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Infant formulas (IFs), the sole adequate substitute to human milk, undergo several thermal treatments during production that can damage milk proteins and promote the formation of Maillard reaction products, modifying nutritional and sensory properties. The aim of this study was to determine the impact of a minimally processing route based on membrane filtration associated with different levels of heat treatment, on the odor, taste, texture and color attributes of IFs, then to compare with those of commercial milks. Three experimental IFs (produced with membrane filtration associated with low - T-, medium - T+, or high thermal treatments - T+++) were evaluated. Triangular tests conducted with a panel of 50 adults highlighted clear disparities between all the IFs. The same panel applied the Check-All-That-Apply method to evaluate the IFs: the range of variability between T- and T+++ was similar to that between the 2 commercial IFs, and the sensory characteristics of the experimental IFs were not far from the commercial brands for flavor and texture attributes. Analysis performed on the citation frequencies for each descriptor differentiated T-/T+ from T+++, but all the experimental IFs were described with positive sensory characteristics, unlike one commercial IF. Volatile organic compounds (VOCs) content of IFs with low and high thermal treatments were analyzed. Forty VOCs were identified by gas chromatography-mass spectrometry. T- contained a higher quantity of VOCs than T+++, except for benzaldehyde (Maillard reaction product), and aldehydes (oxidation-related products) were the most represented compounds. In conclusion, the processing was associated with sensory differences among IFs, but no marked difference in flavors was found according to CATA and physicochemical analysis. Additionally, no unpleasant sensory descriptors were noted. This shows that the minimally processed route leads to IFs that could fit well within the market from a sensory point of view.
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BACKGROUND: Infant formulas (IFs), the only adequate substitute to human milk, are complex matrices that require numerous ingredients and processing steps that may impact protein digestion and subsequent amino acid (AA) absorption. OBJECTIVES: The objective was to understand the impact of the protein ingredient quality within IFs on postprandial plasma AA profiles. METHODS: Four isonitrogenous and isocaloric IFs were produced at a semi-industrial scale using whey proteins from different origins (cheese compared with ideal whey) and denaturation levels (IF-A, -B, -C), and caseins with different supramolecular organizations (IF-C, -D). Ten Yucatan minipiglets (12- to 27-d-old) were used as a human infant model and received each IF for 3 d according to a Williams Latin square followed by a 2-d wash-out period. Jugular plasma was regularly sampled from 10 min preprandial to 4 h postprandial on the third day to measure free AAs, urea, insulin, and glucose concentrations. Data were statistically analyzed using a mixed linear model with diet (IFs), time, and sex as fixed factors and piglet as random factor. RESULTS: IFs made with cheese whey (IF-A and -B) elicited significantly higher plasma total and essential AA concentrations than IFs made with ideal whey (IF-C and -D), regardless of the pre- and postprandial times. Most of the differences observed postprandially were explained by AA homeostasis modifications. IFs based on cheese whey induced an increased plasma concentration of Thr due to both a higher Thr content in these IFs and a Thr-limiting degrading capability in piglets. The use of a nonmicellar casein ingredient led to reduced plasma content of AA catabolism markers (IF-D compared with IF-C). CONCLUSIONS: Overall, our results highlight the importance of the protein ingredient quality (composition and structure) within IFs on neonatal plasma AA profiles, which may further impact infant protein metabolism.
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Aminoácidos , Animais Recém-Nascidos , Fórmulas Infantis , Porco Miniatura , Proteínas do Soro do Leite , Animais , Suínos , Aminoácidos/sangue , Fórmulas Infantis/química , Masculino , Feminino , Período Pós-Prandial , Glicemia/análise , Insulina/sangue , Caseínas , Proteínas AlimentaresRESUMO
During industrial processing, heat treatments applied to infant formulas may affect protein digestion. Recently, innovative processing routes have been developed to produce minimally heat-processed infant formula. Our objective was to compare the in vivo protein digestion kinetics and protein quality of a minimally processed (T−) and a heat-treated (T+++) infant formula. Sixty-eight male Wistar rats (21 d) were fed with either a diet containing 40 % T− (n 30) or T+++ (n 30), or a milk protein control diet (n 8) during 2 weeks. T− and T+++ rats were then sequentially euthanised 0, 1, 2, 3 or 6 h (n 6/time point) after ingestion of a meal containing their experimental diet. Control rats were euthanised 6 h after ingestion of a protein-free meal to determine nitrogen and amino acid endogenous losses. Nitrogen and amino acid true caecal digestibility was high for both T− and T+++ diets (> 90 %), but a tendency towards higher nitrogen digestibility was observed for the T− diet (96·6 ± 3·1 %) compared with the T+++ diet (91·9 ± 5·4 %, P = 0·0891). This slightly increased digestibility led to a greater increase in total amino acid concentration in plasma after ingestion of the T− diet (P = 0·0010). Comparable protein quality between the two infant formulas was found with a digestible indispensable amino acid score of 0·8. In conclusion, this study showed that minimal processing routes to produce native infant formula do not modify protein quality but tend to enhance its true nitrogen digestibility and increase postprandial plasma amino acid kinetics in rats.
