ABSTRACT
We previously demonstrated that postruminal casein infusion and exogenous glucagon-like peptide 2 (GLP-2) administration independently stimulated growth and carbohydrase activity of the pancreas and jejunal mucosa in cattle. The objective of the current study was to profile the jejunal mucosal transcriptome of cattle using next-generation RNA sequencing in response to postruminal casein infusion and exogenous GLP-2. Twenty-four Holstein steers [250 ± 23.1 kg body weight (BW)] received a continuous abomasal infusion of 3.94 g raw corn starch/kg of BW combined with either 0 or 1.30 g casein/kg of BW for 7 d. Steers received subcutaneous injections at 0800 and 2000 h to provide either 0 or 100 µg GLP-2/kg of BW per day. At the end of the 7-d treatment period, steers were slaughtered for collection of the jejunal mucosa. Total RNA was extracted from jejunal mucosal tissue, strand-specific cDNA libraries were prepared, and RNA sequencing was conducted to generate 150-bp paired-end reads at a depth of 40 M reads per sample. Differentially expressed genes (DEG), KEGG pathway enrichment, and gene ontology enrichment were determined based on the FDR-corrected P-value (padj). Exogenous GLP-2 administration upregulated (padj < 0.05) 667 genes and downregulated 1,101 genes of the jejunal mucosa. Sphingolipid metabolism, bile secretion, adherens junction, and galactose metabolism were among the top KEGG pathways enriched with upregulated DEG (padj < 0.05) in response to exogenous GLP-2 administration. The top gene ontologies enriched with upregulated DEG (padj < 0.05) in response to exogenous GLP-2 administration included nutrient metabolic processes, brush border and bicellular tight junction assembly, and enzyme and transporter activities. Exogenous GLP-2 administration increased or tended to increase (padj < 0.10) brush border carbohydrase (MGAM, LCT, TREH), hexose transporter (SLC5A1, SLC2A2), and associated transcription factor (HNF1, GATA4, KAT2B) mRNA expression of the jejunal mucosa. Gene ontologies and KEGG pathways that were downregulated (padj < 0.05) in response to exogenous GLP-2 were related to genetic information processing. Postruminal casein infusion downregulated (padj < 0.05) 7 jejunal mucosal genes that collectively did not result in enriched KEGG pathways or gene ontologies. This study highlights some of the transcriptional mechanisms associated with increased growth, starch assimilation capacity, and barrier function of the jejunal mucosa in response to exogenous GLP-2 administration.
Subject(s)
Caseins , Glucagon-Like Peptide 2 , Intestinal Mucosa , Jejunum , Transcriptome , Animals , Cattle , Glucagon-Like Peptide 2/administration & dosage , Glucagon-Like Peptide 2/pharmacology , Jejunum/metabolism , Jejunum/drug effects , Caseins/genetics , Caseins/administration & dosage , Transcriptome/drug effects , Intestinal Mucosa/metabolism , Intestinal Mucosa/drug effects , Male , Abomasum/drug effects , Abomasum/metabolism , Gene Expression ProfilingABSTRACT
This study used the dip-coating method to develop a new biocompatible coating composed of polylactide (PLA) and casein for ZnMg1.2 wt% alloy implants. It evaluated its impact on the alloy's degradation in a simulated body fluid. After 168 h of immersion in Ringer's solution, surface morphology analysis showed that the PLA-casein coatings demonstrated uniform degradation, with the corrosion current density measured at 48 µA/cm2. Contact angle measurements indicated that the average contact angles for the PLA-casein-coated samples were below 80°, signifying a hydrophilic nature that promotes cell adhesion. Fourier-transform infrared spectroscopy (FTIR) revealed no presence of lactic acid on PLA-casein coatings after immersion, in contrast to pure PLA coatings. Pull-off adhesion tests showed tensile strength values of 7.6 MPa for pure PLA coatings and 5 MPa for PLA-casein coatings. Electrochemical tests further supported the favorable corrosion resistance of the PLA-casein coatings, highlighting their potential to reduce tissue inflammation and improve the biocompatibility of ZnMg1.2 wt% alloy implants.
Subject(s)
Alloys , Caseins , Coated Materials, Biocompatible , Polyesters , Alloys/chemistry , Caseins/chemistry , Coated Materials, Biocompatible/chemistry , Polyesters/chemistry , Corrosion , Materials Testing , Surface Properties , Spectroscopy, Fourier Transform Infrared , Tensile StrengthABSTRACT
It has been reported that the modification of immobilized glyoxyl-ficin with aldehyde dextran can promote steric hindrances that greatly reduce the activity of the immobilized protease against hemoglobin, while the protease still maintained a reasonable level of activity against casein. In this paper, we studied if this effect may be different depending on the amount of ficin loaded on the support. For this purpose, both the moderately loaded and the overloaded glyoxyl-ficin biocatalysts were prepared and modified with aldehyde dextran. While the moderately loaded biocatalyst had a significantly reduced activity, mainly against hemoglobin, the activity of the overloaded biocatalyst was almost maintained. This suggests that aldehyde dextran was able to modify areas of the moderately loaded enzyme that were not available when the enzyme was overloaded. This modification promoted a significant increase in biocatalyst stability for both biocatalysts, but the stability was higher for the overloaded biocatalyst (perhaps due to a combination of inter- and intramolecular crosslinking).
