Preventative Effects of Milk Fat Globule Membrane Ingredients on DSS-Induced Mucosal Injury in Intestinal Epithelial Cells.

The goblet cells of the gastrointestinal tract (GIT) produce glycoproteins called mucins that form a protective barrier from digestive contents and external stimuli. Recent evidence suggests that the milk fat globule membrane (MFGM) and its milk phospholipid component (MPL) can benefit the GIT through improving barrier function. Our objective was to compare the effects of two digested MFGM ingredients with or without dextran sodium sulfate (DSS)-induced barrier stress on mucin proteins. Co-cultured Caco-2/HT29-MTX intestinal cells were treated with in vitro digests of 2%, 5%, and 10% (w/v) MFGM or MPL alone for 6 h or followed by challenge with 2.5% DSS (6 h). Transepithelial electrical resistance and fluorescein isothiocyanate (FITC)-dextran (FD4) permeability measurements were used to measure changes in barrier integrity. Mucin characterization was performed using a combination of slot blotting techniques for secreted (MUC5AC, MUC2) and transmembrane (MUC3A, MUC1) mucins, scanning electron microscopy (SEM), and periodic acid Schiff (PAS)/Alcian blue staining. Digested MFGM and MPL prevented a DSS-induced reduction in secreted mucins, which corresponded to the prevention of DSS-induced increases in FD4 permeability. SEM and PAS/Alcian blue staining showed similar visual trends for secreted mucin production. A predictive bioinformatic approach was also used to identify potential KEGG pathways involved in MFGM-mediated mucosal maintenance under colitis conditions. This preliminary in silico evidence, combined with our in vitro findings, suggests the role of MFGM in inducing repair and maintenance of the mucosal barrier.

Reduction in the antigenicity of beta-lactoglobulin in whole milk powder via supercritical CO2 treatment.

Cows' milk allergy (CMA) is a common phenomenon experienced in early childhood (<5 years of age) with an average occurrence rate of roughly 2.5%. The most prevalent allergen in cows' milk is believed to be ß-lactoglobulin (ß-LG). The objective of this study was to evaluate the use of hydrophobic supercritical CO2 (ScCO2) to modify the chemical structure ß-LG thus impairing its recognition by antibodies. Whole milk powder was selected because of its closest compositional resemblance to bovine fluid milk and its applications in reconstitution and in the beverage (infant, toddler, and adult), confectionary, bakery, and meat industries. For this study, whole milk powder was treated with food-grade CO2 at temperatures of 50, 63, and 75 °C under operating pressures of 100, 150, 200, 250, and 300 bar. Proteins in whole milk powder were examined using SDS-PAGE, Western blot, and ELISA. Orbitrap Fusion LC/MS-MS and periodic staining was performed to confirm post-translational modifications in ß-LG. Functional properties of whole milk powder before and after treatment were assessed by its solubility index, oil holding capacity, emulsion capacity and stability, zeta-potential, particle size, and color analysis. SDS-PAGE of treated samples yielded fuzzy bands (variable mobility of molecules due to different MW results in ill-defined bands) indicative of an increase in molecular weight, presumably due to chemical change in the protein, and demonstrated a maximum of 71.13 ± 0.29% decrease in the band intensity of ß-LG under treatment conditions of 75 °C/300 bar for 30 min (P < 0.05). These changes were small with samples treated with heat only. Lighter, diffused bands were observed using Western blot analysis. ELISA tests proved that ScCO2 treatment specifically and significantly affected the antigenicity of ß-LG with a reduction of 42.9 ± 2.83% and 54.75 ± 2.43% at 63 °C/200 bar and 75 °C/300 bar, respectively. Orbitrap fusion detected the presence of fatty acids and sugar moieties bound to ß-LG and the latter was confirmed by periodic staining. Functional properties of ScCO2-treated milk powder yielded a decrease in solubility index and an increase in emulsion capacity of whole milk powder was observed under ScCO2 treatment at 75 °C/300 bar (P < 0.05), with small and insignificant changes at other treatments producing a decrease in antigenicity. Color changes were small for most samples, except at 63 °C/200 bar, where a significant increase in yellowness was observed. Zeta-potential and particle size measurements indicated that most changes were temperature driven. This study demonstrates 2 approaches to mitigate ß-LG antigenicity via fatty acid binding and lactosylation using hydrophobic ScCO2.

