scholarly journals Heat and Pressure Treatments on Almond Protein Stability and Change in Immunoreactivity after Simulated Human Digestion

Nutrients ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 1679 ◽  
Author(s):  
Elisabetta De Angelis ◽  
Simona Bavaro ◽  
Graziana Forte ◽  
Rosa Pilolli ◽  
Linda Monaci
Keyword(s):  
Protein Stability ◽  
Total Protein ◽  
Protein Pattern ◽  
Protein Quantification ◽  
Protein Profiling ◽  
Enzyme Linked Immunosorbent Assay ◽  
Total Protein Content ◽  
Bioinformatic Tools ◽  
Sds Page ◽  
Allergenic Proteins

Almond is consumed worldwide and renowned as a valuable healthy food. Despite this, it is also a potent source of allergenic proteins that can trigger several mild to life-threatening immunoreactions. Food processing proved to alter biochemical characteristics of proteins, thus affecting the respective allergenicity. In this paper, we investigated the effect of autoclaving, preceded or not by a hydration step, on the biochemical and immunological properties of almond proteins. Any variation in the stability and immunoreactivity of almond proteins extracted from the treated materials were evaluated by total protein quantification, Enzyme Linked Immunosorbent Assay (ELISA), and protein profiling by electrophoresis-based separation (SDS-PAGE). The sole autoclaving applied was found to weakly affect almond protein stability, despite what was observed when hydration preceded autoclaving, which resulted in a loss of approximately 70% of total protein content compared to untreated samples, and a remarkable reduction of the final immunoreactivity. The final SDS-PAGE protein pattern recorded for hydrated and autoclaved almonds disclosed significant changes. In addition, the same samples were further submitted to human-simulated gastro-intestinal (GI) digestion to evaluate potential changes induced by these processing methods on allergen digestibility. Digestion products were identified by High Pressure Liquid Chromatography-High Resolution Tandem Mass Spectrometry (HPLC-HRMS/MS) analysis followed by software-based data mining, and complementary information was provided by analyzing the proteolytic fragments lower than 6 kDa in size. The autoclave-based treatment was found not to alter the allergen digestibility, whereas an increased susceptibility to proteolytic action of digestive enzymes was observed in almonds subjected to autoclaving of prehydrated almond kernels. Finally, the residual immunoreactivity of the GI-resistant peptides was in-silico investigated by bioinformatic tools. Results obtained confirm that by adopting both approaches, no epitopes associated with known allergens survived, thus demonstrating the potential effectiveness of these treatments to reduce almond allergenicity.

scholarly journals Investigation of Heat and Pressure Treatments on Almond Protein Stability and Immunoreactivity after Simulated Human Digestion

2018 ◽  
Author(s):  
Elisabetta De Angelis ◽  
Simona L. Bavaro ◽  
Graziana Forte ◽  
Rosa Pilolli ◽  
Linda Monaci
Keyword(s):  
Total Protein ◽  
Protein Pattern ◽  
Protein Quantification ◽  
Protein Profiling ◽  
Total Protein Content ◽  
Bioinformatic Tools ◽  
Sds Page ◽  
Allergenic Proteins ◽  
The Stability

Almond is worldwide consumed and renowned as a valuable healthy food. In spite of this, it is also a potent source of allergenic proteins able to trigger several mild to life-threatening immunoreactions. Food processing proved to alter biochemical characteristics of proteins, thus affecting the respective allergenicity. In this paper we investigated the effect of autoclaving, preceded or not by a hydration step, on the biochemical and immunological properties of almond proteins. Any variation in the stability and immunoreactivity of almond proteins extracted from the treated materials, were evaluated by total protein quantification, ELISA assay and protein profiling by electrophoresis-based separation (SDS-PAGE). The autoclaving alone was found to weakly affect almond proteins stability, despite what observed for the combination of hydration and autoclaving, which resulted in a loss of approximately 70% of total protein content compared to untreated sample, and in a final negligible immunoreactivity, as well. The final SDS-PAGE protein pattern recorded for almonds hydrated and autoclaved disclosed significant changes. In addition, the same samples were further submitted to in vitro simulated gastro-duodenal (GI) digestion to evaluate potential changes induced by these processing on allergens digestibility. Digestion products were identified by HPLC-HRMS/MS analysis followed by software-based data mining, and complementary information were provided by analyzing the proteolytic fragments lower that 6 kDa in size. The autoclave based treatment was found not to alter the allergens digestibility, whereas an increased susceptibility to proteolytic action of digestive enzymes was observed in almonds subjected to the combination of prehydration and autoclaving. Finally, the residual immunoreactivity of the GI resistant peptides was investigated in-silico by bioinformatic tools, confirming that by following both approaches, no epitopes survived the almond digestion, thus demonstrating the potential effectiveness of these treatments to reduce almond allergenicity.

