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The present work was aimed at identifying the IgE-binding epitopic regions on the surface of the Cup s 3 allergen from the common cypress Cupressus sempervirens, that are possibly involved in the IgE-binding cross-reactivity reported between Cupressaceae species. Three main IgE-binding epitopic regions were mapped on the molecular surface of Cup s 3, the PR-5 thaumatin-like allergen of common cypress Cupressus sempervirens. They correspond to exposed areas containing either electropositive (R, K) or electronegative (D, E) residues. A coalescence occurs between epitopes #1 and #2, that creates an extended IgE-binding regions on the surface of the allergen. Epitope #3 contains a putative N-glycosylation site which is actually glycosylated and could therefore comprise a glycotope. However, most of the allergenic potency of Cup s 3 depends on non-glycosylated epitopic peptides. The corresponding regions of thaumatin-like allergens from other closely related Cupressaceae (Cryptomeria, Juniperus, Thuja) exhibit a very similar conformation that should account for the IgE-binding cross-reactivity observed among the Cupressaceae allergens.

Plantings surrounding kindergartens are an integral part of urban green spaces. These open areas play a very important role in the lives of children, in their development, their eating habits, and environmental awareness. For this reason, it is extremely important to select adequate plant species, which are not dangerous for the children. There are 10 public and 9 private preschool institutions in the entire area of the two Novi Zagreb (East and West) city districts. Only public kindergartens have been included in this research. The aim was to determine the size of green open space surrounding public kindergartens, analyse woody plants on site, determine the taxonomic affiliation of plant species, and detect the presence of poisonous and allergenic plant taxa. In green space surrounding kindergartens in the Novi Zagreb – East city district, 49 taxa of trees, and 18 taxa of shrubs and woody climbers from a total of 22 plant families have been identified. Similary, in the Novi Zagreb – West city district, 49 taxa of trees, and 24 taxa of shrubs and woody climbers from a total of 26 plant families have been determined in kindergarten plantings. A total of 32 poisonous taxa, and 36 allergenic taxa have been catalogued in the Novi Zagreb – East city district, while a total of 34 poisonous taxa, and 42 allergenic taxa have been listed in the Novi Zagreb – West city district. The most poisonous species were: Taxus baccata, Prunus laurocerasus and Sambucus nigra. The species with the highest allergenic potential were: Thuja occidentalis, Betula pendula and Acer platanoides., Vrtovi u sklopu vrtića sastavni su dio gradskih zelenih površina. Ovi vrtovi imaju vrlo značajnu ulogu u životu djece, potenciranju njihovih radnih i prehrambenih navika te razvijanju ekološke svijesti. Iz tog razloga, izuzetno je važno da biljne vrste u sklopu vrtića budu primjereno odabrane. Na cijelom području Novog Zagreba (Istok i Zapad) postoji 10 gradskih i 9 privatnih predškolskih ustanova, a u istraživanje je bilo uključeno 10 gradskih vrtića. Ciljevi ovog istraživanja bili su ustanoviti postojanje vrtova uz vrtiće na području gradskih četvrti (GČ) Novog Zagreba, odrediti njihovu veličinu, inventarizirati dendrološku floru, odrediti taksonomsku pripadnost te analizirati prisutnost otrovnih i alergenih biljnih svojti. U vrtovima vrtića GČ Novi Zagreb – istok utvrđeno je 49 svojti stabala i 18 svojti grmlja i penjačica iz ukupno 22 porodice. U vrtovima vrtića GČ Novi Zagreb – zapad determinirano je 49 svojti stabala te 24 svojti grmlja i penjačica iz ukupno 26 porodica. U vrtovima GČ Novi Zagreb – istok ukupno su zabilježene 32 otrovne svojte i 36 alergenih svojti dok je u vrtovima GČ Novi Zagreb – zapad ukupno utvrđeno 34 otrovnih svojti te 42 alergene svojte. Najotrovnije vrste bile su: Taxus baccata, Prunus laurocerasus i Sambucus nigra. Vrste s najvećim alergenim potencijalom bile su: Thuja occidentalis, Betula pendula i Acer platanoides.

Background: Methods and approaches that can be used in food and nutrition research are changing at a faster pace than ever. Whereas animal methods are mostly known for their use in food safety analysis (see Part I), they also play in important role in proof-of-concept and mechanistic studies of products, as well as studying potency, efficacy, and tolerance of foods and food ingredients. Members of the International Life Sciences Institute (ILSI) Europe have formed an expert group to review possibilities, opportunities, and challenges for the potential use of alternative testing strategies in nutrition research and regulatory requirements, supporting the 3Rs principle of Replacement, Reduction, Refinement of animal research, which can ultimately be used in support of regulatory submissions for pre-market authorisation. Scope and approach: For the different areas of food for specific groups and health claims, the acceptability of nonanimal approaches is evaluated in comparison to legislative requirements in Europe. The alternative approaches considered cover emerging tools and methodologies such as organoids, organs-on-a-chip or human in vitro gastrointestinal simulators. Conclusions: In nutrition research, there has been a long tradition of following a certain experimental trajectory for grounding scientific hypotheses starting from in vitro data moving on to in vivo verification in a preferred animal model and finally proving this in a human setting. From a regulatory perspective there is no specific requirement for animal experimentation that justifies the use of the majority of animal experiments in the assessment of nutritional content and value of food products. However, animal data are mostly considered as the standard, and guidance for alternative approaches that would be accepted is lacking. It is therefore important to further build evidence and offer validation for the adequacy of already existing in vitro tools to ensure their suitability for substantiating dose levels and further planning clinical trials. What are we waiting for?

In the past decades, numerous landscaping efforts in the urban and suburban areas of Skopje have provided a diversified presence of allochthonous woody species. From the registered 206 allochthonous woody plants, in the past 25 years, 65 species (19 Gymnospermae and 46 Angiospermae) have been the focus of various research and monitoring efforts, including seed propagation and analysis of the generative propagation potential. Considering the variability in the experimental approach, we have performed an extensive literature revision and combined the results from the two in a summary of species that could be of potential interest in the green infrastructure in Skopje, due to their benefits (ornamental use and air pollution remediation) or risks (invasive and allergenic potential). We have also underlined the potential of using urban species as seed banks and species conservation, along with various services they could provide. To the best of our knowledge, this is the first attempt that summarizes the allochthonous woody species in Skopje along with the experience regarding their generative potential and possible use in urban forestry. As such, it serves as a base for future experimental research that could provide more information about the seed quality and the species’ benefits in Skopje and the surrounding areas.

Presently, many studies assess allergenicity via IgE immunoblotting and IgE binding tests; however, IgE detection does not always signal the manifestation of a clinical allergy. However, the capacity of food allergens to trigger basophils makes it possible to use the in vitro functional basophil activation test (BAT) to assess allergenicity. The effect of the Maillard reaction (MR) on the allergenic potential of processed soy proteins was evaluated by two IgE binding tests (Competitive ELISA and Inhibition ImmunoCAP), a Western Blot and a functional BAT; with the aim to analyze whether the sIgE binding results correspond to the functional assay results. The results between the IgE binding tests and the functional assay were in-line for 2 of the 6 studied patients. For one patient there was no correlation between any of the results. For the raw soy protein extract heated with glucose for 10 and 30 min (SH SPE + Glu and LH SPE + Glu, respectively), the results were in-line for only 3 out of the 6 patients. Thus, the present study shows a discrepancy between IgE binding tests and basophil stimulation when assessing the effect of soy processing on its allergenicity. Since IgE-binding capacity does not always correlate to IgE cross-linking capacity, the conclusions of the allergenic potential based on the IgE binding tests alone should be drawn with care and further studies on this matter would benefit from the inclusion of a functional assay such as the BAT.

In the last decade, soybean allergies have been on the increases to such an extent that they have now become a public health issue thus prompting more studies and researches on the topic. The allergenicity of soybean is attributed to its protein fraction. The best way to prevent hypersensitive patients from ingesting allergenic compounds is to exclude such soybean allergens from their diet. As a result, it is essential to provide detailed and reliable knowledge of food ingredients. Therefore, precise and reliable approaches for detecting soybean allergens found in various food products must be used. The main way to reduce allergy risk is the identification of allergenic sites in food and their inactivation by various food-processing methods. It has been reported that food processing may lead to the modification of conformational structure of the protein or protein distortion that inhibit the binding of immunoglobulin E (IgE) to epitopes on food allergens and also the mechanism of allergic reactions. Food processing technologies employed for inactivating allergenic epitopes used thermal and nonthermal techniques. Currently, several detection methods including protein-based and DNA-based approaches using analytical techniques such as enzyme-linked immunosorbent assay (ELISA), enzyme allergosorbent test (EAST), radioallergosorbent test (RAST), Lateral flow immunoassay (LFIA), Immunoblotting, real-time polymerase chain reaction (PCR), mass spectrometry and biosensors have been improved for identifying and quantifying these epitopes. This research focused on allergenic proteins of soybean, the most modern approaches for detecting and quantifying these allergens, and finally, the various methods used to inactivate these proteins and their effects on soy allergenicity.