Subject(s)
Aldehydes , Dextrans , Enzymes, Immobilized , Ficain , Dextrans/chemistry , Enzymes, Immobilized/chemistry , Enzymes, Immobilized/metabolism , Ficain/chemistry , Ficain/metabolism , Aldehydes/chemistry , Hemoglobins/chemistry , Hemoglobins/metabolism , Biocatalysis , Substrate Specificity , Caseins/chemistry , Caseins/metabolism , Enzyme StabilityABSTRACT
Pasta filata-style cheese products are among the world's most famous cheese varieties. Thermo-mechanical processing of cheese curd results in stringy, fibrous, and anisotropic structures with pleasing texture attributes. A recent area of research focuses on improving yield during the manufacturing of pasta filata-type cheese products by homogenizing the milk. This process reduces the size of fat droplets, leading to better retention of milk fat during curd plasticization. As this sometimes results in texture deficits, this study aims to investigate the impact of thermo-mechanical processing on curd from homogenized and non-homogenized milk. The hypothesis is that increased thermo-mechanical processing, leading to more anisotropic structural elements, may offset texture deficits caused by homogenization. To assess textural and structural changes due to homogenization and thermo-mechanical processing, mechanical tests including rheology and texture analysis were conducted, along with confocal-laser-scanning microscopy. Additionally, sensory evaluation involving panelists consuming the samples and recording mastication properties such as muscle activity and jaw movement was carried out. Dynamic data modeling was used to derive connections between structure and texture. Results showed that homogenization alone did not yield significant differences between the samples, but plasticization and texturization properties differed significantly. Non-homogenized samples developed a distinct fibrous structure, and muscle activities and jaw movements increased significantly (p < 0.01) with longer thermo-mechanical processing.
Subject(s)
Caseins , Cheese , Food Handling , Gels , Mastication , Milk , Cheese/analysis , Food Handling/methods , Animals , Mastication/physiology , Milk/chemistry , Gels/chemistry , Caseins/chemistry , Rheology , Humans , Hot TemperatureABSTRACT
Much attention has been given to studying the translational diffusion of globular proteins, whereas the translational diffusion of intrinsically disordered proteins (IDPs) is less studied. In this study, we investigate the translational diffusion and how it is affected by the self-association of an IDP, κ-casein, using pulsed-field gradient nuclear magnetic resonance and time-resolved Förster resonance energy transfer. Using the analysis of the shape of diffusion attenuation and the concentration dependence of κ-casein diffusion coefficients and intermolecular interactions, we demonstrate that κ-casein exhibits continuous self-association. When the volume fraction of κ-casein is below 0.08, we observe that κ-casein self-association results in a macroscopic phase separation upon storage at 4 °C. At κ-casein volume fractions above 0.08, self-association leads to the formation of labile gel-like networks without subsequent macroscopic phase separation. Unlike α-casein, which shows a strong concentration dependence and extensive gel-like network formation, only one-third of κ-casein molecules participate in the gel network at a time, resulting in a more dynamic and less extensive structure. These findings highlight the unique association properties of κ-casein, contributing to a better understanding of its behavior under various conditions and its potential role in casein micelle formation.
