Neuroprotective benefits of grape seed and skin extract in a mouse model of Parkinson's disease.

Parkinson's disease is a neurodegenerative disorder characterized by the progressive loss of midbrain dopaminergic (mDA) neurons in the substantia nigra pars compacta, and it involves oxidative stress. Our goal was to evaluate the neuroprotective effect of Vitis vinifera red grape seed and skin extract (GSSE) in a model of Parkinson's disease. GSSE is very rich in phenolic compounds, such as flavonoids, anthocyanins, catechins and stilbenes, which are present in the pulp, seeds, and leaves of the fruit. GSSE is known for its antioxidant properties and has shown beneficial effects against oxidative injury in different organs, such as the kidneys, liver, heart and brain. In this study, we revealed the neuroprotective effect of GSSE on midbrain dopaminergic neurons both in vitro and in vivo. We used the neurotoxin 6-hydroxydopamine (6-OHDA), which induces oxidative damage and mimics the degeneration of dopaminergic neurons observed in Parkinson's disease. We found that GSSE was effective in protecting dopamine neurons from 6-OHDA toxicity by reducing apoptosis, the level of reactive oxygen species (ROS) and inflammation. Furthermore, we found that GSSE treatment efficiently protected against neuronal loss and improved motor function in an in vivo 6-OHDA model of Parkinson's disease (PD). Altogether, our results show that GSSE acts at multiple levels to protect dopamine neurons from degeneration in a model of PD.

Occurrence of Peptide-Peptide Interactions during the Purification of Self-Assembling Peptide f1-8 from a ß-Lactoglobulin Tryptic Hydrolysate.

Self-assembling peptides have gained attention because of their nanotechnological applications. Previous work demonstrated that the self-assembling peptide f1-8 (Pf1-8) that is generated from the tryptic hydrolysis of ß-lactoglobulin can form a hydrogel after several purification steps, including membrane filtration and consecutive washes. This study evaluates the impact of each processing step on peptide profile, purity, and gelation capacity of each fraction to understand the purification process of Pf1-8 and the peptide-peptide interactions involved. We showed that peptide-peptide interactions mainly occurred through electrostatic and hydrophobic interactions, influencing the fraction compositions. Indeed, the purity of Pf1-8 did not correlate with the number of wash steps. In addition to Pf1-8, two other hydrophobic peptides were identified, peptide f15-20, and peptide f41-60. The gelation observed could be induced either through peptide-peptide interactions or through self-assembling, both being driven by non-covalent bond and more specifically hydrophobic interactions.

Symposium review: The dairy matrix-Bioaccessibility and bioavailability of nutrients and physiological effects.

Several studies have linked food structure and texture to different kinetics of nutrients delivery. Changes in some nutrients' release rate, such as proteins and lipids, could induce different physiological effects (e.g., satiety effect, reduction of postprandial lipemia). Recently, experts are proposing to consider the food as a whole instead of looking at specific nutrients, as the combination of food components and the way they are structured could change their physiological effects. This review highlights recent knowledge linking the different levels of structure of dairy products to their digestion, absorption, and physiological effects. Two examples, yogurt and cheese, will be presented to showcase the contributions of dairy food structure to nutrient release rates. One study aimed to validate whether changes in the casein:whey protein ratio or addition of fiber could influence the digestion kinetics of protein and, subsequently, satiety. A static in vitro digestion model has been used on experimental yogurts differing by their casein:whey protein ratio or dietary fiber content. A human trial with healthy men (n = 20) consuming 5 isocaloric and isoproteinemic yogurt snacks before monitoring lunch intake revealed that the yogurt formulation with increased whey protein content significantly reduced subsequent energy intake compared with its control. This result was linked to slower in vitro disintegration rate and soluble protein release for yogurts with increased whey protein, whereas no difference was observed for yogurts with fiber. A second study allowed discrimination between the effects of cheese attributes on lipid release and absorption. Nine commercial cheeses were digested in vitro, and 2 were selected for the in vivo study, in which plasma concentrations of triglycerides (TAG) were followed before and after meal consumption. The in vivo study revealed that cream cheese, but not cheddar, induced a greater increase in TAG concentrations at 2 h than did butter; this difference was linked to their in vitro disintegration. These studies demonstrate that the dairy food matrix per se modulates foods' nutritional properties. Other studies recently published on this topic will also be included, to put in perspective the important role of the dairy food matrix on release of nutrients and their physiological effects, and how this can be compared with other foods.

