Search Results (330)

The prediction of polypeptide chain fragmentation during digestion (proteolysis) of protein substrates by trypsin was carried out for globular β-lactoglobulin (β-LG) and micellar β-casein (β-CN). Despite significant differences in the protein structures of these substrates, the concentrations of peptide fragments are calculated as functions of time or degree of hydrolysis using the same equations derived from the general proteolysis model. This model considers the opening of protein substrates in the course of proteolysis, the so-called demasking process, and the subsequent hydrolysis of specific peptide bonds at different rates determined by the amino acid sequence of hydrolyzed sites. The use of this model for in silico prediction of proteolysis is discussed. An algorithm for calculating demasking rate constants based on the experimental distribution of peptide fragments is presented. The calculated concentration dependence on the degree of hydrolysis of peptide bonds was compared with the experimental data for the intermediate and final peptide fragments of β-LG and β-CN. The predicted and experimental concentration curves for the final products were compared based on their curvatures. For both substrates, the predicted redistribution of peptide fragments in the course of proteolysis was found to be consistent with the experimental one. Full article

Background/Objectives: The composition and function of gut microbiome is significantly influenced by dietary factors. Growing evidence suggests that A1-type and A2-type beta casein (β-CN) may exert distinct effects on the gut environment, with implications for digestive discomfort and broader health outcomes. This review summarizes current evidence on how milk-derived A1 and A2 β-CN affect the gut microbiota. Methods: We conducted a literature search using PubMed, Web of Science, and Scopus to identify studies examining effects of milk β-CN on gut microbiota. Results: A total of eight studies were included. Results show inconsistencies within the limited number of studies. However, compared to A2, A1 β-CN was more frequently associated with dysbiosis and an increased abundance of potentially pathogenic species. Conversely, A2 β-CN promoted microbial diversity, which is linked to improved gut integrity and metabolic health. Conclusions: These findings suggest that β-CN variants distinctly influence the gut microbiota composition, and results were more significant in immunosuppressed subjects or those with other underlying health conditions, indicating that dairy products rich in A2 β-CN may offer advantages in personalized dietary management. However, well-designed human studies are essential to translate findings from rodent models to clinically relevant outcomes and future research should focus on mechanistic exploration and population-specific responses. Full article

(1) Lifestyle changes to modify unhealthy dietary patterns with the goal of preventing MASLD have proven challenging. Here, dietary proteins and their modification with ammonium hydroxide enhancement (AHE) provide molecular evidence that this novel approach may attenuate the development of MASLD without undue dietary adjustments, potentially bypassing non-compliance. (2) High-fat diets containing dietary beef (HFB) or casein (HFC) + AHE (HFBN and HFCN, respectively) were fed to 256 C3H/HeJ female and male mice long term. At 6, 12, or 18 months, hepatic samples were analyzed with targeted metabolomics (glucose, lactate, alanine, glutamine, carnitine) and Western analysis (β-catenin, glutamine synthetase, CYP3A4). RNA sequencing was performed on samples collected at 18 months (n = 3; male HFC n = 2). (3) Metabolomics results showed that at 18 months, hepatic glutamine was greater in HFBN versus HFCN in females, whereas in males, hepatic glutamine, glucose and lactate were lower in HFBN versus HFCN. Additionally, diets with AHE decreased β-catenin and CYP3A4 protein expression in males. Ingenuity pathway analysis (IPA) of RNA-seq data predicted that HFBN activates PPARα signaling in the liver in both sexes compared to HFCN. Inflammatory activity showed predicted activation for females in the HFBN:HFCN comparison. In males, the inflammatory pathway molecular mechanisms of cancer was predicted as deactivated in HFBN:HFCN. (4) Dietary protein source impacts outcomes, and these outcomes improved with AHE. The HFBN diet improves signaling associated with lipid utilization for females and males, and improved inflammatory signaling for males compared with HFCN. Further exploration of AHE as a dietary intervention in high-fat diets is warranted. Full article

