Food Allergen

Kutatási eredményeink "terméke" egy egyedülálló prototípus egy vegán golyó és hamburger készítő automata berendezés. A fejlesztés technológiai eredménye egy új, tesztelt prototípus. Intelligens gyártási innovációt tartalmaz a prototípus egésze, de külön kiemelnénk az új típusú keverő egységet, mely az alapanyagot vákuum alatt főzi, homogenizálja. Kiemelendő még a formázó egység, ahol a súlyadagolás után megfelelően szép alakot formázunk. Így speciálisan a növényi anyag súlyának és textúrájának megfelelő módszerrel végezzük el a termék formázását, úgy, hogy utána azt a technológia a sütési, sokkolási, hűtési és csomagolási folyamaton a lehető leggyorsabban átviszi. Technológia és termék szinten is arra kutatási eredményre jutottunk, hogy olyan technológia szükséges, ami a termék eltarthatóságát növelni tudja. Így a hűtés és vákuumos-védőgázas csomagolás (beadagolókkal) helyett a terméket fagyasztjuk, ezáltal az eltarthatóság 3 hónapról 1 évre lett növelhető. 2018.év vége és 2019 év vége között közötti időszakban folyamatosan zajlottak a prototípus kísérletek, eltarthatósági és beltartalmi vizsgálatokkal együtt. Az eredetileg tervezett növényi burger fejlesztési irány a mai divatos trendeknek megfelelően mind ízben, mind állagban egy húsimitációsabb irányt vett. Az eredetileg tervezett köleses és hajdinás irány megmaradt, illetve további receptek is születtek a fejlesztések során. A kísérleti gyártások eredményeképpen kifejlesztésre kerültek a vegán termékek első körös, tesztelésre alkalmas receptúrái, melyek működőképesek voltak. A technológia kapcsán viszont megállapítást nyert, hogy a piac egyre jobban elhatárolódik az olajban történő kisütéstől, ezért kontakt grill technológia irányába fordultunk. Valamint ahhoz, hogy a napjainkban divatos, jelenleg nagyon felkapott hús jellegű textúrát elérjük (beyond meat) a receptúrában szereplő vizet, zsírt, keményítőt és egyéb adalékanyagokat emulgeáltuk, így és egy magas szintű homogenizációt értünk el. A technológiai eljárás során az anyag előkészítésben a vákuumos keverés helyett emulziós vákuumos keverés történik a húsjellegűbb textúra elérése érdekében

Lysozyme, a muramidase enzyme from egg whites (EC 3.2.1.17), is widely used in soluble form to control lactic acid bacteria in different foods. Moreover, hen egg white lysozyme is a hydrolytic enzyme that can be used to the control malolactic fermentation (MLF) during winemaking. MLF is only desirable in red and in some white wine, this suggest that MLF is at fault and needs to be controlled in all other types of wine. Lysozyme exhibits selective antimicrobial activity based on the hydrolysis of peptidoglycan cell wall constituents in lactic acid bacteria. In the last decade, several studies identified allergic reactions due to the presence of lysozyme in food. Given this relatively high incidence of lysozyme sensitization, and in accordance with the recently changed EC food legislation (1266/2010/CE), the use of lysozyme as an additive has to be declared on the ingredient label. To overcome this problem, the immobilization of the enzyme on insoluble supports, which allows the enzyme to be removed, has been the preferred strategy. In this context, this article offers a review of reports on lysozyme enological use over the last decade. It surveys the immobilization techniques and support materials used for lysozyme food preservation. This study attempts to provide useful guidance from the wealth of available immobilization data in the literature and, more importantly, to develop an integrated perspective on how to customize lysozyme for future enological uses.

