Human milk proteins differentiate over the sex of newborns and across stages of lactation.

BACKGROUND & AIMS: Human milk (HM) is a complete food that meets the nutritional and energy demands of the newborns. It contains numerous bioactive components, including functional proteins. Variations in HM energy and lipid content have already been reported related to the newborn's sex, but differences between protein profiles are still scarce. This work aimed to identify differences between HM proteins produced by mothers of female and male newborns, in the lactation stages of colostrum and mature milk, and the metabolic pathways involved. METHODS: A total of 98 HM samples were collected from 39 lactating women and classified according to the newborn's sex, stages of lactation, and three mothers' age groups, and evaluated about protein concentration and one-dimensional electrophoretic profile. Next, to assess samples with the greatest differences, the HM proteins regarding the newborn's sex and the stages of lactation were compared using nano-LC-MS/MS, in 24 HM samples randomly rearranged into four groups: female and male infants, and colostrum and mature milk. Functional classification, metabolic pathways, and protein interaction networks were analyzed by Gene Ontology, KEGG, and STRING, respectively. RESULTS: The soluble protein content of HM decreased throughout lactation, with differences regarding isolated factors, such as mothers' age group, child's sex and stages of lactation, and also in terms of their interactions. A total of 146 proteins were identified, 42 of which showed different abundances over the sexes of newborns and 53 between the stages of lactation. In general, proteins related to metabolic processes were up-regulated for mothers of male infants and in the mature stage of lactation, while proteins related to defense were up-regulated in mothers of female infants and in the colostrum phase. CONCLUSION: This study indicated that there are differentiated and specific nutritional and defense needs of newborns, by sex and by lactation phase, which is highly relevant for a more appropriate supply of food to infants receiving HM from donor mothers.

Mice Regulate Dietary Amino Acid Balance and Energy Intake by Selecting between Complementary Protein Sources.

BACKGROUND: A balanced intake of protein and constituent amino acids (AAs) requires adjustments to total food intake (protein leverage [PL]) and food selection to balance deficits and excesses (complementary feeding). We provided mice with choices of casein and whey, 2 protein sources that are complementary in AA balance, across a range of protein concentrations (P%) of digestible energy (DE). OBJECTIVES: We aimed to determine if: 1) PL operates similarly for casein and whey; 2) one protein source is preferred at control P%; 3) the preference changes as P% falls; and 4) AA intakes under control and low P% levels identify AAs that drive changes in protein selection. METHODS: Food intake and plasma fibroblast growth factor-21 (FGF21) concentrations were measured in mice at various P% (P7.5%-P33%). For direct comparisons, defined diets were used in which the protein source was either casein or whey. In food choice studies, mice had access to foods in which both casein and whey were provided at the same P% level at the same time. RESULTS: PL operated at different P% thresholds in casein (13%)- and whey (10%)-based diets, and the magnitude of PL was greater for casein. Although mice preferred casein under control conditions (P23%), a pronounced preference shift to whey occurred as P% fell to P13% and P10%. At low P%, increases in food intake were accompanied by increases in plasma FGF21, a protein hunger signal. Among AAs deficient in casein and enriched in whey, the intake of Cys was the most invariant as P% changed between P23% and P10%, appearing to drive the switch in protein preference. CONCLUSIONS: Mice selected between complementary protein sources, casein and whey, achieving stable total energy intake and regulated intake of AAs as P% varied. Supplementation of low P% casein diets with one whey-enriched AA, Cys, suppressed plasma FGF21 and total food intake.

Out-of-pocket expenses and parent reported quality of life in children with cow's milk protein allergy in Bogotá, Colombia.