Animal protein plays a key role in the human diet as the most balanced amino acid composition; however, its consumption often causes allergic reactions. Plant protein serves as a substitute for animal protein. The most promising sources of plant protein are the seeds of cereals, pulses, oilseeds and cereals. Research aim: selection of conditions for obtaining protein isolates and enzymatic hydrolysates having the desired functional properties from different types of vegetable raw materials.Pea, corn and oat flour LLC "FavoritT"; linseed flour LLC NGO "Compass Health". Enzyme preparations: chymotrypsin LLC "Samson-Med"; Protex 40E Genencor; protosubtilin G3x produced by PO "Sibbiofarm" LLC; pancreatin PJSC "Biosintez"; trypsin LLC "Diaem"; beef pepsin OJSC "MHSF". Crude protein content was determined by Kjeldahl method, protein substances - by modified Lowry method. Fat-holding, water-holding, emulsifying and foaming capacities, as well as allergenicity of protein isolates and hydrolysates were determined. Conditions for protein substances extraction from flax, corn, oat and pea flour with the yield of high-molecular protein fraction not less than 70 % of raw protein content were selected. The conditions of protein isolates precipitation to produce preparations containing not less than 85% of protein have been selected. The type of enzyme preparation for hydrolysis - pancreatin - was selected. It was found that in order to increase water- and fat-holding capacity of pea isolate, hydrolysis is possible with duration not exceeding 15 min, for all other isolates hydrolysis is undesirable. The best emulsifying and foam-forming capacities are possessed by linseed hydrolysates after 60 and 90 min of hydrolysis, respectively. Enzymatic hydrolysis was shown to reduce the allergenicity of plant proteins. The obtained hydrolysates of vegetable proteins can be used as ingredients for functional products, as well as for obtaining products with reduced allergenicity., Ключевую роль в питании человека играет животный белок как наиболее сбалансированный по аминокислотному составу, однако его потребление часто вызывает аллергические реакции. Заменой животному белку служит растительный. Наиболее перспективными источниками растительного белка являются семена зерновых, бобовых, масличных и злаковых культур. Цель исследования – подбор условий получения белковых изолятов и ферментативных гидролизатов, обладающих заданными функциональными свойствами, из различных видов растительного сырья. Подобраны условия извлечения белковых веществ из льняной, кукурузной, овсяной и гороховой муки с выходом высокомолекулярной фракции белка не менее 70% от содержания сырого протеина в сырье. Подобраны условия осаждения белковых изолятов с получением препаратов, содержащих не менее 85% белка. Подобран тип ферментного препарата для проведения гидролиза – панкреатин. Установлено, что для повышения водо- и жироудерживающей способности для горохового изолята возможен гидролиз продолжительностью не более 15 мин, для всех остальных изолятов гидролиз нежелателен. Наилучшими эмульгирующей и пенообразующей способностями обладают льняные гидролизаты после 60 и 90 мин гидролиза соответственно. Показано, что ферментативный гидролиз способствует снижению аллергенности растительных белков. Полученные гидролизаты растительных белков могут быть использованы в качестве ингредиентов для функциональных продуктов, а также для получения продуктов с пониженной аллергенностью.

ZusammenfassungÄpfel besitzen mehrere Allergene, die beim Essen innerhalb von 5–10 min zu Symptomen im Mundbereich führen – und deshalb von Apfelallergikern nicht gegessen werden können. In Deutschland haben rund 7,5 Mio. Menschen spezifische Antikörper gegen das Hauptallergen (Mal d 1) in Äpfeln entwickelt und sind damit sensibilisiert. Mindestens 3,5 Mio. von ihnen entwickeln die teilweise erheblichen allergischen Symptome als Ausdruck eines Oralen Allergie-Syndroms. Es gibt bisher keine medikamentöse Therapie gegen diese Allergie.Apfelallergiker können daher nur auf Äpfel ganz verzichten, oder vorher erhitzte Äpfel essen oder Sorten suchen, die wenig Allergene enthalten und deshalb als allergikerfreundliche Apfelsorten bezeichnet werden können.Alleinige Bestimmungen von Allergenen im Labor können nicht voraussagen, ob ein Apfel ohne allergische Symptome von Apfelallergikern gegessen werden kann; dazu bedarf es klinischer Prüfungen.Wir beschreiben eine standardisierte klinische, orale Provokationstestung, die zur Charakterisierung eines allergenarmen, allergikerfreundlichen Apfels bzw. Apfelsorte benutzt werden kann.. Die Ergebnisse solcher mindestens dreijährigen Tests können zur Verleihung des ECARF-Siegels für allergikerfreundliche Produkte genutzt werden

This article belongs to the Special Issue Role of Food Digestion and Digestive System in the Nutritional, Functional and Health Properties of Food Bioactives., Dairy foods are essential in the diet, although in some susceptible individuals they may cause allergy to cow’s milk proteins. Therefore, alternative methods are sought to reduce their allergenicity. Transglutaminase (TG) is widely used in dairy products mainly to improve texture. Although it has been claimed that TG can be used to modify the digestibility and allergenicity of foods, its impact within a real matrix has been rarely studied. The aim of this work was to assess the allergenic potential of crosslinked skim milk (SM), milk casein fraction (CN), and whey protein (WP). To this purpose, inhibition ELISA with sera from milk allergic patients, in vitro activation tests of mouse mast cells and splenocytes, and simulated gastrointestinal digestion assays were performed. The results showed that cross-linking increased the binding of IgE to WP, but decreased IgE-binding to SM and CN. However, no differences were observed in the ability of cross-linked proteins to induce mast cell degranulation compared to native proteins. The cross-linking of SM and CN reduced Th2 cytokine release from the splenocytes of sensitized mice. All TG-treated samples exhibited more resistance to in vitro digestion than the untreated proteins and the human IgE binding capacity after digestion was higher. In conclusion, TG treatment of milk proteins does not reduce the risk of eliciting allergic symptoms in cow’s milk allergic patients., This work was supported by the Ministerio de Ciencia, Innovación y Universidades (project AGL2017-88964-R) and Ministerio de Ciencia e Innovación through a project (PID2021-123236OB-I00) and a Juan de la Cierva Incorporación contract (SB) (IJCI-2017-31345 B).

Allergenic pollen protein can be released from pollen grains and suspended in the air to cause allergenic reactions. However, the allergenic protein and its relationship with redox trace elements in ambient size-resolved particles has not been reported. Ambient size-resolved particles in Shanghai’s atmosphere were sampled during the Platanus pollen season in the spring season of 2017. Planatus pollen protein 3 (Pla a3) and redox trace elements in the ambient particles were investigated and their relationship was analyzed. Our data demonstrated that the mass level of the Pla a3 in the size-resolved particles ranged from 0.41 ± 0.28 to 7.46 ± 2.23 pg/m3, and decreased with the size range. Mass concentrations (ppb) of crustal elements (Fe, Al, Ca, Mg, Na) in the size-resolved particles ranged from 20.11 ± 9.87 to 1126.22 ± 659.51, while trace elements (V, Cr, Mn, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Cd, Cs, Ba, Pb) varied from 0.05 ± 0.03 to 57.53 ± 19.7. Mass levels of these trace elements decreased according to particle size. The Abundance of redox trace elements, including Fe (R2 = 0.82), Mn (R2 = 0.54), Cu (R2 = 0.61), Ba (R2 = 0.82), and Pb (R2 = 0.82) in the size-resolved particles was significantly related to that of Pla a3, and our data implied redox trace elements might take syngenetic effects on the allergenicity induced by Pla a3 protein.

Whey is a by-product of the production of various types of cottage cheese and cheese, casein, and coprecipitates. Conditions of milk coagulation directly affect the physico-chemical properties of whey and the formation of its protein profile. This fact makes it difficult to standardize the protein profile of milk whey for its further processing. Whey proteins have a great potential to release a wide range of bioactive peptides (BAP), capable of reducing the risk of a number of chronic food-related diseases. Computer modeling of an enzymatic hydrolysis of proteins is one of the ways to increase the efficiency of BAP release studies and to reduce the number of labor consuming experiments. This research is aimed at generating a digital model of the peptide complex of different whey types with predicted bioactivity, safety, and sensory properties using bioinformatic modeling approaches. The study was performed with the use of the proteomic databases tools according to the algorithm of hybrid strategy of bioinformatic modeling developed earlier. As a result of the study, taking into account the ranking of the proteins ratio in the protein profile, the hydrolysis by the protease complex chymotrypsin C-subtilisin was characterized as the maximum efficacy method to release peptides with both antioxidant and ACE-inhibitory activity. It was also observed that the bioactive peptides obtained as a result of in silico hydrolysis after GI digestion simulation can be considered safe in terms of allergic reactions and toxicological effects.