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Digestão , Guanidinas , Fórmulas Infantis , Humanos , Masculino , Ratos , Animais , Ratos Wistar , Proteínas/metabolismo , Aminoácidos/metabolismo , Dieta , Nitrogênio/metabolismo , Íleo/metabolismo , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição AnimalRESUMO
Protein digestibility, a key indicator of dietary protein quality for human nutrition, can be estimated using an in vitro digestion model, however its definition and determination remain variable across studies. The present study aimed to determine the contribution of the endogenous nitrogen (N) to the plant and animal protein digestibility values obtained in vitro. 15N-labelled gluten and caseins (4, 8 and 16 % of the model meal) were used to differentiate dietary and endogenous N and were digested using the INFOGEST in vitro digestion model with no oral phase. The dietary and endogenous N were measured before and during digestion after centrifugation and 10 kDa ultrafiltration. The proteolysis degree was measured by the OPA method. The endogenous and dietary N were determined by elemental analyser coupled with isotopic ratio mass spectrometry. Apparent and true digestibility were determined and values of 135, 92 and 71 % for apparent vs. 78, 69, 60 % for true digestibility were obtained for 4, 8 and 16 % dietary protein level, respectively, with a significant effect of protein level. Differences between apparent and true digestibility pointed out the important contribution of the endogenous nitrogen. Our results showed that 40 % of the N below 10 kDa, i.e., the digestible fraction, were from endogenous origin (i.e. from the pancreatin) and was even present before digestion. An average value of 27 % for pancreatin N autolysis was estimated independently of the protein levels or sources. The use of 15N-labelled protein to evaluate in vitro protein digestibility highlighted the important contribution of the endogenous N, in particular when low dietary protein solution (4 %) are digested. This gives new keys to overcome drawbacks of in vitro models for determining protein digestibility.
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Aminoácidos , Nitrogênio , Animais , Humanos , Nitrogênio/análise , Aminoácidos/análise , Pancreatina , Digestão , Proteínas Alimentares/metabolismoRESUMO
Infant formula (IF) is a complex matrix requiring numerous ingredients and processing steps. The objective was to understand how the quality of protein ingredients impacts IF structure and, in turn, their kinetics of digestion. Four powdered IFs (A/B/C/D), based on commercial whey protein (WP) ingredients, with different protein denaturation levels and composition (A/B/C), and on caseins with different supramolecular organisations (C/D), were produced at a semi-industrial level after homogenization and spray-drying. Once reconstituted in water (13 %, wt/wt), the IF microstructure was analysed with asymmetrical flow field-flow fractionation coupled with multi-angle light scattering and differential refractometer, transmission electron microscopy and electrophoresis. The rehydrated IFs were subjected to simulated infant in vitro dynamic digestion (DIDGI®). Digesta were regularly sampled to follow structural changes (confocal microscopy, laser-light scattering) and proteolysis (OPA, SDS-PAGE, LC-MS/MS, cation-exchange chromatography). Before digestion, different microstructures were observed among IFs. IF-A, characterized by more denatured WPs, presented star-shaped mixed aggregates, with protein aggregates bounded to casein micelles, themselves adsorbed at the fat droplet interface. Non-micellar caseins, brought by non-micellar casein powder (IF-D) underwent rearrangement and aggregation at the interface of flocculated fat droplets, leading to a largely different microstructure of IF emulsion, with large aggregates of lipids and proteins. During digestion, IF-A more digested (degree of proteolysis + 16 %) at 180 min of intestinal phase than IF-C/D. The modification of the supramolecular organisation of caseins implied different kinetics of peptide release derived from caseins during the gastric phase (more abundant at G80 for IF-D). Bioactive peptide release kinetics were also different during digestion with IF-C presenting a maximal abundance for a large proportion of them. Overall, the present study highlights the importance of the structure and composition of the protein ingredients (WPs and caseins) selected for IF formulation on the final IF structure and, in turn, on proteolysis. Whether it has some physiological consequences remains to be investigated.
