The effect of feeding on different hosts on the egg proteins in Haemaphysalis qinghaiensis tick.

The majority of ixodid ticks display host-specificity to varying extents. Feeding on different hosts affects their development and reproduction. Consequences can be analyzed at the level of the egg, as it is the initial stage of tick development. Tick egg proteins are abundant and diverse, providing nutrients for embryonic development. However, studies on tick egg profiles are scarce. In this study, we aimed to analyze whether feeding Haemaphysalis qinghaiensis ticks on the yaks (Bos grunniens) and domestic sheep (Ovis aries) has an impact on the variety and variability of the egg proteome. Detached engorged females were used to lay eggs, which were then collected, dewaxed, and subjected to protein extraction. The extracted egg proteins were enzymatically digested using Filter-Aided Sample Preparation (FASP), and the unique peptides were separated and detected by Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS). The MS data were searched against the previously constructed whole tick transcriptome library of H. qinghaiensis, and the UniProt database for the identification of tick-derived egg proteins. The analysis revealed 49 and 53 high-confidence proteins identified in eggs collected from B. grunniens (EggBg) and O. aries (EggOa), respectively. Of these, 46 high-confidence proteins were common to both egg types, while three were unique to EggBg and seven to EggOa. All the identified proteins mainly belonged to enzymes, enzyme inhibitors, transporters, and proteins with unknown functions. The differential abundance analysis showed that nine proteins were significantly more present in EggBg, while six were significantly more present in EggOa. Overall, enzymes were the most diverse group, while vitellogenin (Vg) was the most abundant. Blood meal uptake on different hosts has a certain effect on the egg proteome composition and the abundance of some proteins, but it may also lead to compensation of protein roles.

Advances in Understanding the Antioxidant and Antigenic Properties of Egg-Derived Peptides.

Pepsin, trypsin and proteinase K were used in the present study to hydrolyse the proteins from whole eggs, yolks or whites, and the resulting hydrolysates were characterised in terms of antioxidant and IgE-binding properties, using a combination of in vitro and in silico methods. Based on the degree of hydrolysis (DH) results, the egg yolk proteins are better substrates for all the tested enzymes (DH of 6.2-20.1%) compared to those from egg whites (DH of 2.0-4.4%). The SDS-PAGE analysis indicated that pepsin and proteinase K were more efficient compared to trypsin in breaking the intramolecular peptide bonds of the high molecular weight egg proteins. For all the tested substrates, enzyme-assisted hydrolysis resulted in a significant increase in antioxidant activity, suggesting that many bioactive peptides are encrypted in inactive forms in the parent proteins. The hydrolysates obtained with proteinase K exhibited the highest DPPH radical scavenging activity (124-311 µM Trolox/g protein) and the lowest residual IgE-binding capacity. The bioinformatics tools revealed that proteinase K is able to break the integrity of the main linear IgE-binding epitopes from ovalbumin and ovomucoid. It can be concluded that proteinase K is a promising tool for modulating the intrinsic properties of egg proteins.

The Impact of Processing and Extraction Methods on the Allergenicity of Targeted Protein Quantification as Well as Bioactive Peptides Derived from Egg.

This review article discusses advanced extraction methods to enhance the functionality of egg-derived peptides while reducing their allergenicity. While eggs are considered a nutrient-dense food, some proteins can cause allergic reactions in susceptible individuals. Therefore, various methods have been developed to reduce the allergenicity of egg-derived proteins, such as enzymatic hydrolysis, heat treatment, and glycosylation. In addition to reducing allergenicity, advanced extraction methods can enhance the functionality of egg-derived peptides. Techniques such as membrane separation, chromatography, and electrodialysis can isolate and purify specific egg-derived peptides with desired functional properties, improving their bioactivity. Further, enzymatic hydrolysis can also break down polypeptide sequences and produce bioactive peptides with various health benefits. While liquid chromatography is the most commonly used method to obtain individual proteins for developing novel food products, several challenges are associated with optimizing extraction conditions to maximize functionality and allergenicity reduction. The article also highlights the challenges and future perspectives, including optimizing extraction conditions to maximize functionality and allergenicity reduction. The review concludes by highlighting the potential for future research in this area to improve the safety and efficacy of egg-derived peptides more broadly.