The recurrent and recent global outbreak of the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has turned into a global concern which has infected more than 42 million people all over the globe, and this number is increasing in hours. Unfortunately, no vaccine or specific treatment is available, which makes it more deadly. A vaccine-informatics approach has shown significant breakthrough in peptide-based epitope mapping and opens the new horizon in vaccine development. In this study, we have identified a total of 15 antigenic peptides [including thymus cells (T-cells) and bone marrow or bursa-derived cells] in the surface glycoprotein (SG) of SARS-CoV-2 which is nontoxic and nonallergenic in nature, nonallergenic, highly antigenic and non-mutated in other SARS-CoV-2 virus strains. The population coverage analysis has found that cluster of differentiation 4 (CD4+) T-cell peptides showed higher cumulative population coverage over cluster of differentiation 8 (CD8+) peptides in the 16 different geographical regions of the world. We identified 12 peptides ((LTDEMIAQY, WTAGAAAYY, WMESEFRVY, IRASANLAA, FGAISSVLN, VKQLSSNFG, FAMQMAYRF, FGAGAALQI, YGFQPTNGVGYQ, LPDPSKPSKR, QTQTNSPRRARS and VITPGTNTSN) that are \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} }{}$80\hbox{--} 90\%$\end{document} identical with experimentally determined epitopes of SARS-CoV, and this will likely be beneficial for a quick progression of the vaccine design. Moreover, docking analysis suggested that the identified peptides are tightly bound in the groove of human leukocyte antigen molecules which can induce the T-cell response. Overall, this study allows us to determine potent peptide antigen targets in the SG on intuitive grounds, which opens up a new horizon in the coronavirus disease (COVID-19) research. However, this study needs experimental validation by in vitro and in vivo.

Background:Multi-valent based vaccines have advantage over conventional vaccines because of its multi-faceted action targeted at antigen; thereby raising hope of a more sustained actions against allergens. Escherichia coli(E. coli)is a bacterium that is commonly found in the gut of humans and warm-blooded animals. An increasing number of outbreaks are associated with the consumption of fruits and vegetables (including sprouts, spinach, lettuce, coleslaw, and salad) thereby contamination may be due to contact with faeces from domestic or wild animals at some stages during cultivation or handling. Due to the reported increase in resistance to antibiotics used forEscherichia colicontrol; an effective vaccine is a would-be alternative of proven interest. Hence, a need for a rational, strategic, and efficient vaccine candidate againstE.coliis of paramount necessity by the use of the most current bioinformatics tools to achieve this task.Method:In this study, immunoinformatics tools mined from diverse molecular databases were used for a novel putative epitope based oral vaccine againstE.coli. The prospective vaccine proteins were carefully screened and validated to achieve a high thorough-put three-dimensional protein structure. The eventual propsective vaccine candidate proteins was evaluated for its non-allergenicity, antigenicity, solubility, appropriate molecular weight testing and isoelectric point evaluation.Conclusion:The resultant vaccine candidate could serve as a promisinganti-E.colivaccine candidate. Immunoinformatics is a new field over pharmaco-therapeutics; this newest technology should continue to be a rescue from age-long traditional approach in vaccine developments.

Tropomyosin (TM) is an important crustacean (Scylla paramamosain) allergen. This study aimed to assess Maillard-reacted TM (TM-G) induction of allergenic responses with cell and mouse models. We analyzed the difference of sensitization and the ability to induce immune tolerance between TM and TM-G by in vitro and in vivo models, then we compared the relationship between glycation sites of TM-G and epitopes of TM. In the in vitro assay, we discovered that the sensitization of TM-G was lower than TM, and the ability to stimulate mast cell degranulation decreased from 55.07 ± 4.23% to 27.86 ± 3.21%. In the serum of sensitized Balb/c mice, the level of specific IgE produced by TM-G sensitized mice was significantly lower than TM, and the levels of interleukins 4 and interleukins 13 produced by Th2 cells in spleen lymphocytes decreased by 82.35 ± 5.88% and 83.64 ± 9.09%, respectively. In the oral tolerance model, the ratio of Th2/Th1 decreased from 4.05 ± 0.38 to 1.69 ± 0.19. Maillard reaction masked the B cell epitopes of TM and retained some T cell epitopes. Potentially, Maillard reaction products (MRPs) can be used as tolerance inducers for allergen-specific immunotherapy.

The addition of soy proteins, currently classified as a food allergen, into meat products is a commonly used practice due to its functional properties and low cost. Its addition to meat products can cause health problems for individuals allergic to these proteins. Allergic individuals can be affected by the ingestion of low amounts of the allergen. In Brazil, limits are set for the addition of soy proteins in meat products in order to avoide fraud. Starting in 2015 reporting the name of the added component became mandatory for all food labelling. Some studies have reported that food processing can reduce the allergenicity, either by irreversible removal of allergens or by modifying the allergen structure. However, the technological approach to decrease allergenicity has largely been empirical. This review describes the use of soy protein in meat products and the health risk for allergic individuals and consumers of these products. Finally, appropriate methodologies for the detection and quantification of these proteins must be further explored and established to avoid fraud and to preserve consumer health.

Introduction Allergen‐specific immunotherapy (AIT) represents a curative approach for treating allergies. In the tropical and subtropical regions of the world, Blomia tropicalis (Blo t 5 and Blo t 21) is the likely dominant source of indoor allergens. Aim To generate a hypoallergenic Blo t 5/Blo t 21 hybrid molecule that can treat allergies caused by B tropicalis. Methods Using in silico design of B tropicalis hybrid proteins, we chose two hybrid proteins for heterologous expression. Wild‐type Blo t 5/Blo t 21 hybrid molecule and a hypoallergenic version, termed BTH1 and BTH2, respectively, were purified by ion exchange and size exclusion chromatography and characterized by physicochemical, as well as in vitro and in vivo immunological, experiments. Results BTH1, BTH2 and the parental allergens were purified to homogeneity and characterized in detail. BTH2 displayed the lowest IgE reactivity that induced basophil degranulation using sera from allergic rhinitis and asthmatic patients. BTH2 essentially presented the same endolysosomal degradation pattern as the shortened rBlo t 5 and showed a higher resistance towards degradation than the full‐length Blo t 5. In vivo immunization of mice with BTH2 led to the production of IgG antibodies that competed with human IgE for allergen binding. Stimulation of splenocytes from BTH2‐immunized mice produced higher levels of IL‐10 and decreased secretion of IL‐4 and IL‐5. In addition, BTH2 stimulated T‐cell proliferation in PBMCs isolated from allergic patients, with secretion of higher levels of IL‐10 and lower levels of IL‐5 and IL‐13, when compared to parental allergens. Conclusions and Clinical Relevance BTH2 is a promising hybrid vaccine candidate for immunotherapy of Blomia allergy. However, further pre‐clinical studies addressing its efficacy and safety are needed.

The present study aims to investigate the effects of ultrasound on the non-covalent interaction of β-lactoglobulin (β-LG) and luteolin (LUT) and to investigate the relationship between allergenicity and human intestinal microbiota. After treatment, the conformational structures of β-LG were changed, which reflected by the decrease in α-helix content, intrinsic fluorescence intensity and surface hydrophobicity, whereas the β-sheet content increased. Molecular docking studies revealed the non-covalent interaction of β-LG and LUT by hydrogen bond, van der Walls bond and hydrophobic bond. β-LG-LUT complex treated by ultrasound has a lower IgG/IgE binding ability and inhibits the allergic reaction of KU812 cells, depending on the changes in the conformational epitopes of β-LG. Meanwhile, the β-LG-LUT complex affected the composition of human intestinal microbiota, such as the relative abundance of Bifidobacterium and Prevotella. Therefore, ultrasound improved the non-covalent interaction of β-LG with LUT, and the reduction in allergenicity of β-LG depends on conformational epitopes and human intestinal microbiota changes.

Both protein consumption and the world’s population are increasing, which leads to an urgent demand for new sustainable food protein sources. However, introduction of novel protein sources is not without risk. Novel protein sources may induce new food sensitizations and allergies and/or can elicit cross-reactive allergies in already allergic individuals. It is therefore pivotal that we can accurately assess and predict the allergenicity of novel proteins. Predictive methods that can measure the allergenicity accurately can be developed more effectively when it is agreed upon which measure should be used to characterize allergenicity. Regulators should make this decision to direct the development of predictive test methods. Several parameters have been proposed in the past that can be used to assess and express allergenicity and regulate upon. In this thesis, we evaluated the sensitizing potency of a protein, that we think has the highest potential to accurately characterize the allergenicity of novel food proteins. Additionally, we provided a framework which includes multiple steps that can be followed to allow regulators to establish acceptance or rejection criteria for novel food proteins. We also investigated if certain methods have the potential to predict the allergenicity of proteins and how these may be improved. The gene expression of blood cells exposed to proteins can potentially be used in this regard. However, we also found that digestion and uptake of proteins can affect the allergenicity of proteins. This should be taken into account when developing the much needed methods that can assess allergenicity.