Evaluation of Kefir Grain Microbiota, Grain Viability, and Kefir Bioactivity from Fermenting Dairy Processing By-Products.

Four dairy foods processing by-products (acid whey permeate (AWP), buttermilk (BM), sweet whey permeate (SWP), and sweet whey permeate with added milk fat globule ingredient (SWP+MFGM)) were fermented for 4 weeks and compared with traditional kefir milks for production of novel kefir-like dairy products. AU: Sweet whey permeates and SWP supplemented with 1.5% milk fat globule membrane (MFGM) showed to be the most viable by-products for kefir grain fermentation, exhibiting diverse abundance of traditional kefir microorganisms and positive indicators of bioactive properties. Grain viability was assessed with shotgun metagenomics, texture profile analysis, live cell counts, and scanning electron microscopy. Assessed bioactivities of the kefir-like products included antibacterial, antioxidant, potential anticancerogenic properties, and membrane barrier effect on human colorectal adenocarcinoma Caco-2 cells. All kefir grains were most abundant in Lactobacillus kefiranofaciens when analyzed with shotgun metagenomics. When analyzed with live cell counts on selective media, AWP kefir-like product had no countable Lactococcus spp. indicating suboptimal conditions for kefir grain microbiota survival and application for fermented dairy starter culture bacterium. Live cell counts were affirmed with kefir grain surface scanning electron microscopy images. SWP had the most adhesive kefir grain surface while SWP+MFGM had the largest exopolysaccharide (EPS) yield from grain extraction. All kefir and kefir-like products were able to achieve a 6-log reduction against Listeria innocua and Escherichia coli. Traditional milk kefirs had the highest antioxidant capacity for 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay. AWP had a significantly higher DPPH antioxidant activity and SWP had the lowest Trolox equivalence concentration in the ABTS assay. Sweet whey and supplemented milk fat sweet whey had upregulation of Cldn-1 and Ocln-1 gene expression, which correspond with a significant increase in transepithelial electrical resistance.

Milk phospholipids protect Bifidobacterium longum subsp. infantis during in vitro digestion and enhance polysaccharide production.

Bifidobacterium longum subsp. infantis is associated with the gut microbiota of breast-fed infants. Bifidobacterium infantis promotes intestinal barrier and immune function through several proposed mechanisms, including interactions between their surface polysaccharides, the host, and other gut microorganisms. Dairy foods and ingredients are some of the most conspicuous food-based niches for this species and may provide benefits for their delivery and efficacy in the gut. Milk phospholipid (MPL)-rich ingredients have been increasingly recognized for their versatile benefits to health, including interactions with the gut microbiota and intestinal cells. Therefore, our objective was to investigate the capacity for MPL to promote survival of B. infantis during simulated digestion and to modulate bacterial polysaccharide production. To achieve these aims, B. infantis was incubated with or without 0.5% MPL in de Man, Rogosa, and Sharpe (MRS) media at 37°C under anaerobiosis. Survival across the oral, gastric, and intestinal phases using in vitro digestion was measured using plate count, along with adhesion to goblet-like intestinal cells. MPL increased B. infantis survival at the end of the intestinal phase by at least 7% and decreased adhesion to intestinal cells. The bacterial surface characteristics, which may contribute to these effects, were assessed by ζ-potential, changes in surface proteins using comparative proteomics, and production of bound polysaccharides. MPL decreased the surface charge of the bifidobacteria from -17 to -24 mV and increased a 50 kDa protein (3-fold) that appears to be involved in protection from stress. The production of bound polysaccharides was measured using FTIR, HPLC, and TEM imaging. These techniques all suggest an increase in bound polysaccharide production at least 1.7-fold in the presence of MPL. Our results show that MPL treatment increases B. infantis survival during simulated digestion, induces a stress resistance surface protein, and yields greater bound polysaccharide production, suggesting its use as a functional ingredient to enhance probiotic and postbiotic effects.

Invited review: Properties of ß-galactosidases derived from Lactobacillaceae species and their capacity for galacto-oligosaccharide production.

ß-galactosidase (enzymatic class 3.2.1.23) is one of the dairy industry's most important and widely used enzymes. The enzyme is part of a large family known to catalyze hydrolysis and transglycosylation reactions. Its hydrolytic activity is commonly used to decrease lactose content in dairy products, while its transglycosylase activity has recently been used to synthesize galacto-oligosaccharides (GOS). During the past couple of years, researchers have focused on studying ß-galactosidase isolated and purified from lactic acid bacteria. This review will focus on ß-galactosidase purified and characterized from what used to be the Lactobacillus genera. Furthermore, particular emphasis is given to its kinetics, biochemical characteristics, GOS production, market, and utilization by Lactobacilllaceae species.