Soybean and Lupine Addition in Hen Nutrition-Influence on Egg Immunoreactivity.

Modifying hen fodder is a common way of changing eggs composition today. However, there is no information on the effect of the source of protein in the fodder replacement on egg allergenicity. This research aimed to detect potential differences in the immunoreactivity and protein composition of eggs from hens fed with fodder containing legume. The aim of the first step of the study was to select the proper solvent for extracting allergenic proteins from hen eggs. Two of them (containing Tween 20 and Triton 100) were selected, based on protein profile and concentration analysis. Egg-white- and egg-yolk-proteins extracts prepared with them were checked for potential differences, using SDS-PAGE electrophoresis, and then the Western-blot method, using sera from children allergic to eggs and soy. Preliminary studies on the influence of fodder composition on the composition of egg proteins suggest that the addition of soy and lupine to fodder modifies the expression of egg proteins. The observed differences in the immunoreactivity of proteins contained in hen egg-white samples do not seem to be as significant as the appearance of protein with a molecular weight of ~13 kDa in the yolk of eggs obtained from soybean-fed hens. This protein may increase the immunoreactivity of eggs for children allergic solely to soy.

Changes in stability and in vitro digestion of egg-protein stabilized emulsions and ß-carotene gels in the presence of sodium tripolyphosphate.

BACKGROUND: Egg proteins are effective emulsifiers and gelators in food systems. However, the physicochemical stability and control release properties of egg-protein stabilized emulsions and gels need to be further improved. The potential of sodium tripolyphosphate (St) to improve the functionality of egg proteins was evaluated. RESULTS: The emulsions with St had smaller particle sizes and higher zeta potential, leading to better physical stability. Furthermore, the oxidation stability increased with increasing St contents, possibly due to its metal chelating capacity and the improved emulsifying activity of whole-egg dispersions. Phosphate had a positive impact on the chemical stability of ß-carotene in whole-egg liquids and gels, decreasing the degradation during thermal treatment. The gel made with St was firm and broke down slowly, leading to a low rate of digestion and ß-carotene release in simulated gastric fluid. CONCLUSION: This study shows that St is useful to improve the egg proteins stabilized emulsions and gels, which is applicable in the development of emulsion-based food grade gel products. © 2021 Society of Chemical Industry.

Effect of processing technologies on the digestibility of egg proteins.

Egg and egg products are a rich source of highly bioavailable animal proteins. Several processing technologies can affect the structural and functional properties of these proteins differently and can influence their fate inside the gastrointestinal tract. The present review examines some of the processing technologies for improving egg protein digestibility and discusses how different processing conditions affect the digestibility of egg proteins under gastrointestinal digestion environments. To provide up-to-date information, most of the studies included in this review have been published in the last 5 years on different aspects of egg protein digestibility. Digestibility of egg proteins can be improved by employing some processing technologies that are able to improve the susceptibility of egg proteins to gastrointestinal proteases. Processing technologies, such as pulsed electric field, high-pressure, and ultrasound, can induce conformational and microstructural changes that lead to unfolding of the polypeptides and expose active sites for further interactions. These changes can enhance the accessibility of digestive proteases to cleavage sites. Some of these technologies may inactivate some egg proteins that are enzyme inhibitors, such as trypsin inhibitors. The underlying mechanisms of how different technologies mediate the egg protein digestibility have been discussed in detail. The proteolysis patterns and digestibility of the processed egg proteins are not always predictable and depends on the processing conditions. Empirical input is required to tailor the optimization of processing conditions for favorable effects on protein digestibility.

Gel properties of salty liquid whole egg as affected by preheat treatment.

Heat treatment is an indispensable processing step of seasoned liquid egg. The effects of preheat treatment (60-75 °C) on gel properties of liquid whole egg (LWE) at different NaCl concentrations (0-3%, w/w) were investigated to provide guidance for the production of salty LWE. Results showed that LWE exhibited higher particle size after heating, with coincidental increases in surface hydrophobicity and decreases in protein solubility. While LWE with NaCl added exhibited increase in protein solubility and decrease in particle size of aggregates. Electrophoresis and optical microscopy showed that NaCl would induce the transformation of egg granules from insoluble form to soluble form, inhibiting the aggregation of LWE proteins during preheat treatment, reflected by the reduced particle size. The analysis of gel aggregated force and texture indicated that NaCl addition and preheat treatment can improve gelling properties of LWE synergistically by strengthening the hydrophobic interaction and hydrogen bonds.