Subject(s)
Caseins , Fluorescence Resonance Energy Transfer , Intrinsically Disordered Proteins , Caseins/chemistry , Caseins/metabolism , Intrinsically Disordered Proteins/chemistry , Intrinsically Disordered Proteins/metabolism , Diffusion , Nuclear Magnetic Resonance, BiomolecularABSTRACT
OBJECTIVES: To synthesize casein enzymatic hydrolysate (CEH)-laden gelatin methacryloyl (GelMA) fibrous scaffolds and evaluate the cytocompatibility and anti-inflammatory effects on dental pulp stem cells (DPSCs). MATERIALS AND METHODS: GelMA fibrous scaffolds with 10%, 20%, and 30% CEH (w/w) and without CEH (control) were obtained via electrospinning. Chemo-morphological, degradation, and mechanical analyses were conducted to evaluate the morphology and composition of the fibers, mass loss, and mechanical properties, respectively. Adhesion/spreading and viability of DPSCs seeded on the scaffolds were also assessed. The anti-inflammatory potential on DPSCs was tested after the chronic challenge of cells with lipopolysaccharides (LPS), followed by treatment with extracts obtained after immersing the scaffolds in α-MEM. The synthesis of the pro-inflammatory cytokines IL-6, IL-1α, and TNF-α was measured by ELISA. Data were analyzed by ANOVA/post-hoc tests (α = 5%). RESULTS: CEH-laden electrospun fibers had a larger diameter than pure GelMA (p ≤ 0.036). GelMA scaffolds laden with 20% and 30% CEH had a greater mass loss. Tensile strength was reduced for the 10% CEH fibers (p = 0.0052), whereas no difference was observed for the 20% and 30% fibers (p ≥ 0.6736) compared to the control. Young's modulus decreased with CEH (p < 0.0001). Elongation at break increased for the 20% and 30% CEH scaffolds (p ≤ 0.0038). Over time, DPSCs viability increased across all groups, indicating cytocompatibility, with CEH-laden scaffolds exhibiting greater cell viability after seven days (p ≤ 0.0166). Also, 10% CEH-GelMA scaffolds decreased the IL-6, IL-1α, and TNF-α synthesis (p ≤ 0.035). CONCLUSION: CEH-laden GelMA scaffolds facilitated both adhesion and proliferation of DPSCs, and 10% CEH provided anti-inflammatory potential after chronic LPS challenge. CLINICAL RELEVANCE: CEH incorporated in GelMA fibrous scaffolds demonstrated the potential to be used as a cytocompatible and anti-inflammatory biomaterial for vital pulp therapy.
Subject(s)
Anti-Inflammatory Agents , Caseins , Cell Survival , Dental Pulp , Gelatin , Tissue Scaffolds , Gelatin/chemistry , Dental Pulp/cytology , Dental Pulp/drug effects , Tissue Scaffolds/chemistry , Humans , Anti-Inflammatory Agents/pharmacology , Cell Survival/drug effects , Methacrylates/chemistry , Materials Testing , Enzyme-Linked Immunosorbent Assay , Tensile Strength , Cells, Cultured , Stem Cells/drug effects , Cell Adhesion/drug effects , Biocompatible Materials/pharmacology , Biocompatible Materials/chemistry , Cytokines/metabolism , Surface PropertiesABSTRACT
This study was conducted to determine the best combination of protein sources in diets for jundiá, based on growth, metabolism, and nutrient deposition. Five protein combinations were tested: casein + fish meal (control), casein + gelatin, casein + albumin, casein + albumin + fish meal, and albumin + fish meal, in diets containing 370 g Kg-1 of crude protein and 13.4 MJ Kg-1 of digestible energy. The fish (9.38 ± 0.12 g) were allocated in a water recirculation system at a density of 3.35 g L-1 per experimental unit and fed until apparent satiety for 40 days with the diets. The fish fed with the control diet had the highest final weight, specific growth rate, protein and feed efficiency ratio, protein retention, and best apparent feed conversion. On the other hand, fish that received casein + albumin and albumin + fish meal diets showed worse results in growth and body protein retention, low trypsin and chymotrypsin activity, and high intestinal amylase activity. Therefore, the combination referred to as control (casein + fish meal) conclusively provides the best rhythm for nutrient digestion and metabolism processes, enabling fish to reach greater growth and retention of body protein with low whole-fish fat content.
Subject(s)
Animal Feed , Dietary Proteins , Animals , Animal Feed/analysis , Dietary Proteins/administration & dosage , Dietary Proteins/metabolism , Caseins/administration & dosage , Caseins/metabolism , Digestion/physiology , Digestion/drug effects , Animal Nutritional Physiological PhenomenaABSTRACT
