Low Allergenicity Research Articles

Enhancement of extraction efficiency and functional properties of chickpea protein isolate using pulsed electric field combined with ultrasound treatment

Chickpea protein isolate (CPI) is a promising dietary protein with the advantages of low allergenicity, easy digestion and balanced composition of essential amino acids. However, due to the thick skin of chickpeas, the extraction of CPI is challenging, resulting in lower efficiency of the alkaline extraction-isoelectric precipitation (AE-IEP) method. Therefore, the present study investigated the effect of pulsed electric field combined with ultrasound (PEF-US) treatment on the extraction efficiency of CPI and the functional properties was characterized. Parameter optimization was carried out using response surface methodology (RSM), with the following optimized conditions: pulse duration of 87 s, electric field intensity of 0.9 kV/cm, ultrasonic time of 15 min, and ultrasonic power of 325 W. Under the optimized conditions, the yield of CPI after combined (PEF-US) treatment was 13.52 ± 0.13 %, which was a 47.28 % improvement over the AE-IEP method. This yield was better than that obtained with either individual PEF or US treatment. Additionally, the functional properties (solubility, emulsification, and foaming) of CPI were significantly enhanced compared to AE-IEP. However, the stability of emulsification and foaming did not show significant differences among the four methods. The PEF-US method efficiently extracts CPI with excellent functional properties, enabling the production of proteins as desired functional additives in the food industry.

A comprehensive review on composition to application of pea protein and its components

Pea protein, a valuable plant-based protein source, is notable for its nutritional value, essential amino acids, and low allergenicity, making it widely applicable in food, medicine, and materials. It consists mainly of globulin and albumin, which influence its functional properties and applications. However, there is a lack of comprehensive reviews on its extraction methods, functional properties, modification techniques, and applications in food. This paper aims to fill these gaps by detailing pea protein composition, extraction methods, functional properties, and modification impacts while summarizing its food applications and proposing future research directions. The goal is to enhance pea protein's functionality and expand its applications through optimized extraction and advanced technology. By improving extraction techniques and adapting pea protein for better functionality, we aim to develop high-quality market applications, ensuring the growth and sustainability of the pea protein industry globally. This approach promises a flourishing future for pea protein, meeting global competition demands and driving industry advancement.

Effect of enzymatic hydrolysis and ultrasound pretreatment on allergenicity, functional properties and bioactivity of whey protein isolates

Whey protein isolate (WPI) is an important food ingredient and a major allergen in food. Enzymatic hydrolysis is an important method for WPI modification, but there is limited information on the effect of ultrasound pretreatment on WPI hydrolysates (WPH). The aim of this study was to analyze the effect of enzymatic hydrolysis and ultrasound pretreatment on the allergenicity, functional properties and bioactivity of WPH by multispectroscopy and peptidomics combined with bioinformatics. The results showed that ultrasound pretreatment contributed to the production of WPH with low allergenicity (IgE binding capacity is 55.59%) and good functional properties (emulsifying activity index is 27.02%, foaming characteristic is 50.00%) and bioactivity (total antioxidant capacity is 65.94%). This was attributed to the synergistic hydrolysis of WPI by endopeptidases and exopeptidases, and ultrasound pretreatment that altered the accessibility of WPI to proteases and the peptide profiles and main proteins of WPH, affecting the changes in their spatial and primary structure. This research will advance the application of WPI in hypoallergenic foods.

Effects of four kinds of improver on fermentation characteristics of gluten-free rice dough.