The antioxidant activity of rosemary (Rosmarinus officinalis) extract from different raw materials has been studied. Extracts were prepared from
wild or drip-irrigated plants, as well as from the by-product resulting from the distillation of the aromatic essential oil. The radical scavenging
activity of rosemary extracts was compared with that of antioxidants widely used in food, such as BHT and
-tocopherol, using an optimization
of the method based on the reduction of the radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). The results pointed the excellent antioxidant activity
of the crude fresh rosemary extracts, which was almost identical to that of pure
-tocopherol, and higher than that of BHT; extracts prepared from
distilled rosemary showed the lowest activity, although they are also of interest due to the low cost of the raw material. High performance liquid
chromatography (HPLC) combined with diode array (DAD) and electrospray (ESI)-ion trap-MS detection was used to separate and identify the
compounds present in the rosemary extracts. Rosmarinic acid, carnosic acid and seven of their terpene-type metabolites, and seven flavones were
identified. The drying and/or distillation treatments used with the plant material strongly affected the content of the two compounds of higher
antioxidant activity: rosmarinic acid and carnosic acid.

ABSTRAK Pendahuluan: Reaksi hipersensitivitas tipe I merupakan reaksi alergi yang dipicu oleh aktivitas IgE. Hipersensitivitas tipe I dapat berupa reaksi anafilaktik sistemik atau reaksi lokal yang ditandai oleh inflamasi dan kerusakan epitel mukosa. Udang dikenal sebagai makanan yang sering memicu alergi akibat kandungan protein di dalamnya merangsang reaksi system imun secara berlebihan pada pasien tertentu. Tujuan: Laporan kasus ini bertujuan membahas penatalaksanaan stomatitis alergika yang disebabkan alergi udang. Kasus: Seorang wanita berusia 36 tahun datang ke RSGM Trisakti dengan keluhan terasa gatal pada bibir atas dan bawah bagian dalam sejak 1 hari sebelum kedatangan. Keluhan tersebut dirasakan setelah memakan udang sehari sebelumnya.Tatalaksana: Pasien diberikan Aloclair gel 3x1 hari, dan kontrol dilakukan satu minggu setelah kunjungan pertama. Kata kunci :Alergi makanan, stomatitis alergika, alergi, hipersensitivitas tipe I

Background: Challenge tests for food-dependent exercise-induced anaphylaxis (FDEIA) carry some risk and have a high rate of false negatives. Our aim was to explore the usefulness of an in vitro immunodepletion assay and an allergen microarray test in the identification of IgE-mediated cross-reactive food allergens in patients with suspected FDEIA or food-dependent exercise-induced urticaria and panallergen sensitization. Methods: Three patients with a history of food dependent exercise induced urticaria/anaphylaxis and food panaller-gen sensitization in whom a food-exercise challenge was not feasible were selected: a 25-year-old man with choliner-gic urticaria who experienced generalized urticaria and angioedema during a soccer match after drinking a peach-based soft drink; a 19-year-old woman with allergic rhinitis and controlled asthma who experienced anaphylactic shock while playing soccer, having eaten walnuts in the previous 90 min; and a 57-year-old man with baker's asthma who experienced four episodes of anaphylaxis during exercise after ingesting wheat-containing food. All individuals underwent a diagnostic work-up with skin prick tests, specific IgE (sIgE) and ImmunoCAP ISAC test. For the in vitro immunodepletion procedure, patients' serum was pre-incubated with the suspected native allergen (peach, walnut, or wheat) in solid phase (ImmunoCAP). The eluted serum, containing unbound IgE, was collected and samples were re-tested using Immunocap ISAC 112 and compared with baseline results. Results: All individuals were sensitized to lipid transfer proteins. The first patient was sensitized to Pru p 3, Cor a 8, Jug r 3, and Ara h 9; after pre-incubation with peach there was 100% depletion of sIgE to all components. The second patient was sensitized to Pru p 3, Cor a 8, Jug r 3, and Ara h 9; immunodepletion with walnut depleted sIgE to Ara h 9 by 67%, Pru p 3 and Pla a 3 (60%), Art v 3 (75%), Jug r 3 (88%), and Cor a 8 (100%). The third patient was sensitized to Pru p 3, Jug r 3, Ara h 9, and Tri a 14; immunodepletion with wheat depleted Tri a 14 only (100%). Conclusions: In vitro immunodepletion might be a useful diagnostic tool in food dependent exercise induced urticaria/anaphylaxis with panallergen sensitization, particularly for identifying the culprit allergen and guiding dietary elimination recommendations.