BACKGROUND: In the realm of sports science, nutrition is a well-established pillar for athletes' training, performance, and post-workout recovery. However, the role of gut microbiota, often overlooked, is a novel and intriguing aspect that can significantly impact athletic performance. With this in mind, our study ventures into uncharted territory, investigating the effect of probiotic and casein supplementation on the aerobic capacity of male soccer players. METHOD: A double-blinded and placebo-controlled study was conducted with 44 male soccer players (Age: 22.81 ± 2.76 yr, Height: 177.90 ± 6.75 cm, Weight: 67.42 ± 8.44 kg). The participants were subjected to the Bruce test in the beginning; then, they were randomly divided into four groups, each consisting of 11 people: probiotics (PRO), casein (CAS), probiotics with casein (PRO+CAS), and placebo (PLA). PRO group was given one probiotic capsule (containing strains of Lactiplantibacillus plantarum BP06, Lacticaseibacillus casei BP07, Lactobacillus acidophilus BA05, Lactobacillus delbrueckii BD08 bulgaricus, Bifidobacterium infantis BI04, Bifidobacterium longum BL03, Bifidobacterium breve BB02 and Streptococcus salivarius thermophilus BT01, with a total dose of 4.5 × 1011 CFU) during dinner, while the CAS group consumed 20 grams of casein powder 45 minutes before bed. The PRO+CAS group was given one probiotic capsule during dinner and 20 grams of casein powder 45 minutes before bed. The participants in the PLA group were given one red capsule (containing 5 grams of starch) during dinner. All participants were instructed to take the supplements only on training days, three times a week for four weeks. The maximal oxygen consumption (VO2max), Ventilatory Threshold (VT), Time-to-exhaustion (TTE), Respiratory Compensation Point (RCP), Isocapnic area Time (Time-IC), Isocapnic area oxygen consumption (VO2-IC), and Hypocapnic Hyperventilation area Time (Time-HHV), after the Bruce test were Measured. All data were analyzed using SPSS Windows software, mixed repeated measure ANOVA, and Bonferroni post hoc test at p < 0.05 level. RESULTS: The current study's findings illustrated that, after the intervention, TTE (p = 0.01) and RCP (p = 0.01) were significantly improved in PRO+CAS compared to the PLA group. No significant difference was observed between PRO and PLA (p = 0.52), PRO and CAS (p = 0.999), PRO and PRO+CAS (p = 0.9), CAS and PLA (p = 0.65), CAS and PRO+CAS (p = 0.73) in TTE. In addition, no significant difference was observed between PRO and CAS (p = 0.999), PRO and PLA (p = 0.40), PRO and PRO+CAS (p = 0.999), CAS and PLA (p = 0.263), CAS and PRO+CAS (p = 0.999) in RCP. Time-HHV was significantly higher in PRO+CAS (p = 0.000) and CAS (p = 0.047) compared to the PLA group. However, no significant difference was observed in the Time-HHV between PRO and CAS (p = 0.999), PRO and PRO+CAS (p = 0.25), PRO and PLA (p = 0.12), and CAS and PRO+CAS (p = 0.57). Additionally, all the groups had no significant differences in VO2max, VT1, VO2-IC and Time-IC. CONCLUSION: The findings showed that consuming probiotics and casein could relatively improve the aerobic capacity of male soccer players. Nevertheless, simultaneous consumption of probiotics and casein had a more pronounced effect on aerobic capacity indicators, especially TTE and Time-HHV.
Subject(s)
Athletic Performance , Caseins , Dietary Supplements , Probiotics , Soccer , Humans , Male , Probiotics/administration & dosage , Probiotics/pharmacology , Double-Blind Method , Soccer/physiology , Caseins/administration & dosage , Caseins/pharmacology , Young Adult , Athletic Performance/physiology , Sports Nutritional Physiological Phenomena , Oxygen Consumption , Gastrointestinal Microbiome/drug effects , AdultABSTRACT
The constantly diverse demand scenarios for rapid on-site analysis have put forward high requirements for developing low-cost and user-friendly visual detection methods. Therefore, developing a visual detection method with simple operation and intuitive results has important practical value in the field of analysis and detection, but it is also challenging. In this work, we propose a microsyringe-assisted visual volume detection method based on phase separation, and apply it to analyze the milk-clotting activity of chymosin. Chymosin can cause phase separation of milk with whey in the mobile phase and curd in the gel state. The network structures of casein in curd can trap water molecules, resulting in separation of whey from curd gradually. Therefore, the analysis of chymosin milk-clotting activity can be realized according to the volume of whey measured using a portable microsyringe. This method shows a good linear correlation when the concentration of chymosin ranges from 1.02 U L-1 to 1020 U L-1 and the limit of detection of this method for chymosin is calculated to be 0.03 U mL-1. This work successfully realizes the visual analysis of chymosin milk-clotting activity based on the enzyme-triggered phase separation. It also shows great promise to be applied in other phase separation-based detection systems with the advantages of high accuracy, great portability and user-friendliness.