scholarly journals Naringenin Regulates Lipopolysaccharide-Induced Abnormal Airway Surface Liquid Secretion

2021 ◽  
Vol 16 (9) ◽  
pp. 1934578X2110403
Author(s):  
Rui Shi ◽  
Min-Yi Guan ◽  
Wei-Yang Fan ◽  
Pei-Bo Li ◽  
Zhong-Yi Yang ◽  
...  
Keyword(s):  
Total Protein ◽  
Cultured Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Total Protein Content ◽  
Airway Surface Liquid ◽  
Airway Epithelial ◽  
Aquaporin 5 ◽  
Factors Affecting ◽  
Aqp5 Expression ◽  
Surface Liquid

Airway surface liquid (ASL) is one of the key factors affecting the respiratory system's physiological function. Abnormal ASL secretion can increase the incidence of various respiratory diseases. Lipopolysaccharide (LPS) stimulation can damage the airway epithelial barrier, affect the concentration of ASL contents, and down-regulate ion channel expression, which in turn causes abnormal ASL secretion. Naringenin, which exists in many Citrus foods, has the ability to promote airway surface liquid secretion. This work is designed to investigate the regulatory mechanism of naringenin on LPS-induced abnormal ASL secretion. The effects of naringenin and LPS on the viability of Calu-3 cells were measured by CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS). ASL secretion volume was measured by a micropipette on air–liquid interface cultured cells. The concentration of Cl−, Na+, lysozyme, and total protein in ASL were respectively measured by assay kits. The mRNA expressions were determined by quantitative real-time polymerase chain reaction, and proteins were measured by enzyme-linked immunosorbent assay. The results indicated that LPS could affect ASL secretion and regulate cystic fibrosis transmembrane conductance regulator (CFTR), aquaporin 1 (AQP1) and aquaporin 5 (AQP5) expression. Naringenin had the ability to regulate the ASL secretion by increasing secretion volume, and Cl− and Na+ concentrations, reducing lysozyme and total protein content, and regulating CFTR, AQP1, and AQP5 expression. This study indicated that naringenin had regulating effects to attenuate LPS-induced abnormal ASL secretion.

Further Observations on in vivo Radioprotection of Rats by Selenourea

1974 ◽  
Vol 29 (9-10) ◽  
pp. 647-649
Author(s):  
Roberto Badiello ◽  
Enrico Gattavecchia ◽  
Mario Mattii ◽  
Maurizio Tamba
Keyword(s):  
Ionizing Radiation ◽  
Gamma Irradiation ◽  
Protein Content ◽  
Total Protein ◽  
Protein Pattern ◽  
Serum Enzymes ◽  
Total Protein Content

Abstract This paper deals with a study of some tests in rats in vivo after gamma irradiation in the presence and in the ab­ sence of selenourea. The factors considered were the total protein content, the protein pattern, and some serum enzymes, like GOT, GPT and AP. The result shows that the preadministration of selenourea modifies fovourably the changes induced by ionizing radiation

Seed protein content and composition of near-isogenic and induced mutant pea lines

1993 ◽  
Vol 3 (3) ◽  
pp. 187-194 ◽  
Author(s):  
M. D. Perez ◽  
S. J. Chambers ◽  
J. R. Bacon ◽  
N. Lambert ◽  
C. L. Hedley ◽  
...  
Keyword(s):  
Differential Scanning Calorimetry ◽  
Protein Content ◽  
Total Protein ◽  
Dry Weight ◽  
Total Protein Content ◽  
Scanning Calorimetry ◽  
Sds Page ◽  
A Chain ◽  
Total Seed ◽  
Mutant Pea