Combination of High Hydrostatic Pressure and Ultrafiltration to Generate a New Emulsifying Ingredient from Egg Yolk.

Egg yolk granule phosvitin (45 kDa) is a phosphoprotein known for its emulsifying properties. Recently, high hydrostatic pressure (HHP) treatment of granule induced the transfer of phosvitin to the soluble plasma fraction. This project evaluated the performance of the ultrafiltration (UF) used to concentrate phosvitin from the plasma fraction to produce a natural emulsifier. Phosvitin was characterized in plasma from a pressure-treated granule (1.73 ± 0.07% w/w) and in its UF retentate (26.00 ± 4.12% w/w). The emulsifying properties of both retentates were evaluated. The emulsion prepared with phosvitin-enriched retentate was more resistant to flocculation and creaming. Confocal laser scanning microscopy showed a network of aggregated protein similar to a gel, which encapsulated oil droplets in emulsions made with UF-retentate of plasma from pressure-treated granule. However, although sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that ß-phosvitin is recovered in the cream, it is difficult to attribute the improved emulsifying properties of the UF-retentate of plasma from pressure-treated granules only to phosvitin.

Effect of high-hydrostatic pressure on the digestibility of egg yolk and granule.

The impact of high hydrostatic pressure (HHP) on protein digestibility of egg yolk and egg yolk granule was evaluated by static in vitro digestion using the standardized INFOGEST 2.0 method. The degree of hydrolysis (DH) and the phospholipid content were determined during digestion, and the protein and peptide profiles were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and reverse phase-high pressure liquid chromatography (RP-HPLC). The results showed that HHP induced protein aggregation in egg yolk and granule, mainly by disulfide bridges, which were not disrupted in the oral phase. Proteolysis during the gastric phase improved egg yolk and granule protein solubility, regardless of whether HHP was applied. However, the extent of the samples' digestibility was not affected, with DH values ranging from 15% to 20%. During the intestinal phase, the DH of egg yolk protein (∼40%) was higher than that of the granule (∼25%), probably due to the denser structure of the granule reducing the accessibility of intestinal enzymes. The DH, peptide, and protein profiles of control and HHP-treated egg yolk showed similar protein digestion behaviors for both gastric and intestinal phases. Among the different proteins, only the digestibility of ß-phosvitin in HHP-treated granule was enhanced. Consequently, applying HHP to granules represents an interesting process that improves the digestibility of phosvitin with the potential to generate bioactive phosvitin-derived phosphopeptides. PRACTICAL APPLICATION: High hydrostatic pressure, mainly used as a preservation process, did not impair the nutritional quality of the egg yolk and granule proteins but improved the susceptibility of phosvitin (protein contained in egg yolk) proteolysis to produce bioactive phosphopeptides. Consequently, applying HHP to granules represents an interesting process that improves the digestibility of phosvitin.

Assessment of protein and phospholipid bioaccessibility in ultrafiltered buttermilk cheese using TIM-1 in vitro gastrointestinal methods.