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Caseínas , Fórmulas Infantis , Humanos , Caseínas/química , Proteólise , Fórmulas Infantis/química , Cromatografia Líquida , Espectrometria de Massas em Tandem , Peptídeos/metabolismo , DigestãoRESUMO
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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Aminoácidos , Leite Humano , Masculino , Recém-Nascido , Lactente , Feminino , Humanos , Animais , Suínos , Aminoácidos/metabolismo , Leite Humano/química , Fórmulas Infantis/química , Triptofano/metabolismo , Nitrogênio/metabolismo , Digestão/fisiologia , Íleo/metabolismo , Dieta , Dieta com Restrição de Proteínas , Ração Animal/análiseRESUMO
Early nutrition plays a dominant role in infant development and health. It is now understood that the infant diet impacts the gut microbiota and its relationship with gut function and brain development. However, its impact on the microbiota-gut-brain axis has not been studied in an integrative way. The objective here was to evaluate the effects of human milk (HM) or cow's milk based infant formula (IF) on the relationships between gut microbiota and the collective host intestinal-brain axis. Eighteen 10-day-old Yucatan mini-piglets were fed with HM or IF. Intestinal and fecal microbiota composition, intestinal phenotypic parameters, and the expression of genes involved in several gut and brain functions were determined. Unidimensional analyses were performed, followed by multifactorial analyses to evaluate the relationships among all the variables across the microbiota-gut-brain axis. Compared to IF, HM decreased the α-diversity of colonic and fecal microbiota and modified their composition. Piglets fed HM had a significantly higher ileal and colonic paracellular permeability assessed by ex vivo analysis, a lower expression of genes encoding tight junction proteins, and a higher expression of genes encoding pro-inflammatory and anti-inflammatory immune activity. In addition, the expression of genes involved in endocrine function, tryptophan metabolism and nutrient transport was modified mostly in the colon. These diet-induced intestinal modifications were associated with changes in the brain tissue expression of genes encoding the blood-brain barrier, endocrine function and short chain fatty acid receptors, mostly in hypothalamic and striatal areas. The integrative approach underlined specific groups of bacteria (Veillonellaceae, Enterobacteriaceae, Lachnospiraceae, Rikenellaceae, and Prevotellaceae) associated with changes in the gut-brain axis. There is a clear influence of the infant diet, even over a short dietary intervention period, on establishment of the microbiota-gut-brain axis.
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Hydrolysis of proteins leads to the release of bioactive peptides with positive impact on human health. Peptides exhibiting antihypertensive properties (i.e., inhibition of angiotensin-I-converting enzyme) are commonly found in whey protein hydrolysates made with enzymes of animal, plant or microbial origin. However, bioactive properties can be influenced by processing conditions and gastrointestinal digestion. In this study, we evaluated the impact of three plant enzymes (papain, bromelain and ficin) in the manufacture of whey protein hydrolysates with varying level of pH, enzyme-to-substrate ratio and time of hydrolysis, based on a central composite design, to determine the degree of hydrolysis and antihypertensive properties. Hydrolysates made on laboratory scales showed great variation in the type of enzyme used, their concentrations and the pH level of hydrolysis. However, low degrees of hydrolysis in papain and bromelain treatments were associated with increased antihypertensive properties, when compared to ficin. Simulated gastrointestinal digestion performed for selected hydrolysates showed an increase in antihypertensive properties of hydrolysates made with papain and bromelain, which was probably caused by further release of peptides. Several peptides with reported antihypertensive properties were found in all treatments. These results suggest plant enzymes used in this study can be suitable candidates to develop ingredients with bioactive properties.