Allergies to cow's milk are very common and can present as life-threatening anaphylaxis. Consequently, food labeling legislation mandates that foods containing milk residues, including casein and/or b-lactoglobulin, provide an indication of such on the product label. Because contamination with either component independent of the other can occur during food manufacturing, effective allergen management measures for containment of milk residues necessitates the use of dual screening methods. To assist the food industry in improving food safety practices, we have developed a rapid lateral flow immunoassay test kit that reliably reports both residues down to 0.01 lg per swab and 0.1 ppm of protein for foods. The assay utilizes both sandwich and competitive format test lines and is specific for bovine milk residues. Selectivity testing using a panel of matrices with potentially interfering substances, including commonly used sanitizing agents, indicated reduction in the limit of detection by one-to fourfold. With food, residues were easily detected in all cow's milk–based foods tested, but goat and sheep milk residues were not detected. Specificity analysis revealed no cross-reactivity with common commodities, with the exception of kidney beans when present at high concentrations (.1%). The development of a highly sensitive and rapid test method capable of detecting trace amounts of casein and/or b-lactoglobulin should aid food manufacturers and regulatory agencies in monitoring for milk allergens in environmental and food samples.

More than 50 years ago food allergy (FA) was suggested as a contributing factor in AD. It was shown that cow’s milk (CM) feeding predisposed to the development of AD while breast fed babies had a prevalence of eczema seven times lower than that of CM-fed babies. Later on, it was reported that children with a family history of allergy and soy formula fed during the first six months of life had less eczema and asthma in comparison with CM-fed children. In a double-blind, cross-over study, Atherton et al. demonstrated a marked improvement of AD in 21 2-8yr old children receiving a CM and egg-free diet. It was noted that 14/20 patients responded more favorably to the antigen avoidance diet employing a soy formula as CM substitute, whereas only one had a more favorable response to a not CM and egg free diet.

Background: The intestinal epithelium forms a barrier that food allergens must cross in order to induce sensitiza-tion. The aim of this study was to evaluate the impact of the plant-derived food cysteine protease — actinidin (Act d1) on the integrity of intestinal epithelium tight junctions (TJs). Methods: Effects of Act d1 on the intestinal epithelium were evaluated in Caco-2 monolayers and in a mouse model by measuring transepithelial resistance and in vivo permeability. Integrity of the tight junctions was analyzed by confocal microscopy. Proteolysis of TJ protein occludin was evaluated by mass spectrometry. Results: Actinidin (1 mg/mL) reduced the transepithelial resistance of the cell monolayer by 18.1% (after 1 h) and 25.6% (after 4 h). This loss of barrier function was associated with Act d 1 disruption of the occludin and zonula occludens (ZO)-1 network. The effect on intestinal permeability in vivo was demonstrated by the significantly higher concentration of 40 kDa FITC-dextran (2.33 μg/mL) that passed from the intestine into the serum of Act d1 treated mice in comparison to the control group (0.5 μg/mL). Human occludin was fragmented, and putative Act d1 cleavage sites were identified in extracellular loops of human occludin. Conclusion: Act d1 caused protease-dependent disruption of tight junctions in confluent Caco-2 cells and increased intestinal permeability in mice. General significance: In line with the observed effects of food cysteine proteases in occupational allergy, these results suggest that disruption of tight junctions by food cysteine proteases may contribute to the process of sensitization in food allergy.