Vaccination Against Measles, Mumps, and Rubella in the Light of Current Epidemic Threats: Unjustified Postponement.

A worrying increase in the number of measles cases has been noted recently in Poland, which may have to do with a decreasing proportion of children vaccinated against measles, mumps, and rubella (MMR) in the second year of life (<95%). For many years, MMR vaccination in children has been associated with a fear of allergy to eggs. This study seeks to define the reason and justification for postponing MMR vaccination in a population of children referred to the outpatient specialist immunization clinic. One hundred and thirty eight (138) children, mean 24.5 ± 26.6 months, with a history of past allergies, in whom the first-time MMR vaccination was delayed by family doctors for fear of allergic reactions, were enrolled into the study. The mean delay in a vaccine shot was 12.3 ± 26.9 months. There were 101 children who displayed a distinct allergy to the egg proteins, among other accompanying types of allergy. All of the 138 children were found eligible to receive MMR vaccine at the visit to the clinic. No early allergic responses were noticed in any of the children. There were negligible delayed allergic responses in six children, all from the egg allergy group. We conclude that MMR vaccination in children with egg allergy is safe and can be conducted on the outpatient basis without any specific precautions or safety measures. Delays in vaccination were unjustified and may jeopardize children's health. There is a need for insightful education of primary care doctors concerning of MMR vaccination safety, particularly when allergy is suspected, to avoid unduly and potentially harmful delays.

Modifying the Functional Properties of Egg Proteins Using Novel Processing Techniques: A Review.

Egg proteins can be used in a wide range of food products, owing to their excellent foaming, emulsifying, and gelling properties. Another important functional property is the susceptibility of egg proteins to enzymatic hydrolysis, as protein digestion is closely related to its nutritional value. These functional properties of egg proteins are likely to be changed during food processing. Conventional thermal processing can easily induce protein denaturation and aggregation and consequently reduce the functionality of egg proteins due to the presence of heat-labile proteins. Accordingly, there is interest from the food industry in seeking novel nonthermal or low-thermal techniques that sustain protein functionality. To understand how novel processing techniques, including high hydrostatic pressure, pulsed electric fields, ionizing radiation, ultraviolet light, pulsed light, ultrasound, ozone, and high pressure homogenization, affect protein functionality, this review introduces the mechanisms involved in protein structure modification and describes the structure-functionality relationships. Novel techniques differ in their mechanisms of protein structure modification and some have been shown to improve protein functionality for particular treatment conditions and product forms. Although there is considerable industrial potential for the use of novel techniques, further studies are required to make them a practical reality, as the processing of egg proteins often involves other influencing factors, such as different pH and the presence of other food additives (for example, salts, sugar, and polysaccharides).

Bioactive peptides derived from egg proteins: A review.

Egg proteins have various functional and biological activities which make them potential precursor proteins for bioactive peptide production. Simulated in vitro gastrointestinal digestion and enzymatic hydrolysis using non-gastrointestinal proteases have been used as tools to produce these peptides. Bioactive peptides derived from egg proteins are reported to display various biological activities, including angiotensin I-converting enzyme (ACE) inhibitory (antihypertensive), antioxidant, antimicrobial, anti-inflammatory, antidiabetic and iron-/calcium-binding activities. More importantly, simulated in vitro gastrointestinal digestion has indicated that consumption of egg proteins has physiological benefits due to the release of such multifunctional peptides. This review encompasses studies reported to date on the bioactive peptide production from egg proteins.

Egg proteins: fractionation, bioactive peptides and allergenicity.