Routine milk-recording data may provide valuable insights into dairy cow welfare, although their ability to accurately reflect herd-level welfare outcomes remains unclear. This study explored the associations between routinely collected milk biomarkers and farm-level welfare status using a comparative machine learning approach. Using the Welfare Quality^® (WQ^®) protocol, 43 commercial dairy farms were classified as Enhanced, Acceptable, or Not Classified. Farm-level milk variables included somatic cell count (SCC), differential somatic cell count (DSCC), fat-to-protein ratio (FPR), fat, protein, casein, lactose, urea, β-hydroxybutyrate (BHB), acetone, total plate count (TPC), and morning milk yield. Kruskal–Wallis tests revealed significant differences among welfare classes for DSCC, SCC, lactose, and milk yield (False Discovery Rate-adjusted p < 0.05). Six machine learning algorithms were trained using 10-fold stratified cross-validation. The Elastic-Net (ENET) model showed the highest mean performance (Accuracy = 0.72 ± 0.19; Kappa = 0.56 ± 0.31), followed by Random Forest and Multilayer Perceptron (Accuracy = 0.70). Model accuracy exhibited substantial variability across cross-validation folds, reflecting the limited sample size and class imbalance. Across models, the most influential variables were SCC, DSCC, lactose, milk yield, FPR, fat, and urea. Overall, the findings provide preliminary and exploratory evidence that routine milk biomarkers capture welfare-relevant patterns at the herd level, supporting their potential role as complementary indicators within data-driven welfare assessment frameworks. Full article

β-casein (β-CN) is the predominant casein fraction in breast milk, while current infant milk formula (IMF) contains substantially lower β-CN levels than breast milk. The impact of β-CN fortification on neonatal digestive characteristic and bioactive peptide release remains an understudied area in vivo. This study investigated the effect of β-CN fortification in milk protein on digestion properties and release of bioactive peptides using a suckling rat pup model. Rat pups were, respectively, gavaged with two milk protein solutions: one with ordinary β-CN content (OBCN) and the other with fortified β-CN content (FBCN). The gastric emptying rate, proteolytic efficiency, and peptidomic profiles of intestinal digesta were evaluated. Results indicated that the FBCN group exhibited accelerated gastric emptying into the intestinal phase and enhanced proteolytic efficiency compared to OBCN group. Furthermore, the FBCN group generated greater peptide diversity in the small intestine, with significantly elevated abundance of bioactive peptide candidates exhibiting broader functional spectra. These findings provide additional evidence for the health effects of β-CN fortification in IMF. Full article

Background and Objectives: Milk with A2/A2 β-casein (A2 milk) is currently the subject of numerous studies on the effects of its consumption on health. Commonly consumed milk contains a mixture of β-casein of different genetic variants (most often A1 and A2). In the polypeptide chain of A2/A2 β-casein, proline occurs at position 67, while in β-casein A1/A2, histidine occurs. The main goal of the dietary intervention was to identify and compare the effects of consuming A2 milk, conventional milk (A1) and oat drink on bone health, cardiometabolic health and immune system function in adults. Methods: The controlled IMPA-CT (Investigating Milk and Plant Alternatives Comparative Trial) Study was a randomized study with three groups (A2 Milk group, A1 Milk group, and Oat Drink group). The study included 162 adults with normal and/or overweight, without coexisting chronic diseases, aged 30–60 years. The intervention study consisted of the consumption of 500 mL of an appropriate product (A2 milk/A1 milk/oat drink) daily for 12 weeks. After qualification of the subjects, before the start of the study (T1′), in the 4th week of the study (T2′), in the 8th week of the study (T3′) and at the end of the study, after 12 weeks (T4’), an assessment of the diet and nutritional status was planned. Body composition, bone mineral density (DEXA) and biochemical tests were done. The primary outcome will be the effect of cow’s milk variants and oat drink consumption on bone health. Secondary outcomes will include changes in nutrient intake and cardiometabolic health as well as the immune system in adults. Expected Results and Contributions: The study design, including extensive follow-up and robust endpoint measures, contributed to understanding the therapeutic potential and safety profile or otherwise of β-casein A2/A2 milk and plant-based drinks. Full article