The aim of the study was to analyse the potential pea-peanut cross-reactivity using the mice BALB/c as a biological in vivo model in the research on immune response to peanut proteins (PnE). BALB/c mice were three-fold sensitised (on days 1, 7, and 21) by oral or intraperitoneal (IP) administration of PnE in 0.5 mg or 1 mg dose, with or without adjuvant -aluminum hydroxide gel (Alum). Serum immunoglobulins (IgE, IgG, IgG1 and IgG2a) and level of cytokines (IL-4, IL-10, IFNγ), secreted by the isolated lymphocytes were examined. The highest increase in total IgE and peanut-specific IgG1 was noted in the group sensitised by IP administration of PnE in the presence of Alum. Lymphocytes from peanut-sensitised (with and without Alum) mice showed a significantly high level of IL-4 and this cytokine was secreted to a much higher extent as compared to IFN-γ. Stimulation of a culture of lymphocytes with pea proteins resulted in high IFN-γ secretion. A weak reaction of peanut-specific IgG1 present in mice serum with pea globulins (vicilin -PV and legumin -PL) can suggest that the cross-reactivity between peanut and pea proteins results from the presence of proteins other than 7S and 11S globulins. Due to the demonstrated low cross-reactivity between peanut proteins and pea globulins, the possibility of applying pea proteins in peanut-allergy immunotherapy may be suggested.

Background The number of individuals with food allergies or intolerances attending catered university residential colleges is increasing, and safe dining options are required to minimize the risk of allergic reactions and food-induced death. Objective This qualitative research study sought to advance professional knowledge of the factors affecting allergen management practices, particularly pertaining to college foodservices. Design Three catered residential colleges affiliated with a major university in New Zealand were selected as research sites. The study used an ethnographic approach and systems-practice theory as a framework for data collection and organizing results. Data collection techniques included document analyses (3 hours per site), observations (6 to 8 hours per site), focus groups with foodservice workers (30 to 45 minutes per site, n¼16), and interviews with foodservice managers (45 to 90 minutes per interview, n¼5). Notes and transcripts were coded through the process of thematic analysis using NVivo for Mac software, version 11.1.1, to identify factors affecting allergen management practices. Results The main factors affecting allergen management practices at college foodservices included information provided by residents about dietary requirements; communication between residents and foodservice staff; systems for allergen management; attitude of foodservice staff; and college size. Conclusions Detailed dietary information, effective communication with residents, sufficient resources, clarification of responsibilities, and thorough systems are required for staff to perform safe allergen management practices. Ultimately, successful implementation was predominantly determined by staff attitude. Foodservice managers are advised to identify motivators and address barriers of staff attitudes toward allergen management practices to promote successful implementation.

BACKGROUND: Actinidin is a cysteine protease and major allergen from kiwi fruit. When purified under specific native conditions, actinidin preparations from fresh kiwi fruit contain both an active and inactive form of this enzyme. In this study, biochemical and immunological properties upon simulated gastro-intestinal digestion, as well as thermal stability, were investigated for both active and E-64-inhibited actinidin. RESULTS: Active actinidin retained its primary structure and proteolytic activity after 2 h of simulated gastric digestion, followed by 2 h of intestinal digestion, as assessed by SDS-PAGE, zymography and mass spectroscopy. Immunological reactivity of active actinidin was also preserved, as tested by immunoelectrophoresis. The E-64 inhibited actinidin was fully degraded after 1 h of pepsin treatment. Differential scanning calorimetry showed that active actinidin has one transition maximum temperature (T m) at 73.9 ∘ C, whereas in the E-64-actinidin complex the two actinidin domains unfolded independently, with the first domain having a T m value of only 61 ∘ C. CONCLUSION: Active actinidin is capable of reaching the intestinal mucosa in a proteolytically active and immunogenic state. Inhibitor binding induces changes in the actinidin molecule that go beyond inhibition of proteolytic activity, also influencing the digestion stability and T m values of actinidin, features important in the characterisation of food allergens.