Xylanase inhibitors (XIs) are plant cell wall proteins largely distributed in monocots that inhibit the hemicellulose degrading activity of microbial xylanases. XIs have been classified into three classes with different structures and inhibition specificities, namely Triticum aestivum xylanase inhibitors (TAXI), xylanase inhibitor proteins (XIP), and thaumatin-like xylanase inhibitors (TLXI). Their involvement in plant defense has been established by several reports. Additionally, these inhibitors have considerable economic relevance because they interfere with the activity of xylanases applied in several agro-industrial processes. Previous reviews highlighted the structural and biochemical properties of XIs and hypothesized their role in plant defense. Here, we aimed to update the information on the genomic organization of XI encoding genes, the inhibition properties of XIs against microbial xylanases, and the structural properties of xylanase-XI interaction. We also deepened the knowledge of XI regulation mechanisms in planta and their involvement in plant defense. Finally, we reported the recently studied strategies to reduce the negative impact of XIs in agro-industrial processes and mentioned their allergenicity potential.

Parvalbumin is considered a major fish allergen. Here, we report the molecular evolution of the parvalbumin genes in bony fishes based on 19 whole genomes and 70 transcriptomes. We found unexpectedly high parvalbumin diversity in teleosts; three main gene types (pvalb-α, pvalb-β1, and pvalb-β2, including oncomodulins) originated at the onset of vertebrates. Teleosts have further multiplied the parvalbumin gene repertoire up to nine ancestral copies—two copies of pvalb-α, two copies of pvalb-β1, and five copies of pvalb-β2. This gene diversity is a result of teleost-specific whole-genome duplication. Two conserved parvalbumin genomic clusters carry pvalb-β1 and β2 copies, whereas pvalb-α genes are located separately in different linkage groups. Further, we investigated parvalbumin gene expression in 17 tissues of the common carp (Cyprinus carpio), a species with 21 parvalbumin genes in its genome. Two pvalb-α and eight pvalb-β2 copies are highly expressed in the muscle, while two alternative pvalb-α copies show expression in the brain and the testes, and pvalb-β1 is dominant in the retina and the kidney. The recent pairs of muscular pvalb-β2 genes show differential expression in this species. We provide robust genomic evidence of the complex evolution of the parvalbumin genes in fishes.

Parvalbumin is considered a major fish allergen. Here, we report the molecular evolution of the parvalbumin genes in bony fishes based on 19 whole genomes and 70 transcriptomes. We found unexpectedly high parvalbumin diversity in teleosts; three main gene types (pvalb-α, pvalb-β1, and pvalb-β2, including oncomodulins) originated at the onset of vertebrates. Teleosts have further multiplied the parvalbumin gene repertoire up to nine ancestral copies—two copies of pvalb-α, two copies of pvalb-β1, and five copies of pvalb-β2. This gene diversity is a result of teleost-specific whole-genome duplication. Two conserved parvalbumin genomic clusters carry pvalb-β1 and β2 copies, whereas pvalb-α genes are located separately in different linkage groups. Further, we investigated parvalbumin gene expression in 17 tissues of the common carp (Cyprinus carpio), a species with 21 parvalbumin genes in its genome. Two pvalb-α and eight pvalb-β2 copies are highly expressed in the muscle, while two alternative pvalb-α copies show expression in the brain and the testes, and pvalb-β1 is dominant in the retina and the kidney. The recent pairs of muscular pvalb-β2 genes show differential expression in this species. We provide robust genomic evidence of the complex evolution of the parvalbumin genes in fishes.

A major part of important mammalian respiratory allergens belongs to the lipocalin family of proteins. By this time, 19 respiratory mammalian lipocalin allergens have been registered in the WHO/IUIS Allergen Nomenclature Database. Originally, lipocalins, small extracellular proteins (molecular mass ca. 20 kDa), were characterized as transport proteins but they are currently known to exert a variety of biological functions. The three-dimensional structure of lipocalins is well-preserved, and lipocalin allergens can exhibit high amino acid identities, in several cases more than 50%. Lipocalins contain an internal ligand-binding site where they can harbor small principally hydrophobic molecules. Another characteristic feature is their capacity to bind to specific cell-surface receptors. In all, the physicochemical properties of lipocalin allergens do not offer any straightforward explanations for their allergenicity. Allergic sensitization begins at epithelial barriers where diverse insults through pattern recognition receptors awaken innate immunity. This front-line response is manifested by epithelial barrier-associated cytokines which together with other components of immunity can initiate the sensitization process. In the following, the crucial factor in allergic sensitization is interleukin (IL)-4 which is needed for stabilizing and promoting the type 2 immune response. The source for IL-4 has been searched widely. Candidates for it may be non-professional antigen-presenting cells, such as basophils or mast cells, as well as CD4+ T cells. The synthesis of IL-4 by CD4+ T cells requires T cell receptor engagement, i.e., the recognition of allergen peptides, which also provides the specificity for sensitization. Lipocalin and innate immunity-associated cell-surface receptors are implicated in facilitating the access of lipocalin allergens into the immune system. However, the significance of this for allergic sensitization is unclear, as the recognition by these receptors has been found to produce conflicting results. As to potential adjuvants associated with mammalian lipocalin allergens, the hydrophobic ligands transported by lipocalins have not been reported to enhance sensitization while it is justified to suppose that lipopolysaccharide plays a role in it. Taken together, type 2 immunity to lipocalin allergens appears to be a harmful immune response resulting from a combination of signals involving both the innate and adaptive immunities.

Ovalbumin (OVA) is one of the major allergenic proteins in egg that contain numerous functional properties. Here, oxidation-induced changes to the structure, functional properties, and allergenicity of OVA were evaluated after 2,2′-azobis (2-amidinopropane) dihydrochloride (AAPH) and acrolein treatment. Oxidation complexes were prepared by the lipid peroxidation, which caused structural changes, and increased digestibility, foaming, and emulsifying properties. The IgE-binding capacity of OVA decreased after oxidation. KU812 cell assay showed that the histamine and IL-4 levels decreased upon oxidation with AAPH and acrolein. A mouse model study showed that oxidation caused a reduction in the allergenicity by reducing IgE and IgG1 levels and the production of Th2 cytokines after acrolein (1 mmol/L) and AAPH (5 mmol/L) treatment. Thus, OVA could cause changes to structural and functional properties, modulating Th1/Th2 immunobalance, and reducing potential allergenicity after oxidation by lipid peroxidation, which might provide an effective and potential method to process hypoallergenic OVA.

The different effects on animals of the thrombolytic protease complex of the new producer S. strictum 203 were studied. The tests of acute toxicity, immunotoxicity and allergenicity should conclude that the studied proteolytic complex is safe for medical usage. For the intravenous and the intraperitoneal routes of administration, the maximum tolerated dose and the median lethal dose were not determined.

There have been many attempts to identify common biophysical properties which differentiate allergens from their non-immunogenic counterparts. This review will focus on recent studies which examine two such factors: abundance and stability. Anecdotal accounts have speculated that the elevated abundance of potential allergens would increase the likelihood of human exposure and thus the probability of sensitization. Similarly, the stability of potential allergens dictates its ability to remain a viable immunogen during the transfer from the source to humans. This stability could also increase the resilience of potential allergens to both gastric and endosomal degradation, further skewing the immune system toward allergy. Statistical analyses confirm both abundance and stability as common properties of allergens, while epidemiological surveys show a correlation between exposure levels (abundance) and allergic disease. Additional studies show that changes in protein stability can predictably alter gastric/endosomal processing and immunogenicity, providing a mechanistic link between stability and allergenicity. However, notable exceptions exist to both hypotheses which highlight the multifaceted nature of immunological sensitization, and further inform our understanding of some of these other factors and their contribution to allergic disease.

In this study, we assessed the effects of microwave irradiation of wheat gluten proteins as a pretreatment performed in a microwave reactor that could accurately control process parameters as a function of power and temperature, as well as comparing it with conventional heat treatment. The aim was to identify suitable combinations of partial enzymatic hydrolysis and microwave pretreatment parameters to produce gluten hydrolysates with reduced allergenicity and conserved techno-functional features for food application. FTIR analysis, and total and reactive SH group contents confirmed that the microwave-controlled heating can significantly change the secondary structure and conformation of gluten protein. The microwave treatment had the largest effect at 200 W and 100 °C, at which the content of gluten has been reduced by about 2.5-fold. The microwave pretreatment also accelerated the enzymatic hydrolysis of gluten, changing the kinetic profile. The apparent hydrolysis rate constants (k2) were 1.00, 3.68, 3.48, 4.64 and 4.17 min−1 for untreated gluten, and those pretreated with microwave power of 200, 400, 600 and 800 W, respectively. Compared to the heat treatment, it appeared that microwave specific non-thermal effects had a significant influence on the gluten structure and allergenicity and, in combination with the enzymatic hydrolysis, ultimately yielded protein hydrolysates with enhanced antioxidant and functional properties.