Rice is commonly utilized as a wheat bread substitute due to its low allergenicity. However, rice bread faces challenges in processing efficiency and the formation of a cohesive gel network structure, resulting in suboptimal taste Hence, this study compared four improvers-trypsin, whey protein (WPC), hydroxypropyl methyl cellulose (HPMC), and molecularly distilled monoglycerides (GMSs). The impacts of the four improvers on the processing attributes of rice dough were comprehensively assessed across fermentation, moisture content analysis, rheology, heat stability, and pasting characteristics. The findings indicated that the incorporation of trypsin, HPMC, and WPC resulted in 107%, 61%, and 1% increases in gas production of fermented rice dough, respectively, while reducing the regrowth values to 564.00±7.21, 176.67±0.58, and 611.00±3.61cP. Notably, the air-holding capacity of HPMC-fermented rice dough exhibited a 7% enhancement. All four types of improvers raised the enthalpy of melting (ΔH) and the difference in melting point (ΔT) of fermented rice doughs, with trypsin enhancing ΔH by 44% and ΔT by 40%. GMS, HPMC, and WPC increased the degree of water incorporation in fermented doughs. This study could serve as a benchmark for enhancing the fermentation attributes of rice dough and establish a groundwork for the future advancement of gluten-free dietary options. PRACTICAL APPLICATION: The thorough analysis conducted in this experiment provides a theoretical framework for rice dough preparation during the fermentation process, addressing the dietary needs of individuals with coeliac disease and those following a gluten-free diet. This study also paves the way for the development of improved gluten-free rice products in future research pursuits.

Anti-Inflammatory, Cytotoxic, and Genotoxic Effects of Soybean Oligopeptides Conjugated with Mannose.

Soy protein is considered to be a high-quality protein with a range of important biological functions. However, the applications of soy protein are limited due to its poor solubility and high level of allergenicity. Its peptides have been of interest because they exert the same biological functions as soy protein, but are easier to absorb, more stable and soluble, and have a lower allergenicity. Moreover, recent research found that an attachment of chemical moieties to peptides could improve their properties including their biodistribution, pharmacokinetic, and biological activities with lower toxicity. This study therefore aimed to acquire scientific evidence to support the further application and safe use of the soybean oligopeptide (OT) conjugated with allulose (OT-AL) or D-mannose (OT-Man). The anti-inflammation, cytotoxicity, and genotoxicity of OT, OT-AL, and OT-Man were investigated. The results showed that OT, AL, Man, OT-AL, and OT-Man at doses of up to 1000 µg/mL were not toxic to HepG2 (liver cancer cells), HEK293 (kidney cells), LX-2 (hepatic stellate cells), and pre- and mature-3T3-L1 (fibroblasts and adipocytes, respectively), while slightly delaying the proliferation of RAW 264.7 cells (macrophages) at high doses. In addition, the oligopeptides at up to 800 µg/mL were not toxic to isolated human peripheral blood mononuclear cells (PBMCs) and did not induce hemolysis in human red blood cells (RBCs). OT-Man (200 and 400 µg/mL), but not OT, AL, Man, and OT-AL, significantly reduced the production of NO and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX2) stimulated by lipopolysaccharide (LPS) in RAW 264.7 cells, suggesting that the mannose conjugation of soy peptide had an inhibitory effect against LPS-stimulated inflammation. In addition, the secretion of interleukin-6 (IL-6) stimulated by LPS was significantly reduced by OT-AL (200 and 400 µg/mL) and OT-Man (400 µg/mL). The tumor necrosis factor-α (TNF-α) level was significantly decreased by OT (400 µg/mL), AL (400 µg/mL), OT-AL (200 µg/mL), and OT-Man (200 and 400 µg/mL) in the LPS-stimulated cells. The conjugation of the peptides with either AL or Man is likely to be enhance the anti-inflammation ability to inhibit the secretion of cytokines. As OT-Man exhibited a high potential to inhibit LPS-induced inflammation in macrophages, its mutagenicity ability was then assessed in bacteria and Drosophila. These findings showed that OT-Man did not trigger DNA mutations and was genome-safe. This study provides possible insights into the health advantages and safe use of conjugated soybean peptides.

Allergenicity assessment of β-lactoglobulin ferulic acid–glucose conjugates

We prepared the β-lactoglobulin (BLG)–ferulic acid (FA)–glucose (Glu) conjugates by alkaline method and Maillard reaction to assess the allergenicity. FA and Glu can form a ternary covalent conjugate with BLG, as evidenced by the shortening of SEC retention time, upward migration of SDS-PAGE protein bands, considerable decrease in free amino and sulfhydryl content, and changes in multistructure. BLG–Glu–FA conjugates weakly bound to immunoglobulin E in allergic sera was weak, reduced interleukin 4 and tumor necrosis factor α levels in RBL-2H3 cells and histamin and interleukin 6 secretion levels in KU812 cells, and inhibited the nuclear factor-κB signaling pathway. In vivo experiments showed that the conjugates regulated T-cell homeostasis in mouse splenic and mesenteric lymphocytes and attenuated splenic and duodenal immune injury. Therefore, the conjugates of BLG with FA combined with Glu altered the epitope structure and exhibited low allergenicity.

