High-Moisture Extrusion of a Dietary Protein Blend Impairs In Vitro Digestion and Delays In Vivo Postprandial Plasma Amino Acid Availability in Humans.

BACKGROUND: Industrial processing can alter the structural complexity of dietary proteins and, potentially, their digestion and absorption upon ingestion. High-moisture extrusion (HME), a common processing method used to produce meat alternative products, affects in vitro digestion, but human data are lacking. We hypothesized that HME of a mycoprotein/pea protein blend would impair in vitro digestion and in vivo postprandial plasma amino acid availability. METHODS: In Study A, 9 healthy volunteers completed 2 experimental trials in a randomized, double-blind, crossover design. Participants consumed a beverage containing 25 g protein from a "dry" blend (CON) of mycoprotein/pea protein (39%/61%) or an HME content-matched blend (EXT). Arterialized venous blood samples were collected in the postabsorptive state and regularly over a 5-h postprandial period to assess plasma amino acid concentrations. In Study B, in vitro digestibility of the 2 beverages were assessed using bicinchoninic acid assay and optical fluorescence microscopy at baseline and during and following gastric and intestinal digestion using the INFOGEST model of digestion. RESULTS: Protein ingestion increased plasma total, essential (EAA), and branched-chain amino acid (BCAA) concentrations (time effect, P < 0.0001) but more rapidly and to a greater magnitude in the CON compared with the EXT condition (condition × time interaction, P < 0.0001). This resulted in greater plasma availability of EAA and BCAA concentrations during the early postprandial period (0-150 min). These data were corroborated by the in vitro approach, which showed greater protein availability in the CON (2150 ± 129 mg/mL) compared with the EXT (590 ± 41 mg/mL) condition during the gastric phase. Fluorescence microscopy revealed clear structural differences between the 2 conditions. CONCLUSIONS: These data demonstrate that HME delays in vivo plasma amino acid availability following ingestion of a mycoprotein/pea protein blend. This is likely due to impaired gastric phase digestion as a result of HME-induced aggregate formation in the pea protein. This trial was registered at clinicaltrials.gov as NCT05584358.

Role of calcium on lipid digestion and serum lipids: a review.

Calcium is an essential nutrient for humans that can be taken as supplement or in a food matrix (e.g. dairy products). It is suggested that dietary calcium may have a beneficial effect on cardiovascular risk but the mechanism is not clear. In this review, the main mechanisms of the possible cholesterol-lowering effect of calcium, i.e. interaction with fatty acids and bile acids, are described and clinical evidences are presented. The observations from interventional studies of the possible cholesterol-lowering effect in terms of the main related mechanisms are variable and do not seem to fulfill all the related aspects. It seems that the interplay of calcium in blood lipid metabolism might be due to its complex and multiple roles in the lipid digestion in the small intestine. The interactions between calcium and, fatty acids and bile may lead to impaired mixed micelle formation and solubilization, which is crucial in the lipid absorption and metabolism. In addition, the calcium source and its surrounding matrix will have an influence over the physiological outcome. This research is important for the delivery and formulation of calcium, particularly with the move toward plant-based diets.

An evolving view on food viscosity regulating gastric emptying.

Viscosity is a property of most foods. The consumption of the high-viscosity food is associated with a variety of physiological responses, one of which is their ability to regulate gastric emptying and modulate postprandial glycemic response. Gastric emptying has been proven to be a key step affecting the digestion and absorption of food, whereas, the relationship between viscosity and gastric emptying is still far away from being understood. Here, we reviewed the factors that influence food viscosity and food viscosity changes during digestion. Besides, the effect of food viscosity on gastric emptying and food-viscosity-physiological response were highlighted. Finally, "quantitative relationship" of viscosity and gastric emptying was discussed. This review can contribute to the understanding that how food viscosity affects gastric emptying, and help for developing foods that could control satiety and manage body weight for the specific populations.

Anti-Inflammatory Activity of an In Vitro Digested Anthocyanin-Rich Extract on Intestinal Epithelial Cells Exposed to TNF-α.