Eggs are an important source of macro and micronutrients within the diet, comprised of proteins, lipids, vitamins, and minerals. They are constituted by a shell, the white (containing 110 g kg-1 proteins: ovalbumin, ovotransferrin, ovomucoid, lysozyme and ovomucin), and the yolk (containing 150-170 g kg-1 proteins: lipovitellins, phosvitin, livetins, and low-density lipoproteins). Owing to their nutritional value and biological characteristics, both the egg white and yolk proteins are extensively fractionated using different techniques (e.g., liquid chromatography, ultrafiltration, electrophoresis, and chemical precipitation), in which liquid chromatography is the most commonly used technique to obtain individual proteins with high protein recovery and purity to develop novel food products. However, concerns over allergenic responses induced by certain egg proteins (e.g., ovomucoid, ovalbumin, ovotransferrin, lysozyme, α-livetin, and lipoprotein YGP42) limit their widespread use. As such, processing technologies (e.g., thermal processing, enzymatic hydrolysis, and high-pressure treatment) are investigated to reduce the allergenicity by conformational changes. In addition, biological activities (e.g., antioxidant, antimicrobial, antihypertensive, and anticancer activities) associated with egg peptides have received more attention, in which enzyme hydrolysis is demonstrated as a promising way to break polypeptides sequences and produce bioactive peptides to provide nutritional and therapeutic benefits for human health. © 2018 Society of Chemical Industry.

Allergenic proteins in enology: a review on technological applications and safety aspects.

Proteinaceous products are widely used as fining agents during winemaking to remove unwanted insoluble particles and undissolved microscopic particles (colloidal material) from the must or wine to improve stability. Some of them (egg white, caseinates, and fish gelatine) have allergenic potential and the presence of their residues in the final product could represent a risk for allergic individuals. Moreover, lysozyme (an egg allergen) is included among wine additives to control the fermentation processes and avoid spoiling during winemaking. The aim of this paper is to review the experimental/clinical data on the use of allergenic products in enology and the measurement of relative risk for sensitized subjects. In addition, methods developed specifically for the quantification of allergenic residues in must and wine are described.

Improving gelling properties of diluted whole hen eggs with sodium chloride and sodium tripolyphosphate: Study on intermolecular forces, water state and microstructure.

Individual and synergistic effects of sodium chloride (NaCl) and sodium tripolyphosphate (STPP) on the physicochemical and gelling properties of highly diluted liquid whole eggs were studied. Results showed that NaCl and STPP acted differently whereby NaCl addition increased the surface hydrophobicity of the egg proteins and STPP addition increased the protein solubility and the negative surface charge. When combined together, these changes led to a significant increase in a number of intermolecular forces after heat treatment, including hydrophobic interactions, disulfide bonds, and hydrogen bonds, contributing to the best structure and texture of the whole egg gels enriched with binary salts. The amount of the free water in the heat-induced gel products with the addition of both NaCl and STPP were the least as compared to systems with single salt addition, which was related to the coarse and dense network microstructure.

Differences in allergic symptoms after the consumption of egg yolk and egg white.

Hen's eggs are one of the most common causes of food allergy. Although hen's eggs are known to cause more gastrointestinal symptoms than other foods, it is not known whether there is a difference in organ-specific symptoms between egg yolk (EY) and egg white (EW). The present study aimed to determine whether there are organ-specific differences in the immediate symptoms of EY and EW in patients with hen's egg allergies. We retrospectively investigated the immediate symptoms and treatment contents of those who had a positive result in an oral food challenge (OFC) of boiled whole EY or 10 g of boiled EW in our hospital from January 2013 to July 2019. We compared 80 patients in the EY-OFC-positive group with 106 patients in the EW-OFC-positive group. The EY-OFC-positive group had significantly fewer respiratory symptoms and significantly more gastrointestinal symptoms than the EW-OFC-positive group and had significantly more gastrointestinal symptoms only. In terms of treatment, significantly fewer patients in the EY-OFC-positive group required beta 2-agonist inhalation, and a significantly higher proportion of patients did not require treatment. Compared to EW, EY is more likely to cause gastrointestinal symptoms and less likely to cause respiratory symptoms. It may be necessary to discriminate between EY and EW allergy during diagnosis.

Nutritional composition and proteomic analysis of soft-shelled turtle (Pelodiscus sinensis) egg and identification of oligopeptides with alpha-glucosidase inhibitory activity.