Subject(s)
Chymosin , Milk , Chymosin/chemistry , Chymosin/metabolism , Milk/chemistry , Animals , Whey/chemistry , Caseins/analysis , Caseins/chemistry , Phase SeparationABSTRACT
BACKGROUND: The milk's nutritional value is determined by its constituents, including fat, protein, carbohydrates, and minerals. The mammary gland's ability to produce milk is controlled by a complex network of genes. Thereby, the fat, protein, and lactose synthesis must be boost in milk to increase milk production efficiency. This can be accomplished by fusing genetic advancements with proper management practices. Therefore, this study aimed to investigate the association between the Lipoprotein lipase (LPL), kappa casein CSN3, and Glucose transporter 1 (GLUT1) genes expression levels and such milk components as fat, protein, and lactose in different dairy breeds during different stages of lactation. METHODS: To achieve such a purpose, 94 milk samples were collected (72 samples from 36 multiparous black-white and red-white Holstein-Friesian (HF) cows and 22 milk samples from 11 Egyptian buffaloes) during the early and peak lactation stages. The milk samples were utilized for milk analysis and genes expressions analyses using non- invasive approach in obtaining milk fat globules (MFGs) as a source of Ribonucleic acid (RNA). RESULTS: LPL and CSN3 genes expressions levels were found to be significantly higher in Egyptian buffalo than Holstein-Friesian (HF) cows as well as fat and protein percentages. On the other hand, GLUT1 gene expression level was shown to be significantly higher during peak lactation than early lactation. Moreover, lactose % showed a significant difference in peak lactation phase compared to early lactation phase. Also, fat and protein percentages were significantly higher in early lactation period than peak lactation period but lactose% showed the opposite pattern of Egyptian buffalo. CONCLUSION: Total RNA can be successfully obtained from MFGs. The results suggest that these genes play a role in glucose absorption and lactose synthesis in bovine mammary epithelial cells during lactation. Also, these results provide light on the differential expression of these genes among distinct Holstein-Friesian cow breeds and Egyptian buffalo subspecies throughout various lactation phases.
Subject(s)
Caseins , Glycolipids , Glycoproteins , Lactation , Lipid Droplets , Mammary Glands, Animal , Milk , RNA, Messenger , Animals , Cattle/genetics , Lactation/genetics , Female , Lipid Droplets/metabolism , Milk/chemistry , Milk/metabolism , Glycolipids/metabolism , Caseins/genetics , Caseins/metabolism , Glycoproteins/genetics , Glycoproteins/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Mammary Glands, Animal/metabolism , Lipoprotein Lipase/genetics , Lipoprotein Lipase/metabolism , Glucose Transporter Type 1/genetics , Glucose Transporter Type 1/metabolism , Buffaloes/genetics , Buffaloes/metabolism , Lactose/metabolism , Lactose/analysis , Milk Proteins/analysis , Milk Proteins/metabolism , Milk Proteins/genetics , Gene Expression RegulationABSTRACT
This study investigates the dynamic changes in milk nutritional composition and microbial communities in Tibetan sheep and goats during the first 56 days of lactation. Milk samples were systematically collected at five time points (D0, D7, D14, D28, D56) post-delivery. In Tibetan sheep, milk fat, protein, and casein contents were highest on D0, gradually decreased, and stabilized after D14, while lactose and galactose levels showed the opposite trend. Goat milk exhibited similar initial peaks, with significant changes particularly between D0, D7, D14, and D56. 16S rRNA gene sequencing revealed increasing microbial diversity in both species over the lactation period. Principal coordinates analysis identified distinct microbial clusters corresponding to early (D0-D7), transitional (D14-D28), and mature (D56) stages. Core phyla, including Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria, dominated the milk microbiota, with significant temporal shifts. Core microbes like Lactobacillus, Leuconostoc, and Streptococcus were common in both species, with species-specific taxa observed (e.g., Pediococcus in sheep, Shewanella in goats). Furthermore, we observed a highly shared core microbiota in sheep and goat milk, including Lactobacillus, Leuconostoc, and Streptococcus. Spearman correlation analysis highlighted significant relationships between specific microbial genera and milk nutrients. For instance, Lactobacillus positively correlated with total solids, non-fat milk solids, protein, and casein, while Mannheimia negatively correlated with protein content. This study underscores the complex interplay between milk composition and microbial dynamics in Tibetan sheep and goats, informing strategies for livestock management and nutritional enhancement. KEY POINTS: ⢠The milk can be classified into three types based on the microbiota composition ⢠The changes of milk microbiota are closely related to the variations in nutrition ⢠Filter out microbiota with species specificity and age specificity in the milk.