Goat Milk Allergy and a Potential Role for Goat Milk in Cow's Milk Allergy.

In many parts of the world, goat milk has been part of the human diet for millennia. Allergy to goat's milk, not associated with allergy to cow's milk, is a rare disorder, although some cases have been described. Goat milk proteins have substantial homology with cow's milk proteins and even show cross-reactivity; therefore, they are not advised as an alternative to cow's milk for infants with IgE-mediated cow's milk allergies. However, there are indications that, due to the composition of the goat milk proteins, goat milk proteins show lower allergenicity than cow's milk due to a lower αS1-casein content. For this reason, goat milk might be a better choice over cow's milk as a first source of protein when breastfeeding is not possible or after the breastfeeding period. Additionally, some studies show that goat milk could play a role in specific types of non-IgE-mediated cow milk allergy or even in the prevention of sensitization to cow's milk proteins. This review discusses a possible role of goat milk in non-IgE mediated allergy and the prevention or oral tolerance induction of milk allergy.

Virus-like Particles as Vaccines for Allergen-Specific Therapy: An Overview of Current Developments.

Immune engineering and modulation are the basis of a novel but powerful tool to treat immune diseases using virus-like particles (VLPs). VLPs are formed by the viral capsid without genetic material making them non-infective. However, they offer a wide variety of possibilities as antigen-presenting platforms, resulting in high immunogenicity and high efficacy in immune modulation, with low allergenicity. Both animal and plant viruses are being studied for use in the treatment of food allergies. These formulations are combined with adjuvants, T-stimulatory epitopes, TLR ligands, and other immune modulators to modulate or enhance the immune response toward the presented allergen. Here, the authors present an overview of VLP production systems, their immune modulation capabilities, and the applicability of actual VLP-based formulations targeting allergic diseases.

Hierarchical biocatalytic membranes embedded with trypsin–inorganic hybrid nanoflowers for effective β-lactoglobulin hydrolysis

β-lactoglobulin (β-LG) is a significant allergen in milk. Enzymatic hydrolysis of β-LG produces peptides with low allergenicity and improved functionality. The biocatalytic membranes combine enzyme function and membrane separation in one step, but is still challenging for proteolysis. Herein, the biocatalytic membranes embedded with trypsin-inorganic hybrid nanoflowers are developed by a facile and green method for β-LG hydrolysis. Ethylene vinyl alcohol copolymer (EVAL) nanofibers are fabricated as 3D scaffolding on the membrane surface and trypsin-copper phosphate nanoflowers are in-situ grown in the porous structure of membranes. The nanoflowers are uniformly entrapped in the EVAL nanofiber layer and preferentially grow in the EVAL nanofibers layer, leading to the hierarchical structure of membranes. The trypsin loading of the nanoflowers membrane is approximate to 282.1 µg/cm2. Lower Michaelis-Menten constants Km prove the better affinity of Cu3(PO4)2-TR nanoflower membrane with substrates during filtration than that in static mode, even than free trypsin. The hierarchical structure of the membranes ensures both the enhanced trypsin-substrate contacts and low mass transfer resistance during the β-LG filtration. This work not only presents a novel concept of enzyme immobilized biocatalytic membranes, but also offers a sustainable solution for high-performance proteolysis.

Generation of New β-Conglycinin-Deficient Soybean Lines by Editing the lincRNA lincCG1 Using the CRISPR/Cas9 System.