AbstractTotal protein content and amounts of albumin, legumin and vicilin have been determined for pea seeds from lines near-isogenic except for genes at the rugosus loci, r and rb (RR/RbRb; rr/RbRb; RR/rbrb; rr/rbrb). Seeds with the wildtype, round-seeded phenotype (RR/RbRb) had less protein on a total seed dry-weight basis than any of the wrinkled-seeded lines and this protein had a lower proportion of albumin. The lines which had recessive alleles at both r and rb loci had the highest proportion of protein and the highest proportion of albumin. The roundseeded peas possessed nearly two-fold more legumin than the double recessive line, with proportions for the two single recessive lines falling in between these extremes. Vicilin levels were similarfor all four near-isogenic lines. SDS-PAGE analysis of the isolated albumin, legumin and vicilin fractions revealed no significant differences between the four lines. Differential scanning calorimetry of protein extracts showed that all the wrinkled-seeded near-isolines possessed legumin fractions with diminished thermal stability relative to that from the roundseeded, wild-type line.Chemically-induced mutants were also analysed for protein content and composition. These mutants have previously been shown to display great variation in starch and lipid levels. Total protein varied from 20.3% to 37.9%; however, relative proportions of albumin, legumin and vicilin were similar in all mutant lines. SDS-PAGE analysis identified two mutant pea lines which possessed a legumin A-chain of 65 000 Mr as well as the typical 45 000 Mr form. Differential scanning calorimetry of protein extracts indicated that the legumin in all mutants had lower enthalpies of denaturation than the legumin in the round-seeded parent.The mutant pea lines possess exceptional variation with respect to starch, lipid and protein which raises opportunities for their use in the food and animal feedstuff industries.

scholarly journals Evaluation of Non-climacteric Senescence of Gladiolus sp. Flowers

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 760A-760
Author(s):  
G.H. Pemberton ◽  
Terril A. Nell ◽  
James E. Barrett
Keyword(s):  
Total Protein ◽  
Acc Synthase ◽  
Total Protein Content ◽  
Protein Profiles ◽  
Flower Longevity ◽  
Sds Page ◽  
Specific Proteins ◽  
Postharvest Life ◽  
Relationship Of ◽  
The Relationship

Senescence of gladiolus flowers, like many geophytes, does not involve a climacteric burst of ethylene. Eleven gladiolus cultivars were screened and all were non-climacteric (NC) for both respiration and ethylene production. Average ethylene levels for individual flowers were 0.5 μl C2H4/kg per h or less. As in other NC flowers, protein synthesis may be linked to senescence. Our goal was to identify specific proteins that were involved in the senescence process that could be used as indicators of postharvest longevity. SDS-PAGE protein profiles of cut gladiolus flowers were determined from a tight bud stage to senescence. Both increases and decreases were observed in major polypeptides that may be connected to postharvest flower longevity. Total protein content of gladiolus flower petals decreased by ≈70% during the profile period. This could explain the relatively short postharvest life of 3 to 5 days for individual gladiolus flowers. Total protein profiles were probed with an ACC synthase antibody to establish the relationship of this enzyme in NC senescence.

Assessment of total protein and soy protein contents in some heated meat products in Tehran, Iran

2020 ◽  
Author(s):  
Masoomeh Fekri ◽  
Gholamreza Jahed Khaniki ◽  
Mohaddeseh Pirhadi ◽  
Mahdieh Abbassi
Keyword(s):  
Protein Content ◽  
Soy Protein ◽  
Total Protein ◽  
Meat Products ◽  
Soya Protein ◽  
National Standard ◽  
Enzyme Linked Immunosorbent Assay ◽  
Total Protein Content ◽  
Food Stores ◽  
Standard Limit

