In Vitro Bioaccessibility of Proteins and Bioactive Compounds of Wild and Cultivated Seaweeds from the Gulf of Saint Lawrence.

Despite the increased interest in macroalgae protein and fibers, little information is available on their bioaccessibility. The application of an in vitro gastrointestinal digestion model to study the degree of disintegration and release of proteins with expressed bioactivities from wild and cultivated Palmaria palmata and Saccharina latissima was proposed in this study. Macroalgae from the Gulf of St Lawrence, Canada, were submitted to digestive transit times of 2 (oral), 60 (gastric) and 120 (duodenal) minutes. Among wild samples, P. palmata had a higher percentage of disintegration, protein release and degree of hydrolysis than S. latissima. While the least digested sample, wild S. latissima, was the sample with the highest antioxidant activity (210 µmol TE g-1), the most digested sample, cultivated P. palmata, presented the highest ability to inhibit the angiotensin-converting enzyme (ACE), reaching 32.6 ± 1.2% at 3 mg mL-1. ACE inhibitory activity increased from 1 to 3 mg mL-1, but not at 5 mg mL-1. Wild samples from both species showed an ACE inhibition around 27.5%. Data suggested that the disintegration of the samples was influenced by their soluble and insoluble fiber contents. Further information on the bioaccessibility and bioactivity of these macroalgae should consider the characterization of digestion products other than protein, as well as the effects of previous product processing.

Correlating in vitro digestion viscosities and bioaccessible nutrients of milks containing enhanced protein concentration and normal or modified protein ratio to human trials.

This work compared in vitro and in vivo digestion of breakfast cereal with milks containing high protein concentration (9.3 wt%) and the normal protein ratio (80 casein : 20 whey) or a modified ratio (40 casein : 60 whey) and with a water-permeate control. The in vivo study indicated that high protein concentration and modified ratio in milks delays the postprandial appearance of blood glucose (BG) and amino acids due to delayed gastric emptying and hormonal responses. However, the role of viscosity and/or protein structure during digestion was not examined. Therefore, milks and the control were digested in vitro (oral (O, 2 min), gastric (G, 62 min) and duodenal (D, 92 min)) to determine viscosity, particle disintegration, protein solubility and hydrolysis, and the bioaccessibilities of sugars and total amino acids (TAA). The normal ratio (80 : 20) treatment demonstrated higher structural viscosity during digestion (P < 0.05) due to the formation of casein aggregates and interaction with cereal and greater TAA (mg per g protein of undigested breakfast) caused by gastric hydrolysis (DH%; P = 0.01). Overall, there were no treatment differences for disintegration, solubility and d-glucose. Protein-containing treatments inhibited amylolysis and lowered reducing sugars (mg g-1 available carbohydrates of undigested breakfast) compared to the control. Similar trends were observed between higher viscosity (Pa s) during gastric stage and slower in vivo gastric emptying (paracetamol, mmol L-1). Also, protein treatments inhibited amylolysis and lowered reducing sugar (mg g-1 of carbohydrates) and may have contributed to lowered BG (mmol L-1 g-1 of carbohydrates) after the duodenal phase. However, increased viscosity, elicited by higher proportions of casein, did not inhibit starch hydrolysis and appearance of BG. In vitro systems provide similar trends in biomarkers to in vivo studies and can be used to answer specific physiological questions.

Identification of texture parameters influencing commercial cheese matrix disintegration and lipid digestion using an in vitro static digestion model.

Cheese characteristics, such as composition or textural properties, can impact the matrix degradation rate which could modulate the bioaccessibility of fatty acids during digestion. The aim of this study was to identify texture parameters influencing cheese degradation in a gastrointestinal environment. A static in vitro digestion model has been used on nine commercial cheeses: young and aged cheddar, regular and light cream cheese, parmesan, feta, camembert, mozzarella, and sliced processed cheese. At the end of gastric digestion, camembert and mozzarella presented the lowest matrix disintegration whereas aged cheddar, regular and light cream cheeses showed the highest. For all cheeses, the fatty acid release was fast during the first 30 min of duodenal digestion and slowed down afterwards. A partial least square regression revealed that springiness, cohesiveness, and hardness were negatively correlated to the rate of cheese disintegration during gastric digestion. In addition, textural parameters were not correlated with free fatty acid release. By modulating cheese texture, it could be possible to influence matrix disintegration during gastrointestinal digestion which could have an impact on lipids release.