OBJECTIVE: To identify the out-of-pocket expenses and parent-reported quality of life (QoL) of children with a diagnosis of cow's milk protein allergy between the ages of 0 and 5 using the Food Allergy Quality of Life Questionnaire - Parent Form. METHODS: A cross-sectional study was conducted in two tertiary care centers in Bogotá. Demographic, medical information, and QoL scores were collected by parental interview. We carried out a cost-of-illness analysis based on self-reported out-of-pocket expenses attributed to the treatment as a whole and the family's monthly income. Exploratory analyses used the QoL scores and the percentage of out-of-pocket expenses attributable to treatment as outcomes. RESULTS: 122 families were analyzed. Median subject age was 17 months (Q1-Q3: 11-26.75 months) and female subjects made up 71% of the sample. The median QoL score was 3.21 points (Q1-Q3: 2.43-4.34) and only differed by age groups and personal history of other food allergies. The median out-of-pocket treatment related costs was 300,000 Colombian pesos (COP) (Q1-Q3: 280,000-340,000 COP). About 17% of the families had to pay over 15% of their monthly income to purchase food and dietary products. Out-of-pocket treatment related costs differed depending on whether the treatment included formulas (Mann-Whitney test p < 0.001). Out-of-pocket treatment expenses were uncorrelated with the QoL scores. CONCLUSION: Food allergy related QoL scores were not associated with out-of-pocket expenses as a whole or as a fraction of monthly income but were higher in children with additional food allergies and in older age groups, suggesting a lower QoL.

Genome-Wide Analysis of Milk Production Traits and Selection Signatures in Serbian Holstein-Friesian Cattle.

To improve the genomic evaluation of milk-related traits in Holstein-Friesian (HF) cattle it is essential to identify the associated candidate genes. Novel SNP-based analyses, such as the genetic mapping of inherited diseases, GWAS, and genomic selection, have led to a new era of research. The aim of this study was to analyze the association of each individual SNP in Serbian HF cattle with milk production traits and inbreeding levels. The SNP 60 K chip Axiom Bovine BovMDv3 was deployed for the genotyping of 334 HF cows. The obtained genomic results, together with the collected phenotypic data, were used for a GWAS. Moreover, the identification of ROH segments was performed and served for inbreeding coefficient evaluation and ROH island detection. Using a GWAS, a polymorphism, rs110619097 (located in the intron of the CTNNA3 gene), was detected to be significantly (p < 0.01) associated with the milk protein concentration in the first lactation (adjusted to 305 days). The average genomic inbreeding value (FROH) was 0.079. ROH islands were discovered in proximity to genes associated with milk production traits and genomic regions under selection pressure for other economically important traits of dairy cattle. The findings of this pilot study provide useful information for a better understanding of the genetic architecture of milk production traits in Serbian HF dairy cows and can be used to improve lactation performances in Serbian HF cattle breeding programs.

Food peptidomic analysis of bovine milk fermented by Lacticaseibacillus casei LBC 237: In silico prediction of bioactive peptides and anticancer potential.

Bioactive peptides, which exhibited diverse biological activities such as anti-cancer, anti-inflammatory, bactericidal, antiviral, and quorum sensing properties, were considered promising alternative therapeutic agents. Sourced from various raw materials, particularly foods, these peptides garnered significant interest. In this context, the study focused on exploring bioactive peptides derived from bovine whole milk fermentation by Lacticaseibacillus casei LBC 237. Comprehensive peptidomic analysis and in silico predictions, with a specific emphasis on anti-cancer properties, were conducted. The study categorized peptides into BP-LBC, originating from the metabolism of L. casei LBC 237 and not matching any sequence in the Bos taurus database, and BP-MILK, matching a sequence in the Bos taurus database. Among the 143 identified peptides with potential biological activity, 33.56% were attributed to BP-LBC, while 66.43% originated from BP-MILK, demonstrating the important contribution of proteins in bovine milk in the generation of bioactive peptides. Hydrophobic peptides, enriched in Leucine, Lysine, and Proline, dominated both fractions, significantly influencing their functional properties. Pearson correlation analysis revealed inverse relationships between bioactive peptides, molecular weight, and anti-tumor activity in BP-MILK. The DGKVWEESLK peptide exhibited in silico activity against 10 different cancer cell lines. Studying the bioactive properties of peptides from familiar sources enhances the connection between food science and human health. In addition, in silico studies have been crucial in deepening our understanding of the bioactive potential of these peptides and their mode of action.

Effect of lactic acid bacteria fermentation on cow milk allergenicity and antigenicity: A review.

