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1.
Biochimie ; 215: 12-23, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37062468

RESUMO

Pancreatic lipase related-protein 2 (PLRP2) exhibits remarkable galactolipase and phospholipase A1 activities, which depend greatly on the supramolecular organization of the substrates and the presence of surfactant molecules such as bile salts. The objective of the study was to understand the modulation of the adsorption mechanisms and enzymatic activity of Guinea pig PLRP2 (gPLRP2), by the physical environment of the enzyme and the physical state of its substrate. Langmuir monolayers were used to reproduce homogeneous and heterogeneous photosynthetic model membranes containing galactolipids (GL), and/or phospholipids (PL), and/or phytosterols (pS), presenting uncharged or charged interfaces. The same lipid mixtures were also used to form micrometric liposomes, and their gPLRP2 catalyzed digestion kinetics were investigated in presence or in absence of bile salts (NaTDC) during static in vitro, so called "bulk", digestion. The enzymatic activity of gPLRP2 onto the galactolipid-based monolayers was characterized with an optimum activity at 15 mN/m, in the absence of bile salts. gPLRP2 showed enhanced adsorption onto biomimetic model monolayer containing negatively charged lipids. However, the compositional complexity in the heterogeneous uncharged model systems induced a lag phase before the initiation of lipolysis. In bulk, no enzymatic activity could be demonstrated on GL-based liposomes in the absence of bile salts, probably due to the high lateral pressure of the lipid bilayers. In the presence of NaTDC (4 mM), however, gPLRP2 showed both high galactolipase and moderate phospholipase A1 activities on liposomes, probably due to a decrease in packing and lateral pressure upon NaTDC adsorption, and subsequent disruption of liposomes.


Assuntos
Lipase , Lipossomos , Animais , Cobaias , Hidrólise , Fosfolipases A1 , Adsorção , Lipase/química , Fosfolipídeos/metabolismo , Galactolipídeos , Ácidos e Sais Biliares
2.
Crit Rev Food Sci Nutr ; 63(20): 4655-4674, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-34839771

RESUMO

Most lipids in our diet come under the form of triacylglycerols that are often redispersed and stabilized by surfactants in processed foods. In plant however, lipid assemblies constitute interesting sources of natural bioactive and functional ingredients. In most photosynthetic sources, polar lipids rich in ω3 fatty acids are concentrated. The objective of this review is to summarize all the knowledge about the physico-chemical composition, digestive behavior and oxidative stability of plant polar lipid assemblies to emphasize their potential as functional ingredients in human diet and their potentialities to substitute artificial surfactants/antioxidants. The specific composition of plant membrane assemblies is detailed, including plasma membranes, oil bodies, and chloroplast; emphasizing its concentration in phospholipids, galactolipids, peculiar proteins, and phenolic compounds. These molecular species are hydrolyzed by specific digestive enzymes in the human gastrointestinal tract and reduced the hydrolysis of triacylglycerols and their subsequent absorption. Galactolipids specifically can activate ileal break and intrinsically present an antioxidant (AO) activity and metal chelating activity. In addition, their natural association with phenolic compounds and their physical state (Lα state of digalactosyldiacylglycerols) in membrane assemblies can enhance their stability to oxidation. All these elements make plant membrane molecules and assemblies very promising components with a wide range of potential applications to vectorize ω3 polyunsaturated fatty acids, and equilibrate human diet.


Assuntos
Galactolipídeos , Fosfolipídeos , Humanos , Galactolipídeos/metabolismo , Triglicerídeos/metabolismo , Oxirredução , Antioxidantes/metabolismo , Estresse Oxidativo
3.
Colloids Surf B Biointerfaces ; 217: 112646, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35763897