To meet the high consumer demand, butter production has increased over the last few years. As a result, the buttermilk (BM) co-produced volumes require new ways of adding value, such as in cheese manufacturing. However, BM use in cheese milk negatively influences the cheesemaking process (e.g., altered coagulation properties) and the product's final quality (e.g., high moisture content). The concentration of BM by ultrafiltration (UF) could potentially facilitate its use in cheese manufacturing through an increased protein content while maintaining the milk salt balance. Simultaneously, little is known about the digestion of UF BM cheese. Therefore, this study aimed to characterize the impact of UF BM on cheese manufacture, its structure, and its behavior during in vitro digestion. A 2-fold UF concentrated BM was used for cheese manufacture (skim milk [SM] - control). Compositional, textural, and microstructural analyses of cheeses were first conducted. In a second step, the cheeses were fed into an in vitro TNO gastrointestinal digestion model (TIM-1) of the stomach and small intestine and protein and phospholipid (PL) bioaccessibility was studied. The results showed that UF BM cheese significantly differed from SM cheese regarding its composition, hardness (p < 0.05) and microstructure. However, in TIM-1, UF BM and SM cheeses showed similar digestion behavior as a percentage of protein and PL intake. Despite relatively more non-digested and non-absorbed PL in the ileum efflux of UF BM cheese, the initially higher PL concentration contributes to an enhanced nutritional value compared to SM cheese. To our knowledge, this study is the first to compare the bioaccessibility of proteins and PL from UF BM and SM cheeses.

Impact of physicochemical changes in milk ultrafiltration permeate concentrated by reverse osmosis on calcium phosphate precipitation.

This study aimed to characterize and compare the mechanisms of calcium phosphate precipitation in skimmed milk ultrafiltration permeate (MP) and MP preconcentrated by reverse osmosis (ROMP). The effects of different physicochemical parameters such as the pH (8.0), the heating time (60 or 120 min at 60 °C) and the seeding of samples with dicalcium phosphate (DCP) were tested. The concentration of salts (K, Ca, Na, Mg, and P) in the freeze-dried precipitates was measured using inductively coupled plasma (ICP). The amount of remaining ionic calcium was also monitored. Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis were used to characterize the type of calcium phosphate precipitates that formed. The morphological structure of particles was determined using scanning electron microscopy (SEM). The chemical analyses showed that RO increased the rate of precipitated ions, especially Ca and P in MP, while alkalinization to pH 8.0 and heating at 60 °C significantly increased the precipitation of salts, with the calcium phosphate structure changing into complex forms such as hydroxyapatite (HAP) and whitlockite. MP preconcentration by RO paves the way for improving the precipitation yield of milk salts in the form of HAP for Ca fortification in various foods. It offers an original way to valorize the milk salts contained in the high volumes of MP generated by the cheese industry.

Impact of emulsifiers for the nanoencapsulation with maltodextrin of cannabis oil by spray drying on the physicochemical properties and bioaccessibility of cannabinoids.

Our study aimed to investigate the impact of various emulsifiers, namely whey protein isolate (WPI), soy protein isolate (SPI), and Tween 80 (Tw), on their ability to encapsulate cannabis oil with maltodextrin as the wall material. The physicochemical properties of the powder, the stability of the cannabinoids, and their bioaccessibility during static in vitro digestion were examined. The average diameter of fat globules in liquid nanoemulsions was 170, 259, and 95 nm for WPI, SPI, and Tw, respectively. The encapsulation efficiency was high for protein emulsifiers (>95%) compared to Tw (∼16%). Upon powder reconstitution in water, the emulsified fat droplets remained stable for WPI (176 nm); however, higher fat globule size (diameters of 346 nm and 210 nm) was observed for SPI and Tw powders, respectively. All oil powders had high solubility (>97%). The peroxide value (PV) showed nearly a fourfold increase for the oil extracted from the powder than the initial PV of bulk oil (5.2 mEq). However, UPLC-TUV analysis of the main cannabinoids (CBD, THC, and CBN) indicated that there is no significant difference between the various formulations and the bulk oil, except for lower Tw. The in vitro digestion model results showed higher bioaccessibility of the cannabinoids for Tw (∼53%) than for proteins (WPI ∼ 7% and SPI ∼ 10%). These findings suggest that the emulsifiers used for spray drying nanoencapsulation of cannabis oil have an impact on the encapsulation efficiency and cannabinoid bioaccessibility, highlighting the importance of choosing adequate emulsifiying agents for their optimal oral delivery.