Fermented mare’s milk, or koumiss, has been consumed for centuries across Central Asia for its nutritional and therapeutic value. Mare’s milk differs from bovine milk by its near 1:1 casein-to-whey ratio, high lysozyme and lactoferrin, abundant immunoglobulins, and low β-lactoglobulin, which enhance digestibility, reduce allergenicity, and increase antimicrobial activity. During fermentation, lactic acid bacteria and yeasts transform this substrate into a reservoir of bioactive proteins, peptides, and metabolites. Multi-omics profiling has identified more than 2300 peptides and over 350 metabolites, including sequences with angiotensin-converting enzyme (ACE)-inhibitory, antioxidant, antimicrobial, and immunomodulatory activities. Preclinical and limited clinical data indicate potential benefits for lipid metabolism, cardiovascular function, and gut health. Mechanistically, these effects appear to arise from synergistic actions of native proteins, fermentation-derived peptides, and probiotic consortia. Technological advances such as rational starter culture design, controlled proteolysis, and microencapsulation offer strategies to enhance bioactive yield and stability. However, standardized fermentation protocols and clinical validation remain necessary to position koumiss as a scientifically supported functional food. Full article

Bioactive peptides (BAPs) from dairy products have garnered increasing attention as natural agents with health-promoting properties, including antihypertensive, antioxidant, antimicrobial, immunomodulatory, opioid, and antidiabetic activities. This systematic review synthesizes research published between 2014 and 2024, retrieved from Scopus and PubMed, and selected according to PRISMA guidelines. A total of 192 studies met the inclusion criteria, collectively reporting over 3200 distinct peptides, with antihypertensive sequences, predominantly angiotensin-converting enzyme (ACE) inhibitors, constituting the largest category (n = 1237). β-casein was the principal precursor across bioactivities, followed by αs1-casein, β-lactoglobulin, and α-lactalbumin. Peptides were primarily produced via enzymatic hydrolysis, microbial fermentation, and gastrointestinal digestion, with peptide profiles influenced by the type of milk, microbial strains, and processing conditions. While cow’s milk remained the dominant source, investigations into goat, sheep, camel, buffalo, and donkey milk revealed species-specific biopeptides. Recent advances in proteomics have enhanced peptide identification and bioactivity prediction, enabling the discovery of novel sequences. These findings underscore the significant potential of dairy-derived BAPs as functional food components and nutraceutical ingredients, while highlighting the need for further in vivo validation, bioavailability studies, and broader exploration of underrepresented milk sources. Full article

Background/Objectives: Endothelial dysfunction plays a central role in the development of cardiovascular diseases. β-Casomorphin-7 (BCM-7), a biologically active peptide generated during the digestion of A1 β-casein, is presumed to contribute to this process; however, its direct effects on endothelial cells have not been previously investigated. Here, we aimed to assess whether BCM-7 treatment induces endothelial cell dysfunction through inflammatory cytokines and reactive oxygen species (ROS). Methods: In our study, we analyzed the effects of BCM-7 (5 µg/mL) in combination with lipopolysaccharide (LPS, 100 ng/mL) on human umbilical vein endothelial cells (HUVECs/TERT2). The cell viability, apoptosis, necrosis, and intracellular reactive oxygen species were measured. Furthermore, proinflammatory cytokines and enzymes involved in the regulation of inflammation were assessed with quantitative real-time PCR. The gene and protein expression of enzymes that regulate inflammation and vascular function, thus maintaining endothelial homeostasis were assessed. Results: BCM-7 enhanced intracellular ROS production p ≤ 0.001, increased the expression of interleukin-6 (IL-6) and interleukin-8 (IL-8) p ≤ 0.001, and was more effective when used in combination with LPS p ≤ 0.001. It decreased the expression of cyclooxygenase-1 (COX-1) p ≤ 0.05, during 4 h of exposure, whereas it increased the expression of cyclooxygenase-2 (COX-2) p ≤ 0.001, lipoxygenase-5 (LOX-5) p ≤ 0.01, and nitric oxide synthase 3 (NOS3) p ≤ 0.001; prostaglandin D2 synthase (PTGDS) (p ≤ 0.05), expression was also increased after short treatment. Conclusions: Our results suggest that BCM-7 may contribute to the development of endothelial dysfunction, especially in the presence of LPS, by enhancing oxidative stress and inflammatory response. Full article