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Fortification of human milk (HM) is often necessary to meet the nutritional requirements of preterm infants. This study sought to establish whether HM supplemented with an experimental donkey milk-derived fortifier (DMF) or a commercial bovine milk-derived fortifier (BMF) affected digestion, using an in vitro dynamic system at the preterm stage. Particle size in gastric phase was higher in DMF than in BMF, due to protein aggregates surrounding lipid globules. Before digestion, BMF, with its extensively hydrolysed proteins, had a higher degree of proteolysis (30%) than DMF (11%), which contained intact proteins. After digestion, this difference was reduced concomitantly to a similar net degree of proteolysis (33%). DMF, with a higher proportion of ω3, resulted in a lower ω6/ω3 free PUFA ratio than BMF throughout digestion, although the final degree of lipolysis was similar (54%). In summary, DMF could represent a better source of proteins and lipids for the preterm infant.
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Recém-Nascido Prematuro , Leite Humano , Animais , Digestão , Equidae , Alimentos Fortificados , Humanos , Lactente , Recém-Nascido , Lipólise , Leite Humano/química , ProteóliseRESUMO
High Temperature-Short Time (HTST) pasteurization was proposed as an alternative to Holder pasteurization (HOP) to increase the retention of specific human milk (HM) bioactive proteins. The present study explored whether HTST and HOP differently affect peptide release during simulated preterm infant gastrointestinal digestion. Raw (RHM), HOP- and HTST- pasteurized HM were digested using an in vitro dynamic system, and the identified peptides were analyzed by mass spectrometry and multivariate statistics. Before digestion, 158 peptides were identified in either RHM, HTST- or HOP- HM, mostly (84.4%) originating from ß-casein (CASB). During gastric digestion, HOP-HM presented a greater number and more abundant specific CASB peptides. A delayed release of peptides was observed in RHM during the intestinal phase, with respect to both pasteurized HM. Although limited to gastric digestion, the HM peptidomic profile differed according to the pasteurization type, and the pattern of the HTST peptides showed a greater similarity with RHM.
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Leite Humano , Pasteurização , Animais , Digestão , Humanos , Lactente , Recém-Nascido , Recém-Nascido Prematuro , Leite , Peptídeos , TemperaturaRESUMO
Breast milk is the gold standard in neonatal nutrition, but most infants are fed infant formulas in which lipids are usually of plant origin. The addition of dairy lipids and/or milk fat globule membrane extracts in formulas improves their composition with beneficial consequences on protein and lipid digestion. The probiotic Lactobacillus fermentum (Lf) was reported to reduce transit time in rat pups, which may also improve digestion. This study aimed to investigate the effects of the addition of dairy lipids in formulas, with or without Lf, on protein and lipid digestion and on gut physiology and metabolism. Piglets were suckled from postnatal days 2 to 28, with formulas containing either plant lipids (PL), a half-half mixture of plant and dairy lipids (DL), or this mixture supplemented with Lf (DL+Lf). At day 28, piglets were euthanized 90 min after their last feeding. Microstructure of digesta did not differ among formulas. Gastric proteolysis was increased (P < 0.01) in DL and DL+Lf (21.9 ± 2.1 and 22.6 ± 1.3%, respectively) compared with PL (17.3 ± 0.6%) and the residual proportion of gastric intact caseins decreased (p < 0.01) in DL+Lf (5.4 ± 2.5%) compared with PL and DL (10.6 ± 3.1% and 21.8 ± 6.8%, respectively). Peptide diversity in ileum and colon digesta was lower in PL compared to DL and DL+Lf. DL and DL+Lf displayed an increased (p < 0.01) proportion of diacylglycerol/cholesterol in jejunum and ileum digesta compared to PL and tended (p = 0.07) to have lower triglyceride/total lipid ratio in ileum DL+Lf (0.019 ± 0.003) as compared to PL (0.045 ± 0.011). The percentage of endocrine tissue and the number of islets in the pancreas were decreased (p < 0.05) in DL+Lf compared with DL. DL+Lf displayed a beneficial effect on host defenses [increased goblet cell density in jejunum (p < 0.05)] and a trophic effect [increased duodenal (p = 0.09) and jejunal (p < 0.05) weights]. Altogether, our results demonstrate that the addition of dairy lipids and probiotic Lf in infant formula modulated protein and lipid digestion, with consequences on lipid profile and with beneficial, although moderate, physiological effects.