This study aimed to explore nutritional compositions and proteomics of soft-shelled turtle (SST) egg, as well as identify potential antidiabetic oligopeptides with α-glucosidase inhibitory property. Results revealed that SST egg is a promising source of highly nutritious proteins and minerals (54.64% and 5.81% of dry matter, respectively). Further proteomic analysis showed SST egg proteins contained at least 9 protein families, such as transferrin/iron binding protein and immunoregulation-related protein. Hydrolysis by different enzymes, especially papain, remarkably increased α-glucosidase inhibitory activity and scavenging activity for ABTS, DPPH, hydroxyl and oxygen radicals of SST egg proteins. Peptides from papain hydrolysate were fractionated using ultrafiltration followed by reverse phase chromatography, and 16 peptides were identified in the most active fraction by LC-QTOF-MS/MS. Molecular docking revealed that 14 of these peptides could easily dock into the substrate-binding pocket and/or inhibitor binding sites of α-glucosidase with the docking score below -150 kcal/mol, indicating their potential α-glucosidase inhibitory properties. The five most abundant oligopeptides with potent interaction with α-glucosidase were further synthesized, and oligopeptides HNKPEVEVR, ARDASVLK and SGTLLHK strongly inhibited the activity of α-glucosidase (IC50 of 56, 195 and 289 µmol/L, respectively). Therefore, oligopeptides from enzymatic hydrolysate of SST egg protein exhibit potential antidiabetic activity, making it a promising functional food ingredient.

A heuristic, computer-driven and top-down approach to identify novel bioactive peptides: A proof-of-principle on angiotensin I converting enzyme inhibitory peptides.

Bioactive peptides are short peptides (3-20 amino acid residues in length) endowed of specific biological activities. The identification and characterization of bioactive peptides of food origin are crucial to better understand the physiological consequences of food, as well as to design novel foods, ingredients, supplements, and diets to counteract mild metabolic disorders. For this reason, the identification of bioactive peptides is also relevant from a pharmaceutical standpoint. Nevertheless, the systematic identification of bioactive sequences of food origin is still challenging and relies mainly on the so defined "bottom-up" approaches, which rarely results in the total identification of most active sequences. Conversely, "top-down" approaches aim at identifying bioactive sequences with certain features and may be more suitable for the precise identification of very potent bioactive peptides. In this context, this work presents a top-down, computer-assisted and hypothesis-driven identification of potent angiotensin I converting enzyme inhibitory tripeptides, as a proof of principle. A virtual library of 6840 tripeptides was screened in silico to identify potential highly potent inhibitory peptides. Then, computational results were confirmed experimentally and a very potent novel sequence, LMP was identified. LMP showed an IC50 of 15.8 and 6.8 µM in cell-free and cell-based assays, respectively. In addition, a bioinformatics approach was used to search potential food sources of LMP. Yolk proteins were identified as a possible relevant source to analyze in further experiments. Overall, the method presented may represent a powerful and versatile framework for a systematic, high-throughput and top-down identification of bioactive peptides.

Effects of yolkin on the immune response of mice and its plausible mechanism of action.

Yolkin is a product of proteolytic degradation of vitellogenin, a protein contained in eggs' yolk, with already described procognitive properties. Here, we investigated effects of yolkin on the humoral and cellular immune response in mice, phenotype of cells from lymphoid organs and function of innate immunity cells. In vitro studies included effects of yolkin on mitogen-induced thymocyte proliferation, percentage of CD19 cells in bone marrow cells culture, expression of signaling molecules in Jurkat cells, interleukin 2 receptor (IL-2R) subunits in WEHI 231 cells and susceptibility of these cells to anti-Ig-induced cell death. The results showed that repeatable i.p. injections of yolkin stimulated the humoral immune response to sheep red blood cells (SRBC) irrespective of the time of the treatment. On the other hand, yolkin inhibited contact sensitivity to oxazolone. Treatment of mice with yolkin diminished the percentage of double positive cells and increasing the content of single positive CD4+ and CD8+ cells in the thymus. At the same time an increase of percentage of CD19 + B cells in the spleen and mesenteric lymph nodes was observed. In addition, the protein, given i.p., diminished ex vivo ability to synthesize nitric oxide by resident, peritoneal macrophages, stimulated with lipopolisaccharide (LPS). In vitro studies showed that yolkin increased CD19+ cell content in bone marrow cell population. The protein also enhanced proliferation of thymocytes to concanavalin A and stimulated expression of MAP kinases in Jurkat cells. In WEHI 231 B cell line yolkin caused a loss of IL-2R gamma chain expression, correlated with an increased resistance of these cells to proapoptotic action of anti-Ig antibodies. In conclusion, this is a first demonstration of immunotropic properties of yolkin in in vitro and in vivo tests. The results provide evidence for induction of maturation and stimulatory signals in immature T and B cells by the protein, suggesting its potential role in the development of an embryo's immune system.