To enhance the high-value utilization of camellia oil and innovation in functional foods, this study developed a stable emulsion template using xanthan gum (XG) and sodium caseinate (CAS) for the preparation of camellia oil microcapsules via spray drying. Employing scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and thermogravimetric analysis (TGA), alongside additional analytical methods, this study systematically examined the influence of drying temperature (145 °C, 165 °C, and 185 °C) and XG concentration (0.2%, 0.3%, and 0.4%) on the physicochemical properties and functional attributes of the microcapsules. Results indicated that 0.3% XG was the optimal concentration, enabling uniform emulsion droplet dispersion while balancing microcapsule bulk density and solubility, thereby optimizing processing and dissolution properties. 165 °C was identified as the optimal drying temperature, yielding the highest microcapsule yield (53.68%), moisture content (<2.84%) meeting storage standards, and optimal β-carotene encapsulation efficiency (89.6%) and DPPH radical scavenging rate (74.80 ± 0.34%). FTIR analysis confirmed successful encapsulation of camellia oil within microcapsules. TGA and in vitro digestion experiments demonstrated excellent thermal stability and digestive characteristics of the microcapsules. In summary, this study identified the most favorable preparation conditions for camellia oil microcapsules, providing theoretical support and technical reference for expanding camellia oil applications in the food industry. Full article

Background: β-casomorphin-10 (CM-10), a peptide fragment derived from milk casein with the sequence YPFPGPIPNS, has demonstrated notable anxiolytic activity in BALB/c mice. Yet, its cellular responses and mechanistic pathways remain largely uncharacterized. Methods: We performed RNA-seq analysis to profile gene expression changes in δ-opioid receptor-expressing HEK293 cells (DOR-HEK), comparing CM-10-treated and untreated conditions. Results: CM-10 exposure led to differential expression of 1714 genes in DOR-HEK cells, with 34 upregulated (>1.4-fold) (1.9%) and 1680 downregulated (<0.71-fold) (98.1%), based on a predicted p-value threshold of <0.05. Notably, we identified 10 clusters that were associated with reduced cyclic AMP (cAMP) in DOR-HEK cells following CM-10 treatment. These clusters particularly involved genes related to regulatory subunits of cAMP-dependent protein kinases, such as PRKAR2A, cAMP-responsive element-binding pathway, circadian rhythms, such as CLOCK, ARNT1, CRY2, PER1, and PER2, and anxiety and depression, such as NOTCH1, NOTCH2 and ANK2. A network with these selected genes was confirmed by STRING analysis. Conclusions: These findings indicate that CM-10 may activate DOR-mediated signaling by suppressing cAMP levels, implicating a distinct molecular cascade in HEK293 cells. Full article

Food-derived bioactive peptides are known to possess immunomodulatory properties, although their molecular mechanisms remain incompletely characterized. In this study, we investigated the immunoregulatory effects and underlying mechanisms of YPFPGPIH, a peptide derived from bovine β-casein, using the RAW264.7 macrophage model. Our results demonstrate that YPFPGPIH enhanced macrophage proliferation and phagocytosis in a dose-dependent manner and promoted chemotactic migration through the upregulation of monocyte chemoattractant proteins MCP-1 and MCP-3. Under lipopolysaccharide (LPS)-induced inflammatory conditions, YPFPGPIH significantly reduced the levels of pro-inflammatory mediators, including interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and nitric oxide (NO), while increasing the production of the anti-inflammatory cytokine interleukin-10 (IL-10), thereby reestablishing cytokine balance. Mechanistic studies revealed that YPFPGPIH inhibited LPS-induced activation of the NF-κB and MAPK pathways, as indicated by reduced nuclear translocation of p65 and decreased phosphorylation of ERK, JNK, and p38. Molecular docking analysis indicated strong binding affinities between YPFPGPIH and Toll-like receptors TLR2 and TLR4, suggesting the involvement of TLR-mediated signaling. Notably, YPFPGPIH downregulated inducible nitric oxide synthase (iNOS) expression and upregulated chemokine mRNA levels, reflecting its dual role in modulating inflammatory and migratory responses. These findings highlight YPFPGPIH as a multifunctional immunomodulatory peptide that fine-tunes macrophage activity through crosstalk between TLR, NF-κB, and MAPK signaling pathways. This study provides new insights for developing peptide-based therapeutics and functional foods aimed at managing inflammatory diseases. Full article