Soybean β-conglycinin is a major allergen that adversely affects the nutritional properties of soybean. Soybean deficient in β-conglycinin is associated with low allergenicity and high nutritional value. Long intergenic noncoding RNAs (lincRNAs) regulate gene expression and are considered important regulators of essential biological processes. Despite increasing knowledge of the functions of lincRNAs, relatively little is known about the effects of lincRNAs on the accumulation of soybean β-conglycinin. The current study presents the identification of a lincRNA lincCG1 that was mapped to the intergenic noncoding region of the β-conglycinin α-subunit locus. The full-length lincCG1 sequence was cloned and found to regulate the expression of soybean seed storage protein (SSP) genes via both cis- and trans-acting regulatory mechanisms. Loss-of-function lincCG1 mutations generated using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system led to the deficiency of the allergenic α'-, α-, and β-subunits of soybean β-conglycinin as well as higher content of proteins, sulfur-containing amino acids, and free arginine. The dominant null allele LincCG1, and consequently, the β-conglycinin-deficient phenotype associated with the lincCG1-gene-edited line was stably inherited by the progenies in a Mendelian fashion. The dominant null allele LincCG1 may therefore be exploited for engineering/developing novel hypoallergenic soybean varieties. Furthermore, Cas9-free and β-conglycinin-deficient homozygous mutant lines were obtained in the T1 generation. This study is the first to employ the CRISPR/Cas9 technology for editing a lincRNA gene associated with the soybean allergenic protein β-conglycinin. Moreover, this study reveals that lincCG1 plays a crucial role in regulating the expression of the β-conglycinin subunit gene cluster, besides highlighting the efficiency of employing the CRISPR/Cas9 system for modulating lincRNAs, and thereby regulating soybean seed components.

Development of a low allergenic product for patients with milk allergy and assessment of its specific IgE reactivity.

Milk oral immunotherapy is the riskiest and most unpredictable form of oral immunotherapy. We aimed to produce a low allergenic product than conventional once baked-cake/muffin, to develop indirect in-house ELISA to check the tolerance status with milk products and evaluate IgE reactivity of patients' sera via western blotting (WB) and indirect in-house ELISA. A low allergenic product named biscotti-twice baked-cake was developed, and the total protein concentration was determined. The protein content was studied by SDS-PAGE and proteomics. Milk-specific IgE (sIgE) binding assays were performed by WB and indirect in-house ELISA by using patients' sera. Casein band intensity was observed to be lower in the biscotti-twice baked-cake than in the once baked-cake (p = .014). Proteomics analysis and αS1-casein measurement showed that the lowest intensity of casein was found in biscotti. The low binding capacity of milk sIgE to biscotti compared with once baked-cake was shown by WB (p = .0012) and by indirect in-house ELISA (p = .0001). In the ROC analysis, the area under the curve (AUC) of the in-house ELISA IgE was comparable with Uni-CAP milk and casein sIgE. The AUC of the in-house ELISA IgE for cake (0.96) and biscotti (1) was slightly better than Uni-CAP milk sIgE (0.94; 0.97) and casein sIgE (0.96; 0.97), respectively. The low allergenicity of the newly developed low allergenic product "biscotti-twice baked-cake" has been demonstrated by invitro experiments. Biscotti could be a safe treatment option than once baked-cake/muffin in patients who are reactive to once baked-milk products.