Impact of Ultra-High Pressure Homogenization on the Structural Properties of Egg Yolk Granule.

Ultra-high pressure homogenization (UHPH) is a promising method for destabilizing and potentially improving the techno-functionality of the egg yolk granule. This study's objectives were to determine the impact of pressure level (50, 175 and 300 MPa) and number of passes (1 and 4) on the physico-chemical and structural properties of egg yolk granule and its subsequent fractions. UHPH induced restructuration of the granule through the formation of a large protein network, without impacting the proximate composition and protein profile in a single pass of up to 300 MPa. In addition, UHPH reduced the particle size distribution up to 175 MPa, to eventually form larger particles through enhanced protein-protein interactions at 300 MPa. Phosvitin, apovitellenin and apolipoprotein-B were specifically involved in these interactions. Overall, egg yolk granule remains highly stable during UHPH treatment. However, more investigations are needed to characterize the resulting protein network and to evaluate the techno-functional properties of UHPH-treated granule.

Impact of Ultra-High-Pressure Homogenization of Buttermilk for the Production of Yogurt.

Despite its nutritional properties, buttermilk (BM) is still poorly valorized due to its high phospholipid (PL) concentration, impairing its techno-functional performance in dairy products. Therefore, the objective of this study was to investigate the impact of ultra-high-pressure homogenization (UHPH) on the techno-functional properties of BM in set and stirred yogurts. BM and skimmed milk (SM) were pretreated by conventional homogenization (15 MPa), high-pressure homogenization (HPH) (150 MPa), and UHPH (300 MPa) prior to yogurt production. Polyacrylamide gel electrophoresis (PAGE) analysis showed that UHPH promoted the formation of large covalently linked aggregates in BM. A more particulate gel microstructure was observed for set SM, while BM gels were finer and more homogeneous. These differences affected the water holding capacity (WHC), which was higher for BM, while a decrease in WHC was observed for SM yogurts with an increase in homogenization pressure. In stirred yogurts, the apparent viscosity was significantly higher for SM, and the pretreatment of BM with UHPH further reduced its viscosity. Overall, our results showed that UHPH could be used for modulating BM and SM yogurt texture properties. The use of UHPH on BM has great potential for lower-viscosity dairy applications (e.g., ready-to-drink yogurts) to deliver its health-promoting properties.

High hydrostatic pressure induced extraction and selective transfer of ß-phosvitin from the egg yolk granule to plasma fractions.

Egg yolk phosvitin is of particular interest due to its functional and biological properties. Recently, it was demonstrated that high hydrostatic pressure (HHP) (400 MPa for 5 min) induced the transfer of folic acid and phosvitin from the egg yolk granule to the plasma fraction. A granule fraction (Gin) produced by egg yolk centrifugation was pressure-treated at 400 and 600 MPa for 5 and 10 min, and centrifuged to generate granule fractions (GP1 to GP4) and plasmas (PP1 to PP4). Iron and phosphorus contents were also increased in PP1 to PP4 fractions, confirming the transfer of phosvitins from pressure-treated granule to plasma. Pressurization drastically improved phosvitin recovery in PP fractions, specifically at 600 MPa for 10 min, which had the highest value of phosvitin/100 mg of dry plasma at 33.3 ± 4.39 mg. Consequently, HHP represents an alternative approach for phosvitin transfer and recovery in the egg yolk soluble fraction.

Evaluation of casein as a binding ligand protein for purification of alpha-lactalbumin from beta-lactoglobulin under high hydrostatic pressure.

