-
Food Chemistry Jun 2021The interaction between ovalbumin (OVA) and isoflavonoid glabridin (GB) was investigated using spectroscopic and molecular docking techniques. Fluorescence spectroscopy...
The interaction between ovalbumin (OVA) and isoflavonoid glabridin (GB) was investigated using spectroscopic and molecular docking techniques. Fluorescence spectroscopy revealed that GB was bound to OVA mainly due to hydrogen bonding and hydrophobic forces. FT-IR spectroscopy showed that the combination of GB and OVA resulted in a decrease in the β-sheet content of OVA and an increase in the α-helix and extended-chain content. All these experimental results were supported and clarified by molecular docking simulations. GB binding was able to inhibit chemical denaturant-induced structural changes in OVA as observed by intrinsic tryptophan and ANS fluorescence. Moreover, GB-OVA complex increased the aqueous solubility of GB by about 4.45 times at pH 7.0. These results provided insights into the interaction between GB and OVA that contributes to the utilization of GB in the food and pharmaceutical industries.
Topics: Binding Sites; Hydrogen Bonding; Hydrogen-Ion Concentration; Hydrophobic and Hydrophilic Interactions; Isoflavones; Molecular Docking Simulation; Nanostructures; Ovalbumin; Particle Size; Phenols; Protein Binding; Protein Conformation, alpha-Helical; Protein Denaturation; Urea
PubMed: 33444886
DOI: 10.1016/j.foodchem.2020.128981 -
Journal of the Science of Food and... Feb 2021This study investigates the complexation of a pea albumin-rich fraction and ovalbumin with pectin of different degrees of esterification (DE) and blockiness (DB) as a...
BACKGROUND
This study investigates the complexation of a pea albumin-rich fraction and ovalbumin with pectin of different degrees of esterification (DE) and blockiness (DB) as a function of pH and biopolymer mixing ratio by turbidimetric titration and isothermal titration calorimetry (ITC).
RESULTS
Turbidimetric analysis found maximum complexation occurred at a mixing ratio of 4:1 for pea albumin with high methoxy pectin, 8:1 for pea albumin with low methoxy pectin, and 8:1 for ovalbumin with low methoxy pectin. In the case of ovalbumin with high methoxy pectin, interactions were very weak. The pectin with high levels of esterification and blockiness displayed greater interactions with the pea albumin in both turbidimetry and ITC. However, low methoxy pectin imparted better interactions with ovalbumin and displayed higher optical density values than high methoxy pectin.
CONCLUSIONS
The current study indicated that the different thermodynamic parameters of PA-pectin complexes can be tuned by controlling the structural characteristics (DB, DE, and d-galacturonic acid) of the pectin. © 2020 Society of Chemical Industry.
Topics: Albumins; Biopolymers; Calorimetry; Nephelometry and Turbidimetry; Ovalbumin; Pisum sativum; Pectins; Plant Proteins; Thermodynamics
PubMed: 32789852
DOI: 10.1002/jsfa.10733 -
PLoS Neglected Tropical Diseases Oct 2023Excretory/secretory products (ESPs) derived from helminths have been reported to effectively control allergic inflammation, which have better therapeutic prospects than...
INTRODUCTION
Excretory/secretory products (ESPs) derived from helminths have been reported to effectively control allergic inflammation, which have better therapeutic prospects than live parasite infections. However, it remains unknown whether ESPs from schistosome eggs can protect against allergies, despite reports alleging that schistosome infection could alleviate disordered allergic inflammation.
METHOD
In the present study, we investigated the protective effects of ESPs from Schistosoma japonicum eggs (ESP-SJE) on asthmatic inflammation. Firstly, we successfully established an allergic airway inflammation model in mice by alum-adjuvanted ovalbumin (OVA) sensitization and challenge. ESP-SJE were administered intraperitoneally on days -1 and 13 (before sensitization), on day 20 (before challenge), and on days 21-24 (challenge phase).
RESULTS
The results showed that ESP-SJE treatment significantly reduced the infiltration of inflammatory cells, especially eosinophils into the lung tissue, inhibited the production of the total and OVA-specific IgE during OVA-sensitized and -challenged phases, respectively, and suppressed the secretion of Th2-type inflammatory cytokines (IL-4). Additionally, ESP-SJE treatment significantly upregulated the regulatory T cells (Tregs) in the lung tissue during OVA challenge. Furthermore, using liquid chromatography-mass spectrometry analysis and Treg induction experiments in vitro, we might identify nine potential therapeutic proteins against allergic inflammation in ESP-SJE. The targets of these candidate proteins included glutathione S-transferase, egg protein CP422 precursor, tubulin alpha-2/alpha-4 chain, actin-2, T-complex protein 1 subunit beta, histone H₄, whey acidic protein core region, and molecular chaperone HtpG.