Draft genome sequence of Lactobacillus helveticus OSU-BDGOAK2 and Lactobacillus kefiranofaciens OSU-BDGOA1, kefir grain isolates with potential antibacterial activity.

We present the draft genome sequence and assembly of Lactobacillus helveticus OSU-BDGOAK2 and Lactobacillus kefiranofaciens OSU-BDGOA1 isolated from kefir grains that exhibited in vitro antibacterial activity against Escherichia coli ATCC 25922, Listeria innocua ATCC 51742, and Staphylococcus epidermidis ATCC 1222. Genome analysis of both strains revealed gene clusters encoding bacteriocins.

Enhanced probiotic potential of Lactobacillus kefiranofaciens OSU-BDGOA1 through co-culture with Kluyveromyces marxianus bdgo-ym6.

Introduction: Due to the increasing consumer demand for the development and improvement of functional foods containing probiotics, new probiotic candidates need to be explored as well as novel means to enhance their beneficial effects. Lactobacillus kefiranofaciens OSU-BDGOA1 is a strain isolated from kefir grains that has demonstrated probiotic traits. This species is the main inhabitant of kefir grains and is responsible for the production of an exopolysaccharide (EPS) whit vast technological applications and potential bioactivities. Research has shown that interkingdom interactions of yeast and lactic acid bacteria can enhance metabolic activities and promote resistance to environmental stressors. Methods: Comparative genomic analyses were performed to distinguish OSU-BDGOA1 from other strains of the same species, and the genome was mined to provide molecular evidence for relevant probiotic properties. We further assessed the cumulative effect on the probiotic properties of OSU-BDGOA1 and Kluyveromyces marxianus bdgo-ym6 yeast co-culture compared to monocultures. Results: Survival during simulated digestion assessed by the INFOGEST digestion model showed higher survival of OSU-BDGOA1 and bdgo-ym6 in co-culture. The adhesion to intestinal cells assessed with the Caco-2 intestinal cell model revealed enhanced adhesion of OSU-BDGOA1 in co-culture. The observed increase in survival during digestion could be associated with the increased production of EPS during the late exponential and early stationary phases of co-culture that, by enhancing co-aggregation between the yeast and the bacterium, protects the microorganisms from severe gastrointestinal conditions as observed by SEM images. Immune modulation and barrier function for recovery and prevention of flagellin-mediated inflammation by Salmonella Typhimurium heat-killed cells (HKSC) in Caco-2 cells were also measured. OSU-BDGOA1 in mono- and co-culture regulated inflammation through downregulation of pro-inflammatory cytokine expression and increased membrane barrier integrity assessed by TEER, FD4 permeability, and expression of tight junctions. Discussion: The results of the study warrant further research into the application of co-cultures of yeast and LAB in functional probiotic products and the potential to increase EPS production by co-culture strategies.

Partition of milk phospholipids during ice cream manufacturing.

The distribution of phospholipids (PL) within the fat and serum phase of ice cream manufacturing was evaluated through partition coefficients (KPL) after mixing, pasteurization, freezing, and hardening. Ice creams containing about 40.41 ± 3.45 (± standard deviation; control formulation) and 112.29 ± 9.06 (enriched PL formulation) mg of PL per g of fat were formulated with nonfat dry milk and ß-serum, respectively. Overall, the KPL were lower than 1, indicating that the PL were predominantly found in the fat phase, and only a small amount was left in the serum and sediment. Confocal micrographs visually confirmed this generalization. The addition of PL significantly increased the viscosity of the mixes between 4- and 9-fold, depending on the shear rate. Additionally, mixes containing high PL exhibited higher yield stress than those formulated with low PL (0.15 ± 0.09 and 0.016 ± 0.08 Pa, respectively). Ice creams with high PL delayed the onset of meltdown and exhibited a slower rate of a meltdown than low-PL ice creams (18.53 ± 0.57 and 14.83 ± 0.85 min, and 1.01 ± 0.05 and 0.71 ± 0.04% min-1, respectively). This study provides useful guidelines for manufacturing ice cream enriched in milk PL. Additionally, the use of ß-serum, a byproduct stream, as a source of PL is illustrated. The development will require studying the sensorial description of the product as well as consumer acceptance.