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As plant proteins are increasingly used as a source of amino acids in the diet, studies on in vitro digestion of plant proteins are key to understand the different factors affecting proteolysis, with the ultimate goal of optimising the nutritional composition/intake of plant protein-rich products. More realistic scenarios including the most likely food matrix and physiologically relevant gastrointestinal (GI) conditions should be considered when assessing the in vitro digestion of proteins. The research described here compares the extent of hydrolysis of proteins from peanuts and wheat bread, in particular the vicilin-like 7S globulin (Ara h 1) and gliadin, respectively, with three GI scenarios simulating either infant, early phase adult (fed state) or late phase adult (fasted state) conditions. The digestibility of these proteins, in isolation or when naturally present in the respective food matrix, has been evaluated with SDS-PAGE, LC-MS/MS and a spectrophotometric assay. Results from the food matrices showed lower extent of total protein GI digestion under simulated infant conditions, intermediate behaviour under fed state adult conditions and larger extent under fasted state adult conditions. This was also the case for isolated gliadin. However, isolated Ara h 1 only showed lower extent of proteolysis in the gastric phase under infant conditions, reaching a similar extent to both adult conditions over the course of the intestinal phase. The food matrix seems to have delayed the proteolysis. Choosing an appropriate GI scenario as well as the matrix of the end food product is paramount when assessing in vitro protein digestion.
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Arachis/química , Pão , Digestão , Trato Gastrointestinal/metabolismo , Proteínas de Plantas/química , Adulto , Cromatografia Líquida , Eletroforese em Gel de Poliacrilamida , Gliadina/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Lactente , Proteínas de Armazenamento de Sementes/metabolismo , Espectrometria de Massas em Tandem , Triticum/químicaRESUMO
Heat treatments induce changes in the protein structure in infant milk formulas (IMFs). The present study aims to investigate whether these structural modifications affect protein digestion. Model IMFs (1.3% proteins), with a bovine or a human whey protein profile, were unheated or heated at 67.5 °C or 80 °C to reach 65% of denaturation, resulting in six protein structures. IMFs were submitted to in vitro static gastrointestinal digestion simulating infant conditions. During digestion, laser light scattering was performed to analyze IMF destabilization and SDS-PAGE, OPA assay and cation exchange chromatography were used to monitor proteolysis. Results showed that, during gastric digestion, α-lactalbumin and ß-lactoglobulin were resistant to hydrolysis in a similar manner for all protein structures within IMFs (p > 0.05), while the heat-induced denaturation of lactoferrin significantly increased its susceptibility to hydrolysis. Casein hydrolysis was enhanced when the native casein micelle structure was modified, i.e. partially disintegrated in the presence of lactoferrin or covered by heat-denatured whey proteins. The IMF destabilization at the end of the gastric digestion varied with protein structures, with larger particle size for IMF containing native casein micelles. During intestinal digestion, the kinetics of protein hydrolysis varied with the IMF protein structures, particularly for IMFs containing denatured lactoferrin, exhibiting higher proteolysis degree (67.5 °C and 80 °C vs. unheated) and essential amino acid bioaccessibility (67.5 °C vs. unheated). Overall, the protein structures, generated by modulating the whey protein profile and the heating conditions, impacted the IMF destabilization during the gastric phase and the proteolysis during the entire simulated infant digestion.
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Digestão/efeitos dos fármacos , Temperatura Alta/efeitos adversos , Fórmulas Infantis/química , Proteólise/efeitos dos fármacos , Proteínas do Soro do Leite/farmacocinética , Animais , Caseínas/farmacocinética , Bovinos , Eletroforese em Gel de Poliacrilamida , Humanos , Hidrólise/efeitos dos fármacos , Lactente , Lactalbumina/efeitos dos fármacos , Lactoglobulinas/efeitos dos fármacos , Micelas , Leite/química , Tamanho da Partícula , Desnaturação Proteica/efeitos dos fármacosRESUMO
Donor human milk, pasteurised for safety reasons, is the first alternative for feeding preterm infants when mothers' own milk is unavailable. Breastmilk pasteurisation impact on lipid digestion and absorption was evaluated by a static in vitro digestion model for preterm infants coupled with intestinal absorption using Caco-2/TC7 cells. Lipid absorption was quantified by digital image analysis of lipid droplets, by measurement of basolateral triglyceride concentration and by analysing the expression of major genes involved. After in vitro digestion, lipolysis extent was 13% lower in pasteurised human milk (PHM) than in raw human milk (RHM). In Caco-2/TC7 cells, the number of lipid droplets was identical for both milk types, while the mean droplet area was 17% smaller with PHM. Altogether, pasteurisation decreased the pre-lipolysis of human milk. This initial difference in free fatty acid amount was only partially buffered by the subsequent processes of in vitro digestion and cellular lipid absorption.