Influence of food structure on dairy protein, lipid and calcium bioavailability: A narrative review of evidence.

Beyond nutrient composition matrix plays an important role on food health potential, notably acting on the kinetics of nutrient release, and finally on their bioavailability. This is particularly true for dairy products that present both solid (cheeses), semi-solid (yogurts) and liquid (milks) matrices. The main objective of this narrative review has been to synthesize available data in relation with the impact of physical structure of main dairy matrices on nutrient bio-accessibility, bioavailability and metabolic effects, in vitro, in animals and in humans. Focus has been made on dairy nutrients the most studied, i.e., proteins, lipids and calcium. Data collected show different kinetics of bioavailability of amino acids, fatty acids and calcium according to the physicochemical parameters of these matrices, including compactness, hardness, elasticity, protein/lipid ratio, P/Ca ratio, effect of ferments, size of fat globules, and possibly other qualitative parameters yet to be discovered. This could be of great interest for the development of innovative dairy products for older populations, sometimes in protein denutrition or with poor dentition, involving the development of dairy matrices with optimized metabolic effects by playing on gastric retention time and thus on the kinetics of release of the amino acids within bloodstream.

Differential impact of the cheese matrix on the postprandial lipid response: a randomized, crossover, controlled trial.

Background: In a simulated gastrointestinal environment, the cheese matrix modulates dairy fat digestion. However, to our knowledge, the impact of the cheese matrix on postprandial lipemia in humans has not yet been evaluated.Objective: In healthy subjects, we compared the impact of dairy fat provided from firm cheese, soft cream cheese, and butter on the postprandial response at 4 h and on the incremental area under the curve (iAUC) of plasma triglycerides.Design: Forty-three healthy subjects were recruited to this randomized, crossover, controlled trial. In random order at intervals of 14 d and after a 12-h fast, subjects ingested 33 g fat from a firm cheese (young cheddar), a soft cream cheese (cream cheese), or butter (control) incorporated into standardized meals that were matched for macronutrient content. Plasma concentrations of triglycerides were measured immediately before the meal and 2, 4, 6, and 8 h after the meal.Results: Cheddar cheese, cream cheese, and butter induced similar increases in triglyceride concentrations at 4 h (change from baseline: +59%, +59%, and +62%, respectively; P = 0.9). No difference in the triglyceride iAUC0-8 h (P-meal = 0.9) was observed between the 3 meals. However, at 2 h, the triglyceride response caused by the cream cheese (change from baseline: +44%) was significantly greater than that induced by butter (change from baseline: +24%; P = 0.002) and cheddar cheese (change from baseline: +16%; P = 0.0004). At 6 h, the triglyceride response induced by cream cheese was significantly attenuated compared with that induced by cheddar cheese (change from baseline: +14% compared with +42%; P = 0.0004).Conclusion: This study demonstrates that the cheese matrix modulates the impact of dairy fat on postprandial lipemia in healthy subjects. This trial was registered at clinicaltrials.gov as NCT02623790.

Low-Temperature Blanching as a Tool to Modulate the Structure of Pectin in Blueberry Purees.

Blueberry composition was characterized for 6 cultivars. It contains a good amount of dietary fiber (10% to 20%) and pectin (4% to 7%) whose degree of methylation (DM) is sensitive to food processing. A low temperature blanching (LTB: 60 °C/1 h) was applied on blueberry purees to decrease pectin DM, in order to modulate puree properties and functionalities (that is, viscosity and stability), and to enhance pectin affinity toward other components within food matrices. Fiber content, viscosity, pectin solubility, DM, and monosaccharide composition were determined for both pasteurized, and LTB+pasteurized blueberry purees. The results showed that neither the amount of fiber, nor the viscosity were affected by LTB, indicating that this treatment did not result in any significant pectin depolymerization and degradation. LTB caused a decrease both in pectin DM from 58-67% to 45-47% and in the amount of water-soluble pectin fraction, the latter remaining the major fraction of total pectin at 52% to 57%. A LTB is a simple and mild process to produce blueberry purees with mostly soluble and low-methylated pectin in order to extend functionality and opportunities for interactions with other food ingredients.

Formation and functional properties of protein-polysaccharide electrostatic hydrogels in comparison to protein or polysaccharide hydrogels.