Milk-Fat-Globule-Membrane-Enriched Dairy Milk Compared with a Soy-Lecithin-Enriched Beverage Did Not Adversely Affect Endotoxemia or Biomarkers of Gut Barrier Function and Cardiometabolic Risk in Adults with Metabolic Syndrome: A Randomized Controlled Crossover Trial.

Full-fat dairy milk may protect against cardiometabolic disorders, due to the milk fat globule membrane (MFGM), through anti-inflammatory and gut-health-promoting activities. We hypothesized that a MFGM-enriched milk beverage (MEB) would alleviate metabolic endotoxemia in metabolic syndrome (MetS) persons by improving gut barrier function and glucose tolerance. In a randomized crossover trial, MetS persons consumed for two-week period a controlled diet with MEB (2.3 g/d milk phospholipids) or a comparator beverage (COMP) formulated with soy phospholipid and palm/coconut oil. They then provided fasting blood and completed a high-fat/high-carbohydrate test meal challenge for evaluating postprandial metabolism and intestinal permeability. Participants had no adverse effects and achieved high compliance, and there were no between-trial differences in dietary intakes. Compared with COMP, fasting endotoxin, glucose, incretins, and triglyceride were unaffected by MEB. The meal challenge increased postprandial endotoxin, triglyceride, and incretins, but were unaffected by MEB. Insulin sensitivity; fecal calprotectin, myeloperoxidase, and short-chain fatty acids; and small intestinal and colonic permeability were also unaffected by MEB. This short-term study demonstrates that controlled administration of MEB in MetS persons does not affect gut barrier function, glucose tolerance, and other cardiometabolic health biomarkers, which contradicts observational evidence that full-fat milk heightens cardiometabolic risk. Registered at ClinicalTrials.gov (NCT03860584).

Effects of Fructose and Palmitic Acid on Gene Expression in Drosophila melanogaster Larvae: Implications for Neurodegenerative Diseases.

One of the largest health problems worldwide is the development of chronic noncommunicable diseases due to the consumption of hypercaloric diets. Among the most common alterations are cardiovascular diseases, and a high correlation between overnutrition and neurodegenerative diseases has also been found. The urgency in the study of specific damage to tissues such as the brain and intestine led us to use Drosophila melanogaster to study the metabolic effects caused by the consumption of fructose and palmitic acid in specific tissues. Thus, third instar larvae (96 ± 4 h) of the wild Canton-S strain of D. melanogaster were used to perform transcriptomic profiling in brain and midgut tissues to test for the potential metabolic effects of a diet supplemented with fructose and palmitic acid. Our data infer that this diet can alter the biosynthesis of proteins at the mRNA level that participate in the synthesis of amino acids, as well as fundamental enzymes for the dopaminergic and GABAergic systems in the midgut and brain. These also demonstrated alterations in the tissues of flies that may help explain the development of various reported human diseases associated with the consumption of fructose and palmitic acid in humans. These studies will not only help to better understand the mechanisms by which the consumption of these alimentary products is related to the development of neuronal diseases but may also contribute to the prevention of these conditions.

Food matrix impacts bioaccessibility and assimilation of acid whey-derived milk fat globule membrane lipids in Caco-2 cells.

