantified. The amounts of endogenous amino acids in ileal digesta can be determined after feeding an animal or human a protein-free diet (traditional approach) or by various methods after giving a protein-containing diet. When the protein-free method has been applied with adult human subjects an overall mean value (three separate studies) for endogenous ileal nitrogen flow of 800 mg N/d has been reported. This value is considerably lower than a comparable value obtained after feeding protein of 1852 mg N/d (mean of four separate studies), and thus endogenous ileal N and amino acids should be measured under conditions of protein alimentation. There is some confusion concerning the terminology used to define digestibility, with the term “true” digestibility having different adopted meanings. Here, true amino acid digestibility is defined as apparent amino acid digestibility corrected for the basal amino acid losses determined after giving either a protein-free or a protein-containing diet. Basal losses should be determined at a defined dry-matter and protein intake. The protein-free diet approach to determining endogenous amino acids is considered unphysiological and basal losses refer to ileal endogenous amino acid flows associated with digesta dry-matter flow, and not including “specific” effects of dietary factors such as non starch polysaccharides and anti nutritional factors. Arguments are advanced that the enzyme hydrolysed protein/ultra filtration method may be suitable for routine application with a cannulated pig model, to obtain physiologically-valid basal estimates of ileal endogenous amino acids to allow calculation of true ileal amino acid digestibility in the pig, and then prediction (via statistical relationships) of true coefficients of amino acid digestibility in humans.