While seeking novel food sources to feed the increasing population of the globe, several alternatives have been discussed, including algae, fungi or in vitro meat. The increasingly propagated usage of farmed insects for human nutrition raises issues regarding food safety, consumer information and animal protection. In line with law, insects like any other animals must not be reared or manipulated in a way that inflicts unnecessary pain, distress or harm on them. Currently, there is a great need for research in the area of insect welfare, especially regarding species-specific needs, health, farming systems and humane methods of killing. Recent results from neurophysiological, neuroanatomical and behavioral sciences prompt caution when denying consciousness and therefore the likelihood of presence of pain and suffering or something closely related to it to insects. From an animal protection point of view, these issues should be satisfyingly solved before propagating and establishing i...

Historically, food allergy diagnosis was a tedious procedure, characterized by extensive physician involvement and regular checkups. Recently, home testing kits for food allergies have been introduced to the public, largely due to the rising popularity of self-administered genetic testing. Extensive research has been conducted on the validity and reliability of various food allergy diagnostics. In this review, we will synthesize research related to various testing schemes, including that of immunoglobulin, genetics, along with unvalidated alternative methods, such as applied kinesiology and electrodermal tests. Recent findings indicate a possible connection between genetic markers and food allergy susceptibilities, a current focus of precision medicine.

Gluten derived from wheat and related triticeae cereals possesses distinct amino acid sequences that provoke the immunopathogenic features of celiac disease (CD) in genetically susceptible individuals. However, the role of oat-derived gluten, or avenins, in CD pathogenesis remains a disputed matter, as evidenced by a lack in harmonized legislation regarding gluten classification in relation to gluten-free labeling. In this study, we have analyzed a panel of pure oat cultivars using a sandwich ELISA based on the R5 monoclonal antibody (mAb), which binds to canonical epitopes occurring within celiagenic peptides present in triticeae-derived gluten but reportedly not present in avenins. We have identified three varieties of oats that reproducibly bind R5 antibodies and levels indicating the presence of gluten at more than the 20 ppm gluten regulatory threshold. Nested assessment using Western blot analysis and alternative gluten detection systems corroborated these results. Collectively, these data suggest that select oat varieties may prove problematic to patients with CD and to food companies and regulatory agencies and will extend our basic understanding of current gluten detection systems.

Three tomato allergens (Lyc e 1, Lyc e 2 and Lyc e 3) have been characterised until now. Previous studies have demonstrated the existence of other proteins with capacity to bind IgE. The objective of this study was to identify new tomato allergens by molecular biology techniques. A tomato peel cDNA library was screened using a specific pool of sera from sensitised individuals. A clone was obtained and its sequence compared to other described allergens. This clone contained 34 aminoacids from the C-terminal region of an acidic ribosomal protein 60S. This protein showed more than 85% of homology with fungal allergens and 93.8% of homology with the almond allergen Pru du 5. The clone also contains 169 base pairs without homology with other known genes and could correspond to non-coding sequences. The IgE detection with a pool of sera was positive which confirms that is a new tomato allergen.

Gluten derived from wheat and related Triticeae can induce gluten sensitivity as well as celiac disease. Consequently, gluten content in foods labeled " gluten-free " is regulated. Determination of potential contamination in such foods is achieved using immunoassays based on monoclonal antibodies (mAbs) that recognize specific epitopes present in gluten. However, food-processing measures can affect epitope recognition. In particular, preparation of wheat protein isolate through deamidation of glutamine residues significantly limits the ability of commercial gluten testing kits in their ability to recognize gluten. Adding to this concern, evidence suggests that deamidated gluten imparts more pathogenic potential in celiac disease than native gluten. To address the heightened need for antibody-based tools that can recognize deamidated gluten, we have generated a novel mAb, 2B9, and subsequently developed it as a rapid lateral flow immunoassay. Herein, we report the ability of the 2B9-based lateral flow device (LFD) to detect gluten from wheat, barley, and rye and deamidated gluten down to 2 ppm in food as well as its performance in food testing.