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Lipídeos/química , Leite Humano/química , Linhagem Celular , Digestão , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Mucosa Intestinal , Intestinos , Lipólise , PasteurizaçãoRESUMO
The high-temperature short-time (HTST, 72 °C, 15 s) pasteurization of human milk (HM) has been proposed as an alternative to the Holder method (HoP, 62.5 °C, 30 min), to increase the preservation of bioactive compounds. We have investigated the impact of HTST and HoP pasteurization on the gastrointestinal kinetics of human milk, using a dynamic in vitro system in a preterm newborn model. An increased protein aggregation on the surface of fat globules following pasteurization, albeit to a lesser extent in HTST than in HoP, was observed. Despite relevant differences in the undigested milk samples, both pasteurization methods led to similar proteolytic patterns, while raw HM presented a higher native lactoferrin content throughout digestion. The slightly decreased amino acid release following HoP, with respect to HTST and raw HM, indicated that peptidomic analysis, which is currently underway, might provide interesting insights on the differential digestive kinetics of differently pasteurized HM.
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Leite Humano/química , Pasteurização/métodos , Cromatografia Líquida de Alta Pressão , Temperatura Alta , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Lactoferrina/química , Espectrometria de Massas , Proteólise , Fatores de TempoRESUMO
Evaluating the gastrointestinal (GI) fate of proteins is part of the assessment to determine whether proteins are safe to consume. In vitro digestion tests are often used for screening purposes in the evaluation of potential allergenicity. However, the current pepsin resistant test used by the European Food Safety Authority, only corresponds to fasted gastric conditions representative of a late phase adult stomach. In addition, these tests are performed on isolated proteins and the effect of the food matrix and processing are not systematically considered. The aim of this research is to compare three different static in vitro GI scenarios that are physiologically relevant. Namely, an infant, early phase (fed state) adult and late phase (fasted state) adult model. These protocols are applied to well-characterised isolated dairy (ß-lactoglobulin and ß-casein) and egg (lysozyme and ovalbumin) proteins and the impact of food matrix/processing on their proteolysis is also investigated. A combination of SDS-PAGE, LC-MS/MS and spectrophotometric assay was used for the evaluation of the proteolysis. Results highlight differences across the three GI scenarios whether on isolated proteins or within food matrices. The infant model led to incomplete digestion, leaving intact egg proteins, either isolated or in the food matrix, and intact ß-lactoglobulin in the milk. In addition, peptides greater than 9 amino acids were found throughout the intestinal phase for all proteins studied, regardless of the scenario. This reinforces the difficulty of linking protein digestibility to potential allergenicity because many other factors are involved that need further investigation.
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Digestão , Proteínas Dietéticas do Ovo/metabolismo , Trato Gastrointestinal/metabolismo , Proteínas do Leite/metabolismo , Peptídeos/metabolismo , Adulto , Caseínas/metabolismo , Hipersensibilidade a Ovo , Jejum , Humanos , Técnicas In Vitro , Lactente , Lactoglobulinas/metabolismo , Hipersensibilidade a Leite , Modelos Biológicos , Muramidase/metabolismo , Ovalbumina/metabolismo , Pepsina A/metabolismo , ProteóliseRESUMO
Infant formulas (IFs) are used as substitutes for human milk and are mostly based on cow milk proteins. For sustainability reasons, animal protein alternatives in food are increasingly being considered, as plant proteins offer interesting nutritional and functional benefits for the development of innovative IFs. This study aimed to assess how a partial substitution (50%) of dairy proteins with faba bean and pea proteins influenced the digestibility of IFs under simulated dynamic in vitro digestion, which were set up to mimic infant digestion. Pea- and faba bean-based IFs (PIF and FIF, respectively) have led to a faster aggregation than the reference milk-based IF (RIF) in the gastric compartment; that did not affect the digesta microstructure at the end of digestion. The extent of proteolysis was estimated via the hydrolysis degree, which was the highest for FIF (73%) and the lowest for RIF (50%). Finally, it was apparent that in vitro protein digestibility and protein digestibility-corrected amino acid score (PDCAAS)-like scores were similar for RIF and FIF (90% digestibility; 75% PDCAAS), but lower for PIF (75%; 67%). Therefore, this study confirms that faba bean proteins could be a good candidate for partial substitution of whey proteins in IFs from a nutritional point of view, provided that these in vitro results are confirmed in vivo.