Protein and polysaccharide mixed systems have been actively studied for at least 50years as they can be assembled into functional particles or gels. This article reviews the properties of electrostatic gels, a recently discovered particular case of associative protein-polysaccharide mixtures formed through associative electrostatic interaction under appropriate solution conditions (coupled gel). This review highlights the factors influencing gel formation such as protein-polysaccharide ratio, biopolymer structural characteristics, final pH, ionic strength and total solid concentration. For the first time, the functional properties of protein-polysaccharide coupled gels are presented and discussed in relationship to individual protein and polysaccharide hydrogels. One of their outstanding characteristics is their gel water retention. Up to 600g of water per g of biopolymer may be retained in the electrostatic gel network compared to a protein gel (3-9g of water per g of protein). Potential applications of the gels are proposed to enable the food and non-food industries to develop new functional products with desirable attributes or new interesting materials to incorporate bioactive molecules.

Acute effects of protein composition and fibre enrichment of yogurt consumed as snacks on appetite sensations and subsequent ad libitum energy intake in healthy men.

The objective of the study was to assess the impact of protein composition and/or fibre enrichment of yogurt on appetite sensations and subsequent energy intake. In this double-blind crossover study, 20 healthy men (aged 32.4 ± 9.1 years) were submitted to 5 randomized testing sessions, during which they had to consume 5 isocaloric and isonproteinemic yogurt snacks (120-g servings, ∼230 kJ, ∼4.5 g protein) differing by their casein-to-whey protein ratio (C:W) or dietary fibre content: (i) control C:W = 2.8:1; (ii) high whey (HW) C:W = 1.5:1, and fibre-enriched formulations using control; (iii) 2.4 g of inulin; (iv) 1.9 g of inulin and 0.5 g of ß-glucan (+IN-ßG); and (v) 0.5 g of ß-glucan. Appetite sensations were assessed using 150-mm visual analog scales. Plasma variables (glucose, insulin, ghrelin) were measured at 30-min intervals post-yogurt consumption for 2 h. Finally, energy intakes during ad libitum lunches offered 2 h after yogurt snacks were recorded. None of the yogurts impacted appetite sensations. Ad libitum energy intake was significantly different only between HW and control yogurts (-812 kJ; p = 0.03). Regarding post-yogurt plasma variables, a significant difference was found only between ghrelin area under the curve of the +IN-ßG and the HW yogurts (-15 510 pmol/L per 120 min, p = 0.04). In conclusion, although appetite sensations were not influenced by variations in yogurts' protein compositions, a reduced energy intake was observed during the ad libitum lunch after the HW yogurt that may be attributable to its lower C:W. Surprisingly, the fibre enrichments studied did not exert effect on appetite sensations and energy intake.

Low-Molecular-Weight Peptides from Salmon Protein Prevent Obesity-Linked Glucose Intolerance, Inflammation, and Dyslipidemia in LDLR-/-/ApoB100/100 Mice.

BACKGROUND: We previously reported that fish proteins can alleviate metabolic syndrome (MetS) in obese animals and human subjects. OBJECTIVES: We tested whether a salmon peptide fraction (SPF) could improve MetS in mice and explored potential mechanisms of action. METHODS: ApoB(100) only, LDL receptor knockout male mice (LDLR(-/-)/ApoB(100/100)) were fed a high-fat and -sucrose (HFS) diet (25 g/kg sucrose). Two groups were fed 10 g/kg casein hydrolysate (HFS), and 1 group was additionally fed 4.35 g/kg fish oil (FO; HFS+FO). Two other groups were fed 10 g SPF/kg (HFS+SPF), and 1 group was additionally fed 4.35 g FO/kg (HFS+SPF+FO). A fifth (reference) group was fed a standard feed pellet diet. We assessed the impact of dietary treatments on glucose tolerance, adipose tissue inflammation, lipid homeostasis, and hepatic insulin signaling. The effects of SPF on glucose uptake, hepatic glucose production, and inducible nitric oxide synthase activity were further studied in vitro with the use of L6 myocytes, FAO hepatocytes, and J774 macrophages. RESULTS: Mice fed HFS+SPF or HFS+SPF+FO diets had lower body weight (protein effect, P = 0.024), feed efficiency (protein effect, P = 0.018), and liver weight (protein effect, P = 0.003) as well as lower concentrations of adipose tissue cytokines and chemokines (protein effect, P ≤ 0.003) compared with HFS and HFS+FO groups. They also had greater glucose tolerance (protein effect, P < 0.001), lower activation of the mammalian target of rapamycin complex 1/S6 kinase 1/insulin receptor substrate 1 (mTORC1/S6K1/IRS1) pathway, and increased insulin signaling in liver compared with the HFS and HFS+FO groups. The HFS+FO, HFS+SPF, and HFS+SPF+FO groups had lower plasma triglycerides (protein effect, P = 0.003; lipid effect, P = 0.002) than did the HFS group. SPF increased glucose uptake and decreased HGP and iNOS activation in vitro. CONCLUSIONS: SPF reduces obesity-linked MetS features in LDLR(-/-)/ApoB(100/100) mice. The anti-inflammatory and glucoregulatory properties of SPF were confirmed in L6 myocytes, FAO hepatocytes, and J774 macrophages.