Subject(s)
Goats , Microbiota , Milk , RNA, Ribosomal, 16S , Animals , Goats/microbiology , Milk/microbiology , Milk/chemistry , Sheep/microbiology , RNA, Ribosomal, 16S/genetics , Tibet , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Female , Lactation , Caseins , Milk Proteins/analysisABSTRACT
The effects of oil type, emulsifier type, and emulsion particle size on the texture, gel strength, and rheological properties of SPI emulsion-filled gel (SPI-FG) and TFSP emulsion-filled gel (TFSP-FG) were investigated. Using soybean protein isolate or sodium caseinate as emulsifiers, emulsions with cocoa butter replacer (CBR), palm oil (PO), virgin coconut oil (VCO), and canola oil (CO) as oil phases were prepared. These emulsions were filled into SPI and TFSP gel substrates to prepare emulsion-filled gels. Results that the hardness and gel strength of both gels increased with increasing emulsion content when CBR was used as the emulsion oil phase. However, when the other three liquid oils were used as the oil phase, the hardness and gel strength of TFSP-FG decreased with the increasing of emulsion content, but those of SPI-FG increased when SPI was used as emulsifier. Additionally, the hardness and gel strength of both TFSP-FG and SPI-FG increased with the decreasing of mean particle size of emulsions. Rheological measurements were consistent with textural measurements and found that compared with SC, TFSP-FG, and SPI-FG showed higher G' values when SPI was used as emulsifier. Confocal laser scanning microscopy (CLSM) observation showed that the distribution and stability of emulsion droplets in TFSP-FG and SPI-FG were influenced by the oil type, emulsifier type and emulsion particle size. SPI-stabilized emulsion behaved as active fillers in SPI-FG reinforcing the gel matrix; however, the gel matrix of TFSP-FG still had many void pores when SPI-stabilized emulsion was involved. In conclusion, compared to SPI-FG, the emulsion filler effect that could reinforce gel networks became weaker in TFSP-FG.
Subject(s)
Emulsifying Agents , Emulsions , Gels , Particle Size , Rheology , Soybean Proteins , Soybean Proteins/chemistry , Emulsions/chemistry , Emulsifying Agents/chemistry , Gels/chemistry , Plant Oils/chemistry , Palm Oil/chemistry , Rapeseed Oil/chemistry , Coconut Oil/chemistry , Hardness , Caseins/chemistry , Dietary FatsABSTRACT
Human milk reduces risk for necrotizing enterocolitis in preterm infants. Necrotizing enterocolitis occurs in the ileocecal region where thousands of milk protein-derived peptides have been released from digestion. Digestion-released peptides may exert bioactivity, such as antimicrobial and immunomodulatory activities, in the gut. In this study, we applied mass spectrometry-based peptidomics to characterize peptides present in colostrum before and after in vitro digestion. Sequence-based computational modeling was applied to predict peptides with antimicrobial activity. We identified more peptides in undigested samples, yet the abundances were much higher in the digested samples. Heatmapping demonstrated highly different peptide profiles between undigested and digested samples. Four peptides (αS1-casein [157-163], αS1-casein [157-165], ß-casein [153-159] and plasminogen [591-597]) were selected, synthesized and tested against common pathogenic bacteria associated with necrotizing enterocolitis. All four exhibited bacteriostatic, though not bactericidal, activities against Klebsiella aerogenes, Citrobacter freundii and Serratia marcescens, but not Escherichia coli.
Subject(s)
Colostrum , Enterocolitis, Necrotizing , Milk, Human , Humans , Colostrum/chemistry , Infant, Newborn , Enterocolitis, Necrotizing/prevention & control , Milk, Human/chemistry , Antimicrobial Peptides/pharmacology , Peptides/pharmacology , Female , Caseins/pharmacology , Anti-Bacterial Agents/pharmacology , Digestion , Milk Proteins/pharmacologyABSTRACT
In the present study, different oligosaccharides (fructooligosaccharide (FOS), galactooligosaccharide (GOS), isomaltooligosaccharide (IMO), and xylooligosaccharide (XOS)) were modified on casein (CN) via Maillard reaction. The CN-oligosaccharide conjugates were evaluated for modifications to functional groups, fluorescence intensity, water- and oil-holding properties, emulsion foaming properties, as well as general emulsion properties and stability. The results demonstrated that the covalent combination of CN and oligosaccharides augmented the spatial repulsion and altered the hydrophobic milieu of proteins, which resulted in a diminution in water-holding capacity, an augmentation in oil-holding capacity, and an enhancement in the emulsification properties of proteins. Among them, CN-XOS exhibited the most pronounced changes, with the emulsification activity index and emulsion stability index increasing by approximately 72% and 84.3%, respectively. Furthermore, CN-XOS emulsions have smaller droplet sizes and higher absolute potential values than CN emulsions. Additionally, CN-XOS emulsions demonstrate remarkable stability when ion concentration and pH are varied. These findings indicate that oligosaccharides modified via Maillard reaction can be used as good natural emulsifiers. This provides a theoretical basis for using oligosaccharides to modify proteins and act as natural emulsifiers.