RESUMO

The structural behavior of model assemblies composed of monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG), the two main galactolipids found in plants, was investigated at the air/water interface and in aqueous dispersion. To approach the composition of the natural photosynthetic membranes, tunable Langmuir model membrane of galactolipids (GL) were used, and were complexified to form either heterogenous binary or ternary assemblies of GL, phospholipids (PL), and phytosterols (pS). The impact of pS, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) or both on the structural properties of GL membrane was studied. The nature of the interactions between the different molecules was investigated using biophysical characterizations (ellipsometry, tensiometry, atomic force microscopy). In addition, the phase behavior was determined by SAXS analysis on the model assemblies in aqueous dispersions. Results revealed the good interfacial stability of these specific plant membrane lipids. The morphology of the GL film was characteristic of a fluid phase, with an interfacial roughness induced by the intercalation of monogalactosyl and digalactosyl polar heads of MGDG and DGDG, respectively. A phase heterogeneity in the monolayer was induced by the addition of DPPC and/or pS, which resulted in the modification of galactolipid organization and headgroup interactions. These structural changes were confirmed by SAXS analysis, showing more favorable interactions between MGDG and DPPC than between DGDG and DPPC in aqueous dispersion. This phenomenon was exacerbated in the presence of pS.


Assuntos
Galactolipídeos , Fitosteróis , Galactolipídeos/química , Plantas , Espalhamento a Baixo Ângulo , Água , Difração de Raios X
4.
Food Chem ; 360: 129880, 2021 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-33989883

RESUMO

Oil bodies (OB), the form of triacylglycerol storage in seeds, are interesting natural assemblies for nutritional applications. In walnuts, OB contain an important amount of polyunsaturated fatty acids that could be interesting food ingredients but may be prone to oxidation. The oxidative and interfacial behavior of walnut OB, either minimally-processed or after processing, were compared with processed complex walnut juice. The good oxidative stability of minimally-processed OB over 10 days (PV ≤ 8.4 meq O2/kg, TBARS = 1.4 mmol eq MDA/kg) and of processed walnut complex matrixes over 20 days (PV ≤ 4.8 meq O2/kg, TBARS = 1.4 mmol eq MDA/kg) was evidenced. In comparison, processing of OB promoted their oxidation. The interfacial studies led to the proposition of a new model of adsorption for minimally-processed OB that will be useful to design functional emulsion or foam in which OB act as emulsifiers.


Assuntos
Juglans/química , Gotículas Lipídicas/química , Óleos de Plantas/química , Adsorção , Emulsões , Nozes/química , Oxirredução , Água/química
5.
Foods ; 10(1)2020 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-33375011

RESUMO

The aim of the present study was to determine to what extent the food matrix could affect the release of docosahexaenoic acid (DHA) during digestion and its incorporation into systemic circulation. In this aim, three DHA-enriched egg products having the same composition but different structure were developed: omelet, hard-boiled egg, and mousse. Then, nine pigs fitted with T-shape cannulas at duodenal level and a jugular venous catheter were fed with the DHA-enriched egg products, and duodenal effluents and plasma were collected throughout the postprandial period. Results highlighted an undeniable effect of the food matrix on digestion parameters and DHA bioavailability. The transit of DHA and protein through the duodenum was faster after the ingestion of the mousse than after the ingestion of the omelet and hard-boiled egg. While most of the DHA and protein ingested under the form of mousse had already passed through the duodenum 4.5 h after its ingestion, significantly higher quantities were still present in the case of the omelet and hard-boiled egg. In terms of bioavailability, the omelet was the most efficient vector for delivering DHA into systemic circulation. It supplied 56% and 120% more DHA than the hard-boiled egg and the mousse, respectively.

6.
Curr Opin Clin Nutr Metab Care ; 23(6): 451-459, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32889824

RESUMO

PURPOSE OF REVIEW: The impact of dietary lipids on cardiometabolic health was mainly studied considering their fatty acid composition. This review aims to present the recent change in paradigm whereby the food matrix, the molecular and supramolecular structures of dietary lipids modulate their digestive fate and cardiometabolic impact. RECENT FINDINGS: Epidemiological studies have reported that the metabolic impact of full-fat dairy products is better than predictable upon saturated fatty acid richness. Milk polar lipid supplementation reduced adiposity and inflammation in rodents by modulating gut microbiota and barrier, and decreased lipid markers of cardiovascular disease risk in humans by lowering cholesterol absorption. The metabolic importance of the structure of lipid molecules carrying omega-3 (molecular carrier) has also been documented. Plant lipids exhibit specific assemblies, membrane and molecular structures with potential health benefits. Lipid emulsifiers used to stabilize fats in processed foods are not mere bystanders of lipid effects and can induce both beneficial and adverse health effects. SUMMARY: These findings open new clinical research questions aiming to further characterize the cardiometabolic fate of lipids, from digestion to bioactive metabolites, according to the food source or molecular carrier. This should be useful to elaborate food formulations for target populations and personalized dietary recommendations.