Assessment of the Allergenic Potential of Urban Woody Flora of Rostov-on-Don

Introduction. Plant pollen causes various allergic reactions in humans, including respiratory diseases, immune system disorders, bronchitis, conjunctivitis, dermatitis, and hay fever. These diseases affect up to 30% of the world's population. In large cities, trees and shrubs used in landscaping are significant sources of allergenic dust. Despite this, the greening of cities worldwide often occurs without considering the allergenic properties of plants. With the development of proteomics, it has become possible to assess the degree of allergenicity of proteins that make up plant pollen in detail. Based on this information, a scale of potential allergenicity for woody plants has been developed. The aim of this study is to assess the allergenic potential of woody plants in the urban flora of Rostov-on-Don.Materials and Methods. The object of the study was trees and shrubs used in the landscaping of Rostov-on-Don. The analysis of floristic data was based on the materials obtained during field work in 2023 on the territory of Rostov-on-Don. The author also used the lists of the city's dendroflora compiled between 2007 and 2022. The assessment of the potential allergenicity of woody plant species was conducted on a five-point scale, with 0 indicating plants that did not pose an allergic hazard, 1 indicating a low allergenicity level, 2 indicating a medium class, 3 indicating a high level, and 4 indicating a very high level of allergenicity.Results. In the flora of woody plants in Rostov-on-Don, 61 species of potentially allergenic plants were identified, posing varying levels of danger to human health. The share of all types of potentially allergenic woody plants was 30% of the total number of urban woody flora species in the city. The most powerful sources of allergenic pollen included nine species (Fraxinus Excelsior, Betula Pendula, B. Verrucosa, Platycladus Orientalis, etc.), which posed the greatest threat of hay fever and other allergic reactions. As a rule, these were typically wind-pollinated plants that produced maximum amounts of pollen. The list of potentially allergenic species included a significant number of adventitious species (24 species), which made it difficult to control their spread. A taxonomic analysis of potentially allergenic species was carried out at the order level, for which specific protein reactions were identified and detailed approaches to the prevention and treatment of hay fever were developed. The orders Pinales and Fagales form the bulk of allergenic pollen in the winter-spring period.Discussion and Conclusion. For the first time, studies were conducted on the allergenic activity of urban woody flora in the southern regions of Russia. An assessment of their allergenic potential allowed us to determine the level of threat of allergic reactions in humans. The greatest danger comes from both allergenic and invasive species that can spread actively and increase in numbers. Representatives of the Pinales and Fagales orders have proven to be significant sources of allergenic pollen, as they often have high ornamental qualities and play a prominent role in design projects. In some cases, it may be possible to replace these cultures with less allergenic alternatives, such as representatives of the Rosales order, to reduce the risk of allergic reactions.

Analysis of proteomics and in silico allergenicity prediction of soluble proteins in selenium-enriched peanut leaves

Selenium (Se) is one of the nutrients contained in plants and an essential trace element for human growth. Peanut (Arachis hypogaea L.) is a good material for dietary Se enrichment. To investigate the effects of Se on the soluble proteins from peanut leaves, a comparative proteomics study was performed on soluble peanut leaf proteins (SPLPs), which were extracted from Se-enriched peanut leaves with different Se contents. Then, the allergenicity of the SPLPs was analyzed using in silico approaches. Bioinformatics analysis revealed that low Se treatment activated the antioxidative system and enhanced photosynthesis, glutathione metabolism and primary metabolism. Furthermore, the Bet v 1 domain-containing protein was affected the most by Se enrichment and can be classified as an allergen by docking simulation. ELISA confirmed that the allergenicity of SPLPs was significantly lower than that of the peanut protein, and this lower allergenicity could give the SPLPs a wider range of applications.

Effect of Ball-Milling Treatment Combined with Glycosylation on the Structure and Functional Properties of Litopenaeus vannamei Protein.

Litopenaeus vannamei protein (LVP) is a high-quality protein. However, its functional properties do not fully meet the needs of food processing. In this study, LVP-xylose conjugates were prepared by conventional wet heat method (GLVP) and ball-milling-assisted wet heat method (GBLVP), respectively. The changes in structure and functional properties of the glycosylated LVP were explored. The findings revealed that ball-milling pretreatment increased the grafting degree to 35.21%. GBLVP had a sparser surface structure and lower particle size than GLVP. FTIR spectra showed that xylose was grafted onto LVP successfully and GBLVP had the lowest α-helix content. Compared with GLVP, GBLVP had a decrease in intrinsic fluorescence intensity and surface hydrophobicity, and an increase in UV absorption intensity. Moreover, GBLVP had higher foaming capacity, solubility and water-holding capacity, and lower allergenicity than GLVP. However, ball-milling pretreatment had a negative impact on the vitro digestibility and oil-holding capacity of GBLVP. In conclusion, ball-milling-assisted treatment of glycosylation could effectively improve the functional properties of LVP, benefiting the broader application of LVP in the food industry.

Molecular Characterization of the Allergenic Arginine Kinase from the Edible Insect Hermetia illucens (Black Soldier Fly).