Integrated Proteomic and N-Glycoproteomic Analyses of Chicken Egg during Embryonic Development.

To better appreciate the alterations of egg proteins and their modifications during embryonic development, a comparative and quantitative study was performed aimed at chicken egg white and yolk proteome and N-glycoproteome after 12 days of incubation using tandem mass tag (TMT)-labeling technology in conjunction with reversed-phase high-performance liquid chromatography (RP-HPLC). A total of 334 unique N-glycosite-containing peptides from 153 N-glycoproteins were identified, of which 82 N-glycosite-containing peptides showed significant changes after 12 days of incubation. The varied proteome was mainly involved with antibacterial, ionic binding, cell proliferation, and embryonic development, while the different degrading and/or absorbing priorities of egg proteins were proposed. This study provides substantial insight into the effects of N-glycoprotein variations on the utilization of egg proteins by chicken embryo during incubation.

Alterations in apoptotic markers and egg-specific protein gene expression in the chicken oviduct during pause in laying induced by tamoxifen.

The aim of this study was to examine the cell apoptosis, gene expression and activity of caspases 2, 3, 8 and 9, and the mRNA expression of selected egg-specific proteins in the chicken oviduct during pause in egg laying induced by tamoxifen (TMX) treatment. The experiment was carried out on Hy-Line Brown laying hens. The control birds were treated subcutaneously with vehicle (ethanol) and the experimental ones with TMX at a dose of 6 mg/kg of body weight. Hens were treated daily until a pause in egg laying occurred and sacrificed on Day 7 of the experiment. Within the oviductal wall, the highest number of apoptotic cells (TUNEL-positive) was found in the luminal epithelium and the lowest in the stroma. The administration of TMX increased the percentage of apoptotic cells in the magnum, isthmus, and shell gland as well as immunoreactivity for caspases 3 and 9. Real-time PCR analysis revealed the segment-dependent mRNA expression of caspases 2, 3, 8 and 9. Treatment of hens with TMX elevated the level of caspase-2 transcript in the infundibulum, caspases 2, 3 and 8 in the isthmus, and caspase-9 in the shell gland (P < 0.05 - P < 0.001). As shown by fluorometric method TMX caused an increase in the activity of caspases 3 and 8 in the magnum, isthmus and shell gland, and the activity of caspases 2 and 9 in the isthmus and shell gland (P < 0.05 - P < 0.01). The expression of ovalbumin, avidin and ovocleidin-116 mRNAs was decreased (P < 0.05 - P < 0.001), ovocalyxin-36 mRNA level tended to increase, and ovocalyxin-32 expression was not affected by TMX. The results obtained indicate that caspases are involved in the chicken oviduct regression during a pause in laying induced by TMX, and estrogen is involved in the regulation of examined caspase expression and activity. The changes in mRNA transcript levels of some examined egg-specific proteins after TMX treatment suggest that there is a relationship between estrogen action and the expression of these genes.

Molecular forces and gelling properties of heat-set whole chicken egg protein gel as affected by NaCl or pH.

Effects of NaCl concentrations and pH on the intermolecular forces and gel properties of whole chicken egg protein dispersions were studied via solubility, surface hydrophobicity, intermolecular forces, texture analysis, low-field nuclear magnetic resonance (LF-NMR) and colour analysis. Results showed that the intermolecular forces involved in the formation of egg gel were regulated by NaCl/pH. The results of gel fracture analysis suggested that the changes of fracture strength and strain were closely related with the internal balance of gel molecular forces. Moreover, a negative/positive correlation existed in the free water/bound water relaxation proportion and fracture strength. These findings provide an important theoretical basis for the innovation of heat-induced egg gel products.