Cow milk is a major allergenic food. The potential prevention and treatment effects of lactic acid bacteria (LAB)-fermented dairy products on allergic symptoms have garnered considerable attention. Cow milk allergy (CMA) is mainly attributed to extracellular and/or cell envelope proteolytic enzymes with hydrolysis specificity. Numerous studies have demonstrated that LAB prevents the risk of allergies by modulating the development and regulation of the host immune system. Specifically, LAB and its effectors can enhance intestinal barrier function and affect immune cells by interfering with humoral and cellular immunity. Fermentation hydrolysis of allergenic epitopes is considered the main mechanism of reducing CMA. This article reviews the linear epitopes of allergens in cow milk and the effect of LAB on these allergens and provides insight into the means of predicting allergenic epitopes by conventional laboratory analysis methods combined with molecular simulation. Although LAB can reduce CMA in several ways, the mechanism of action remains partially clarified. Therefore, this review additionally attempts to summarize the main mechanism of LAB fermentation to provide guidance for establishing an effective preventive and treatment method for CMA and serve as a reference for the screening, research, and application of LAB-based intervention.

Rapid and green discrimination of bovine milk according to fat content, thermal treatment, brand and manufacturer via colloidal fingerprinting.

Addressing food safety and detecting food fraud while fulfilling greenness requisites for analysis is a challenging but necessary task. The use of sustainable techniques, with limited pretreatment, non-toxic chemicals, high throughput results, is recommended. A combination of Field Flow Fractionation (FFF), working in saline carrier and with minimal preprocessing, and chemometrics was for the first time applied to bovine milk grouping. A set of 47 bovine milk samples was analyzed: a single analysis yielded a characteristic multidimensional colloidal dataset, that once processed with multivariate tools allowed simultaneously for different discriminations: fat content, thermal treatment, brand and manufacturing plant. The analytical methodology is fast, green, simple, and inexpensive and could offer great help in the field of quality control and frauds identification. This work represents also the first attempt to identify milk sub-typologies based on colloidal profiles, and the most complete study concerning multivariate analysis of FFF fingerprint.

Enzymatic debittering of cheese flavoring and bitterness characterization of peptide mixture using sensory and peptidomics approach.

Peptides in cheese flavoring produced through proteolysis plus fermentation generated bitterness. Bitterness of individual peptide can be quantified using quantitative structure-activity relationship, where molecular mass (M), hydrophobicity, residues, C-terminal hydrophobic amino acids (C-HAAs), and N-terminal basic ones (N-BAAs) are crucial. However, their accumulative influence on the overall bitterness of peptide mixture remains unknown. This study delved into extensive proteolysis to debitter and to correlate the multi-influencing factors of peptides and the collective bitterness. As hydrolysis increased from 7.5 % to 28.0 %, bitterness reduced from 5.0 to 0.3-2.7 scores, contingent on proteases used, in which FU was optimal. The overall bitterness cannot be predicted through the summation of individual peptide bitterness, which depended on M (0.5-3 kDa) and 5-23 residues, followed by N-BAAs and C-HAAs. Analysis of enzymatic cleavage sites and substrate characteristics revealed, to more effectively debitter bovine milk protein hydrolysates, proteases specifically cleaving Pro, Leu, Phe, and Val were desired.

Non-thermal techniques as an approach to modify the structure of milk proteins and improve their functionalities: a review of novel preparation.

BACKGROUND: Milk proteins (MPs) have been widely used in the food industry due to their excellent functionalities. However, MPs are thermal-unstable substances and their functional properties are easily affected by heat treatment. Emerging non-thermal approaches (i.e., high-pressure homogenization (HPH), ultrasound (US), pulsed electric field (PEF)) have been increasingly popular. A detailed understanding of these approaches' impacts on the structure and functionalities of MPs can provide theoretical guidance for further development to accelerate their industrialization. SCOPE AND APPROACH: This review assesses the mechanisms of HPH, US and PEF technologies on the structure and functionalities of MPs from molecular, mesoscopic and macroscopic levels, elucidates the modifications of MPs by these theologies combined with other methods, and further discusses their existing issues and the development in the food filed. KEY FINDINGS AND CONCLUSIONS: The structure of MPs changed after HPH, US and PEF treatment, affecting their functionalities. The changes in these properties of MPs are related to treated-parameters of used-technologies, the concentration of MPs, as well as molecular properties. Additionally, these technologies combined with other methods could obtain some outstanding functional properties for MPs. If properly managed, these theologies can be tailored for manufacturing superior functional MPs for various processing fields.