Arginine kinase (AK) is an important enzyme for energy metabolism of invertebrate cells by participating in the maintenance of constant levels of ATP. However, AK is also recognized as a major allergen in insects and crustaceans capable of cross-reactivity with sera of patients sensitized to orthologous proteins. In the perspective of introducing insects or their derivatives in the human diet in Western world, it is of primary importance to evaluate possible risks for allergic consumers. This work reports the identification and characterization of AK from Hermetia illucens commonly known as the black soldier fly, a promising insect for human consumption. To evaluate allergenicity of AK from H. illucens, putative linear and conformational epitopes are identified by bioinformatics analyses, and Dot-Blot assays are carried out by using sera of patients allergic to shrimp or mites to validate the cross-reactivity. Gastrointestinal digestion reduces significantly the linear epitopes resulting in lower allergenicity, while the secondary structure is altered at increasing temperatures supporting the possible loss or reduction of conformational epitopes. The results indicate that the possible allergenicity of AK should be taken in consideration when dealing with novel foods containing H. illucens or its derivatives.

Computational Approaches To Design Multi Epitope-Based Vaccine Designing of Dengue virus -2 Enveloped Protein For Dengue Virus

Dengue Fever (DF) is a viral disease transmitted by mosquitoes and is a global concern. A successful vaccine for dengue should induce both neutralizing antibodies and cell-mediated immunity. However, no vaccine currently exists for DF. A multi-epitope vaccine offers a promising strategy for preventing such infections. Objective: To create a dengue virus-2 strain multi-epitope vaccine that is safe, non-allergic, and stimulates a strong immune response. Methods: Leveraging in silico tools, we retrieved and analyzed Dengue virus-2 protein sequences, determining antigenicity using VaxiJen version 2.0 and assessing allergenicity using AllerTop. T-cell epitopes were identified via Immune Epitope Database (IEBD) server for Major Histocompatibility Complex -I (MHC-I) and Major Histocompatibility Complex -II (MHC-II) binding and B-cell epitopes were anticipated through IEBD Linear Epitope Prediction Tool v2.0. Analysis of population coverage estimated the prevalence of MHC alleles interacting with the identified epitopes. A multi-epitope vaccine construct integrated adjuvants, universal linkers, and epitopes, evaluated for physicochemical properties, toxicity, secondary, and tertiary structures. Results: Antigenicity analysis identified highly antigenic Dengue virus-2 protein sequences with low allergenicity. T-cell epitopes revealed multiple epitopes with diverse MHC-I and MHC-II affinity, encompassing conserved regions for potential universal vaccine development. Nine non-toxic, non-allergenic B-cell epitopes were identified. Population coverage analysis demonstrated over 71% prevalence of MHCs binding to identified epitopes across diverse populations. Physicochemical assessments revealed favorable characteristics, including immunogenicity and stability. Tertiary structure prediction illustrated the vaccine's 3D arrangement, validated through Ramachandran plots, exhibiting high-quality protein structure. Conclusions: This multieiptope based vaccine is more immunogenic but further in-vitro and in-vivo study is required for its clinical use

The Effect of Dietary Protein Hydrolysate from Black Soldier Fly Larvae and Schizochytrium on Palatability, Nutrient Metabolites and Health Status in Beagle Dogs.

Protein hydrolysate from black soldier fly larvae (BSFP) has garnered great attention with its lower allergenicity, high amount of essential amino acids, and small bioactive peptides. Schizochytrium is a promising alternative source of n-3 FUFA because it has enriched docosahexaenoic acid (DHA, C22: 6). The aim of this study was to assess palatability, the presence of diarrhea, plasma biochemistry panels, anti-oxidative and anti-inflammatory effects, and immune function in beagle dogs when supplementing a mixture of protein hydrolysate from black soldier fly larvae and schizochytrium (BSFPs) into their diets. Experiment I: 24 young beagle dogs (16 males and 8 females; 4-5 months; BW: 6.40 ± 0.15 kg) were randomly divided into four groups: (1) control (CON), (2) 5% BSFPs, (3) 10% BSFPs, (4) 15% BSFPs. Their body weights and fecal scores were recorded, and blood samples were collected for analysis. Experiment II: three diets containing 5%, 10%, and 15% BSFPs were evaluated by comparing them with a basal diet (CON) to evaluate palatability. These results suggested that a lower presence of diarrhea existed in the BSFP diet than the CON diet (p < 0.05). Three treatment groups remarkably increased their total protein (TP) and albumin (ALB) contents and decreased their concentrations of triglyceride (TG) and total cholesterol (TC) in plasma (p < 0.05). Moreover, the 5% and 15% BSFPs groups had a higher calcium (CA) content in plasma, and the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and contents of creatinine (CREA) and urea nitrogen (BUN) were significantly reduced by supplementing BSFP in their diets (p < 0.05). Their anti-oxidative enzyme activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) were dramatically enhanced, and their malondialdehyde (MDA) concentrations were remarkably reduced (p < 0.05). Immunoglobulin A and G (IgA and IgG) concentrations in the plasma in the 10% and 15% BSFPs groups were significantly increased (p < 0.05). Furthermore, lower interleukin-8 (IL-8) contents were shown in the BSFP diets than the CON diet (p < 0.05). Similarly, the diets supplemented with BSFPs exhibited a positive effect on palatability (p < 0.05). To sum up, the diets supplemented with BSFPs significantly enhanced palatability, immune function, and anti-oxidative and anti-inflammatory capacity to alleviate diarrhea and improve the general health of the beagle dogs.