Razvoj rekombinantne DNA tehnologije je odprl nove možnosti na številnih področjih, še posebno velik vpliv pa ima na kmetijstvo. Nove GM poljščine namreč predstavljajo številne prednosti in rešitve za težave, ki nas pestijo v 21. stoletju. Obenem pa se pojavljajo vprašanja glede morebitnih tveganj, povezanih z GM poljščinami, tako za človeka kot tudi za druge organizme in okolje. Regulativa, ki ureja GMO, se zelo razlikuje med državami. Evropska unija je do GM poljščin zelo zadržana, medtem pa so ZDA vodilna država v razvoju in komercializaciji GM pridelkov. GM rastline so predmet številnih raziskav in odkritja le teh se zelo razlikujejo. Tudi zaradi tega je javno mnenje do GM rastlin še vedno odklonilno. Kljub javnemu nasprotovanju pa se obseg gojenja GM rastlin povečuje, s prihodom tehnik genskega preurejanja pa se bo število aplikacij verjetno še povečalo. Velik potencial ima recimo urejanje genoma s CRISPR metodo, saj omogoča natančnejše modifikacije z manjšimi denarnimi vložki. Uporaba novih genskih tehnik pa bo odvisna tudi od javnega mnenja, saj ima slednje velik vpliv na regulativo. The development of recombinant DNA technology has opened new possibilities in many areas with a particularly large impact on agriculture. The new GM crops represent many advantages and solutions to the problems that concern us in the 21st century. At the same time, questions have arisen regarding potential risks associated with GM crops, both for humans and for other organisms, as well as the environment. The regulations governing GMOs vary greatly between countries. While the European Union is very reticent about GM crops, the US is a leading country in development and commercialization of GM crops. GM plants are the subject of many researches and the discoveries of these differ greatly. This is one of the reasons why the public opinion on GM plants is still unfavorable. Despite public opposition, the cultivation of GM plants is increasing, and with the introduction of genome editing techniques, the number of applications is likely to increase even further. For example, genome editing with the CRISPR method has great potential, as it allows for more precise modifications with less money invested. However, the use of new genetic techniques will also depend heavily on public opinion, as it alone has a major impact on regulation of GMOs.

The goal of this study is to investigate the probable intermediate hosts and the allergenicity of the notorious virus SARS-CoV-2 to understand how this virus emerged. The phylogenetic analysis of the virus spike proteins indicates that SARS-CoV-2 falls into various small subclades that include a bat coronavirus RaTG13, suggesting bats as a likely natural origin. Refined alignment of the spike protein in NCBI found several fragments that are specific to SARS-CoV-2 and/or SARS-CoV are specific to Rattus norvegicus and/or Mus musculus, suggesting that rodents are the intermediate reservoir of SARS-CoV-2 and SARS-CoV. To evaluate the allergenicity values, the binding affinities of human leukocyte antigen (HLA) class I or II molecules with the spike proteins were calculated, and the results showed that both SARS-CoV-2 and SARS-CoV are predicted to bind to fourteen HLA class I and II molecules with super-high HLA allele-peptide affinities. The infection rate of individuals who have HLA alleles with very high binding affinities who might become infected and develop into refractory patients if there were no medical or non-medical interventions is about 7.36% and 4.78% of Chinese and Americans, respectively. Extremely high temperature and exceptionally low precipitation, the common climate factors between the outbreak sites of COVID-19 in Wuhan in 2019 and SARS in Guangdong in 2002, might have promoted coronavirus evolution into more virulent forms. Our hypothesis suggests that early immunization with an allergenically-engineered virus, in combination with continued surveillance of meteorological factors and viral mutations, may be one of the most powerful prophylactic modalities to fight this virus.

Various weight-of-evidence risk-assessment frameworks have been used for regulatory decision making. In this policy commentary, we critically assess the value of including gastric and/or intestinal digestion results in the weight of evidence designed to inform the allergenicity assessment of newly expressed proteins in genetically engineered crops. This example highlights a general concept: specifically, that there is negligible value in considering a factor as a component of the risk assessment when differing outcomes for this factor do not result in different risk decisions under any reasonable hypothetical scenario regarding results from the other components of the risk assessment. We conclude that equitable and science-based regulatory guidance should include examples of weight-of-evidence scenarios illustrating how differing outcomes for each individual component within the weight of evidence would result in a different conclusion on the acceptability of the potential risk under consideration. Critically assessing each component of the weight of evidence in this manner helps avoid inclusion of components that can only distract risk assessors from the critical decision-making process, possibly resulting in inconsistent risk assessment outcomes for the same datasets. https://doi.org/10.21423/jrs-v09i2rod, Journal of Regulatory Science, Vol. 9 No. 2 (2021)

It is obvious that social, biogenic, and anthropogenic environmental factors, as well as nutrition contribute to the development and course of atopic eczema. Social deprivation and stress have a negative impact on atopic eczema symptoms, and social change in recent decades has led to a "westernized" lifestyle associated with high prevalence of atopic eczema in industrialized countries. Urbanization leads to an increase in air pollution and a decrease in biodiversity, which negatively affects atopic eczema. Climate change alters the allergenicity of pollen, which increases atopic eczema symptoms in some patients during the pollen season. Protective natural and social factors for the prevention of atopic eczema and for the promotion of "climate resilience" should be given greater consideration in future research.

Thehigh-temperaturewatertreatmentis oneofthe methods used to reduce the molecular weightofproteins. In this study, in order to establish a practical method for preparinghypoallergenicmaterials using thehigh-temperaturewatertreatment, we investigated the effectsofprocessing temperature on the antigenicity and allergenicityofa food allergen. Additionally, the foaming abilityofthe samples was also evaluated as a function desired in the food industry. We usedovalbuminas a model allergen. As a result, although there was no significant difference among the samples treated with different processing temperatures, all the antigens treated withhigh-temperaturewatershowed a decrease in antigenicity and allergenicity. In addition, whenovalbuminwas treated at a temperatureof130 degrees C or higher, there was a significant improvement in foaming properties. These findings indicate thathigh-temperaturewatertreatmentis a potential strategy for preparing practicalhypoallergenicmaterials., Article, Bioscience, Biotechnology, and Biochemistry. 85(12): 2442–2449 (2021)

There is an increasing demand for alternative and sustainable protein sources, such as vegetables, insects and microorganisms, that can meet the nutritional and sensory pleasantness needs of consumers. This emergent interest for novel protein sources, allied with “green” and cost-effective processing technologies, such as high hydrostatic pressure, ohmic heating and pulsed electric fields, can be used as strategies to improve the consumption of proteins from sustainable sources without compromising food security. In addition to their nutritional value, these novel proteins present several technological-functional properties that can be used to create various protein systems in different scales (i.e., macro, micro and nano scale), which can be tailored for a specific application in innovative food products. However, in order for these novel protein sources to be broadly used in future food products, their fate in the human gastrointestinal tract (e.g., digestion and bioavailability) must be assessed, as well as their safety for consumers must be clearly demonstrated. In particular, these proteins may become novel allergens triggering adverse reactions and, therefore, a comprehensive allergenicity risk assessment is needed. This review presents an overview of the most promising alternative protein sources, their application in the production of innovative food systems, as well as their potential effects on human health. In addition, new insights on sustainable processing strategies are given.

Background Benzoyl peroxide is a topical antiacne drug, which also acts as a strong irritant and a weak allergen. Only a few acne patients with allergic contact dermatitis due to benzoyl peroxide gel have been diagnosed by patch testing in Japan. Therefore, the number of such patients is probably underestimated. Objectives To correctly diagnose such cases by patch testing and to determine their characteristics and frequency. Patients and methods Twenty acne patients that were diagnosed with allergic contact dermatitis between April 2015 and April 2018 were enrolled in this study. Patch tests were performed with acne gels containing benzoyl peroxide and 1% benzoyl peroxide in petrolatum. The patients' profiles and the frequency of dermatitis were analyzed. Results All of the patients were female, and their mean age was 24.1 ± 9.3 years. Two patients were suffering from atopic dermatitis. The onset of allergic contact dermatitis occurred at 1 to 2 days, 9 to 28 days, and >30 days (longest: up to 24 months) after the initial application of the causative substance in 3 patients, 9 patients, and 8 patients, respectively. The frequency of such cases was 4.5% at our clinic. Conclusions Benzoyl peroxide gels for acne were demonstrated to often act as allergic contact allergens, and thus, dermatologists should be aware of their allergenicity and be apprehensive about markedly increasing the use of such gels in the future.

Aim. The purpose of this study is to create the technology of preparation of the polymeric allergoid received by chemical modification of Dermatophagoides farinae extract by a formaldehyde and a monomeric allergoid of Dermatophagoides pteronyssinus received by succinilation and studying of their physical, chemical and immunologic properties. Materials and methods . In the modification species determined the protein content, isoelectric points of protein components by a method of isoelectrofocusing (IEF), esterase activity. Antigenic properties of allergoid studied by methods of an immunodiffusion and an immunoelectrophoresis. Applied microdot immunoenzyme assay to characterize the specific activity of a resulting allergoid. Received by a succinilation allergoid was also studied on physical and chemical indicators. Results. It is established That the formalinization is resulted by the expressed depression of allergenic activity of an allergoid. It is also shown that succinilation leads to essential depression of allergenicity of a monomeric allergoid due to blockade of B-cellular epitopes and to conservation of an adjuvanticity helps to T-cellular epitopes. Conclusion. Thanks to the proved depression of allergenicity and rising of an adjuvanticity monomeric and polymeric allergoid can be recommended for carrying out allergenspecific immunotherapy (ASIT).