Worldwide research on the health effects of bovine milk containing A1 and A2 ß-casein: Unraveling the current scenario and future trends through bibliometrics and text mining.

The possible adverse effect of consuming bovine milk with A1 ß-casein (but not with A2 ß-casein) on health aspects due to the release of ß-casomorphin-7 (BCM-7) is currently under debate. The aim of this study was to perform a bibliometric analysis of studies extracted from Scopus to explore the relationship between BCM-7, A1 or A2 bovine milk with different aspects of health. Over time, several research groups were formed that are no longer active and although some authors have returned to the field of study, they have focused their efforts mainly on conducting reviews that show the same imprecise conclusions due to the few original articles. Research is concentrated in Europe and Asia, where New Zealand, China and Germany are the countries with the most publications, records and citations on the subject, respectively. On the other hand, no country in Africa or South America has scientific production, which opens the possibility of building collaborations between countries and exploring areas that lack scientific studies. Based on conflicting information from primarily in vitro and animal studies, and limited clinical trials with poor designs, A1 milk presents pro-inflammatory and oxidative activity, but the evidence is insufficient to associate its consumption with negative health effects. However, A2 milk may be better tolerated by the digestive system of some individuals, suggesting its possible modulating role in the intestinal microbiota. Stronger scientific evidence is needed to reach a consensus on whether the presence of ß-casein A1 can significantly negatively affect health. The information shown will allow a better understanding of the subject and consumers will be able to make their own decisions regarding A1 or A2 milk.

Tackling food allergens-The role of food processing on proteins' allergenicity.

This chapter examines how innovative and emerging food processing technologies, such as those that use heat, electricity, electromagnetic waves, and pressure, can modify protein denaturation, aggregation, and intermolecular interactions pathways, which can result in varying immunoreactive responses. It emphasizes the need to understand how these processing methods affect the protein epitopes recognized by antibodies and their respective priming pathways, especially during the sensitization stage that precedes an allergic response. Although traditional processing methods have been investigated, the impact of novel technologies on food protein allergenicity remains largely unknown. The chapter specifically focuses on milk proteins, which have clinical significance and are associated with cow's milk allergy, one of the most common food allergies in young children. Additionally, it examines potential scientific advancements that novel processing methods may bring to this field.

Food Allergens: When Friends Become Foes-Caveats and Opportunities for Oral Immunotherapy Based on Deactivation Methods.

Food allergies represent a serious health concern and, since the 1990s, they have risen gradually in high-income countries. Unfortunately, the problem is complex because genetic, epigenetic, and environmental factors may be collectively involved. Prevention and diagnoses have not yet evolved into efficacious therapies. Identification and control of allergens present in edible substances hold promise for multi-purpose biomedical approaches, including oral immunotherapy. This review highlights recent studies and methods to modify the otherwise innocuous native proteins in most subjects, and how oral treatments targeting immune responses could help cancel out the potential risks in hypersensitive individuals, especially children. We have focused on some physical methods that can easily be conducted, along with chemo-enzymatic modifications of allergens by means of peptides and phytochemicals in particular. The latter, accessible from naturally-occurring substances, provide an added value to hypoallergenic matrices employing vegetal wastes, a point where food chemistry meets sustainable goals as well.

Stability of milk proteins subjected to UHT treatments: challenges and future perspectives.

Ultra-high temperature (UHT) treatments are of high economic relevance for food industries because they contribute to extending the shelf life of food products and facilitating their distribution. In the dairy segment, UHT treatments are applied to a wide range of products containing variable protein amounts. In this sense, the changes in the molecular structure of milk proteins induced by the severity of UHT treatments may lead to fouling in equipment during processing or sedimentation and/or gelation during storage. Nowadays, these concerns are even more relevant due to the increasing demand for UHT-treated high-protein beverages. This review will discuss the two main strategies used by industries to increase the stability of milk proteins during and/or after UHT treatments: (i) addition of chelating agents and (ii) use of polysaccharides. Moreover, the challenges and opportunities associated with promising strategies to improve the stability of milk proteins during and/or after UHT treatments will be covered in this review. The information compiled will be useful to guide researchers and industries in developing more stable UHT dairy products in harmony with consumers' demands.