CONCLUSION
Taken together, the results discussed herein demonstrated that ESP-SJE could significantly alleviate OVA-induced asthmatic inflammation in a murine model, which might be mediated by the upregulation of Treg in lung tissues that may be induced by the potential modulatory proteins. Therefore, potential proteins in ESP-SJE might be the best candidates to be tested for therapeutic application of asthma, thus pointing out to a possible new therapy for allergic airway inflammation.
Topics: Animals; Mice; Ovalbumin; Schistosoma japonicum; Egg Hypersensitivity; Asthma; Lung; Cytokines; Inflammation; Mice, Inbred BALB C; Bronchoalveolar Lavage Fluid; Disease Models, Animal
PubMed: 37788409
DOI: 10.1371/journal.pntd.0011625 -
Methods in Molecular Biology (Clifton,... 2021The in vivo killing assay allows the quantification of the antigen-specific killing capacity of Cytotoxic CD8 T Lymphocytes (CTLs) in mice. CTLs are indeed known for the...
The in vivo killing assay allows the quantification of the antigen-specific killing capacity of Cytotoxic CD8 T Lymphocytes (CTLs) in mice. CTLs are indeed known for the lysis of cells expressing foreign or modified antigen peptides on their MHC class I molecules. Here we describe the detailed protocol used for the in vivo specific lysis of cells expressing the H-2 K immunodominant CD8 T-cell epitope of the OVA protein: an 8 amino acid peptide corresponding to the 257-264 region of OVA (SIINFEKL).
Topics: Animals; CD8-Positive T-Lymphocytes; Cytotoxicity, Immunologic; Flow Cytometry; Immunization; Immunodominant Epitopes; Mice; Mice, Inbred C57BL; Ovalbumin; Peptide Fragments; Spleen; T-Lymphocytes, Cytotoxic
PubMed: 34053050
DOI: 10.1007/978-1-0716-1507-2_4 -
Journal of Controlled Release :... Dec 2022Drug delivery systems (DDS) for oral delivery of peptide drugs contain excipients that facilitate and enhance absorption. However, little knowledge exists on how DDS...
Drug delivery systems (DDS) for oral delivery of peptide drugs contain excipients that facilitate and enhance absorption. However, little knowledge exists on how DDS excipients such as permeation enhancers interact with the gastrointestinal mucus barrier. This study aimed to investigate interactions of the permeation enhancer sodium 8-[(2-hydroxybenzoyl)amino]octanoate (SNAC) with ex vivo porcine intestinal mucus (PIM), ex vivo porcine gastric mucus (PGM), as well as with in vitro biosimilar mucus (BM) by profiling their physical and barrier properties upon exposure to SNAC. Bulk mucus permeability studies using the peptides cyclosporine A and vancomycin, ovalbumin as a model protein, as well as fluorescein-isothiocyanate dextrans (FDs) of different molecular weights and different surface charges were conducted in parallel to mucus retention force studies using a texture analyzer, rheological studies, cryo-scanning electron microscopy (cryo-SEM), and single particle tracking of fluorescence-labelled nanoparticles to investigate the effects of the SNAC-mucus interaction. The exposure of SNAC to PIM increased the mucus retention force, storage modulus, viscosity, increased nanoparticle confinement within PIM as well as decreased the permeation of cyclosporine A and ovalbumin through PIM. Surprisingly, the viscosity of PGM and the permeation of cyclosporine A and ovalbumin through PGM was unaffected by the presence of SNAC, thus the effect of SNAC depended on the regional site that mucus was collected from. In the absence of SNAC, the permeation of different molecular weight and differently charged FDs through PIM was comparable to that through BM. However, while bulk permeation of neither of the FDs through PIM was affected by SNAC, the presence of SNAC decreased the permeation of FD4 and increased the permeation of FD150 kDa through BM. Additionally, and in contrast to observations in PIM, nanoparticle confinement within BM remained unaffected by the presence of SNAC. In conclusion, the present study showed that SNAC altered the physical and barrier properties of PIM, but not of PGM. The effects of SNAC in PIM were not observed in the BM in vitro model. Altogether, the study highlights the need for further understanding how permeation enhancers influence the mucus barrier and illustrates that the selected mucus model for such studies should be chosen with care.
Topics: Swine; Animals; Intestinal Absorption; Excipients; Caprylates; Ovalbumin; Sodium; Cyclosporine; Permeability; Pharmaceutical Preparations; Mucus; Peptides
PubMed: 36314534
DOI: 10.1016/j.jconrel.2022.09.034 -
The Clinical Respiratory Journal Dec 2023Many asthmatic patients are exposed to cigarette smoke actively or passively, which contributes to asthma exacerbation and poor control. This study is to explore the...