BACKGROUND: The consumption of foods rich in anthocyanins (ACN) have been associated with beneficial properties in chronic inflammatory disorders such as intestinal bowel diseases (IBD). These effects were attributed not only to a direct antioxidant mechanism but also to the modulation of cell redox-dependent signaling. However, ACN bioavailability is low for their poor stability in the digestive tract, so ACN gastrointestinal digestion should be considered. METHODS: To have a more realistic knowledge of the effects of ACN, we performed an in vitro simulated gastrointestinal digestion of an ACN-rich purified and standardized bilberry and blackcurrant extract (BBE), followed by an evaluation of ACN composition modification (HPLC-DAD and pH differential method) and antioxidant activity (FRAP assay). Then, we studied the effects of BBE gastrointestinal extract on Caco-2 exposed to TNF-α. RESULTS: The results confirmed the high instability of ACN in the mild alkaline environment of the small intestine (17% recovery index). However, the digested BBE maintained part of its bioactivity. Additionally, BBE gastrointestinal extract inhibited the TNF-α-induced NF-κB pathway in Caco-2 and activated the Nrf2 pathway. CONCLUSIONS: Although ACN stability is affected by gastrointestinal digestion, the anti-inflammatory and antioxidant activity of digested extracts were confirmed; thus, the loss of ACN can probably be counterweighed by their metabolites. Then, ACN introduced by diet or food supplements could represent an approach for IBD prevention.

Mechanisms, physiology, and recent research progress of gastric emptying.

Gastric emptying refers to a process in which the stomach discharges its contents into the small intestine to further digest and absorb nutrients. Understanding the mechanisms of gastric emptying and relationships between food and individuals is of paramount importance for the design and manufacture of novel and healthy foods. For ethical and cost reasons, in vivo tests are not always possible. In vitro digestion models therefore play a key role in current exploration of gastric emptying. This review outlines the mechanisms and physiology of gastric emptying, including calories, viscosity, composition of the food, age and gender of the individual. In addition, recent progress on in vitro static and dynamic gastric digestion models and future research trends are included in this review.

Dairy structures and physiological responses: a matter of gastric digestion.

Digestion and health properties of food do not solely rely on the sum of nutrients but are also influenced by food structure. Dairy products present an array of structures due to differences in the origin of milk components and the changes induced by processing. Some dairy structures have been observed to induce specific effects on digestion rates and physiological responses. However, the underlying mechanisms are not fully understood. Gastric digestion plays a key role in controlling digestion kinetics. The main objective of this review is to expose the relevance of gastric phase as the link between dairy structures and physiological responses. The focus is on human and animal studies, and physiological relevant in vitro digestion models. Data collected showed that the structure of dairy products have a profound impact on rate of nutrient bioavailability, absorption and physiological responses, suggesting gastric digestion as the main driver. Control of gastric digestion can be a tool for delivering specific rates of nutrient digestion. Therefore, the design of food structure targeting specific gastric behavior could be of great interest for particular population needs e.g. rapid nutrient digestion will benefit elderly, and slow nutrient digestion could help to enhance satiety.

Structural and technological characterization of pectin extracted with sodium citrate and nitric acid from sunflower heads.

An optimization of temperature, time, and extracting agent concentration of pectin extraction from sunflower heads using sodium citrate and nitric acid (SP-SC and SP-NA) was carried out. At optimal conditions, the yield of extraction with nitric acid (SPO-NA) was twofold greater than the corresponding with sodium citrate (SPO-SC) (14.3 versus 7.7%, respectively). Regarding pectin structure, the galacturonic acid (GalA) content in both, SPO-SC and SPO-NA, was similar (∼85%). However, SPO-NA showed lower molecular weight (Mw) (88.9 kDa) and neutral sugar content (4%) than SPO-SC (464 kDa, 9%), indicating that nitric acid deeply degraded pectin structure. These differences derived into dissimilar behavior in their technological functionality. SPO-SC showed higher viscosity and better emulsifying capacity than SPO-NA, although any of them were able to stabilize the oil/water emulsion. Both sunflower pectins formed gels with Ca2+ (75 mg/g of pectin) at pH 3.0. However, when sucrose was added, the gels formed by SP-SC and 20% sucrose presented the same hardness as those of SP-NA with 40% sucrose. These results suggest that the pectin extracted with sodium citrate, an eco-friendly agent, could be a promising ingredient, with good thickening and gelling properties.

Stabilisation of oil-in-water emulsions with non-chemical modified gelatinised starch.