Role of seaweed laminaran from Saccharina longicruris on matrix deposition during dermal tissue-engineered production.

Our laboratory has developed a technique to reconstruct in vitro tissue from human cells using the self-assembly tissue-engineering method, which utilizes the ability of fibroblasts to deposit the matrix they secrete. The time necessary for tissue construction, several weeks, is a drawback for many clinical uses. We hypothesized that the addition of laminaran can increase the deposition of matrix, speeding up the production of the tissue. Laminaran was isolated from the brown seaweed Saccharina longicruris harvested in Canada and its structure was evaluated. Laminaran is a small molecular weight polysaccharide composed of linear glucose chains. Monolayer-cultured human skin fibroblasts were cultured in the presence of laminaran with ascorbate for 7 or 35 days to produce a dermis. Treatment did not induce any variation in the growth rate or alpha smooth muscle actin content but it did increase the deposition of collagen I in a dose-dependent manner. After 35 days, the reconstructed dermal thickness was increased when laminaran was added, and collagen I deposition and MMP activity were also significantly increased. Thus, laminaran can be used to increase the rate of production of reconstructed self-assembled dermis and can also potentially be used in cosmetic or therapeutic creams to stimulate matrix production.

Alpha-amylase and alpha-glucosidase inhibition is differentially modulated by fucoidan obtained from Fucus vesiculosus and Ascophyllum nodosum.

Fucoidan is a water-soluble, negatively charged, biologically active polysaccharide found in great abundance in brown marine algae. However, the inhibition of α-amylase and α-glucosidase by fucoidan derived from two algal species (Ascophyllum nodosum and Fucus vesiculosus) harvested at different periods (accounting for seasonal and yearly variations) has never been investigated. It was found that fucoidans inhibited α-glucosidase differently, depending on the algal species from which it was extracted and the algae's season of harvest. Fucoidan extracted from A. nodosum was a more potent inhibitor of α-glucosidase, with an IC50 ranging from 0.013 to 0.047 mg/mL, than the inhibition by fucoidan extracted from F. vesiculosus (IC50=0.049 mg/mL). In contrast, fucoidan extracted from F. vesiculosus did not inhibit α-amylase activity, while fucoidan from A. nodosum decreased α-amylase activity by 7-100% at 5 mg/mL depending upon the algae harvest period. An IC50 of 0.12-4.64 mg/mL for fucoidan from A. nodosum was found for the α-amylase inhibition. The ability of fucoidan to inhibit α-amylase and α-glucosidase thus varies according to the algae species and harvest period. A. nodosum is more suitable than F. vesiculosus as a source of fucoidan to inhibit α-amylase and α-glucosidase activities. Their potential benefits towards Type 2 diabetes management should be further investigated.

Differential effects of various fish proteins in altering body weight, adiposity, inflammatory status, and insulin sensitivity in high-fat-fed rats.