The allergenic potential of green urban areas in the Macaronesian islands: The case of Funchal City (Madeira)

Public gardens are elements that constitute the urban green infrastructure. Such units provide ecosystem services that are becoming a key concern in urban planning strategies.In this survey the allergenic potential of two public gardens in Funchal, a city exhibiting sub-tropical and Mediterranean climatic features, was assessed. Different allergenic risk scenarios were considered, and the Allergenicity Indexes recalculated. Results show that the Municipal Garden exhibits an IUGZA (Index of Urban Green Zone Allergenicity) of 0,39 and Santa Catarina Park an IUGZA of 0,16, which are considered spaces of moderate and low allergenicity level, respectively. Among the tested scenarios that could aggravate the IUGZA values, the worst would be an extension of the pollen season for all species. This might represent an issue considering the overall trend towards an earlier start and later end of the pollen season from important allergenic plants in Europe. The assessment of the IUGZA should include all plant biotypes of an urban green area in subtropical regions as they allow us to infer more precisely on the real risk population is exposed to. It seems that increasing species diversity can improve the allergenic indexes, but novel invasive species should be studied to assess their allergenicity level.

Effect of rosmarinic acid covalent conjugation on the structure and allergenic capacity of ovalbumin

Ovalbumin (OVA), a major allergen in egg-induced food allergy, is widely used in the food industry. This study investigated the structural and antigenic changes of OVA after covalent conjugation with rosmarinic acid (RA). The SDS-PAGE and RA binding equivalent analysis confirmed that both the radical and alkaline methods successfully prepared the OVA-RA conjugates. LC-MS/MS assay identified 20 and 15 modified sites of RA on OVA in the radical and alkaline OVA-RA conjugates, respectively. The structural changes of OVA were indicated by fluorescence spectroscopy, Fourier transform infrared (FTIR) spectroscopy, circular dichroism (CD), and ultraviolet absorption (UV) spectrometry. Following conjugation with RA, the antioxidant capacities of OVA for scavenging DPPH• and ABTS•+ showed significant enhancement. Subsequently, the impact of RA on the antigenicity of OVA was assessed both in vitro and in vivo. The ELISA analysis revealed a notable reduction in the binding capacity of OVA to IgG and IgE after covalent conjugation with RA. Additionally, the passive cutaneous anaphylaxis (PCA) assay demonstrated lower allergenicity in radical and alkaline OVA-RA conjugates compared to native OVA. These findings collectively suggest that OVA-RA conjugates hold promise for the development of hypoallergenic food.

Effects of Heat Treatment on Structure and Processing Characteristics of Donkey Milk Whey Protein

Donkey milk is nutritionally rich and has low allergenicity; thus, whey protein derived from donkey milk is a suitable hypoallergenic alternative. However, the temperature during pasteurisation considerably influences the structure and processing characteristics of whey proteins. We found that donkey milk has poor thermal stability; the secondary structure of the whey proteins loosened and became disordered as the temperature increased. In addition, exposing tryptophan and tyrosine to an external polar environment changed the tertiary structure. High-temperature and long-duration heat treatments significantly affected the processing properties, such as emulsification, foaming, and rheological properties. However, a short treatment at approximately 85°C improved the processing properties. Therefore, we recommend that donkey milk be processed at temperatures no higher than this temperature to maintain the beneficial qualities of the whey proteins and provide a reference for processing donkey milk products.