The milk fat globule membrane (MFGM) imparts human health benefits ranging from improved immune system, gut, and brain function to improved cardiometabolic health. The industry's growing interest in introducing MFGM-enriched foods requires scientific evidence that the benefits derived from this compound are not affected by the formulation or processes that may alter its function, such as the digestion process. In this study, the impact of food matrices and supplementation levels on the bioaccessibility and assimilation of MFGM lipids in cell culture was investigated. Three food matrices including a protein-rich jelly, carbohydrate-rich cookie, and a carbohydrate- and fat-rich cookie with sunflower oil (SF-cookie) were supplemented with an MFGM ingredient derived from cottage cheese acid whey at 2, 5, and 10% (w/w). Each formulation underwent simulated digestion consisting of oral, gastric, and intestinal phases, and the micellar fraction was collected for both analysis and lipid assimilation in Caco-2 intestinal cells. The micellar fractions were diluted and applied to the cells for 4 h. A lipidomic approach was used to assess the lipid profiles of micellar fractions and intestinal cells. The micelles from digested jellies, cookies, and SF-cookies containing MFGM showed a distinct separation using partial least squares discriminant analysis (PLS-DA). Both correlation loadings and variable importance in projection (VIP) scores demonstrated a tendency of MFGM polar lipids (ceramides, glucosylceramides) for micelles from digested jelly, whereas micelles from digested cookies were associated with MFGM neutral lipids (free fatty acids, cholesterol, etc.). The effect of supplementation level on the micellar lipid profiles reinforced this pattern. The lipid profiles of intestinal cells after incubation with the micellar fractions differed considerably from the corresponding micellar lipid profiles. Specifically, the SF-cookie-treated cells were associated with a greater abundance of PUFA relative to jelly- and cookie-treated cells; however, increasing MFGM supplementation showed irregular patterns and rearrangement of cellular lipid profiles, suggesting the cells' role in regulating lipid metabolism in response to nutritional stimuli. The nature of lipid micellarization and assimilation in intestinal cells from MFGM-containing food formulations echoes the complexity of lipids inherent to the MFGM itself, suggesting the need for application-based MFGM supplementation.

Metagenomic analysis and antibacterial activity of kefir microorganisms.

The microbiota composition of kefir grain and milk kefir was assessed via a metagenomic approach. Significant microorganisms were isolated and identified using molecular methods. A safety assessment was conducted based on antibiotic susceptibility and blood hemolysis. Probiotic traits such as resistance to gastric tract conditions, surface characteristics, adhesion to intestinal cells, and antibacterial activity were also assessed. Metagenomic analysis revealed that kefir grains are a more stable community with clear dominant species as compared to milk kefir. Lactobacillus kefiranofaciens BDGO-A1, Lactobacillus helveticus BDGO-AK2, and Lactobacillu kefiri strains showed tolerance to acidic pH and the presence of bile salts, adhesion capability to Caco-2 cells, in vitro antibacterial activity, and the production of antibacterial proteins. In the metagenomic analysis, contigs associated with these species showed the presence of genes involved in exporting polyketide antibiotics and bacteriocin production. To fully exploit the potential probiotic properties of these microorganisms to help human health, further investigation is necessary to elucidate the mechanisms behind the biological activity and the genotypic characteristics of the isolated strains.

Efficient in vitro digestion of lipids and proteins in bovine milk fat globule membrane ingredient (MFGMi) and whey-casein infant formula with added MFGMi.

The relative immaturity of the infant digestive system has the potential to affect the bioavailability of dietary lipids, proteins, and their digested products. We performed a lipidomic analysis of a commercial bovine milk fat globule membrane ingredient (MFGMi) and determined the profile of lipids and proteins in the bioaccessible fraction after in vitro digestion of both the ingredient and whey-casein-based infant formula without and with MFGMi. Test materials were digested using a static 2-phase in vitro model, with conditions simulating those in the infant gut. The extent of digestion and the bioaccessibility of various classes of neutral and polar lipids were monitored by measuring a wide targeted lipid profile using direct infusion-mass spectrometry. Digestion of abundant proteins in the ingredient and whey-casein infant formula containing the ingredient was determined by denaturing PAGE with imaging of Coomassie Brilliant Blue stained bands. Cholesterol esters, diacylglycerides, triacylglycerides, phosphatidylcholines, and phosphatidylethanolamines in MFGMi were hydrolyzed readily during in vitro digestion, which resulted in marked increases in the amounts of free fatty acids and lyso-phospholipids in the bioaccessible fraction. In contrast, sphingomyelins, ceramides, and gangliosides were largely resistant to simulated digestion. Proteins in MFGMi and the infant formulas also were hydrolyzed efficiently. The results suggest that neutral lipids, cholesterol esters, phospholipids, and proteins in MFGMi are digested efficiently during conditions that simulate the prandial lumen of the stomach and small intestine of infants. Also, supplementation of whey-casein-based infant formula with MFGMi did not appear to alter the profiles of lipids and proteins in the bioaccessible fraction after digestion.

Impact of ultra-shear technology on quality attributes of model dairy-pea protein dispersions with different fat levels.