In this research, stabilisation of oil-in-water emulsions with non-chemically modified gelatinised starch is presented. Thus far only octenyl succinic anhydride (OSA) modified gelatinised starch has been known to adsorb at emulsion droplet interfaces, acting as emulsifiers. Screening a range of commercially available food starches revealed that a non-waxy rice starch, a waxy rice starch and the waxy maize starch PRIMA600 showed oil-in-water emulsifying ability following gelatinisation. The microstructure of emulsions formulated with 20% oil and 1% starch was stable for at least 3 months. Thermal, crystallinity and molecular property analyses as well as amylose and protein content revealed no obvious link to this property. Nevertheless, this research has provided the food industry with exciting results for the formulation of clean label emulsions. Moreover, it presents a concept for oral release food emulsions with destabilisation via salivary amylase digestion of the stabilising starch emulsifier.

Influence of oat components on lipid digestion using an in vitro model: Impact of viscosity and depletion flocculation mechanism.

Depletion flocculation is a well-known instability mechanism that can occur in oil-in-water emulsions when the concentration of non-adsorbed polysaccharide exceeds a certain level. This critical flocculation concentration depends on the molecular characteristics of the polysaccharide molecules, such as their molecular weight and hydrodynamic radius. In this study, a range of analytical methods (dynamic shear rheology, optical microscopy, and static light-scattering) were used to investigate the interaction between lipid droplets and polysaccharides (guar gum and ß-glucans) of varying weight-average molecular weight and hydrodynamic radius, and concentration. The aim of this work was to see if the health benefits of soluble fibers like ß-glucans could be explained by their influence on the structure and digestibility of lipid emulsions. The apparent viscosity of the emulsions increased with increasing polysaccharide concentration, molecular weight, and hydrodynamic radius. Droplet flocculation was observed in the emulsions only at certain polysaccharide concentrations, which was attributed to a depletion effect. In addition, the water-soluble components in oat flakes, flour, and bran were extracted using aqueous solutions, to examine their impact on emulsion stability and properties. Then, the rate and extent of lipolysis of a sunflower oil-in-water emulsion in the presence of these oat extracts were monitored using the pH-stat method. However, the inhibition of lipolysis was not linearly related to the viscosity of the oat solutions. The water-soluble extracts of ß-glucan collected from oat flakes had a significant inhibitory effect on lipolysis. The results of this study increase our understanding of the possible mechanisms influencing the impact of oat constituents on lipid digestion. This work also highlights the importance of considering the molecular properties of polysaccharides, and not just their impact on solution viscosity.

Intrinsic wheat lipid composition effects the interfacial and foaming properties of dough liquor.

Doughs were prepared from a single variety breadmaking flour (cv. Hereward), from three successive harvests (years; 2011, 2012 and 2013). A preparation of the aqueous phase from dough, known as dough liquor (DL), was prepared by ultracentrifugation and its physico-chemical properties were investigated. Surface tension and interfacial rheology, showed that the interface of DL was lipid-dominated and that 2013 DL had a different type of interface to 2011 and 2012 DL. This data was consistent with the improved foam stability observed for 2013 DL and with the types of lipids identified. All foams collapsed quickly, but the most stable foam was from 2013 DL with 89.2% loss in foam, followed by 2011 DL with 91.7% loss and 2012 had the least stable foam with a loss of 92.5% of the foam structure. Glycolipids (DGDG and MGDG) were enriched in 2013 DL, and were also present in DL foam, contributing towards improved stability. Neutral lipids, such as FFAs, were enriched in DL foams contributing towards instability and rapid foam collapse. Baking trials using 2012 and 2013 flour, showed increased loaf volumes and gas bubble diameter in 2013 bread compared to 2012 bread, highlighting the potential impact that surface active polar lipids, enriched in the aqueous phase of dough, could have on improving breadmaking quality.

Oatmeal particle size alters glycemic index but not as a function of gastric emptying rate.