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Infant formulas (IFs) are the key nutritional source for infants who cannot be breastfed. There is currently a growing interest in these sensitive products in order to control their quality and to design their composition with regard to nutritional balance. In a context of sustainable development and increasing growth of the world population, it seems essential to search for alternative to animal protein in food today. Plant proteins offer interesting nutritional and functional benefits thanks to the latest improvement through research and development. In this context, five model IFs were developed with identical composition, except that 50% of the proteins were either whey proteins in the "milk-reference IF", pea, faba bean, rice or potato proteins in the four "plant IFs" tested. The IFs were evaluated using an in vitro static gastro-intestinal model simulating infant conditions. The protein hydrolysis degree (DH) and the amino acid bioaccessibility (AAB) were used as indicators of protein digestibility. Results showed that both DH and AAB were very similar between the milk-reference IF, pea and faba bean IFs, but significantly lower for the rice and potato IFs. This study provides new insights into the impact of protein sources on IF digestibility.
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Digestão , Proteínas de Plantas/metabolismo , Humanos , Técnicas In Vitro , Lactente , Fórmulas Infantis/químicaRESUMO
BACKGROUND: Endogenous amino acids (AAs) contribute to terminal ileal digesta and must be corrected for in determining coefficients of true ileal digestibility. Such estimates are also needed for the factorial calculation of AA requirements. OBJECTIVE: The effect of the form of delivery of dietary AAs on endogenous nitrogen and AA flows at the end of the ileum was studied. METHODS: Isotopically labeled 15N-casein (33-34 g), in either a hydrolyzed (HC) or intact (C) form, was included as the sole source of nitrogen in a mixed meal (320 mmol N) consumed by healthy adult humans equipped with a triple-lumen sampling tube in the small intestine. Ileal endogenous AA flows were determined by isotope dilution. An additional meal (A) containing a free AA mixture (306 mmol N) simulating the AA composition of casein was included. Serine was omitted from the AA mixture to allow direct determination of its ileal endogenous flow. RESULTS: Endogenous N and AA flows did not differ (P > 0.05) for diets C and HC, with mean respective N flows of 728 and 617 mg/8 h (± pooled SD: 144 mg/8 h). Endogenous serine flow was similar (P > 0.05) for diets C, HC, and A [181, 169, and 191 mg/8 h (± 56 mg/8 h)]. Recycling of the 15N marker was determined to be â¼11%, suggesting that the 15N endogenous loss values could underestimate endogenous protein and AA losses by â¼6% (the proportion of recycled 15N divided by the sum of endogenous N and recycled 15N). CONCLUSIONS: The 3 different forms of dietary AA delivery (free AAs, HC, or C) elicited similar ileal endogenous AA flows in the adult human.This study was registered at clinicaltrials.gov as NCT00873951.
Assuntos
Aminoácidos/metabolismo , Caseínas/metabolismo , Íleo/metabolismo , Adulto , Animais , Caseínas/química , Proteínas Alimentares/metabolismo , Digestão , Feminino , Humanos , Hidrólise , Masculino , Adulto JovemRESUMO
Milk fat globule membrane conditions the reactivity and enzymatic susceptibility of milk lipids. The use of bovine membrane extracts to make infant formulas more biomimetic of human milk has been suggested recently. A comparison of the physico-chemical behavior of human and bovine milk membrane extracts and their interaction with gastric lipase is here undertaken using biophysical tools. Milk membrane extracts (70% of polar lipids) were obtained either pooling of mature human milk (n = 5) or bovine buttermilk. Human extract contained more anionic glycerophospholipids, less phosphatidylethanolamine and more unsaturated fatty acids (57% versus 46%) than bovine extract. Human extract presented a higher compressibility, with slower increase of surface pressure, than bovine extract. Micronic liquid condensed (LC) domains were evidenced in both extracts at 10 mN/m, but the evolution differs upon compression. Upon gastric lipase addition, an adsorption preference for liquid expanded phase (LE) was observed for both extracts. However, insertion was more homogeneous in terms of height level in human extract and impacted less its lipid lateral organization than in bovine extract. Both membrane extracts share close physico-chemical properties, however human membrane higher compressibility may favour gastric lipase insertion and higher interfacial reactivity in gastric conditions.