National audience; Ovalbumin (OVA), a major allergen from hen's egg albumen, tends to aggregate when heated. Depending on the balance of attractive and repulsive interactions, heat-induced OVA aggregates have various morphologies, which differ in digestibility. In the context of food allergy to egg, we investigated the ability of native and thermally aggregated OVA as well as their digests to induce the degranulation of a humanized rat basophil leukemia (RBL) cell line, which was sensitized with a pool of sera from egg-allergic children. Native and two thermally aggregated OVA forms were digested in vitro using a gastrointestinal digestion model based on the INFOGEST harmonized protocol including a final degradation with jejunal brush border membranes (BBM) enzymes. The course of digestion was monitored by the OPA method and by RP-HPLC. Digestibility was OVA small aggregates > OVA large aggregates > > native OVA and BBM peptidases only significantly hydrolyzed small sized peptides from gastro-duodenal digests of the aggregates. The degranulation ability of the native OVA slightly changed during the gastric phase but mostly decreased during the duodenal digestion with no further change with BBM digestion. The degranulation ability of aggregates, which was significantly lower than the ability of native OVA, was not significantly affected by digestion. Digestibility and ability to induce basophil degranulation can thus not be straightforward linked.

Background: Aspirin enhances food allergy symptoms by increasing absorption of ingested allergens. The objective of this study is to elucidate the role of aspirin in facilitating intestinal absorption of the wheat allergen, gliadin, in rats. Methods: Plasma concentrations of gliadin were determined after oral administration by gavage or administration into a closed intestinal loop in rats. We used an in situ intestinal re-circulating perfusion experiment to examine the effect of pepsin on aspirin-facilitated gliadin absorption. Fluorescein isothiocyanate (FITC)-labeled dextran-40 (FD-40) was used as a marker of non-specific absorption. The molecular size of gliadin and its allergenicity in plasma were examined using immunoblot analysis and intradermal reaction tests with Evans blue dye (EBD) extravasation, respectively. Results: Aspirin increased plasma concentrations of gliadin after oral administration but had no effect in the closed intestinal loop study. An in situ intestinal re-circulating perfusion study showed that FITC-labeled gliadin was absorbed similarly to FD-40. Aspirin increased absorption of both intact and pepsin-digested gliadin, with a more significant effect on absorption of pepsin-treated gliadin. Immunoblotting showed that most gliadin was absorbed in intact form. When the gliadin fraction was extracted from rat plasma after gavage and injected intradermally into gliadin-sensitized rats, EBD extravasation was observed at injection sites in a gliadin dose-dependent manner. Conclusions: Aspirin increased the absorption of intact and pepsin-digested gliadin via the paracellular pathway, maintaining their allergenicity. Moreover, the effect of aspirin on gliadin absorption was enhanced by modification and digestion of gliadin in the stomach., This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan (No. 16K08371).

Transglutaminase (TG) is a major food processing enzyme that induces protein cross-linking. Tropomyosin (TM) is a major shellfish allergen that is responsible for at least 80% of the shrimp allergic incidents. The aim of this research was to investigate the effect of TG on TM allergenicity and reveal the involvement of TM structure alternation. After treated with TG for different time, the allergenicity of TM was assessed by indirect ELISA with shrimp allergic patients’ sera, and the structure of TM was characterized by circular dichroism (CD) spectroscopy, ultraviolet (UV) absorption spectroscopy and fluorescence spectroscopy. Results showed that the allergenicity slightly decreased at the early stage of reaction and dramatically decreased at the late stage, which is tightly correlated with the increment of β-turn content and UV absorption. In conclusion, TG modification is an effective and promising food processing method to eliminate TM allergenicity with significant prospect of industrial appli...

A number of studies were devoted to understanding an immunological effect of pressure-treated β-lactoglobulin. In our previous work we have proved that high pressure significantly modifies β-lactoglobulin conformation and consequently its physicochemical properties. Here, structure of β-lactoglobulin complex with myristic acid determined at the highest accepted by the crystal pressure value of 550 MPa is reported. Our results structurally prove that pressure noticeably modifies positions of the major β-lactoglobulin epitopes. Considering the biological impact of observed changes in epitope regions, high pressure β-lactoglobulin structure presents a step forward in understanding the pressure modification of food protein allergenicity. The conformational changes of pressurized β-lactoglobulin did not support the hypothesis that proteolytic digestion facilitated by pressure is caused by an exposure of the digestive sites. Our findings demonstrate that high pressure protein crystallography can potentially identify the most pressure-sensitive fragments in allergens, and can therefore support development of hypoallergenic food products.

House dust mite (HDM) is a globally ubiquitous domestic cause of allergic diseases. There is a pressing demand to discover efficient, harmless, and eco-friendly natural extracts to inhibit HDM allergens that are more likely to trigger allergies and challenging to be prevented entirely. This study, therefore, is aimed at assessing the inhibition of the allergenicity of major HDM allergen Der f 2 by todomatsu oil extracted from residues of Abies Sachalinensis. The inhibition was investigated experimentally (using enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)) and in silico using molecular docking. The results showed that todomatsu oil inhibits the allergenicity of Der f 2 by reducing its amount instead of the IgG binding capacity of a single protein. Moreover, the compounds in todomatsu oil bind to Der f 2 via alkyl hydrophobic interactions. Notably, most compounds interact with the hydrophobic amino acids of Der f 2, and seven substances interact with CYS27. Contrarily, the principal compounds fail to attach to the amino acids forming the IgG epitope in Der f 2. Interestingly, chemical components with the lowest relative percentages in todomatsu oil show high-affinity values on Der f 2, especially β-maaliene (−8.0 kcal/mol). In conclusion, todomatsu oil has been proven in vitro as a potential effective public health strategy to inhibit the allergenicity of Der f 2.

The aim of this work is to research the effect of high pressure on soy protein and its main components: 7S and 11S glycinins; changes of soy protein technological and functional features.Results: soy protein, 7S and 11S glycinins functional features processed by high pressure are analyzed, as well as their emulsion features, ability to retain water, gel features. The impact of pressure, time and temperature of the process on soy protein is researched. Structural changes of 7S and 11S glycinins, conformations, technological and functional features are analyzed. Soy protein isolate processing with high pressure improves its rheological, gel features and moisture content. Due to reasonable parameters soy protein and 7S and 11S glycinins processing with a high pressure increased moisture content; improved gel and emulsion features; influenced on non-covalent and covalent bonds and protein conformation; decreased soy protein's allergenicity in food products, including baby formulas. Despite great efforts, the mechanism of soy protein and 7S and 11S glycinins processing is still insufficiently understood, which makes it difficult to get a clear and unambiguous idea of their behavior.Conclusions. The use of high pressure and soy protein isolate combination can improve functional and consumptional features of soy protein and food safety., Цель. В этой работе было рассмотрено действие соевого белка и его основного ингредиента: глицининов 7S и 11S при воздействии высокого давления. Кратко описаны компоненты и функциональные свойства соевых белков. Полученные результаты. Обсуждались функциональные свойства соевого белка, 7S и 11S глицининов, обработанных обработкой под высоким давлением, такие как свойства эмульсии, водоудерживающая способность, свойства геля. Влияние уровня давления, времени и температуры на соевый белок, глицинины 7S и 11S были проанализированы по изменению их структуры, конформации и технологических свойств. Комбинация обработки под высоким давлением и изолята соевого белка может улучшить реологические свойства, свойства геля и содержание неподвижной воды. Таким образом, правильная обработка под давлением соевого белка, 7S и 11S глицининов увеличивает удерживание воды, свойства геля и эмульсии за счет воздействия на нековалентную связь, ковалентную связь и конформацию белка, а также снижает аллергенность соевых белков в пищевых продуктах, включая детское питание. . Несмотря на большие усилия, механизм обработки соевого белка, 7S и 11S глицина под высоким давлением, все еще недостаточно изучен, что затрудняет получение четкого и однозначного представления об их поведении. Выводы. Использование комбинации обработки под высоким давлением и изолята соевого белка может улучшить функциональные и потребительские свойства соевого белка и безопасность пищевых продуктов., Мета Дослідити вплив високого тиску на соєвий білок та його головні інгредієнти: гліциніни 7S та 11S та зміни технологічних та функціональних властивості соєвих білків. Результати. Проаналізовано функціональні властивості соєвого білка, 7S та 11S гліцинінів, оброблених високим тиском, їх емульсійні властивості, здатність утримувати воду, властивості гелю. Досліджено вплив значень параметрів тиску, часу та температури процесу на соєвий білок, 7S та 11S гліциніни за змінами їх структури, конформацій, технологічних та функціональних властивостей. Обробка високим тиском ізолята соєвого білка покращує їх реологічні властивості, властивості гелю та вміст зв’язаної вологи. Обробка високим тиском при обгрунтованих параметрах процесу соєвого білка, гліцинінів 7S та 11S збільшила вміст зв’язаної вологи, покращила властивості гелю та емульсії, вплинула на нековалентні та ковалентні зв’язки та конформацію білка, а також зменшила алергенність соєвих білків у харчових продуктах, включаючи дитячі суміші. Незважаючи на великі зусилля, механізм обробки високим тиском соєвого білка, 7S та 11S гліцину все ще недостатньо зрозумілий, що ускладнює отримання чіткого і однозначного уявлення про їх поведінку. Висновки. Використання комбінації засобів високого тиску та ізоляту соєвого білка може поліпшити функціональні та споживчі властивості соєвого білка та безпеку харчових продуктів.