Subject(s)
Caseins , Emulsifying Agents , Emulsions , Maillard Reaction , Oligosaccharides , Oligosaccharides/chemistry , Caseins/chemistry , Emulsifying Agents/chemistry , Emulsions/chemistry , Hydrogen-Ion Concentration , Particle Size , Hydrophobic and Hydrophilic Interactions , Water/chemistryABSTRACT
Calcium is the most abundant mineral in the human body and is involved in critical physiological and cellular processes. It is essential for the development, maintenance, and integrity of bone tissue throughout life. Identifying new natural food-grade chelating agents to improve calcium uptake is of increasing interest. Casein phosphopeptides (CPPs), highly phosphorylated peptides obtained after enzymatic hydrolysis of caseins, represent promising calcium-chelating candidates. The aim of this study was to investigate, using cell culture models, the ability of a digested milk matrix enriched in CPPs to regulate calcium transport through the intestinal barrier and elucidate the involved mechanisms. To this end, a CPP-preparation underwent in vitro static digestion and was subsequently incubated with an intestinal barrier model to monitor calcium uptake and transport. Our results demonstrated that the digested CPP preparation enhanced the trans-epithelial calcium transport via paracellular pathways and that CPPs, identified by peptidomics, crossed the intestinal barrier in the same time.
Subject(s)
Calcium , Caseins , Intestinal Mucosa , Phosphopeptides , Caseins/pharmacology , Caseins/metabolism , Caseins/chemistry , Phosphopeptides/pharmacology , Phosphopeptides/metabolism , Phosphopeptides/chemistry , Humans , Calcium/metabolism , Intestinal Mucosa/metabolism , Intestinal Mucosa/drug effects , Caco-2 Cells , Biological Transport , Animals , Digestion , Intestinal Absorption/drug effectsABSTRACT
BACKGROUND: White-spot lesions are considered an initial carious stage characterized by an outer enamel layer with significantly reduced mineralization. This study was conducted to assess the combined effect of Biomin F toothpaste and Diode laser on remineralization of white spot lesions. MATERIALS AND METHODS: An invitro study conducted on a total of 30 premolars divided into three groups; Group A (Biomin F Tooth paste), Group B (Biomin F with laser application for 30 sec), Group C (Negative control). The three groups were submitted to three stages; stage 1:Baseline,stage 2:After demineralization ,and stage 3:After remineralization. In each stage, elemental analysis(calcium, phosphorus, and fluoride)was measured quantitatively using Energy Dispersive X-ray (EDX) analysis and qualitatively by micrographs using scanning electron microscope. The data were tested to find significant difference between mineral changes during stages by using (ANOVA) test and Bonferroni test. RESULTS: Calcium, phosphorus and fluoride ions decreased in all groups after demineralization. In stage 3, after application of remineralizing agents, Calcium ions increased significantly in groups A and B where p<.05. As regards to the phosphorus ions, a significant increase was observed in all groups with group A showed the highest gain as phosphorus level percentage change (%mass) was 56.52±18.02 . Fluoride ions increased significantly in groups A and B (p<0.05) but decreased significantly in group C. There was no statistical significant difference between group A and B (p ≥.05) in calcium, phosphorus, and fluoride level after remineralization. CONCLUSION: Within the limitation of the present study, we concluded that Biomin F toothpaste is promising in the repairing of white spot lesions on the surface of the demineralized enamel. Diode laser did not affect the remineralizing ability of Biomin F toothpaste.
Subject(s)
Calcium , Dental Enamel , Fluorides , Lasers, Semiconductor , Microscopy, Electron, Scanning , Phosphorus , Spectrometry, X-Ray Emission , Tooth Remineralization , Toothpastes , Tooth Remineralization/methods , Toothpastes/therapeutic use , Humans , Phosphorus/analysis , Phosphorus/therapeutic use , Lasers, Semiconductor/therapeutic use , Calcium/analysis , Calcium/therapeutic use , Fluorides/therapeutic use , Dental Enamel/drug effects , In Vitro Techniques , Cariostatic Agents/therapeutic use , Cariostatic Agents/pharmacology , Dental Caries , Tooth Demineralization , Caseins/therapeutic use , Caseins/pharmacology , BicuspidABSTRACT
Cancer is one of the leading causes of death worldwide, with over 10 million fatalities annually. While tumors can be surgically removed and treated with chemotherapy, radiotherapy, immunotherapy, hormonal therapy, or combined therapies, current treatments often result in toxic side effects in normal tissue. Therefore, researchers are actively seeking ways to selectively eliminate cancerous cells, minimizing the toxic side effects in normal tissue. Caseins and its derivatives have shown promising anti-cancer potential, demonstrating antitumor and cytotoxic effects on cells from various tumor types without causing harm to normal cells. Collectively, these data reveals advancements in the study of caseins and their derivative peptides, particularly providing a comprehensive understanding of the molecular mechanism of action in cancer therapy. These mechanisms occur through various signaling pathways, including (i) the increase of interferon-associated STAT1 signaling, (ii) the suppression of stemness-related markers such as CD44, (iii) the attenuation of the STAT3/HIF1-α signaling, (iv) the down-expression of uPAR and PAI-1, (v) the loss of mitochondrial membrane potential and reduced intracellular ATP production, (vi) the increase of caspase-3 activity, and (vii) the suppression of TLR4/NF-кB signaling. Therefore, we conclude that casein could be an effective adjuvant for cancer treatment.