INTRODUCTION
Many asthmatic patients are exposed to cigarette smoke actively or passively, which contributes to asthma exacerbation and poor control. This study is to explore the effects of cigarette smoke on pathological changes in murine surrogate of asthma.
METHODS
C57BL/6 mice were sensitised and challenged with ovalbumin (OVA) to establish a surrogate of asthma and then administered with cigarette smoke extract (CSE). Airways hyperresponsiveness (AHR) was measured using the Flexivent system. Histological staining (haematoxylin-eosin [HE], periodic acid Schiff [PAS], Congo red and Masson's trichrome) was employed to measure pathological changes in sections of lung tissue of experimental mice. Enzyme-linked immunosorbent assay (ELISA) was used to measure the concentrations of total and OVA-specific IgE, cytokines and chemokines (eotaxin-1, IL-13, IL-1β, TNF-α, IL-17A, IL-33) in the lung tissue homogenates. Immunoreactivity for vWF and α-SMA in lung tissue sections was detected by immunohistochemistry.
RESULTS
Exposure of the animals to CSE significantly reduced OVA-induced AHR, the number of eosinophils in bronchoalveolar lavage fluid (BALF) and eosinophils infiltrating into the lung tissue, as well as concentrations of some cytokines in lung homogenate. In contrast, it significantly enhanced the number of macrophages and M2 in BALF, as well as collagen deposition, smooth muscle thickness and alveolar destruction in lung tissue.
CONCLUSION
CSE inhibits OVA-induced AHR, changes inflammation 'phenotypes', while accelerates some aspects of airways remodelling, which might contribute to worse symptoms and be refractory to anti-inflammation therapies for asthmatics.
Topics: Humans; Animals; Mice; Ovalbumin; Cigarette Smoking; Mice, Inbred C57BL; Asthma; Lung; Inflammation; Cytokines; Bronchoalveolar Lavage Fluid; Phenotype; Disease Models, Animal; Mice, Inbred BALB C
PubMed: 37963721
DOI: 10.1111/crj.13718 -
Theranostics 2020: Nanomaterials capable of specifically interacting with proteins are very important for protein storage and vaccine delivery. Supramolecular hydrogels based on peptides...
: Nanomaterials capable of specifically interacting with proteins are very important for protein storage and vaccine delivery. Supramolecular hydrogels based on peptides have emerged as promising vaccine adjuvants because of their good compatibility, ease of antigen incorporation and display, and efficiency in activating immune responses. : We synthesized a self-assembling peptide (Fbp-GFFYK(γE)-NH, ) serving as a supramolecular protein chaperone for protein antigen delivery. The gelation was triggered by simply mixing and proteins. The vaccine adjuvant potential of was demonstrated by using two protein antigens, ovalbumin (OVA) and hepatitis B surface antigen (HBsAg). : The peptide derivative exhibited high protein binding capacity. Upon contacting proteins, rapidly formed coassembled nanofibers/hydrogels with the proteins, which greatly delayed the release of protein antigens. Our supramolecular protein chaperone significantly stimulated specific antibody titers by assisting protein delivery to antigen-presenting cells, promoting dendritic cell (DC) maturation, prolonging antigen accumulation and retention in the lymph nodes, and eliciting the secretion of cytokines. Most importantly, our supramolecular protein chaperone strongly stimulated the cellular immune response and significantly retarded tumor growth. : Our study demonstrated the great potential of the supramolecular protein chaperone in protein storage and delivery, vaccine production and tumor immunotherapy.
Topics: Adjuvants, Immunologic; Animals; Cancer Vaccines; Dendritic Cells; Disease Models, Animal; Female; Hydrogels; Immunity, Cellular; Melanoma, Experimental; Mice; Mice, Inbred BALB C; Nanoparticles; Ovalbumin
PubMed: 31903143
DOI: 10.7150/thno.39132 -
Food Research International (Ottawa,... Nov 2020The aim of this work was to examine the behavior of conjugated linoleic acid (CLA) delivery systems based on ovalbumin (OVA) and their derived nanoparticles (OVA and...
The aim of this work was to examine the behavior of conjugated linoleic acid (CLA) delivery systems based on ovalbumin (OVA) and their derived nanoparticles (OVA and OVA), under static in vitro gastrointestinal digestion model. In addition, potential cytotoxic effect of these inclusion complexes on a human colon cancer cell line (HT-29) was evaluated. OVA was resistant to gastric and intestinal digestion, while OVA nanoparticles were very susceptible to digestive enzymes hydrolysis. Particle size distribution (PDS) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for OVA evidenced the presence of a protein fragment of similar size after simulated digestive process. Conversely, for nanoparticles, partial and total hydrolysis in gastric and intestinal phases, respectively, was evidenced. After in vitro gastrointestinal digestion, released CLA (R) was assayed. In case of OVA, as CLA carrier, R was 37%, while for OVA nanoparticles, lower R values (~10-20%) were obtained. From cytotoxic assays, it was observed that CLA molecule was responsible for cell death, whereas OVA or their derived nanoparticles were not cytotoxic on HT-29 cells. On the other hand, flow cytometry analysis revealed that main death mechanism for CLA, and their inclusion complexes was apoptosis. OVA-CLA and OVA-CLA inclusion complexes displayed the highest potential cytotoxic activity and apoptotic index. Information derived from this work could be relevant for the design of CLA delivery systems as promising nanosupplements for production of new functional and excipient foods for both prevention and control of colon cancer.