The aim of this study was to determine the extent to which oat particle size in a porridge could alter glucose absorption, gastric emptying, gastrointestinal hormone response, and subjective feelings of appetite and satiety. Porridge was prepared from either oat flakes or oat flour with the same protein, fat, carbohydrate, and mass. These were fed to eight volunteers on separate days in a crossover study, and subjective appetite ratings, gastric contents, and plasma glucose, insulin, and gastrointestinal hormones were determined over a period of 3 h. The flake porridge gave a lower glucose response than the flour porridge, and there were apparent differences in gastric emptying in both the early and late postprandial phases. The appetite ratings showed similar differences between early- and late-phase behavior. The structure of the oat flakes remained sufficiently intact to delay their gastric emptying, leading to a lower glycemic response, even though initial gastric emptying rates were similar for the flake and flour porridge. This highlights the need to take food structure into account when considering relatively simple physiological measures and offering nutritional guidance.NEW & NOTEWORTHY The impact of food structure on glycemic response even in simple foods such as porridge is dependent on both timing of gastric emptying and the composition of what is emptied as well as duodenal starch digestion. Thus structure should be accounted for when considering relatively simple physiological measures and offering nutritional guidance.

Correlation between pedicle size and the rate of pedicle screw misplacement in the treatment of thoracic fractures: Can we predict how difficult the task will be?

STUDY DESIGN: A retrospective study. OBJECTIVE: To correlate the incidence of pedicle-screw (PS) misplacement with the dimensions of the pedicles in the treatment of thoracic spine fractures. SUMMARY OF BACKGROUND DATA: The technical challenge of internal fixation with PS in the thoracic spine has been well documented in the literature. However, there are no publications that document the correlation between the pedicle dimensions of the thoracic vertebrae in the preoperative computed tomography scans (CT) and the rate of PS misplacement. METHODS: All patients who had PSs inserted between the T1 and T12 vertebrae during a 24-month period were included in this study. PS position was assessed on high quality CT scans by two independent observers and classified in 2 categories: correct or misplaced. The transverse diameter, craniocaudal diameter and cross-sectional area of the pedicles from T1 to T12 were measured in the pre-operative CT. RESULTS: During the period of this study 36 patients underwent internal fixation with 218 PS. Of the 218 screws, 184 (84.5%) were correct and 34 (15.5%) were misplaced. Misplacement rate was 33% for pedicles with a transverse diameter less than 5 mm, 10.7% for those with a transverse diameter between 5 and 7 mm and 0% for those with a transverse diameter larger than 7 mm. There was a statistically significant difference in the rate of PS misplacement in pedicles with transverse diameter smaller than 5 mm compared with the others. Also, those with transverse diameter between 5.1 and 7 mm compared with those bigger than 7 mm in diameter. The rate of PS misplacement was higher between T3 and T9 (p < 0.05), which in turn correlated with pedicle transverse diameter. CONCLUSION: The rate of PS misplacement in the mid thoracic spine (T4-T9) is high and correlates with pedicle transverse diameter.

Enumerating and indexing many-body intramolecular interactions: a graph theoretic approach.

The central idea observes a recursive mapping of [Formula: see text]-body intramolecular interactions to [Formula: see text]-body terms that is consistent with the molecular topology. Iterative application of the line graph transformation is identified as a natural and elegant tool to accomplish the recursion. The procedure readily generalizes to arbitrary [Formula: see text]-body potentials. In particular, the method yields a complete characterization of [Formula: see text]-body interactions. The hierarchical structure of atomic index lists for each interaction order [Formula: see text] is compactly expressed as a directed acyclic graph. A pseudo-code description of the generating algorithm is given. With suitable data structures (e.g., edge lists or adjacency matrices), automatic enumeration and indexing of [Formula: see text]-body interactions can be implemented straightforwardly to handle large bio-molecular systems. Explicit examples are discussed, including a chemically relevant effective potential model of taurocholate bile salt.

The adsorption-desorption behaviour and structure function relationships of bile salts.

The digestion of dietary components in the human gastrointestinal (GI) tract is a complex, dynamic, inherently heterogeneous process. A key aspect of the digestion of lipid in the GI tract is the combined action of bile salts, lipase and colipase in hydrolysing and solubilising dispersed lipid. The bile salts are a mixture of steroid acid conjugates with surfactant properties. In order to examine whether the different bile salts have different interfacial properties their dynamic interfacial behaviour was characterised. Differences in the adsorption behaviour to solid hydrophobic surfaces of bile salt species were studied using dual polarisation interferometry and atomic force microscopy (AFM) under physiological conditions. Specifically, the cholates adsorbed more slowly and a significant proportion were irreversibly adsorbed following buffer rinsing; whereas the deoxycholates and chenodeoxycholates adsorbed more rapidly and desorbed to a greater extent following buffer rinsing. The conjugating groups (taurine, glycine) did not influence the behaviour. AFM showed that the interfacial structures that remained following buffer rinsing were also different between these two groups. In addition, the adsorption-desorption behaviour affected the adsorption of colipase to a solid surface. This supports the idea that cooperative adsorption occurs between certain bile salts and colipase to facilitate the adsorption and activity of pancreatic lipase in order to restore lipolytic activity in the presence of bile salts. This study provides insights into how differences in bile salt structure could affect lipase activity and solubilisation of lipolysis products and other lipid-soluble bioactive molecules.