Fractionation of ß-lactoglubulin (ß-lg) and α-lactalbumin (α-la) using conventional separation technologies remains challenging mainly due to similar molecular weight. Herein, casein (CN) was used as ligand protein to specifically aggregate ß-lg under high hydrostatic pressure (HHP) in order to separate α-la after acidification to pH 4.6. Specifically, we studied the effect of different concentration of CN on α-la purity and recovery. Model solutions of α-la, ß-lg and CN (from 0 to 5 mg/mL) were pressurized (600 MPa-5 min). After acidification and centrifugation of pressure-treated solutions, purity of α-la was increased up to 78% with a recovery of 88% for solution without CN. In contrast with our initial hypothesis, the presence of CN decreased ß-lg pressure-induced aggregation and co-precipitation upon acidification and significantly reduced purity (∼71%). Therefore, our results suggest a chaperone-like activity of CN on ß-lg pressure-induced aggregation which needs further investigation.

Formation of Stable Supramolecular Structure with ß-Lactoglobulin-Derived Self-Assembling Peptide f1-8 and Bovine Micellar Caseins.

The development of stable macromolecular structures with tailored functional properties in the dairy industry using innovative stabilizers is of great interest. The self-assembling peptide f1-8 (Pf1-8) derived from ß-lactoglobulin was found to interact with whey proteins, consequently changing their physicochemical properties. The objective of the present work was to evaluate the interaction between Pf1-8 and micellar casein (CN) and the changes in their physicochemical properties and stability at different pH values (6.6-2.6) on model solutions containing CN and Pf1-8 at various ratios (1:1, 5:1, and 10:1) using spectrofluorimetry, TEM, SEC-HPLC, and SDS-PAGE analyses. No CN precipitation occurred for the solution at the 1:1 ratio even at pH values below 4.6. In all samples, CN was completely dissociated to primary casein particles (PCP) to form stable supramolecular structures strongly bound to peptide gels via hydrophobic interactions. Thus, a novel milk-protein-derived peptide responsible for stabilizing complex structures composed of CN was discovered.

Transcriptional repression of Plxnc1 by Lmx1a and Lmx1b directs topographic dopaminergic circuit formation.

Mesodiencephalic dopamine neurons play central roles in the regulation of a wide range of brain functions, including voluntary movement and behavioral processes. These functions are served by distinct subtypes of mesodiencephalic dopamine neurons located in the substantia nigra pars compacta and the ventral tegmental area, which form the nigrostriatal, mesolimbic, and mesocortical pathways. Until now, mechanisms involved in dopaminergic circuit formation remained largely unknown. Here, we show that Lmx1a, Lmx1b, and Otx2 transcription factors control subtype-specific mesodiencephalic dopamine neurons and their appropriate axon innervation. Our results revealed that the expression of Plxnc1, an axon guidance receptor, is repressed by Lmx1a/b and enhanced by Otx2. We also found that Sema7a/Plxnc1 interactions are responsible for the segregation of nigrostriatal and mesolimbic dopaminergic pathways. These findings identify Lmx1a/b, Otx2, and Plxnc1 as determinants of dopaminergic circuit formation and should assist in engineering mesodiencephalic dopamine neurons capable of regenerating appropriate connections for cell therapy.Midbrain dopaminergic neurons (mDAs) in the VTA and SNpc project to different regions and form distinct circuits. Here the authors show that transcription factors Lmx1a, Lmx1b, and Otx2 control the axon guidance of mDAs and the segregation of mesolimbic and nigrostriatal dopaminergic pathways.

Characterization of Lactobacillus reuteri interaction with milk fat globule membrane components in dairy products.

A set of methods has been developed to study the adhesion between four Lactobacillus reuteri strains and the milk fat globule membrane (MFGM) components in dairy products. By combining sucrose density gradient (SDG) centrifugation and bacterial DNA quantification it was found which strains of L. reuteri were more strongly associated with the dairy products, and the results were corroborated by direct binding rate and force measurements made with optical tweezers. It was determined that strong binding was associated with hydrophobicity of the bacteria and that this hydrophobicity is correlated with the presence of LiCl-extractable protein on the surface of the bacteria. Confocal laser scanning microscopy (CLSM) allowed for the visualization of interactions between bacteria and MFGM. This study demonstrates that these methods can be used in combination to characterize, both qualitatively and quantitatively, the adhesion of lactic acid bacteria strains in dairy products.