Background: Plant lipid transfer proteins (LTPs) assemble a family of small (7-9 kDa) ubiquitous cationic proteins with an ability to bind and transport lipids as well as participate in various physiological processes including defense against phytopathogens. They also form one of the most clinically relevant classes of plant allergens. Nothing is known to date about correlation between lipid-binding and IgE-binding properties of LTPs. The garden pea Pisum sativum is widely consumed crop and important allergenic specie of the legume family. This work is aimed at isolation of a novel LTP from pea seeds and characterization of its structural, functional, and allergenic properties. Results: Three novel lipid transfer proteins, designated as Ps-LTP1-3, were found in the garden pea Pisum sativum, their cDNA sequences were determined, and mRNA expression levels of all the three proteins were measured at different pea organs. Ps-LTP1 was isolated for the first time from the pea seeds, and its complete amino acid sequence was determined. The protein exhibits antifungal activity and is a membrane-active compound that causes a leakage from artificial liposomes. The protein binds various lipids including bioactive jasmonic acid. Spatial structure of the recombinant uniformly 13 C, 15 N-labelled Ps-LTP1 was solved by heteronuclear NMR spectroscopy. In solution the unliganded protein represents the mixture of two conformers (relative populations~85:15) which are interconnected by exchange process with characteristic time~100 ms. Hydrophobic residues of major conformer form a relatively large internal tunnel-like lipid-binding cavity (van der Waals volume comes up to~1000 Å 3). The minor conformer probably corresponds to the protein with the partially collapsed internal cavity. Conclusions: For the first time conformational heterogeneity in solution was shown for an unliganded plant lipid transfer protein. Heat denaturation profile and simulated gastrointestinal digestion assay showed that Ps-LTP1 displayed a high thermal and digestive proteolytic resistance proper for food allergens. The reported structural and immunological findings seem to describe Ps-LTP1 as potential cross-reactive allergen in LTP-sensitized patients, mostly Pru p 3 + ones. Similarly to allergenic LTPs the potential IgE-binding epitope of Ps-LTP1 is located near the proposed entrance into internal cavity and could be involved in lipid-binding.

BACKGROUND: Actinidin is a cysteine protease and major allergen from kiwi fruit. When purified under specific native conditions, actinidin preparations from fresh kiwi fruit contain both an active and inactive form of this enzyme. In this study, biochemical and immunological properties upon simulated gastro-intestinal digestion, as well as thermal stability, were investigated for both active and E-64-inhibited actinidin.

RESULTS:Active actinidin retained its primary structure and proteolytic activity after 2 h of simulated gastric digestion, followed by 2 h of intestinal digestion, as assessed by SDS-PAGE, zymography andmass spectroscopy. Immunological reactivity of active actinidin was also preserved, as tested by immunoelectrophoresis. The E-64 inhibited actinidin was fully degraded after 1 h of pepsin treatment. Differential scanning calorimetry showed that active actinidin has one transition maximum temperature (Tm) at 73.9∘C, whereas in the E-64–actinidin complex the two actinidin domains unfolded independently,with the first domain having a Tm value of only 61∘C.

CONCLUSION: Active actinidin is capable of reaching the intestinal mucosa in a proteolytically active and immunogenic state. Inhibitor binding induces changes in the actinidin molecule that go beyond inhibition of proteolytic activity, also influencing the digestion stability and Tm values of actinidin, features important in the characterisation of food allergens.

Three tomato allergens (Lyc e 1, Lyc e 2 and Lyc e 3) have been characterised until now. Previous studies have demonstrated the existence of other proteins with capacity to bind IgE. The objective of this study was to identify new tomato allergens by molecular biology techniques. A tomato peel cDNA library was screened using a specific pool of sera from sensitised individuals. A clone was obtained and its sequence compared to other described allergens. This clone contained 34 aminoacids from the C-terminal region of an acidic ribosomal protein 60S. This protein showed more than 85% of homology with fungal allergens and 93.8% of homology with the almond allergen Pru du 5. The clone also contains 169 base pairs without homology with other known genes and could correspond to non-coding sequences. The IgE detection with a pool of sera was positive which confirms that is a new tomato allergen.