Heated meat products are emulsion which have various nutrition materials such as meat as an animal protein and soya as a plant protein. The nutritional value of meat proteins is very high than soya protein but the meat is more expensive than soya which the producers substitute the meat with soya. This study was assessed the soy protein and the total protein contents in some heated meat products collected from food stores in Tehran, Iran. Twenty samples of heated meat products with 40%, 55% and 70% of meat were randomly collected from food stores. The heated meat products samples were transferred to the food analysis lab. The total protein was determined by the macro Kjeldahl method after sample preparation and homogenization. Also, the soy protein content in samples was measured by using the enzyme-linked immunosorbent assay (ELISA) method. Results showed that 4 samples of heated meat products had less total protein content than the standard limit and 16 samples were in accordance with Iranian national standard. Soy protein content in 3 from 8 samples of heated meat products with 40% meat was higher than the standard limit and the others placed in the standard limit (approximate with 4% soy protein). Also, Soy protein content in 6 from 8 samples of heated meat products with 55% meat was higher than the standard limit and only 2 samples were in accordance with the standard limit. All samples of heated meat products with 70% meat set in a standard limit. It was concluded that some heated meat products do not correspond with the Iranian national standard range. The food quality control lab requires doing attention and sensation for correct formulation according to national standard measures.

scholarly journals Proximate analysis and SDS-PAGE protein profiling of cassava leaves: utilization as leafy vegetable in Nigeria

2019 ◽  
Vol 4 (1) ◽  
Author(s):  
Jacob O Popoola
Keyword(s):  
Protein Content ◽  
Proximate Analysis ◽  
Leafy Vegetable ◽  
Protein Profiling ◽  
Leafy Vegetables ◽  
Total Protein Content ◽  
International Institute ◽  
Cassava Leaves ◽  
Sds Page ◽  
Major Cluster

Introduction: Cassava (Manihot esculenta Crantz) is an important starchy food crop whose leaf is yet to be adopted and utilized as leafy vegetable and as potential protein source for monogastric animals in Nigeria. Providing more nutritional information and highlighting health benefits could encourage consumption, hence this study. Methodology: Twelve varieties of cassava were collected from farmers and Genetic Resources Centre of the International Institute of Tropical Agriculture (IITA), Ibadan, Oyo State, Nigeria. Total protein content was estimated in supernatant using BSA as standard. Moisture, dry matter, protein, crude fat, ash, sodium and potassium contents were determined using standard protocols. Results: The proximate analysis show that crude protein percentage ranged from 11.81 in variety OsNo001 to 22.75 in variety IBA30572. Moisture content varied from 8.09 in OsNo001 to 10.90 in IBA011371 while ash content varied from 6.88 in IBA010040 to 9.64 in variety 1089A. Four varieties IBA950289, IBA980505, TMEB91934 and 1089A have richer proximate content in ash, protein, carbohydrate, and fat contents. The result of SDS-PAGE indicates the presence of proteins in all the cassava leaves analyzed. The SDS-PAGE cluster analysis generated four major cluster groups suggesting the presence of four types of protein. Conclusion: The cassava leaves are comparable to known leafy vegetables Amaranthus in nutritional and biochemical contents. It is evident that the protein content might be sufficient enough to adopt cassava leaves as a leafy vegetable based on the SDS-PAGE. Thus, the cassava varieties are good candidates for utilization as leafy vegetable to augment the exotic and known leafy vegetables in Nigeria.

scholarly journals Symbiotic Relationship of Thiothrix spp. with an Echinoderm

1998 ◽  
Vol 64 (9) ◽  
pp. 3491-3495 ◽  
Author(s):  
Robin L. Brigmon ◽  
Chantal De Ridder
Keyword(s):  
Protein Content ◽  
Immunosorbent Assay ◽  
Microscopic Examination ◽  
Total Protein ◽  
Marine Invertebrates ◽  
Enzyme Linked Immunosorbent Assay ◽  
Total Protein Content ◽  
Symbiotic Relationship ◽  
Echinocardium Cordatum ◽  
Relationship Of