Background/Objectives: This study characterized the endogenous peptide profile of human milk from a Chinese multicenter cohort (n = 200 mothers) using the Orbitrap Fusion Lumos LC-MS/MS. Methods: Samples were collected across different lactation stages (2 and 6 months postpartum) and seven geographic regions (Beijing, Chengdu, Guangzhou, Jinhua, Lanzhou, Weihai, and Zhengzhou). Results: In total, 6960 peptides derived from 621 proteins were identified. Peptides from the polymeric immunoglobulin receptor (PIGR) were more abundant in the 2nd month than the 6th month, providing a high antimicrobial activity and immune functions for the infants. Moreover, region-specific variations were observed, with milk from Lanzhou exhibiting significantly higher levels of β-casein (CASB) and butyrophilin subfamily 1 member A1 (BTN1A1) peptides compared to other cities. Conclusions: Furthermore, maternal dietary intake of oils and total fat correlated positively with the intensity of specific antimicrobial peptides, including CASB_199–216, CASB_200–226, and CASB_201–226. Infant growth parameters were inversely correlated with several antimicrobial peptides, although CASB_200–225 demonstrated positive associations. These findings offer novel insights into the dynamics of endogenous peptides in human milk and may guide breastfeeding recommendations and infant formula design. Full article

Stereotypic behaviors are common in farm animals and often signal poor welfare. Tongue-rolling is the most prevalent stereotypic behavior in cows. In this study, we compared the plasma and milk composition of 16 high-frequency tongue-rolling cows (HTR group) and 16 non-stereotypic cows (CON group). All cows were primiparous cows. Biochemical tests, plasma metabolomics, and milk proteomics revealed higher plasma triiodothyronine levels in HTR cows, and lower levels of αs1-casein, β-casein, κ-casein, and lactoferrin in their milk. Multi-omics analyses identified 103 differential metabolites and 73 differential proteins, including various GTP-binding proteins, with the Ras signaling pathway being significantly upregulated in the HTR cows. GO enrichment analysis highlighted significant changes in molecular function, particularly related to GTP/GDP-binding proteins. Additionally, HTR cows exhibited elevated cellular metabolic activity. These findings suggest that high-frequency tongue-rolling is associated with altered endocrine and metabolic profiles, disrupted milk protein synthesis, and impaired immune function potential. The reduction in key milk proteins and lactoferrin may negatively impact milk quality and immune defense. Further research is needed to clarify the causal relationship between these physiological changes and tongue-rolling, providing insights into the underlying mechanisms of stereotypic behaviors in dairy cows and their implications for animal welfare and milk production. Full article

Cheese products worldwide have gained protected designation of origin status in many instances, yet this food group also has the highest reported fraud rates. Quesillo Caquetá is the first Colombian cheese to acquire a protected designation of origin, but still there is a lack of information regarding its composition. In this study, a compositional analysis was performed to establish a set of characteristic parameters to aid the identification of the authenticity of Quesillo Caquetá. Physicochemical analysis, mineral composition determination, carbohydrate, fatty acid, and peptide profiles were conducted on 29 samples of Quesillo Caquetá made with milk from the northern, southern, and central regions of the province of Caquetá. The results revealed 7 minerals, 3 carbohydrates, 19 fatty acids, and 45 peptides (21 peptides from bovine αs1-casein and 24 peptides from bovine β-casein). This suggests that Quesillo Caquetá is a significant source of sodium, calcium, phosphorus, and monounsaturated fatty acids such as oleic acid, omega-3, and omega-6, as well as some peptides that match sequences with antihypertensive, immunomodulatory, antioxidant, and antimicrobial activity reported in the literature. The specificity of the fatty acid and peptide profiles can become a valuable tool for identifying the authenticity of Quesillo Caquetá against possible imitations in the market. Full article