Subject(s)
Antineoplastic Agents , Caseins , Neoplasms , Humans , Neoplasms/drug therapy , Caseins/pharmacology , Antineoplastic Agents/pharmacology , Antineoplastic Agents/therapeutic use , Signal Transduction/drug effects , AnimalsABSTRACT
In recent years, there has been a growing interest in the pure casein fraction of milk protein, particularly ß-casein due to its physicochemical properties as well as its bio- and techno-functional properties. The utilization of self-assembled ß-caseins from bovine origin as nanocarriers for the delivery of nutraceutical compounds or drugs has increased dramatically. Concerning ß-caseins from other milk sources, the use of hypoallergenic donkey ß-caseins as a potential delivery vehicle for nutraceutical hydrophobic compounds is beginning to generate interest. The present review deals with casein micelles models, bovine and donkey ß-casein molecular structures, as well as their physical-chemical properties that account for their exploitation in nutraceutics and pharmaceutics. This review work suggests the possibility of developing delivery systems for hydrophobic bioactive compounds using ß-casein purified from hypoallergenic donkey milk, highlighting the potential of this protein as an innovative and promising vehicle for enhancing the enrichment and bioavailability of various bioactive substances in food products.
Subject(s)
Caseins , Equidae , Micelles , Milk , Animals , Caseins/chemistry , Cattle , Milk/chemistry , Drug Carriers/chemistry , Dietary Supplements/analysis , Hydrophobic and Hydrophilic InteractionsABSTRACT
Essential oils (EOs) represent a pivotal source for developing potent antimicrobial drugs. However, EOs have seldom found their way to the pharmaceutical market due to their instability and low bioavailability. Nanoencapsulation is an auspicious strategy that may circumvent these limitations. In the current study, lemongrass essential oil (LGO) was encapsulated in zein-sodium caseinate nanoparticles (Z-NaCAS NPs). The fabricated nanocomposite was characterized using dynamic light scattering, Fourier-transform infrared spectroscopy, differential scanning calorimetry, and transmission electron microscopy. The antimicrobial activity of LGO loaded NPs was assessed in comparison to free LGO against Staphylococcus epidermidis, Enterococcus faecalis, Escherichia coli, and Klebsiella pneumoniae. Furthermore, their antibacterial mechanism was examined by alkaline phosphatase, lactate dehydrogenase, bacterial DNA and protein assays, and scanning electron microscopy. Results confirmed the successful encapsulation of LGO with particle size of 243 nm, zeta potential of - 32 mV, and encapsulation efficiency of 84.7%. Additionally, the encapsulated LGO showed an enhanced thermal stability and a sustained release pattern. Furthermore, LGO loaded NPs exhibited substantial antibacterial activity, with a significant 2 to 4 fold increase in cell wall permeability and intracellular enzymes leakage versus free LGO. Accordingly, nanoencapsulation in Z-NaCAS NPs improved LGO physicochemical and antimicrobial properties, expanding their scope of pharmaceutical applications.
Subject(s)
Anti-Bacterial Agents , Caseins , Nanocomposites , Oils, Volatile , Zein , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Zein/chemistry , Nanocomposites/chemistry , Oils, Volatile/chemistry , Oils, Volatile/pharmacology , Caseins/chemistry , Plant Oils/chemistry , Plant Oils/pharmacology , Microbial Sensitivity Tests , Particle Size , Staphylococcus epidermidis/drug effects , Klebsiella pneumoniae/drug effects , TerpenesABSTRACT
Functional drugs nano delivery systems manufactured from natural active products are promising for the field of biomedicines. In this study, an anti-ulcerative colitis (UC) curcumin loaded biopolymeric nanocomposite (CZNH) was fabricated and investigated. CZNH nanocomposite was obtained using the anti-solvent precipitation method, wherein curcumin-loaded zein colloidal particles served as the core, while sodium casein (NaCas) and hyaluronic acid (HA) formed the outermost layer of CZNH nanocomposite. Fourier transform infrared (FT-IR) spectrum and transmission electron microscopy (TEM) findings demonstrated that CZNH nanocomposite was a double-layer spherical micelle (250 nm) resulting from the hydrogen bond interactions and electrostatic adsorptions between zein, NaCas, and HA. Furthermore, CZNH nanocomposite exhibited prominent resuspension and storage stability in aqueous solution, which can be stored at 4 °C for approximately 30 days. In vivo anti-UC studies showed that CZNH nanocomposite could effectively alleviate UC symptoms via mediating inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), and IL-6], myeloperoxidase (MPO), and oxidative stress factor [malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px)]. This study suggested that the CZNH nanocomposite showed great promise as an efficient curcumin nanocarrier for UC therapy.