The effect of heat treatment on the lactosylation of milk proteins.

Protein lactosylation is a significant modification that occurs during the heat treatment of dairy products, causing changes in proteins' physical-chemical and nutritional properties. Knowledge of the detailed lactosylation information on milk proteins under various heat treatments is important for selecting appropriate thermo-processing and identifying markers to monitor heat load in dairy products. In the present study, we used proteomics techniques to investigate lactosylated proteins under different heating temperatures. We observed a total of 123 lactosylated lysines in 65 proteins, with lactosylation even occurring in raw milk. The number of lactosylated lysines and proteins increased moderately at 75°C to 130°C, but dramatically at 140°C. We found that 6 out of 10, 9 out of 16, 6 out of 12, and 5 out of 15 lysine residues in κ-casein, ß-lactoglobulin, α-lactalbumin, and αS1-casein, respectively, were lactosylated under the applied heating treatment. Moreover, different lactosylation states of individual lysines and proteins can indicate the intensity of heating processes. Lactosylation of K14 in ß-lactoglobulin could distinguish pasteurized and UHT milk, while lactosylation of lactotransferrin can reflect moderate heat treatment of products.

Food-sensitized pediatric patients show colonic cow's milk protein-specific Th2 cells.

Food allergies have become a health concern worldwide. Around 6% to 10% of children are allergic to cow's milk proteins. We have previously characterized colorectal polyps in patients sensitized to food allergens. These polyps are classified as inflammatory and present a type 2 environment, with elevated interleukin (IL)-13 and IL-4, and are a site of immunoglobulin E synthesis. In this study, we characterized and isolated cow's milk protein-specific T cell lines and T cell clones from the lamina propria of polyps from patients sensitized to these proteins. Isolated T cells responded to cow's milk proteins similarly to peripheral blood T cells, showing antigen-specific cell proliferation and Th2 cytokines release in vitro. T cell clones obtained were all CD4+ T cells and expressed the membrane TCRαß receptor and secreted higher IL-4, IL-5, and IL-13 amounts than unstimulated cells, whereas interferon γ secretion remained unchanged. Remarkably, the gut homing chemokine receptor CCR9 was augmented in cow's milk-specific peripheral and lamina propria T cells, and CCL25 was found to be expressed in the inflammatory polyp tissue and not in the adjacent mucosa. In conclusion, we isolated and characterized cow's milk-specific lamina propria CD4+ Th2 cells from colonic inflammatory polyps. CCR9 expression on these cells, along with increase secretion of CCL25 in the polyp, favors recruitment and cow's milk-specific allergic response within the inflammatory polyp tissue. Our findings may be critical to understand the underlying mechanism that promotes immunoglobulin E synthesis in the colon of cow's milk proteins allergic patients, contributing to the development of novel T cell-targeted immunotherapies.

Proteosomes based on milk phospholipids and proteins to enhance the stability and bioaccessibility of ß-carotene.

Proteosomes (P) based on milk fat globule membrane's phospholipids (MPs), whey protein isolate (WPI) and sodium caseinate (CasNa) were developed by ultrasonication to encapsulate ß-carotene. Entirely milk-ingredients based proteosomes (WPI-MPs-P and CasNa-MPs-P) revealed homogenous distribution with size diameters < 250 nm. WPI-MPs-P depicted positive ζ-potential values (+15.7 ± 0.5 mV), while CasNa-MPs-P demonstrated negative (-32.5 ± 3.4 mV) values of surface charge, respectively and hydrophilic nature of proteosomes was observed by measuring contact-angle (θ). AFM and SEM exhibited spherical to oval and slightly irregular morphology of nanocarriers. For various concentrations of ß-carotene, the highest encapsulation efficiency of ß-carotene was 90 ± 0.2% and 92 ± 0.8% in WPI-MPs-P and CasNa-MPs-P respectively. FTIR analyses confirmed the hydrophobic and electrostatic interactions-based encapsulation of ß-carotene. Beneficial antioxidant-potential of ß-carotene was retained after its encapsulation in the proteosomes. Proteosomes increased the digestive-stability (>50%) and bioaccessibility (>85%) of ß-carotene. Thus, milk-ingredients based proteosomes offer a novel-strategy to develop functional dairy products to overcome widespread vitamin-A-deficiency.