Orientador: Flavia Maria Netto Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos Resumo: O leite bovino contém várias proteínas que são consideradas antigênicas e capazes de induzir resposta imune, dentre elas, a ß-lactoglubulina (ß-Lg) é uma das mais antigênicas. A enzima transglutaminase (TG) é a única enzima utilizada comercialmente que catalisa reação de ligação cruzada inter ou intramolecular em diversas proteínas, formando polímeros de alta massa molar. Até o momento, pouco se conhece sobre a modificação dos epítopos antigênicos na estrutura das proteínas do soro de leite modificada ou polimerizada pela enzima TG. O presente trabalho teve como objetivo estudar o efeito da reação de polimerização induzida pela TG na atividade antigênica da ß-Lg. A modificação da ß-Lg com a TG foi realizada em dois experimentos: (1) a ß-Lg foi tratada termicamente (80 °C/60 min) em diferentes concentrações (3, 5 e 7%) e polimerizada com TG (5, 10, 25 e 50 U/g de proteína) e (2) a ß-Lg (3, 5 e 7%) foi polimerizada com TG 25 U/g de proteína na presença de agente redutor Cys nas concentrações 0,05, 0,1, 0,25 e 0,4 mol/L. A caracterização da ß-Lg polimerizada foi realizada por eletroforese (SDS-PAGE) e cromatografia líquida de alta eficiência de exclusão molecular (CLAE-EM). A antigenicidade da proteína foi avaliada por métodos imunoquímicos, utilizando soro de camundongos BALB/c e técnicas de Immunoblotting e ELISA. Quando a ß-Lg 3% foi tratada termicamente e polimerizada com TG, 40% da ß-Lg permaneceu nas formas monomérica e dimérica. Já as amostras tratadas com maior concentração de proteína (5% e 7%) apresentaram de 70% a 87% de material com massa molar (MM) > 100 kDa, sendo que ß-Lg 7% polimerizada com 25U TG/g foi a que apresentou maior grau de polimerização. Na presença de Cys, observou-se maior percentual de produtos com MM > 100 kDa que o obtido com o tratamento térmico. Nas amostras ß-Lg 5% e ß-Lg 7%, a presença da Cys possivelmente aumentou a susceptibilidade da ß-Lg à polimerização. As amostras selecionadas para avaliação da antigenicidade foram ß-Lg nativa, ß-Lg 7% tratada termicamente e polimerizada com 25U TG/g de proteína (ß-Lg 7% TT 25TG) e ß-Lg 7% polimerizada com 25 U/g de proteína na presença de Cys 0,25 mol/L (ß-Lg 7% 0,25Cys TG). Os resultados mostraram que os animais sensibilizados com ß-Lg polimerizada na presença de agente redutor Cys apresentaram níveis séricos de IgE e IgG menores, comparados aos grupos imunizados com ß-Lg nativa e ß-Lg tratada termicamente e polimerizada com TG, sugerindo que a polimerização na presença de agente redutor Cys pode reduzir o potencial antigênico da proteína ao modificar .e/ou ocultar regiões de epítopos na proteína Abstract: The bovine milk contains several proteins that are considered antigenic and capable of inducing immune responses, among them ß-lactoglubulin (ß-Lg) is one of the most antigenic protein. The enzyme transglutaminase (TG) is the only enzyme commercially used that catalyzes reaction of crosslinking inter or intramolecular in several proteins, forming polymer of high molecular mass. Up to now, little is known about modification of epitopes of whey proteins by the TG. The present study examines the effect of the polymerization reaction induced by TG on the antigenic activity of ß-Lg. The ß-Lg was modified by TG in two experiments: (1) ß-Lg was heat treated (80 °C/60 min) at different concentrations (3, 5 and 7%) and modified with TG (5, 10 , 25 and 50U / g protein), and (2) ß-Lg (3, 5 and 7%) was modified with TG 25U / g of protein in the presence of reducing agent Cys ( 0.05, 0.1, 0.25, 0.4 mol/L). The characterization of the modified ß-Lg was performed by electrophoresis (SDSPAGE) and high performance liquid chromatography molecular exclusion (HPLCMS). The antigenicity of the protein was measured by Immunoblotting and ELISA, using serum from BALB/c mices. When ß-Lg 3% was heat treated and modified by TG, 40% of the ß-Lg remained as monomeric and dimeric forms. However, the samples treated at higher protein concentration (5% and 7%) showed 70% to 87% of material with molecular weigh > 100 kDa. ß-Lg 7% modified with 25U TG / g presented the greatest degree of polymerization. In the presence of Cys, higher percentage of products with MW > 100 kDa was obtained than with the previous heat treatment. The presence of Cys increased the susceptibility of ß-Lg for polymerization, especially those treated at protein 5 and 7% concentrations. The samples selected for evaluation of the antigenicity were native ß-Lg, 7% ß-Lg treated with 25U TG / g of protein (ß-Lg 7% TT 25TG) and 7% ß-Lg treated with 25U TG / g protein in the presence of Cys 0.25 mol / L (ß-Lg 7% 0.25 Cys TG). The results showed that animals sensitized with ß-Lg in the presence of reducing the agent Cys had lower levels of IgG and IgE, compared to the groups immunized with native ß-Lg and ß-Lg treated with high temperature and modified with TG, suggesting that the polymerization in the presence of reducing agent Cys can modify or hide epitopes and reduce the potential antigenic of the protein Mestrado Nutrição Experimental e Aplicada à Tecnologia de Alimentos Mestre em Alimentos e Nutrição

Fish allergy is a life-long food allergy whose prevalence is affected by many demographic factors. Currently, there is no cure for fish allergy, which can only be managed by strict avoidance of fish in the diet. According to the WHO/IUIS Allergen Nomenclature Sub-Committee, 12 fish proteins are recognized as allergens. Different processing (thermal and non-thermal) techniques are applied to fish and fishery products to reduce microorganisms, extend shelf life, and alter organoleptic/nutritional properties. In this concise review, the development of a consistent terminology for studying food protein immunogenicity, antigenicity, and allergenicity is proposed. It also summarizes that food processing may lead to a decrease, no change, or even increase in fish antigenicity and allergenicity due to the change of protein solubility, protein denaturation, and the modification of linear or conformational epitopes. Recent studies investigated the effect of processing on fish antigenicity/allergenicity and were mainly conducted on commonly consumed fish species and major fish allergens using in vitro methods. Future research areas such as novel fish species/allergens and ex vivo/in vivo evaluation methods would convey a comprehensive view of the relationship between processing and fish allergy.

Undirected modifications between food proteins and secondary plant metabolites can occur during food processing. The results of covalent interactions can alter the functional and biological properties of the proteins. The present work studied the extent of which covalent conjugation of the bioactive metabolite benzyl isothiocyanate (BITC, a glucosinolate breakdown product) to the whey protein α-lactalbumin affects the protein’s allergenicity. Additional to the immunological analysis of native untreated and BITC-modified α-lactalbumin, the analysis of antigenic properties of proteolytically digested protein derivatives was also performed by high performance thin layer chromatography and immunostaining. As a result of the chemical modifications, structural changes in the protein molecule affected the allergenic properties. In this process, epitopes are destroyed or inactivated, but at the same time, buried epitopes can be exposed or newly formed, so that the net effect was an increase in allergenicity, in this case. Results from the tryptic hydrolysis suggest that BITC conjugation sterically hindered the cleavage sites for the enzyme, resulting in reduced digestibility and allergenicity. Residual antigenicity can be still present as short peptide fragments that provide epitopes. The desire to make food safer for allergy sufferers and to protect sensitized individuals from an allergenic reaction makes it clear that the detection of food antigens is mandatory, especially by considering protein interactions.