Assuntos
Gorduras na Dieta/farmacocinética , Lipídeos/farmacocinética , Síndrome Metabólica/metabolismo , Fenômenos Fisiológicos da Nutrição/efeitos dos fármacos , Animais , Disponibilidade Biológica , Fatores de Risco Cardiometabólico , Digestão/efeitos dos fármacos , Absorção Gastrointestinal/efeitos dos fármacos , Microbioma Gastrointestinal/efeitos dos fármacos , Humanos , Metabolismo dos Lipídeos , Síndrome Metabólica/etiologia , Relação Estrutura-Atividade
7.
Biotechnol Adv ; 41: 107546, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32275940

RESUMO

Enzymes are essential and ubiquitous biocatalysts involved in various metabolic pathways and used in many industrial processes. Here, we reframe enzymes not just as biocatalysts transforming bioproducts but also as sensitive probes for exploring the structure and composition of complex bioproducts, like meat tissue, dairy products and plant materials, in both food and non-food bioprocesses. This review details the global strategy and presents the most recent investigations to prepare and use enzymes as relevant probes, with a focus on glycoside-hydrolases involved in plant deconstruction and proteases and lipases involved in food digestion. First, to expand the enzyme repertoire to fit bioproduct complexity, novel enzymes are mined from biodiversity and can be artificially engineered. Enzymes are further characterized by exploring sequence/structure/dynamics/function relationships together with the environmental factors influencing enzyme interactions with their substrates. Then, the most advanced experimental and theoretical approaches developed for exploring bioproducts at various scales (from nanometer to millimeter) using active and inactive enzymes as probes are illustrated. Overall, combining multimodal and multiscale approaches brings a better understanding of native-form or transformed bioproduct architecture and composition, and paves the way to mainstream the use of enzymes as probes.


Assuntos
Biodiversidade , Lipase , Enzimas , Glicosídeo Hidrolases , Plantas
8.
Biochimie ; 169: 95-105, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31866313

RESUMO

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.


Assuntos
Fórmulas Infantis/química , Lipase/química , Bicamadas Lipídicas/química , Leite Humano/química , Leite/química , Adsorção , Animais , Bovinos , Colesterol/química , Misturas Complexas/química , Ácidos Graxos Insaturados/química , Glicerofosfolipídeos/química , Glicolipídeos , Glicoproteínas , Humanos , Lactente , Gotículas Lipídicas , Fosfatidiletanolaminas/química , Pressão , Especificidade da Espécie , Esfingomielinas/química , Estômago/química , Estômago/enzimologia , Propriedades de Superfície , Triglicerídeos/química
9.
Adv Drug Deliv Rev ; 142: 3-15, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30926476

RESUMO

The lipids and some surfactants present in oral lipid-based drug delivery systems are potential substrates for the various lipases involved in gastrointestinal (GI) lipolysis. The levels of these enzymes, together with pH and biliairy secretion, are important parameters that condition the fate of lipid-based formulations (LBF) and the dispersion, solubilization and absorption of lipophilic drugs in the GI tract. Since in vitro methods of digestion are now combined with dissolution assays for a better assessment of LBF performance, it is essential to have a basic knowledge on lipase, pH and bile acid (BA) levels in vivo to develop relevant in vitro models. While these parameters and their variations in healthy subjects are today well documented, in vivo data on specific populations (age groups, patients with various diseases, patients with treatment affecting GI tract parameters, …) are scarce and obtaining them from clinical studies is sometimes difficult due to ethical limitations. Here we collected some in vivo data already available on the levels of digestive lipases, gastric and intestinal pH, and BAs at various ages and in patients with exocrine pancreatic insufficiency, a pathological situation that leads to drastic changes in GI tract parameters and impacts pharmacological treatments.