The effects of processing and mastication on almond lipid bioaccessibility using novel methods of in vitro digestion modelling and micro-structural analysis.

A number of studies have demonstrated that consuming almonds increases satiety but does not result in weight gain, despite their high energy and lipid content. To understand the mechanism of almond digestion, in the present study, we investigated the bioaccessibility of lipids from masticated almonds during in vitro simulated human digestion, and determined the associated changes in cell-wall composition and cellular microstructure. The influence of processing on lipid release was assessed by using natural raw almonds (NA) and roasted almonds (RA). Masticated samples from four healthy adults (two females, two males) were exposed to a dynamic gastric model of digestion followed by simulated duodenal digestion. Between 7·8 and 11·1 % of the total lipid was released as a result of mastication, with no significant differences between the NA and RA samples. Significant digestion occurred during the in vitro gastric phase (16·4 and 15·9 %) and the in vitro duodenal phase (32·2 and 32·7 %) for the NA and RA samples, respectively. Roasting produced a smaller average particle size distribution post-mastication; however, this was not significant in terms of lipid release. Light microscopy showed major changes that occurred in the distribution of lipid in all cells after the roasting process. Further changes were observed in the surface cells of almond fragments and in fractured cells after exposure to the duodenal environment. Almond cell walls prevented lipid release from intact cells, providing a mechanism for incomplete nutrient absorption in the gut. The composition of almond cell walls was not affected by processing or simulated digestion.

Structural and compositional changes in the salivary pellicle induced upon exposure to SDS and STP.

Sodium dodecyl sulphate (SDS) and sodium tripolyphosphate (STP) act to remove stained pellicle from dentition and loosen deposits on tooth surfaces that may become cariogenic over time. This study investigated how SDS and STP impact the salivary pellicle adsorbed onto hydroxyapatite and silica sensors using a dual polarisation interferometer and a quartz-crystal microbalance with dissipation. After the pellicle was exposed to SDS and STP the remaining pellicle, although weaker, due to the loss of material, became less dense but with a higher elastic component; suggesting that the viscous component of the pellicle was being removed. This would imply a structural transformation from a soft but dense structured pellicle, to a more diffuse pellicle. In addition, the majority of proteins displaced by both SDS and STP were identified as being acidic in nature; implying that the negatively charged groups of SDS and STP may be responsible for the displacement of the pellicle proteins observed.

Pea protein extraction method impacts the protein (micro)structural organisation and in vitro digestion kinetics.

There is increasing interest in including pulse proteins into food products due to their nutrient-rich and sustainable character. However, little is known regarding the consequences of different extraction approaches on the pulse protein structure and the subsequent protein (micro)structural organization and protein digestion kinetics. Therefore, three green pea protein extracts were created: (i) cooking followed by cotyledon cell isolation, (ii) alkaline extraction followed by isoelectric precipitation, or (iii) salt extraction, and compared to the original pea flour as well as to sodium caseinate. The results showed that encapsulated, denatured protein inside pea cotyledon cells presented the (s)lowest digestion, while accessible and more native protein (e.g., pea flour, pea protein salt extract) presented much faster and higher digestion. Moreover, the alkali extracted pea protein was denatured to some extent, significantly lowering in vitro digestion kinetics. In the second part, three different in vitro approaches were applied to digest the salt extracted pea protein. Semi-dynamic gastric digestion approaches simulate in vivo conditions more closely which especially impacted the rate of digestion.

Understanding the mechanism of high viscosity food delaying gastric emptying.