Endorsement date 21 January 2021 Implementation date 27 March 2021 Following a request from the European Commission, the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) was asked to deliver a scientific opinion on Scientific and technical guidance for the preparation and presentation of applications for exemption from mandatory labelling of food allergens and/or products thereof. This guidance applies to food ingredients or substances with known allergenic potential listed in Annex II of Regulation (EU) No 1169/2011 or products thereof, and aims to assist applicants in the preparation and presentation of well‐structured applications for exemption from labelling. It presents a common format for the organisation of the information to be provided and outlines the information and scientific data which must be included in the application, the hierarchy of different types of data and study designs, reflecting the relative strength of evidence which may be obtained from different study types and the key issues which must be addressed in the application in order to assess the likelihood of a food allergen‐derived preparation/foodstuff(s) triggering adverse reactions in sensitive individuals under the proposed conditions of use. This guidance document was adopted by the NDA Panel in 2013 and updated in 2017 to reflect the application of Regulation (EU) No 1169/2011. Upon request from the European Commission in 2020, it has been revised to inform applicants of new provisions in the pre‐submission phase and submission application procedure set out in Regulation (EC) No 178/2002, as amended by Regulation (EU) 2019/1381 on the transparency and sustainability of the EU risk assessment in the food chain, that are applicable to all applications submitted as of 27 March 2021.

Allergenicity indices are a powerful tool to assess the health hazard posed by urban parks to pollen allergic subjects. Nonetheless, only few indices have been developed and applied to urban vegetation in the last decade, and they were never compared nor standardised over the same dataset. To address this issue, in this paper the two best-known allergenicity indices, the Urban Green Zones Allergenicity Index (IUGZA) and the Specific Allergenicity Index (SAI), have been calculated for the same park (the Botanical Garden of Bologna), collecting vegetation data through both systematic sampling and arboreal census. The results obtained with the two data collection methods were comparable for both indices, indicating systematic sampling as a reliable approximation of the total census. Besides, the allergenic risk resulted moderate to high according to SAI, and very low according to IUGZA. Since SAI does not consider the total volume of the vegetation, it was deemed less reliable than IUGZA in evaluating the allergenicity of an enclosed green space.

Scope: Around 25% of food allergic persons in Central Europe suffer from carrot allergy caused by the major carrot allergen Dau c 1. Three different isoallergens, Dau c 1.01, Dau c 1.02 and Dau c 1.03 are identified. However, information about the qualitative and quantitative composition of natural (n)Dau c 1 is scarce. Methods and Results: The new carrot allergen Dau c 1.0401 is identified on the mRNA and protein level by RT‐PCR and mass spectrometry. It displays only around 60% sequence identity to the other known Dau c 1 isoallergens. NMR and CD‐spectra are typical for a well‐folded protein containing both α‐helices and β‐strands. It showed a poor refolding capacity after incubation at 95 °C. IgE‐binding is impaired in immunoblots, whereas in inhibition assays IgE binding to soluble Dau c 1.0401 is detected and it clearly provoked a response in mediator release assays. Conclusion: Dau c 1.0401 is a new isoallergen which contributes to the allergenicity of carrots. The absence of immunoreactivity in immobilized assays indicates that IgE binding is impaired when the protein is blotted on a solid phase. Altogether, the results point out that its allergenicity can be reduced upon carrot processing.

This review searched for published evidence that could explain how different physicochemical properties impact on the allergenicity of food proteins and if their effects would follow specific patterns among distinct protein families. Owing to the amount and complexity of the collected information, this literature overview was divided in two articles, the current one dedicated to protein families of plant allergens and a second one focused on animal allergens. Our extensive analysis of the available literature revealed that physicochemical characteristics had consistent effects on protein allergenicity for allergens belonging to the same protein family. For example, protein aggregation contributes to increased allergenicity of 2S albumins, while for legumins and cereal prolamins, the same phenomenon leads to a reduction. Molecular stability, related to structural resistance to heat and proteolysis, was identified as the most common feature promoting plant protein allergenicity, although it fails to explain the potency of some unstable allergens (e.g. pollen-related food allergens). Furthermore, data on physicochemical characteristics translating into clinical effects are limited, mainly because most studies are focused on in vitro IgE binding. Clinical data assessing how these parameters affect the development and clinical manifestation of allergies is minimal, with only few reports evaluating the sensitising capacity of modified proteins (addressing different physicochemical properties) in murine allergy models. In vivo testing of modified pure proteins by SPT or DBPCFC is scarce. At this stage, a systematic approach to link the physicochemical properties with clinical plant allergenicity in real-life scenarios is still missing., The authors highly appreciate the support from the European Cooperation in Science and Technology (COST) Office. This article is based upon work from COST Action FA1402, supported by COST (www.cost.eu). This work was also supported by Fundação para a Ciência e Tecnologia under the Partnership Agreement UIDB 50006/2020 and by the projects AlleRiskAssess - PTDC/BAA-AGR/31720/2017 and NORTE-01-0145-FEDER-00001. C.V. is grateful to FCT grants (PD/BD/114576/2016) financed by POPH-QREN (subsidised by FSE and MCTES). T.C.V. is grateful to the Ministry of Education, Science and Technological Development of the Republic of Serbia through grant number OI172024. P.M.R. and D.S. are grateful to FCT through project UIDB/04326/2020 and Mar2020 16–02-01-FMP-0014 – ‘ALLYFISH’. J.K. and A.K. acknowledge the PRIDE program grant (PRIDE/11012546/NEXTIMMUNE) by the Fonds National de la Recherche (FNR), Luxembourg and a translational grant (APSIS, PMC/2017/02) by the Personalised Medicine Consortium (PMC), Luxembourg.

The goal of most studies published on sand contaminants is to gather and discuss knowledge to avoid faecal contamination of water by run-offs and tide retractions. Other life forms in the sand, however, are seldom studied but always pointed out as relevant. The Mycosands initiative was created to generate data on fungi in beach sands and waters, of both coastal and freshwater inland bathing sites. A team of medical mycologists and water quality specialists explored the sand culturable mycobiota of 91 bathing sites, and water of 67 of these, spanning fromthe Atlantic to the EasternMediterranean coasts, including the Italian lakes and the Adriatic, Baltic, and Black Seas. Sydney (Australia) was also included in the study. Thirteen countries took part in the initiative. The present study considered several fungal parameters (all fungi, several species of the genus Aspergillus and Candida and the genera themselves, plus other yeasts, allergenic fungi, dematiaceous fungi and dermatophytes). The study considered four variables that the team expected would influence the results of the analytical parameters, such as coast or inland location, urban and non-urban sites, period of the year, geographical proximity and type of sediment. The genera most frequently found were Aspergillus spp., Candida spp., Fusarium spp. and Cryptococcus spp. both in sand and in water. A site-blind median was found to be 89 Colony-Forming Units (CFU) of fungi per gram of sand in coastal and inland freshwaters, with variability between 0 and 6400 CFU/g. For freshwater sites, that number was 201.7 CFU/g (0, 6400 CFU/g (p = 0.01)) and for coastal sites was 76.7 CFU/g (0,3497.5 CFU/g). For coastalwaters and allwaters, themedianwas 0 CFU/ml (0, 1592 CFU/ml) and for freshwaters 6.7 (0, 310.0) CFU/ml (p < 0.001). The results advocate that beaches should bemonitored for fungi for safer use and better management. Highlights: Fungi are missing from water and sand health protection regulatory parameters; Both sand and water should be monitored for fungi; The median value of 89 CFU/g of all fungi may serve as a reference for sand regulation; Candida albicans, dermatophytes, endemic fungi and other fungi should be considered; Fungal analysis of water needs more data before reference values can be established. Acknowledgements: A. Abdillah, M. C. Esposto, J. Kabtani, K. Sarioglou, P. E. Verweij and F. Vieira for their collaboration in this study; the European Confederation of Medical Mycology for a seeding grant; the International Society for Human and Animal Mycology for some financial support; financial support from CESAM (UID/AMB/50017-POCI-01-0145-FEDER-007638) and CITAB (UID/AGR/04033/2019), via FCT/MCTES, from national funds (PIDDAC), cofounded by FEDER, (PT2020 Partnership Agreement and Compete 2020) and an NHMRC APP1121936 to W. Meyer; National and Kapodistrian University of Athens, Greece, Special Accounts for Research Grants (SARG K.A. 70/3/6915). info:eu-repo/semantics/publishedVersion

Scope Around 25% of food allergic persons in Central Europe suffer from carrot allergy caused by the major carrot allergen Dau c 1. Three different isoallergens, Dau c 1.01, Dau c 1.02 and Dau c 1.03 are identified. However, information about the qualitative and quantitative composition of natural (n)Dau c 1 is scarce. Methods and results The new carrot allergen Dau c 1.0401 is identified on the mRNA and protein level by RT-PCR and mass spectrometry. It displays only around 60% sequence identity to the other known Dau c 1 isoallergens. NMR and CD-spectra are typical for a well-folded protein containing both α-helices and β-strands. It showed a poor refolding capacity after incubation at 95 °C. IgE-binding is impaired in immunoblots, whereas in inhibition assays IgE binding to soluble Dau c 1.0401 is detected and it clearly provoked a response in mediator release assays. Conclusion Dau c 1.0401 is a new isoallergen which contributes to the allergenicity of carrots. The absence of immunoreactivity in immobilized assays indicates that IgE binding is impaired when the protein is blotted on a solid phase. Altogether, the results point out that its allergenicity can be reduced upon carrot processing.