This study investigated the impact of ultra-shear technology (UST) processing on dairy-pea protein dispersions with different fat levels. Raw milk, skim milk, and cream, as well as model dispersions with combinations of dairy products and pea protein (i.e., raw milk with pea protein, skim milk with pea protein, and cream with pea protein) were employed as test samples. UST experiments were conducted at a pressure of 400 MPa and 70 °C shear valve exit temperature. The UST treatment increased the viscosity of the dispersions and the increases depended on the fat level. Dairy-pea protein dispersions from raw milk and skim milk were shear thinning and mathematically described by the power-law model defined by the consistency coefficient, K (Pa·sn) and the flow behavior index, n. UST treated cream + pea protein dispersions produced structures with gel-like characteristics. Microstructure and particle size analysis determined by laser scanning microscope revealed a reduction in particle size after UST treatment in raw milk + pea protein and skim milk + pea protein dispersions up to 7.55 and 8.30 µm, respectively. In contrast, the particle mean diameter of cream + pea protein dispersions increased up to 77.20 µm after the UST treatment. Thus, the effect of UST on the particle size and rheological behavior of the dispersions depended on the fat level. UST-treated dispersions were stable with no visible phase separation or sedimentation upon centrifugation at 4000×g for 30 min (4 °C). Heat treatment and freeze-thaw treatment of UST-treated samples showed stable blends immediately after the treatments, but subsequent centrifugation showed solid separation. Results from the study suggest that UST is a potential technology to produce stable dairy + pea protein liquids foods with different rheological characteristics for diverse applications.

Functional, textural, and rheological properties of mixed casein micelle and pea protein isolate co-dispersions.

In the midst of rising consumer health and environmental concerns, pea protein has increased in popularity as an alternative to animal-origin proteins. However, the use of pea protein in food systems is largely hindered by its poor functionality, including low solubility. The objective of this study was to measure the textural, functional, and rheological properties of a mixed plant- and animal-based protein system. Caseins, the major protein in bovine milk, are a known animal-based protein with optimal functional properties and high sensory acceptability. Through cold-temperature homogenization, insoluble pea proteins were incorporated with casein micelles in a stable, mixed, colloidal dispersion. Three blends with various casein-to-pea ratios (90:10, 80:20, 50:50) were prepared and analyzed. We hypothesized that incorporation with casein micelles would improve the poor functional properties of pea protein, and thus increase its potential uses in the food industry as a functional ingredient. The protein blend successfully underwent chymosin coagulation, a key ability of caseins, and formed protein gels with textures similar to commercial queso fresco and hard tofu. The 50% casein micelle:50% pea protein blend had better emulsification properties than pea protein alone. In contrast, this blend had the same foaming properties as pea protein alone. The mixed protein blends had similar rheological properties to skim milk, thus increasing their potential applications in the food industry. These results serve as a starting point to begin fully understanding the interactions between pea protein isolate and casein micelles combined via low-temperature homogenization and the effect on their techno-functional properties.

Screening and characterization of ß-galactosidase activity in lactic acid bacteria for the valorization of acid whey.

ß-Galactosidase is an enzyme produced by some strains of lactic acid bacteria (LAB) commonly found in dairy products; however, industrial demand for these enzymes is still low. Acid whey (AW), a lactose-rich byproduct, has large output from cottage cheese and remains unexploited. The purpose of this study was to understand the production mechanism of ß-galactosidase from LAB using AW as a culture medium. First, bioinformatics analysis was conducted on 15 species of LAB. Then, 24 strains were selected and inoculated in de Man, Rogosa, and Sharpe (MRS) broth and in AW medium to compare the bacterial kinetic growth and ß-galactosidase production. Bacterial growth and total protein activity were measured using spectrophotometric techniques. ß-Galactosidase activity was determined by 2 methods: following the hydrolysis of o-nitrophenyl-ß-d-galactopyranoside and of 5-bromo-4-chloro-3-indoyl-ß-d-galactopyranoside (X-gal) in tryptic soy agar plates. The relative expression of the ß-galactosidase gene was performed using real-time quantitative PCR. Despite generally lower growth in AW, 18 strains showed higher ß-galactosidase activity when grown in AW compared with MRS medium. The highest ß-galactosidase activity in AW was in Lactobacillus helveticus strain OSU-PECh-4A, which showed almost 5 times higher activity than average. Analysis of 6 selected strains for expression of the bgal-620 gene found higher overexpression in AW than in MRS, regardless of specific ß-galactosidase activity. Strains of LAB such as OSU-PECh-4A could valorize AW through the production of ß-galactosidase (as an aid to lactose digestion) and production of prebiotic galactooligosaccharides.