Assuntos
Envelhecimento/metabolismo , Ácidos e Sais Biliares/metabolismo , Ingestão de Alimentos , Trato Gastrointestinal/metabolismo , Lipase/metabolismo , Animais , Gorduras na Dieta/farmacologia , Digestão , Sistemas de Liberação de Medicamentos , Trato Gastrointestinal/química , Humanos , Concentração de Íons de Hidrogênio , Lipólise , Pâncreas/metabolismo
10.
Food Chem ; 281: 294-303, 2019 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-30658760

RESUMO

Holder pasteurization (62.5 °C, 30 min) of human milk denatures beneficial proteins. The present paper aimed to assess whether this can affect the kinetics of peptide release during digestion at the preterm stage. Raw (RHM) or pasteurized (PHM) human milk were digested in triplicates using an in vitro dynamic system. Mass spectrometry and multivariate statistics were conducted. Pre-proteolysis occurred mostly on ß-casein, for which cumulative peptide abundance was significantly greater in PHM over 28% of the hydrolysed sequence. Eight clusters resumed the kinetics of peptide release during digestion, which differed on seven clusters (69% of the 1134 peptides). Clusters associated to the heat-denaturated proteins, lactoferrin and bile salt-stimulated lipase, presented different kinetics of release during digestion, unlike that for ß-casein. Some bioactive peptides from ß-casein presented significant different abundances between PHM and RHM before digestion (1-18, 185-211) or in during intestinal digestion (154-160, 161-166). Further physiological consequences should be investigated.


Assuntos
Leite Humano/química , Pasteurização , Ácidos e Sais Biliares/análise , Caseínas/análise , Análise por Conglomerados , Digestão , Temperatura Alta , Humanos , Concentração de Íons de Hidrogênio , Recém-Nascido Prematuro/crescimento & desenvolvimento , Lactoferrina/análise , Proteínas do Leite/análise , Peptídeos/análise , Proteólise
11.
Eur J Nutr ; 57(2): 463-476, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27744547

RESUMO

PURPOSE: Although composition of infant formula has been significantly improved during the last decade, major differences with the composition and structure of breast milk still remain and might affect nutrient digestion and gut biology. We hypothesized that the incorporation of dairy fat in infant formulas could modify their physiological impacts by making their composition closer to that of human milk. The effect of milk fat and milk fat globule membrane (MFGM) fragments in infant formulas on gut digestion, mucosal immunity and microbiota composition was evaluated. METHODS: Three formulas containing either (1) vegetable lipids stabilized only by proteins (V-P), (2) vegetable lipids stabilized by a mixture of proteins and MFGM fragments (V-M) and (3) a mixture of milk and vegetable lipids stabilized by a mixture of proteins and MFGM fragments (M-M) were automatically distributed to 42 newborn piglets until slaughter at postnatal day (PND) 7 or 28, and compared to a fourth group of sow's suckling piglets (SM) used as a breast-fed reference. RESULTS: At both PND, casein and ß-lactoglobulin digestion was reduced in M-M proximal jejunum and ileum contents compared to V-P and V-M ones leading to more numerous ß-Cn peptides in M-M contents. The IFNγ cytokine secretion of ConA-stimulated MLN cells from M-M piglets tended to be higher than in V-P ones at PND 7 and PND 28 and was closer to that of SM piglets. No dietary treatment effect was observed on IL-10 MLN cell secretion. Changes in faecal microbiota in M-M piglets resulted in an increase in Proteobacteria and Bacteroidetes and a decrease in Firmicutes phyla compared to V-P ones. M-M piglets showed higher abundances of Parabacteroides, Escherichia/Shigella and Klebsiella genus. CONCLUSIONS: The incorporation of both milk fat and MFGM fragments in infant formula modifies protein digestion, the dynamic of the immune system maturation and the faecal microbiota composition.


Assuntos
Fenômenos Fisiológicos da Nutrição Animal , Microbioma Gastrointestinal/imunologia , Imunidade nas Mucosas , Imunomodulação , Leite/química , Modelos Imunológicos , Óleos de Plantas/administração & dosagem , Animais , Animais Recém-Nascidos , Caseínas/administração & dosagem , Caseínas/metabolismo , Citocinas/metabolismo , Digestão , Fezes/microbiologia , Conteúdo Gastrointestinal/química , Conteúdo Gastrointestinal/microbiologia , Glicolipídeos/administração & dosagem , Glicolipídeos/metabolismo , Glicoproteínas/administração & dosagem , Glicoproteínas/metabolismo , Humanos , Fórmulas Infantis , Fenômenos Fisiológicos da Nutrição do Lactente , Recém-Nascido , Lactoglobulinas/administração & dosagem , Lactoglobulinas/metabolismo , Gotículas Lipídicas , Linfonodos/crescimento & desenvolvimento , Linfonodos/imunologia , Linfonodos/metabolismo , Leite/metabolismo , Óleos de Plantas/metabolismo , Proteínas de Vegetais Comestíveis/administração & dosagem , Proteínas de Vegetais Comestíveis/metabolismo , Sus scrofa/crescimento & desenvolvimento
12.
Food Chem ; 240: 67-74, 2018 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-28946327