Topics: Digestion; HT29 Cells; Humans; Linoleic Acids, Conjugated; Nanoparticles; Ovalbumin
PubMed: 33233083
DOI: 10.1016/j.foodres.2020.109381 -
Food Research International (Ottawa,... Mar 2023In this study, the functional properties of ovalbumin (OVA) were improved through dual modification with succinylation (succinylation degrees of 32.1 % [S1], 74.2 %...
In this study, the functional properties of ovalbumin (OVA) were improved through dual modification with succinylation (succinylation degrees of 32.1 % [S1], 74.2 % [S2], and 95.2 % [S3]) and ultrasonication (ultrasonication durations of 5 min [U1], 15 min [U2], and 25 min [U3]), and the changes in protein structures were explored. Results showed that as the succinylation degree was increased, the particle size and surface hydrophobicity of S-OVA decreased by the maximum values of 2.2 and 2.4 times, respectively, causing emulsibility and emulsifying stability to increase by 2.7 and 7.3 times, respectively. After ultrasonic treatment, the particle size of succinylated-ultrasonicated OVA (SU-OVA) had decreased by 3.0-5.1 times relative to that of S-OVA. Moreover, the net negative charge of S3U3-OVA had increased to the maximum value of - 35.6 mV. These changes contributed to the further enhancement in functional indicators. The unfolding of the protein structure and the conformational flexibility of SU-OVA were illustrated and compared with those of S-OVA via protein electrophoresis, circular dichroism spectroscopy, intrinsic fluorescence spectroscopy, and scanning electron microscopy. The dually modified OVA emulsion (S3U3-E) presented small droplets (243.33 nm), reduced viscosity, and weakened gelation behavior that were indicative of even distribution, which was visually proven by confocal laser scanning microscopy images. Furthermore, S3U3-E exhibited favorable stability, a particle size that was almost unchanged, and a low polydispersity index (<0.1) over 21 days of storage at 4 °C. The above results demonstrated that succinylation combined with ultrasonic treatment could be an effective dual modification method for enhancing the functional performance of OVA.
Topics: Ovalbumin; Microscopy, Confocal; Microscopy, Electron, Scanning; Particle Size; Spectrometry, Fluorescence
PubMed: 36869511
DOI: 10.1016/j.foodres.2023.112511 -
Biomacromolecules Mar 2023Ovalbumin (OVA)/sodium carboxymethylcellulose (CMC) colloidal particles were prepared with different compactness and morphologies by regulating the interaction between...
Ovalbumin (OVA)/sodium carboxymethylcellulose (CMC) colloidal particles were prepared with different compactness and morphologies by regulating the interaction between proteins and polysaccharides during heating. Electrostatic interactions between the amine groups of OVA (-NH) and carboxyl groups of CMC (-COO) enhanced complex formation. The protein conformation change benefited the hydrophobic interaction between the particles. Proteins in colloidal particles were unfolded/folded under thermal induction to form aggregates having more β-sheet structures. When the OVA/CMC ratio was 1:2, the initially loosely connected OVA/CMC aggregation changed into a uniform sphere between 25 and 90 °C. The mass ratio of OVA to CMC within the final colloidal particle (90 °C) was about 1:1.4. The OVA/CMC particle stability was maintained with hydrogen bonding, hydrophobicity, and disulfide bond. When OVA levels were predominant, OVA and CMC developed an approximately hollow sphere. Moreover, the final colloidal particle composition showed the OVA-to-CMC ratio as 3:1 (w/w). OVA bound into colloidal particle pores to increase compactness. Moreover, OVA and CMC bound to the colloidal particle while the particle shrank, thereby increasing the compactness of colloidal particles. There was a significant decrease in ABTS scavenging activity of curcumin compared with that of the particles with a ratio of 1:2. Thus, the rational adjustment of the structure of colloidal particles could effectively enhance their functional characteristics, providing a new way for the controlled release of the active ingredients.
Topics: Ovalbumin; Carboxymethylcellulose Sodium
PubMed: 36908256
DOI: 10.1021/acs.biomac.3c00063