Mounting evidence suggests that the benefits of fish consumption are not limited to the well-appreciated effects of omega-3 fatty acids. We previously demonstrated that cod protein protects against the development of diet-induced insulin resistance. The goal of this study was to determine whether other fish protein sources present similar beneficial effects. Rats were fed a high-fat, high-sucrose diet containing protein from casein or fish proteins from bonito, herring, mackerel, or salmon. After 28 days, oral glucose tolerance tests or hyperinsulinemic-euglycemic clamps were performed; and tissues and plasma were harvested for biochemical analyses. Despite equal energy intake among all groups, the salmon-protein-fed group presented significantly lower weight gain that was associated with reduced fat accrual in epididymal white adipose tissue. Although this reduction in visceral adiposity was not associated with improved glucose tolerance, we found that whole-body insulin sensitivity for glucose metabolism was improved using the very sensitive hyperinsulinemic-euglycemic clamp technique. Importantly, expression of both tumor necrosis factor-α and interleukin-6 was reduced in visceral adipose tissue of all fish-protein-fed groups when compared with the casein-fed control group, suggesting that fish proteins carry anti-inflammatory properties that may protect against obesity-linked metabolic complications. Interestingly, consumption of the salmon protein diet was also found to raise circulating salmon calcitonin levels, which may underlie the reduction of weight gain in these rats. These data suggest that not all fish protein sources exert the same beneficial properties on the metabolic syndrome, although anti-inflammatory actions appear to be common.

Structural characterization of laminaran and galactofucan extracted from the brown seaweed Saccharina longicruris.

Brown seaweed contains several polysaccharides like laminaran, fucoidan and alginate. Laminaran is a beta-glucan that has shown anti-apoptotic and anti-tumoral activities, while galactofucan (fucoidan) is a sulfated polysaccharide that has displayed anticoagulant, anti-tumor, anti-thrombosis, anti-inflammatory and antiviral properties. In this study, crude laminaran and galactofucan (fucoidan) were extracted from the brown seaweed Saccharina longicruris at four harvest periods (M05, A05, N05 and J06). The galactofucan M05 and N05 fractions were depolymerized (RDP) over 2 or 4h to give 4 RDP fractions (M05 RDP 2H, M05 RDP 4H, N05 RDP 2H and N05 RDP 4H) whose molecular weights, monosaccharide compositions and glycosidic linkages were determined by GC-MS. The laminaran fraction gave a molecular weight range from 2900 to 3300 Da and contained between 50.6% and 68.6% d-glucose and an average of 1.3% D-mannitol. The presence of a beta-(1,3) linkage between D-glucose in the main chain was observed, with branching at positions 6 and 2. The M05 fraction contained less branching than other laminaran fractions, which might have influenced its conformation in solution and thus its activity. The crude galactofucan fractions displayed a molecular weight range from 638 to 1529 kDa, whereas the RDP fractions had molecular weights <30 kDa. The structure of the galactofucan fractions remained complex after depolymerization, with these also being more sulfated (30-39%) than the crude fractions (13-20%). The crude and RDP fractions contained 3-linked fucopyranose 4-sulfate and 6-linked galactopyranose 3-sulfate moieties, although the galactofucans isolated from M05 and J06 contained less 6-linked galactopyranose 3-sulfate than the A05 and N05 fractions.

Effect of season on the composition of bioactive polysaccharides from the brown seaweed Saccharina longicruris.

The structural features of laminarans and galactofucans extracted from the brown seaweed Saccharina longicruris were determined for four harvest periods (M05, A05, N05 and J06). Crude laminarans were purified and crude galactofucans were fractionated using DEAE Sepharose anion exchange chromatography with increasing levels of NaCl (0.5, 1 and 2 M). The results showed differences in terms of their monosaccharide compositions. Purified laminaran contained a high proportion of D-glucose, between 45.1% and 69.1%, with a higher amount in M05 and A05, while the amount of D-mannitol remained constant (less than 1.7%). Crude galactofucans from M05, A05, and N05 contained 19.9-21.5% of sulphates, where J06 had only 14.3%. The 2 M fractionated galactofucans contained a higher proportion of sulphate groups, from 27.1% to 36.9%, for each harvest period, while the 1 M fraction contained 9.2% to 15.9% of sulphates. An important variation in the amount of L-fucose and D-galactose was observed for crude and fractionated galactofucans. In M05, a higher content of L-fucose was observed for crude galactofucans compared to that observed for D-galactose (21.5% vs. 11.1%), whereas the opposite was found for A05 (18.5% vs. 36.6%), N05 (20.9% vs. 36.8%), and J06 (12.8% vs. 19.6%). Also, the 0.5 and 2 M fractions were similar to the crude galactofucans. A05, N05, and J06 contained lower amounts of L-fucose than D-galactose, while the M05 fractions showed the opposite behaviour. However, the 1 M fraction showed a higher amount of L-fucose than D-galactose for each harvest period. The next step will be to study the biological activity of the fractions and to attempt to relate this activity to the structure of the galactofucan and laminaran fractions.