RESUMO

Bioactive lipids of the milk fat globule membrane become concentrated in two co-products of the butter industry, buttermilk and butterserum. Their lipid composition is detailed here with special emphasis on sphingolipid composition of nutritional interest, determined using GC, HPLC and tandem mass spectrometry. Butterserum was 2.5 times more concentrated in total fat than buttermilk, with 7.7±1.5vs 19.5±2.9wt% and even more concentrated in polar lipids, with 1.4±0.2vs 8.5±1.1wt%. Both ingredients constitute concentrated sources of sphingomyelin (3.4-21mg/g dry matter) and contained low amounts of bioactive ceramides in a ratio to sphingomyelin of 1:5mol% in buttermilk and 1:10mol% in butterserum. Compared to other natural lecithins, these two co-products are rich in long and saturated fatty acids (C22:0-C24:0), contain cholesterol and could have interesting applications in neonatal nutrition, but also as brain-protective, hepatoprotective and cholesterol lowering ingredients.


Assuntos
Leitelho/análise , Ceramidas/análise , Leite/química , Esfingolipídeos/análise , Animais , Ácidos Graxos , Síndrome de Heterotaxia , Humanos
13.
Clin Nutr ESPEN ; 20: 1-11, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29072162

RESUMO

BACKGROUND & AIMS: It has been suggested that homogenization of Holder-pasteurized human milk (PHM) could improve fat absorption and weight gain in preterm infants, but the impact on the PHM digestive kinetics has never been studied. Our objective was to determine the impact of PHM homogenization on gastric digestion in preterm infants. METHODS: In a randomized controlled trial, eight hospitalized tube-fed preterm infants were their own control to compare the gastric digestion of PHM and of homogenized PHM (PHHM). PHM was obtained from donors and, for half of it, was homogenized by ultrasonication. Over a six-day sequence, gastric aspirates were collected twice a day, before and 35, 60 or 90 min after the start of PHM or PHHM ingestion. The impact of homogenization on PHM digestive kinetics and disintegration was tested using a general linear mixed model. Results were expressed as means ± SD. RESULTS: Homogenization leaded to a six-fold increase in the specific surface (P < 0.01) of lipid droplets. The types of aggregates formed during digestion were different between PHM and PHHM, but the lipid fraction kept its initial structure all over the gastric digestion (native globules in PHM vs. blend of droplets in PHHM). Homogenization increased the gastric lipolysis level (P < 0.01), particularly at 35 and 60 min (22 and 24% higher for PHHM, respectively). Homogenization enhanced the proteolysis of serum albumin (P < 0.05) and reduced the meal emptying rate (P < 0.001, half-time estimated at 30 min for PHM and 38 min for PHHM). The postprandial gastric pH was not affected (4.7 ± 0.9 at 90 min). CONCLUSIONS: Homogenization of PHM increased the gastric lipolysis level. This could be a potential strategy to improve fat absorption, and thus growth and development in infants fed with PHM; however, its gastrointestinal tolerance needs to be investigated further. This trial was registered at clinicaltrials.gov as NCT02112331.