The prevalence of IgG antibodies against milk and milk antigens in patients with multiple sclerosis.

Introduction: Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system (CNS). The pathophysiology of MS is complex and is said to be influenced by multiple environmental determinants, including diet. We and others have previously demonstrated how consumption of bovine milk can aggravate disease severity in MS patients, which can be explained by molecular mimicry between milk antigens and those expressed within the CNS. In this study we set out to identify alternatives to drinking cow milk which might be less detrimental to MS patients who have a genetic predisposition towards developing antibody titers against bovine milk antigens that cross-react with CNS antigens. Methods: To this end, we screened 35 patients with MS and 20 healthy controls for their IgG reactivity against an array of animal-sourced milk, plant-based alternatives as well as individual antigens from bovine milk. Results: We demonstrate that MS patients have a significantly higher IgG response to animal-sourced milk, especially cow milk, in comparison to healthy donors. We also show that the reactivity to cow milk in MS patients can be attributed to reactivity against different bovine milk antigens. Finally, our correlation data indicate the co-existence of antibodies to individual bovine milk antigens and their corresponding cross-reactive CNS antigens. Discussion: Taken together, we suggest screening of blood from MS patients for antibodies against different types of milk and milk antigens in order to establish a personalized diet regimen.

Effects of different steam injection conditions on cappuccino's nutritional profile.

The quality parameters of cappuccinos prepared with pasteurized milk or ultra-high-temperature milk steam-injected at different temperatures by a professional coffee machine have been assessed. In particular, the protein profile, the content of vitamins and lactose, the lipid peroxidation process, and the involvement of milk proteins in the foam formation were evaluated. The nutritional quality of milk seems not affected by the steam injection treatment carried out at a temperature of 60-65 °C, but at higher temperatures a decrement of lactoperoxidase, vitamin B6 and folic acid was observed. The milk used in cappuccino preparation is very important: pasteurized milk can form a more consistent and lasting foam with respect to ultra-high-temperature milk because of the presence of ß-lactoglobulin and lactoferrin, both playing an important role in the foam formation and stability. This work would provide additional information to the coffee industry for the preparation of high nutritional and organoleptic quality cappuccinos.

Recent advances in selective allergies to mammalian milk proteins not associated with Cow's Milk Proteins Allergy.

Cow's milk proteins allergy (CMA) is an atypical immune system response to cow's milk and dairy products. It's one of the most common food allergies in children affecting 8% of the total pediatric population pediatric population. This comprehensive review examines recent studies in CMA, especially regarding mammalian milk allergies such as goat's, sheep's, buffalo's, camel's, mare's and donkey's milk allergies in order to increase awareness of these selective allergies and to reduce allergy risks for those who have them. The consumption of other mammalian milk types is not recommended because of the significant homology between milk proteins from cow, sheep, goat and buffalo resulting in clinical cross-reactivity. However, camel's, mare's or donkey's milk may be tolerated by some allergic patients. Selective mammalian milk allergies are unusual and rare disorders characterized by severe symptoms including angio-oedema, urticaria, respiratory manifestations and anaphylaxis. Based on the reported allergic cases, cheese products including Ricotta, Romano, Pecorino and Mozzarella, are considered as the most common source of allergens especially in goat's, sheep's and buffalo's milk allergies, while the major allergens in donkey's and mare's milk seems to be whey proteins including lysozyme, α-lactalbumin and ß-lactogloblin due to the low casein/whey proteins ratio in equine's milk.

Combination of Milk and Plant Proteins to Develop Novel Food Systems: What Are the Limits?

In the context of a diet transition from animal protein to plant protein, both for sustainable and healthy scopes, innovative plant-based foods are being developing. A combination with milk proteins has been proposed as a strategy to overcome the scarce functional and sensorial properties of plant proteins. Based on this mixture were designed several colloidal systems such as suspensions, gels, emulsions, and foams which can be found in many food products. This review aims to give profound scientific insights on the challenges and opportunities of developing such binary systems which could soon open a new market category in the food industry. The recent trends in the formulation of each colloidal system, as well as their limits and advantages are here considered. Lastly, new approaches to improve the coexistence of both milk and plant proteins and how they affect the sensorial profile of food products are discussed.