ABSTRACT Immunoassay procedures were used to investigate the symbiotic relationship of Thiothrix spp. in the intestinal cecum of the spatangoid species Echinocardium cordatum. Thiothrixspp. were identified in nodule samples from E. cordatum digestive tubes based on microscopic examination, enzyme-linked immunosorbent assay, and indirect immunofluorescence.Thiothrix spp. protein made up as much as 84% of the total protein content of the nodules. This is the first identification ofThiothrix spp. internally symbiotic with marine invertebrates.

scholarly journals Saliva proteomics updates in biomedicine

2019 ◽  
Vol 26 (1) ◽  
Author(s):  
Katerina R. Katsani ◽  
Dimitra Sakellari
Keyword(s):  
Human Physiology ◽  
Precision Medicine ◽  
High Throughput ◽  
Total Protein ◽  
High Efficiency ◽  
Biological Fluids ◽  
Total Protein Content ◽  
Biomedical Sciences ◽  
Biological Specimen ◽  
Bioinformatic Tools

AbstractIn the years of personalized (or precision) medicine the ‘omics’ methodologies in biomedical sciences—genomics, transcriptomics, proteomics and metabolomics—are helping researchers to detect quantifiable biological characteristics, or biomarkers, that will best define the human physiology and pathologies. Proteomics use high throughput and high efficiency approaches with the support of bioinformatic tools in order to identify and quantify the total protein content of cells, tissues or biological fluids. Saliva receives a lot of attention as a rich biological specimen that offers a number of practical and physiological advantages over blood and other biological fluids in monitoring human health. The aim of this review is to present the latest advances in saliva proteomics for biomedicine.

Circulating exosomes from patients with Graves’ disease induce an inflammatory immune response

Endocrinology ◽  
2020 ◽  
Author(s):  
Xuejiao Cui ◽  
Mingshi Huang ◽  
Shiwei Wang ◽  
Na Zhao ◽  
Ting Huang ◽  
...  
Keyword(s):  
Total Protein ◽  
Mononuclear Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Total Protein Content ◽  
Graves Disease ◽  
Peripheral Blood Mononuclear ◽  
Hsp60 Expression ◽  
Myeloid Differentiation Factor ◽  
Serum Exosomes ◽  
Cd63 Expression

Abstract Exosomes are extracellular vesicles that can participate in autoimmune diseases. The purpose of this study was to explore whether circulating exosomes are involved in Graves’ disease (GD) pathogenesis. In this study, serum exosomes were extracted from 26 healthy controls (HC-EXO), 26 GD patients (GD-EXO), and 7 Graves’ ophthalmopathy patients (GO-EXO). For each group, the total protein content was detected, and thyrotropin receptor (TSHR), insulin-like growth factor 1 receptor (IGF-1R), HSP60, and CD63 expression were analyzed by Western blotting (WB). Healthy volunteer-derived peripheral blood mononuclear cells (PBMCs) and HC-EXO or GD-EXO were cocultured for 24 h, and immunofluorescence was used to observe the locations of the exosomes and Toll-like receptor (TLR) 2/3. CD11c+TLR2+ and CD11c+TLR3+ cell percentages were determined by flow cytometry. Myeloid differentiation factor 88 (MyD88), Toll/IL-1 receptor domain-containing adaptor inducing interferon-β (TRIF) and p-P65 expression were analyzed by WB. IL-6 and IL-1β supernatant levels were detected using enzyme-linked immunosorbent assay (ELISA). The results showed that the total protein concentration was similar among GD-EXO, GO-EXO and HC-EXO. IGF-1R and HSP60 expression was significantly higher in GD-EXO and GO-EXO than in HC-EXO. After coculturing PBMCs with GD-EXO or HC-EXO for 24 h, GD-EXO could bind to TLR2/3. GD-EXO significantly increased CD11c+TLR2+ and CD11c+TLR3+ cell percentages; MyD88, TRIF, and p-P65 protein expression; and IL-6 and IL-1β levels. In conclusion, we first demonstrated that GD-EXO and GO-EXO highly expressed IGF-1R and HSP60. GD-EXO may induce an inflammatory response through the TLR/NF-κB signaling pathway and be involved in the pathogenesis of GD.

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