Assuntos
Digestão , Mucosa Gástrica/metabolismo , Recém-Nascido Prematuro/fisiologia , Leite Humano/metabolismo , Feminino , França , Humanos , Fenômenos Fisiológicos da Nutrição do Lactente , Recém-Nascido , Masculino , Leite Humano/química , Pasteurização , Resultado do Tratamento , Ondas Ultrassônicas
14.
Chem Phys Lipids ; 209: 19-28, 2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-29061286

RESUMO

Regardless of the applications: therapeutic vehicle or membrane model to mimic complex biological systems; it is of a great importance to develop simplified, reproducible and rapid model assays allowing for a relevant assessment of the liposomal membrane oxidation and therefore antioxidant activity of selected molecules. Here, we describe a new and high-throughput assay that we called "Vesicle Conjugated Autoxidizable Triene (VesiCAT)". It is based on specific UV absorbance spectral properties of a new phospholipid probe, synthesized with natural conjugated eleostearic acid extracted from Tung oil. The VesiCAT assay has been developed with two different radical generators (2,2'-azobis(2-amidinopropane) dihydrochloride; AAPH and 2,2'-azobis(2,4-dimethylvaleronitrile); AMVN), producing a constant flux of oxidant species, either in membrane or in aqueous phase. This method appears very efficient in assessing the effect of various pure antioxidant molecules in their ability to preserve liposomes from oxidative degradation. In addition, the AAPH- and AMVN-induced oxidations offer the possibility of extracting different but complementary information with respect to the antioxidants efficacy.


Assuntos
Antioxidantes/química , Ácidos Linolênicos/química , Lipossomos/química , Sondas Moleculares/química , Fosfolipídeos/química , Ensaios de Triagem em Larga Escala , Oxirredução , Fosfolipídeos/síntese química , Óleos de Plantas/química
15.
Am J Clin Nutr ; 105(2): 379-390, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-28052887

RESUMO

BACKGROUND: Holder pasteurization has been reported to modify human milk composition and structure by inactivating bile salt-stimulated lipase (BSSL) and partially denaturing some of its proteins, potentially affecting its subsequent digestion. OBJECTIVE: We sought to determine the impact of human milk pasteurization on gastric digestion (particularly for proteins and lipids) in preterm infants who were fed their mothers' own milk either raw or pasteurized. DESIGN: In a randomized controlled trial, 12 hospitalized tube-fed preterm infants were their own control group in comparing the gastric digestion of raw human milk (RHM) with pasteurized human milk (PHM). Over a 6-d sequence, gastric aspirates were collected 2 times/d before and after RHM or PHM ingestion. The impact of milk pasteurization digestive kinetics and disintegration was tested with the use of a general linear mixed model. RESULTS: Despite inactivating BSSL, instantaneous lipolysis was not affected by pasteurization (mean ± SD at 90 min: 12.6% ± 4.7%; P > 0.05). Lipolysis occurred in milk before digestion and was higher for PHM than for RHM (mean ± SD: 3.2% ± 0.6% and 2.2% ± 0.8%, respectively; P < 0.001). Pasteurization enhanced the proteolysis of lactoferrin (P < 0.01) and reduced that of α-lactalbumin (only at 90 min) (P < 0.05). Strong emulsion destabilization was observed, with smaller aggregates and a higher specific surface for PHM (P < 0.05). Pasteurization did not affect gastric emptying (∼30-min half time) or pH (mean ± SD: 4.4 ± 0.8) at 90 min. CONCLUSIONS: Overall, pasteurization had no impact on the gastric digestion of lipids and some proteins from human milk but did affect lactoferrin and α-lactalbumin proteolysis and emulsion disintegration. Freeze-thawing and pasteurization increased the milk lipolysis before digestion but did not affect gastric lipolysis. Possible consequences on intestinal digestion and associated nutritional outcomes were not considered in this study. This trial was registered at clinicaltrials.gov as NCT02112331.


Assuntos
Digestão , Leite Humano/química , Pasteurização , Caseínas/sangue , Carboidratos da Dieta/análise , Gorduras na Dieta/análise , Proteínas na Dieta/análise , Ácidos Graxos/análise , Esvaziamento Gástrico , Mucosa Gástrica/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Lactente , Fenômenos Fisiológicos da Nutrição do Lactente , Recém-Nascido Prematuro , Lactalbumina/sangue , Lactoferrina/sangue , Lipólise , Proteínas do Leite/química , Proteólise , Albumina Sérica/metabolismo , Esterol Esterase/antagonistas & inibidores , Esterol Esterase/metabolismo
16.
Food Chem ; 211: 171-9, 2016 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-27283620