Controlling the structure and viscosity of food can influence the development of diet-related diseases. Food viscosity has been linked with health through its impact on human digestion and gastrointestinal transit, however, there is limited understanding of how the viscosity of food regulates gastric emptying. Here, we used model food preparations with different viscosities using guar gum, to explore the mechanism underlying the influence of viscosity on gastric motility, gastric emptying and postprandial blood glucose. Based on experiments in human volunteers and animals, we demonstrated that high viscosity meals increased gastric antrum area and gastric retention rate. Viscosity also affected gut hormone secretion, reduced the gene expression level of interstitial cells of Cajal, resulting in a delay of gastric emptying and limiting the increase in postprandial glucose. This improved mechanistic understanding of food viscosity during gastric digestion is important for designing new foods to benefit human health.

Postprandial lipid and vascular responses following consumption of a commercially-relevant interesterified palmitic acid-rich spread in comparison to functionally-equivalent non-interesterified spread and spreadable butter: a randomised controlled trial in healthy adults.

Background: Interesterification is an industrial processing technique used widely where hard fats are essential for functionality and consumer acceptability, e.g. margarines and lower fat spreads. Objective: The aim of this study was to compare acute cardiovascular effects of functionally equivalent spreads (similar solid fat content) made with interesterified (IE) or non-IE palm-based fats, or spreadable butter. Methods: A randomised, controlled, 4-armed crossover, double-blind study (25 men, 25 women; 35-75 years; healthy; mean BMI 24.5, SD 3.8), compared effects of mixed nutrient meals containing 50 g fat from functionally equivalent products [IE spread, non-IE spread and spreadable butter (SB), with rapeseed oil (RO) as a reference treatment: with 16.7%, 27.9%, 19.3% and 4% palmitic acid, respectively] on 8 h postprandial changes in plasma triacylglycerol (TAG) and endothelial dysfunction (flow-mediated dilatation; FMD). Circulating reactive oxygen species (estimated using a neutrophil oxidative burst assay), glucose, insulin, NEFA, lipoprotein particle profiles, inflammatory markers (glycoprotein acetylation (Glyc-A) and IL-6), and biomarkers of endotoxemia were measured. Results: Postprandial plasma TAG concentrations after test meals were similar. However following RO versus the 3 spreads, there were significantly higher postprandial apolipoprotein B concentrations, and small HDL and LDL particle concentrations, and lower postprandial extra-large, large, and medium HDL particle concentrations, as well as smaller average HDL and LDL particle sizes. There were no differences following IE compared to the other spreads. Postprandial FMD% did not decrease after high-fat test meals, and there were no differences between treatments. Postprandial serum IL-6 increased similarly after test meals, but RO provoked a greater increase in postprandial concentrations of glycoprotein acetyls (GlycA), as well as 8 h sCD14, an endotoxemia marker. All other postprandial outcomes were not different between treatments. Conclusions: In healthy adults, a commercially-available IE-based spread did not evoke a different postprandial triacylglycerol, lipoprotein subclass, oxidative stress, inflammatory or endotoxemic response to functionally-equivalent, but compositionally-distinct alternative spreads. Clinical trial registry number: NCT03438084 (https://ClinicalTrials.gov).

Cyanidin-3-O-glucoside protects intestinal epithelial cells from palmitate-induced lipotoxicity.

CONTEXT: Increased free fatty acids (FFAs) levels, typical in obesity condition, can contribute to systemic lipotoxicity and inflammation adversely influencing Inflammatory Bowel Disease development and progression. Anthocyanins possess health promoting properties mainly associated to the induction of Nrf2-regulated cytoprotective proteins. OBJECTIVE: Using a novel experimental model, we evaluated the in vitro intracellular mechanisms involved in FFAs modulation of intestinal epithelial lipotoxicity and the protective effects of cyanidin-3-O-glucoside (C3G) in Caco-2 cells. RESULTS: Caco-2 exposed to palmitic acid (PA) in the serosal (basolateral) side showed a combined state of epithelial inflammation, inducing NF-κB pathway and downstream cytokines, that was reverted by C3G apical pre-treatment. In addition, PA altered intracellular redox status and induced reactive oxygen species that were reduced by C3G via the redox-sensitive Nrf2 signalling. DISCUSSION AND CONCLUSION: Results suggest that anti-inflammatory properties of anthocyanins, mediated by Nrf2, could represent an interesting tool for intestinal inflammatory disorders.