High baseline expression of IL-6 and IL-10 decreased CCR7 B cells in individuals with previous SARS-CoV-2 infection during BNT162b2 vaccination.

The current pandemic generated by SARS-CoV-2 has led to mass vaccination with different biologics that have shown wide variations among human populations according to the origin and formulation of the vaccine. Studies evaluating the response in individuals with a natural infection before vaccination have been limited to antibody titer analysis and evaluating a few humoral and cellular response markers, showing a more rapid and intense humoral response than individuals without prior infection. However, the basis of these differences has not been explored in depth. In the present work, we analyzed a group of pro and anti-inflammatory cytokines, antibody titers, and cell populations in peripheral blood of individuals with previous SARS-CoV-2 infection using BNT162b2 biologic. Our results suggest that higher antibody concentration in individuals with an earlier disease could be generated by higher production of plasma cells to the detriment of the presence of memory B cells in the bloodstream, which could be related to the high baseline expression of cytokines (IL-6 and IL-10) before vaccination.

Characterization and Evaluation of Proteolysis Products during the Fermentation of Acid Whey and Fish Waste and Potential Applications.

Reduction of waste in the food industry is critical to sustainability. This work represents one strategy of valorizing waste streams from the dairy (acid whey) and fisheries industries (fish waste) using fermentation. The main approach was to characterize the peptides produced by this fermentation under three conditions: (1) fermentation without adding inoculum; (2) with the addition of a single lactic acid bacterial strain; and (3) the addition of a consortium of lactic acid bacteria. Previous results indicated that the rapid acidification of this fermentation was advantageous for its food safety and microbial activity. This work complements our previous results by defining the rate of peptide production due to protein digestion and using two-dimensional (2D) gel electrophoresis and proteomic analysis to give a more detailed identification of the peptides produced from different waste streams. These results provide important information on this process for eventual applications in industrial fermentation and, ultimately, the efficient valorization of these waste streams.

Characterization and Cellular Uptake of Peptides Derived from In Vitro Digestion of Meat Analogues Produced by a Sustainable Extrusion Process.

Whether proteins in meat analogues (MAs) have the ability to provide equivalent nutrition as those in animal meat remains unknown. Herein, a MA was produced by high-moisture extrusion using soy and wheat proteins. The physicochemical properties, in vitro digestion, and cellular uptake of the released peptides were systematically compared between the MA and the chicken breast (CB). The MA showed a higher hardness but a lower degree of texturization than the CB. After simulated digestion, soluble peptides in the MA had a higher molecular weight and higher hydrophobicity. No observable cytotoxicity or inflammatory response to Caco-2 cells was found for both MA and CB digests. The former exhibited less permeability of peptides across Caco-2 cells. Liquid chromatography with tandem mass spectrometry found that the identified peptides in MA and CB digests contained 7-30 and 7-20 amino acid residues, respectively, and they became shorter after cellular transportation. The amino acid composition showed fewer essential and non-essential amino acids in the MA permeate than in the CB permeate.

Epiregulin as an Alternative Ligand for Leptin Receptor Alleviates Glucose Intolerance without Change in Obesity.

The leptin receptor (LepR) acts as a signaling nexus for the regulation of glucose uptake and obesity, among other metabolic responses. The functional role of LepR under leptin-deficient conditions remains unclear. This study reports that epiregulin (EREG) governed glucose uptake in vitro and in vivo in Lepob mice by activating LepR under leptin-deficient conditions. Single and long-term treatment with EREG effectively rescued glucose intolerance in comparative insulin and EREG tolerance tests in Lepob mice. The immunoprecipitation study revealed binding between EREG and LepR in adipose tissue of Lepob mice. EREG/LepR regulated glucose uptake without changes in obesity in Lepob mice via mechanisms, including ERK activation and translocation of GLUT4 to the cell surface. EREG-dependent glucose uptake was abolished in Leprdb mice which supports a key role of LepR in this process. In contrast, inhibition of the canonical epidermal growth factor receptor (EGFR) pathway implicated in other EREG responses, increased glucose uptake. Our data provide a basis for understanding glycemic responses of EREG that are dependent on LepR unlike functions mediated by EGFR, including leptin secretion, thermogenesis, pain, growth, and other responses. The computational analysis identified a conserved amino acid sequence, supporting an evolutionary role of EREG as an alternative LepR ligand.