RESUMO

Human milk feeding is an important recommendation for preterm newborns considering their vulnerability and digestive immaturity. Holder pasteurization (62.5°C, 30min) applied in milk banks modifies its biological quality and its microstructure. We investigated the impact of pasteurization of preterm human milk on its gastrointestinal kinetics of lipolysis, proteolysis and structural disintegration. An in vitro dynamic system was set up to simulate the gastrointestinal digestion of preterm newborns. A pool of preterm human milk was digested as raw or after Holder pasteurization. Pasteurization impacted the microstructure of undigested human milk, its gastrointestinal disintegration and tended to limit the intestinal lipolysis. Furthermore, the gastrointestinal bioaccessibility of some fatty acids was decreased by pasteurization, while the intestinal bioaccessibility of some amino acids was selectively modulated. The impact of pasteurization on the digestion of human milk may have nutritional relevance in vivo and potentially modulates preterm development and growth.


Assuntos
Ácidos Graxos/análise , Gastroenteropatias/fisiopatologia , Lipólise , Leite Humano/química , Pasteurização/métodos , Disponibilidade Biológica , Cromatografia em Camada Delgada , Digestão , Ácidos Graxos/química , Feminino , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Luz , Lipídeos/química , Microscopia Confocal , Tamanho da Partícula , Proteólise , Espalhamento de Radiação
17.
Electrophoresis ; 37(13): 1839-50, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-26990007

RESUMO

Holder pasteurization (62.5°C, 30 min) ensures sanitary quality of donor's human milk but also denatures beneficial proteins. Understanding whether this further impacts the kinetics of peptide release during gastrointestinal digestion of human milk was the aim of the present paper. Mature raw (RHM) or pasteurized (PHM) human milk were digested (RHM, n = 2; PHM, n = 3) by an in vitro dynamic system (term stage). Label-free quantitative peptidomics was performed on milk and digesta (ten time points). Ascending hierarchical clustering was conducted on "Pasteurization × Digestion time" interaction coefficients. Preproteolysis occurred in human milk (159 unique peptides; RHM: 91, PHM: 151), mostly on ß-casein (88% of the endogenous peptides). The predicted cleavage number increased with pasteurization, potentially through plasmin activation (plasmin cleavages: RHM, 53; PHM, 76). During digestion, eight clusters resumed 1054 peptides from RHM and PHM, originating for 49% of them from ß-casein. For seven clusters (57% of peptides), the kinetics of peptide release differed between RHM and PHM. The parent protein was significantly linked to the clustering (p-value = 1.4 E-09), with ß-casein and lactoferrin associated to clusters in an opposite manner. Pasteurization impacted selectively gastric and intestinal kinetics of peptide release in term newborns, which may have further nutritional consequences.


Assuntos
Digestão , Proteínas do Leite/farmacocinética , Leite Humano , Pasteurização , Peptídeos/farmacocinética , Cromatografia Líquida , Humanos , Recém-Nascido , Proteólise , Espectrometria de Massas em Tandem
18.
Colloids Surf B Biointerfaces ; 143: 97-106, 2016 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-27011347

RESUMO

The enzymatic lipolysis of complex natural lipoproteic assemblies such as milk fat globules is central in neonatal digestion. This process first requires the rapid adsorption of a lipolytic enzyme, gastric lipase, onto the membrane enveloping the triglyceride substrate before the onset of catalytic activity. The interactions governing lipase adsorption onto this complex lipid/water interface are not fully elucidated. This study was designed to unravel the interactions of recombinant dog gastric lipase (rDGL) with model monolayers presenting liquid-liquid phase coexistence and mimicking the outer leaflet of the milk fat globule membrane. Combining biophysical tools (ellipsometry, tensiometry and atomic force microscopy), it was evidenced that rDGL partitions toward liquid expanded phase and at phase boundaries. rDGL gets adsorbed at several levels of insertion suggesting molecular cooperation that may favor insertion and strongly impacts on the lipid phase lateral organization. The addition of phosphatidylserine, negatively charged, reinforced adsorption; hence besides hydrophobic interactions and as further investigated through surface potential modeling, rDGL adsorption is favored by electrostatic interactions.


Assuntos
Glicolipídeos/química , Glicoproteínas/química , Lipase/química , Lipossomas Unilamelares/química , 1,2-Dipalmitoilfosfatidilcolina/química , Adsorção , Animais , Bovinos , Cães , Gotículas Lipídicas , Microscopia de Força Atômica , Fosfatidilcolinas/química , Fosfatidiletanolaminas/química , Fosfatidilserinas/química , Proteínas Recombinantes/química , Eletricidade Estática , Estômago/química , Estômago/enzimologia , Tensão Superficial , Água/química
19.
J Nutr ; 145(8): 1770-7, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26136586

RESUMO

BACKGROUND: Polar lipid (PL) emulsifiers such as milk PLs (MPLs) may affect digestion and subsequent lipid metabolism, but focused studies on postprandial lipemia are lacking. OBJECTIVE: We evaluated the impact of MPLs on postprandial lipemia in mice and on lipid digestion in vitro. METHODS: Female Swiss mice were gavaged with 150 µL of an oil-in-water emulsion stabilized with 5.7 mg of either MPLs or soybean PLs (SPLs) and killed after 1, 2, or 4 h. Plasma lipids were quantified and in the small intestine, gene expression was analyzed by reverse transcriptase-quantitative polymerase chain reaction. Emulsions were lipolyzed in vitro using a static human digestion model; triglyceride (TG) disappearance was followed by thin-layer chromatography. RESULTS: In mice, after 1 h, plasma TGs tended to be higher in the MPL group than in the SPL group (141 µg/mL vs. 90 µg/mL; P = 0.07) and nonesterified fatty acids (NEFAs) were significantly higher (64 µg/mL vs. 44 µg/mL; P < 0.05). The opposite was observed after 4 h with lower TGs (21 µg/mL vs. 35 µg/mL; P < 0.01) and NEFAs (20 µg/mL vs. 32 µg/mL; P < 0.01) in the MPL group compared with the SPL group. This was associated at 4 h with a lower gene expression of apolipoprotein B (Apob) and Secretion Associated, Ras related GTPase 1 gene homolog B (Sar1b), in the duodenum of MPL mice compared with SPL mice (P < 0.05). In vitro, during the intestinal phase, TGs were hydrolyzed more in the MPL emulsion than in the SPL emulsion (decremental AUCs were 1750%/min vs. 180%/min; P < 0.01). MPLs enhance lipid intestinal hydrolysis and promote more rapid intestinal lipid absorption and sharper kinetics of lipemia. CONCLUSIONS: Postprandial lipemia in mice can be modulated by emulsifying with MPLs compared with SPLs, partly through differences in chylomicron assembly, and TG hydrolysis rate as observed in vitro. MPLs may thereby contribute to the long-term regulation of lipid metabolism.


Assuntos
Metabolismo dos Lipídeos/efeitos dos fármacos , Lipídeos/farmacologia , Lipólise/efeitos dos fármacos , Leite/química , Animais , Emulsificantes , Feminino , Regulação da Expressão Gênica , Intestino Delgado/metabolismo , Lecitinas , Lipídeos/química , Camundongos , Período Pós-Prandial
20.
Food Chem ; 182: 224-35, 2015 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-25842331

RESUMO

Milk lipids supply most of the calories necessary for newborn growth in maternal milk or infant formulas. The chemical composition of infant formulas has been optimized but not the structure of the emulsion. There is still a major difference between the native emulsions of milk fat globules and processed submicronic emulsions in infant formulas. This difference may modify the kinetics of digestion of emulsions in newborns and influence lipid metabolism. To check this, semi-dynamic gastric in vitro digestions were conducted on three matrices: a standardized milk emulsion containing native milk fat globules referred to as minimally-processed emulsion and two processed model infant formulas (homogenized or homogenized/pasteurized). Gastric conditions mimicked those reported in newborns. The minimally-processed emulsion was lipolyzed and proteolyzed slower than processed formulas. The difference in initial structure persisted during digestion. The surface of the droplets was the key parameter to control gastric lipolysis kinetics, the pattern of released fatty acids and proteolysis by faster hydrolysis of adsorbed proteins.


Assuntos
Emulsões/química , Ácidos Graxos/química , Fórmulas Infantis/química , Fórmulas Infantis/síntese química , Metabolismo dos Lipídeos/fisiologia , Leite Humano/química , Animais , Digestão , Humanos , Recém-Nascido , Lipídeos/química , Lipólise , Proteólise
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